Search results for: long-term experiments
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3190

Search results for: long-term experiments

880 Construction of Microbial Fuel Cells from Local Benthic Zones

Authors: Maria Luiza D. Ramiento, Maria Lissette D. Lucas

Abstract:

Electricity is said to serve as the backbone of modern technology. Considering this, electricity consumption has dynamically grown due to the continuous demand. An alternative producer of energy concerning electricity must therefore be given focus. Microbial fuel cell wholly characterizes a new method of renewable energy recovery: the direct conversion of organic matter to electricity using bacteria. Electricity is produced as fuel or new food is given to the bacteria. The study concentrated in determining the feasibility of electricity production from local benthic zones. Microbial fuel cells were constructed to harvest the possible electricity and to test the presence of electricity producing microorganisms. Soil samples were gathered from Calumpang River, Palawan Mangrove Forest, Rosario River and Batangas Port. Eleven modules were constructed for the different trials of the soil samples. These modules were made of cathode and anode chambers connected by a salt bridge. For 85 days, the harvested voltage was measured daily. No parameter is added for the first 24 days. For the next 61 days, acetic acid was included in the first and second trials of the modules. Each of the trials of the soil samples gave a positive result in electricity production.There were electricity producing microbes in local benthic zones. It is observed that the higher the organic content of the soil sample, the higher the electricity harvested from it. It is recommended to identify the specific species of the electricity-producing microorganism present in the local benthic zone. Complement experiments are encouraged like determining the kind of soil particles to test its effect on the amount electricity that can be harvested. To pursue the development of microbial fuel cells by building a closed circuit in it is also suggested.

Keywords: microbial fuel cell, benthic zone, electricity, reduction-oxidation reaction, bacteria

Procedia PDF Downloads 377
879 Assessing Two Protocols for Positive Reinforcement Training in Captive Olive Baboons (Papio anubis)

Authors: H. Cano, P. Ferrer, N. Garcia, M. Popovic, J. Zapata

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Positive Reinforcement Training is a well-known methodology which has been reported frequently to be used in captive non-human primates. As a matter of fact, it is an invaluable tool for different purposes related with animal welfare, such as primate husbandry and environmental enrichment. It is also essential to perform some cognitive experiments. The main propose of this pilot study was to establish an efficient protocol to train captive olive baboons (Papio anubis). This protocol seems to be vital in the context of a larger research program in which it will be necessary to train a complete population of around 40 baboons. Baboons were studied at the Veterinary Research Farm of the University of Murcia. Temporally isolated animals were trained to perform three basic tasks. Firstly, they were required to take food prices directly from the researchers’ hands. Then a clicker sound or bridge stimulus was added each time the animal acceded to the reinforcement. Finally, they were trained to touch a target, consisted of a whip with a red ball in its end, with their hands or their nose. When the subject completed correctly this task, it was also exposed to the bridge stimulus and awarded with a food price, such as a portion of banana, orange, apple, peach or a raisin. Two protocols were tested during this experiment. In both of them, there were 6 series of 2min training periods each day. However, in the first protocol, the series consisted in 3 trials, whereas in the second one, in each series there were 5 trials. A reliable performance was obtained with only 6 days of training in the case of the 5-trials protocol. However, with the 3-trials one, 26 days of training were needed. As a result, the 5-trials protocol seems to be more effective than the 3-trials one, in order to teach these three basic tasks to olive baboons. In consequence, it will be used to train the rest of the colony.

Keywords: captive primates, olive baboon, positive reinforcement training, Papio anubis, training

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878 Compost Bioremediation of Oil Refinery Sludge by Using Different Manures in a Laboratory Condition

Authors: O. Ubani, H. I. Atagana, M. S. Thantsha

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This study was conducted to measure the reduction in polycyclic aromatic hydrocarbons (PAHs) content in oil sludge by co-composting the sludge with pig, cow, horse and poultry manures under laboratory conditions. Four kilograms of soil spiked with 800 g of oil sludge was co-composted differently with each manure in a ratio of 2:1 (w/w) spiked soil:manure and wood-chips in a ratio of 2:1 (w/v) spiked soil:wood-chips. Control was set up similar as the one above but without manure. Mixtures were incubated for 10 months at room temperature. Compost piles were turned weekly and moisture level was maintained at between 50% and 70%. Moisture level, pH, temperature, CO2 evolution and oxygen consumption were measured monthly and the ash content at the end of experimentation. Bacteria capable of utilizing PAHs were isolated, purified and characterized by molecular techniques using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), amplification of the 16S rDNA gene using the specific primers (16S-P1 PCR and 16S-P2 PCR) and the amplicons were sequenced. Extent of reduction of PAHs was measured using automated soxhlet extractor with dichloromethane as the extraction solvent coupled with gas chromatography/mass spectrometry (GC/MS). Temperature did not exceed 27.5O°C in all compost heaps, pH ranged from 5.5 to 7.8 and CO2 evolution was highest in poultry manure at 18.78 µg/dwt/day. Microbial growth and activities were enhanced. Bacteria identified were Bacillus, Arthrobacter and Staphylococcus species. Results from PAH measurements showed reduction between 77 and 99%. The results from the control experiments may be because it was invaded by fungi. Co-composting of spiked soils with animal manures enhanced the reduction in PAHs. Interestingly, all bacteria isolated and identified in this study were present in all treatments, including the control.

Keywords: bioremediation, co-composting, oil refinery sludge, PAHs, bacteria spp, animal manures, molecular techniques

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877 Experimental Evaluation of Electrocoagulation for Hardness Removal of Bore Well Water

Authors: Pooja Kumbhare

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Water is an important resource for the survival of life. The inadequate availability of surface water makes people depend on ground water for fulfilling their needs. However, ground water is generally too hard to satisfy the requirements for domestic as well as industrial applications. Removal of hardness involves various techniques such as lime soda process, ion exchange, reverse osmosis, nano-filtration, distillation, and, evaporation, etc. These techniques have individual problems such as high annual operating cost, sediment formation on membrane, sludge disposal problem, etc. Electrocoagulation (EC) is being explored as modern and cost-effective technology to cope up with the growing demand of high water quality at the consumer end. In general, earlier studies on electrocoagulation for hardness removal are found to deploy batch processes. As batch processes are always inappropriate to deal with large volume of water to be treated, it is essential to develop continuous flow EC process. So, in the present study, an attempt is made to investigate continuous flow EC process for decreasing excessive hardness of bore-well water. The experimental study has been conducted using 12 aluminum electrodes (25cm*10cm, 1cm thick) provided in EC reactor with volume of 8 L. Bore well water sample, collected from a local bore-well (i.e. at – Vishrambag, Sangli; Maharashtra) having average initial hardness of 680 mg/l (Range: 650 – 700 mg/l), was used for the study. Continuous flow electrocoagulation experiments were carried out by varying operating parameters specifically reaction time (Range: 10 – 60 min), voltage (Range: 5 – 20 V), current (Range: 1 – 5A). Based on the experimental study, it is found that hardness removal to the desired extent could be achieved even for continuous flow EC reactor, so the use of it is found promising.

Keywords: hardness, continuous flow EC process, aluminum electrode, optimal operating parameters

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876 Analysing the Interactive Effects of Factors Influencing Sand Production on Drawdown Time in High Viscosity Reservoirs

Authors: Gerald Gwamba, Bo Zhou, Yajun Song, Dong Changyin

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The challenges that sand production presents to the oil and gas industry, particularly while working in poorly consolidated reservoirs, cannot be overstated. From restricting production to blocking production tubing, sand production increases the costs associated with production as it elevates the cost of servicing production equipment over time. Production in reservoirs that present with high viscosities, flow rate, cementation, clay content as well as fine sand contents is even more complex and challenging. As opposed to the one-factor at a-time testing, investigating the interactive effects arising from a combination of several factors offers increased reliability of results as well as representation of actual field conditions. It is thus paramount to investigate the conditions leading to the onset of sanding during production to ensure the future sustainability of hydrocarbon production operations under viscous conditions. We adopt the Design of Experiments (DOE) to analyse, using Taguchi factorial designs, the most significant interactive effects of sanding. We propose an optimized regression model to predict the drawdown time at sand production. The results obtained underscore that reservoirs characterized by varying (high and low) levels of viscosity, flow rate, cementation, clay, and fine sand content have a resulting impact on sand production. The only significant interactive effect recorded arises from the interaction between BD (fine sand content and flow rate), while the main effects included fluid viscosity and cementation, with percentage significances recorded as 31.3%, 37.76%, and 30.94%, respectively. The drawdown time model presented could be useful for predicting the time to reach the maximum drawdown pressure under viscous conditions during the onset of sand production.

Keywords: factorial designs, DOE optimization, sand production prediction, drawdown time, regression model

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875 Manodharmam: A Scientific Methodology for Improvisation and Cognition in Carnatic Music

Authors: Raghavi Janaswamy, Saraswathi K. Vasudev

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Music is ubiquitous in human lives. Ever since the fetus hears the sound inside the mother’s womb and later upon birth, the baby experiences alluring sounds, the curiosity of learning emanates and evokes exploration. Music is an education than mere entertainment. The intricate balance between music, education, and entertainment has well been recognized by the scientific community and is being explored as a viable tool to understand and improve human cognition. There are seven basic swaras (notes) Sa, Ri, Ga, Ma, Pa, Da, and Ni in the Carnatic music system that are analogous to C, D, E, F, G, A, and B of the western system. The Carnatic music builds on the conscious use of microtones, gamakams (oscillation), and rendering styles that evolved over centuries and established its stance. The complex but erudite raga system has been designed with elaborate experiments on srutis (musical sounds) and human perception abilities. In parallel, ‘rasa’- the emotions evoked by certain srutis and hence the ragas been solidified along with the power of language in combination with the musical sounds. The Carnatic music branches out as Kalpita sangeetam (pre-composed music) and Manodharma sangeetam (improvised music). This article explores the Manodharma sangeetam and its subdivisions such as raga alapana, swara kalpana, neraval, and ragam-tanam-pallavi (RTP). The intrinsic mathematical strategies in it’s practice methods toward improvising the music have been explored in detail with concert examples. The techniques on swara weaving for swara kalpana rendering and methods on the alapana development are also discussed at length with an emphasis on the impact on the human cognitive abilities. The articulation of the outlined conscious practice methods not only helps to leave a long-lasting melodic impression on the listeners but also onsets cognitive developments.

Keywords: Carnatic, Manodharmam, music cognition, Alapana

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874 Study on the Mechanism of CO₂-Viscoelastic Fluid Synergistic Oil Displacement in Tight Sandstone Reservoirs

Authors: Long Long Chen, Xinwei Liao, Shanfa Tang, Shaojing Jiang, Ruijia Tang, Rui Wang, Shu Yun Feng, Si Yao Wang

Abstract:

Tight oil reservoirs have poor physical properties, insufficient formation energy, and low natural productivity; it is necessary to effectively improve their crude oil recovery. CO₂ flooding is an important technical means to enhance oil recovery and achieve effective CO₂ storage in tight oil reservoirs, but its heterogeneity is strong, which makes CO₂ flooding prone to gas channeling and poor recovery. Aiming at the problem of gas injection channeling, combined with the excellent performance of low interfacial tension viscoelastic fluid (GOBTK), the research on CO₂-low interfacial tension viscoelastic fluid synergistic oil displacement in tight reservoirs was carried out, and the synergy of CO₂ and low interfacial tension viscoelastic fluid was discussed. Oil displacement mechanism. Experiments show that GOBTK has good injectability in tight oil reservoirs (Kg=0.141~0.793mD); CO₂-0.4% GOBTK synergistic flooding can improve the recovery factor of low permeability layers (31.41%) under heterogeneous (gradient difference of 10) conditions the) effect is better than that of CO₂ flooding (0.56%) and 0.4% GOBT-water flooding (20.99%); CO₂-GOBT synergistic oil displacement mechanism includes: 1) The formation of CO₂ foam increases the flow resistance of viscoelastic fluid, forcing the displacement fluid to flow 2) GOBTK can emulsify and disperse residual oil into small oil droplets, and smoothly pass through narrow pores to produce; 3) CO₂ dissolved in GOBTK synergistically enhances the water wettability of the core, and the use of viscosity Elastomeric fluid injection and stripping of residual oil; 4) CO₂-GOBTK synergy superimposes multiple mechanisms, effectively improving the swept volume and oil washing efficiency of the injected fluid to the reservoir.

Keywords: tight oil reservoir, CO₂ flooding, low interfacial tension viscoelastic fluid flooding, synergistic oil displacement, EOR mechanism

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873 An Energy Transfer Fluorescent Probe System for Glucose Sensor at Biomimetic Membrane Surface

Authors: Hoa Thi Hoang, Stephan Sass, Michael U. Kumke

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Concanavalin A (conA) is a protein has been widely used in sensor system based on its specific binding to α-D-Glucose or α-D-Manose. For glucose sensor using conA, either fluoresence based techniques with intensity based or lifetime based are used. In this research, liposomes made from phospholipids were used as a biomimetic membrane system. In a first step, novel building blocks containing perylene labeled glucose units were added to the system and used to decorate the surface of the liposomes. Upon the binding between rhodamine labeled con A to the glucose units at the biomimetic membrane surface, a Förster resonance energy transfer system can be formed which combines unique fluorescence properties of perylene (e.g., high fluorescence quantum yield, no triplet formation) and its high hydrophobicity for efficient anchoring in membranes to form a novel probe for the investigation of sugar-driven binding reactions at biomimetic surfaces. Two glucose-labeled perylene derivatives were synthesized with different spacer length between the perylene and glucose unit in order to probe the binding of conA. The binding interaction was fully characterized by using high-end fluorescence techniques. Steady-state and time-resolved fluorescence techniques (e.g., fluorescence depolarization) in combination with single-molecule fluorescence spectroscopy techniques (fluorescence correlation spectroscopy, FCS) were used to monitor the interaction with conA. Base on the fluorescence depolarization, the rotational correlation times and the alteration in the diffusion coefficient (determined by FCS) the binding of the conA to the liposomes carrying the probe was studied. Moreover, single pair FRET experiments using pulsed interleaved excitation are used to characterize in detail the binding of conA to the liposome on a single molecule level avoiding averaging out effects.

Keywords: concanavalin A, FRET, sensor, biomimetic membrane

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872 Enhancing Mitochondrial Activity and Metabolism in Aging Female Germ Cells: Synergistic Effects of Dual ROCK and ROS Inhibition

Authors: Kuan-Hao Tsui, Li-Te Lin, Chia-Jung Li

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The combination of Y-27632 and Vitamin C significantly enhances the quality of aging germ cells by reducing reactive oxygen species (ROS) production, restoring mitochondrial membrane potential balance, and promoting mitochondrial fusion. The age-related decline in oocyte quality contributes to reduced fertility, increased aneuploidy, and diminished embryo quality, with mitochondrial dysfunction in both oocytes and granulosa cells being a key factor in this decline. Experiments on aging germ cells investigated the effects of the Y-27632 and Vitamin C combination. In vivo studies involved aged mice to assess oocyte maturation and ROS accumulation during culture. The assessment included mitochondrial activity, ROS levels, mitochondrial membrane potential, and mitochondrial dynamics. Cellular energy metabolism and ATP production were also measured. The combination treatment effectively addressed mitochondrial dysfunction and regulated cellular energy metabolism, promoting oxygen respiration and increasing ATP production. In aged mice, this supplement treatment enhanced in vitro oocyte maturation and prevented ROS accumulation in aging oocytes during culture. While these findings are promising, further research is needed to explore the long-term effects and potential side effects of the Y-27632 and Vitamin C combination. Additionally, translating these findings to human subjects requires careful consideration. Overall, the study suggests that the Y-27632 and Vitamin C combination could be a promising intervention to mitigate aging-related dysfunction in germ cells, potentially enhancing oocyte quality, particularly in the context of in vitro fertilization.

Keywords: ovarian aging, supplements, ROS, mitochondria

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871 Developing an Intelligent Table Tennis Ball Machine with Human Play Simulation for Technical Training

Authors: Chen-Chi An, Jun-Yi He, Cheng-Han Hsieh, Chen-Ching Ting

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This research has successfully developed an intelligent table tennis ball machine with human play simulate all situations of human play to take the service. It is well known; an excellent ball machine can help the table tennis coach to provide more efficient teaching, also give players the good technical training and entertainment. An excellent ball machine should be able to service all balls based on human play simulation due to the conventional competitions are today all taken place for people. In this work, two counter-rotating wheels are used to service the balls, where changing the absolute rotating speeds of the two wheels and the differences of rotating speeds between the two wheels can adjust the struck forces and the rotating speeds of the ball. The relationships between the absolute rotating speed of the two wheels and the struck forces of the ball as well as the differences rotating speeds between the two wheels and the rotating speeds of the ball are experimentally determined for technical development. The outlet speed, the ejected distance, and the rotating speed of the ball were measured by changing the absolute rotating speeds of the two wheels in terms of a series of differences in rotating speed between the two wheels for calibration of the ball machine; where the outlet speed and the ejected distance of the ball were further converted to the struck forces of the ball. In process, the balls serviced by the intelligent ball machine were based on the received calibration curves with help of the computer. Experiments technically used photosensitive devices to detect the outlet and rotating speed of the ball. Finally, this research developed some teaching programs for technical training using three ball machines and received more efficient training.

Keywords: table tennis, ball machine, human play simulation, counter-rotating wheels

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870 Impact of Soot on NH3-SCR, NH3 Oxidation and NH3 TPD over Cu/SSZ-13 Zeolite

Authors: Lidija Trandafilovic, Kirsten Leistner, Marie Stenfeldt, Louise Olsson

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Ammonia Selective Catalytic Reduction (NH3 SCR), is one of the most efficient post combustion abatement technologies for removing NOx from diesel engines. In order to remove soot, diesel particulate filters (DPF) are used. Recently, SCR coated filters have been introduced, which captures soot and simultaneously is active for ammonia SCR. There are large advantages with using SCR coated filters, such as decreased volume and also better light off characteristics, since both the SCR function as well as filter function is close to the engine. The objective of this work was to examine the effect of soot, produced using an engine bench, on Cu/SSZ-13 catalysts. The impact of soot on Cu/SSZ-13 in standard SCR, NH3 oxidation, NH3 temperature programmed desorption (TPD), as well as soot oxidation (with and without water) was examined using flow reactor measurements. In all experiments, prior to the soot loading, the fresh activity of Cu/SSZ-13 was recorded with stepwise increasing the temperature from 100°C till 600°C. Thereafter, the sample was loaded with soot and the experiment was repeated in the temperature range from 100°C till 700°C. The amount of CO and CO2 produced in each experiment is used to calculate the soot oxidized at each steady state temperature. The soot oxidized during the heating to next temperature step is included, e.g. the CO+CO2 produced when increasing the temperature to 600°C is added to the 600°C step. The influence of the two factors seem to be of the most importance to soot oxidation: ammonia and water. The influence of water on soot oxidation shift the maximum of CO2 and CO production towards lower temperatures, thus water increases the soot oxidation. Moreover, when adding ammonia to the system it is clear that the soot oxidation is lowered in the presence of ammonia, resulting in larger integrated COx at 500°C for O2+H2O, while opposite results at 600 °C was received where more was oxidised for O2+H2O+NH3 case. To conclude the presence of ammonia reduces the soot oxidation, which is in line with the ammonia TPD results where we found ammonia storage on the soot. Interestingly, during ammonia SCR conditions the activity for soot oxidation is regained at 500°C. At this high temperature the SCR zone is very short, thus the majority of the catalyst is not exposed to ammonia and therefore the inhibition effect of ammonia is not observed.

Keywords: NH3-SCR, Cu/SSZ-13, soot, zeolite

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869 Human-Machine Cooperation in Facial Comparison Based on Likelihood Scores

Authors: Lanchi Xie, Zhihui Li, Zhigang Li, Guiqiang Wang, Lei Xu, Yuwen Yan

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Image-based facial features can be classified into category recognition features and individual recognition features. Current automated face recognition systems extract a specific feature vector of different dimensions from a facial image according to their pre-trained neural network. However, to improve the efficiency of parameter calculation, an algorithm generally reduces the image details by pooling. The operation will overlook the details concerned much by forensic experts. In our experiment, we adopted a variety of face recognition algorithms based on deep learning, compared a large number of naturally collected face images with the known data of the same person's frontal ID photos. Downscaling and manual handling were performed on the testing images. The results supported that the facial recognition algorithms based on deep learning detected structural and morphological information and rarely focused on specific markers such as stains and moles. Overall performance, distribution of genuine scores and impostor scores, and likelihood ratios were tested to evaluate the accuracy of biometric systems and forensic experts. Experiments showed that the biometric systems were skilled in distinguishing category features, and forensic experts were better at discovering the individual features of human faces. In the proposed approach, a fusion was performed at the score level. At the specified false accept rate, the framework achieved a lower false reject rate. This paper contributes to improving the interpretability of the objective method of facial comparison and provides a novel method for human-machine collaboration in this field.

Keywords: likelihood ratio, automated facial recognition, facial comparison, biometrics

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868 Numerical Investigation of Turbulent Inflow Strategy in Wind Energy Applications

Authors: Arijit Saha, Hassan Kassem, Leo Hoening

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Ongoing climate change demands the increasing use of renewable energies. Wind energy plays an important role in this context since it can be applied almost everywhere in the world. To reduce the costs of wind turbines and to make them more competitive, simulations are very important since experiments are often too costly if at all possible. The wind turbine on a vast open area experiences the turbulence generated due to the atmosphere, so it was of utmost interest from this research point of view to generate the turbulence through various Inlet Turbulence Generation methods like Precursor cyclic and Kaimal Spectrum Exponential Coherence (KSEC) in the computational simulation domain. To be able to validate computational fluid dynamic simulations of wind turbines with the experimental data, it is crucial to set up the conditions in the simulation as close to reality as possible. This present work, therefore, aims at investigating the turbulent inflow strategy and boundary conditions of KSEC and providing a comparative analysis alongside the Precursor cyclic method for Large Eddy Simulation within the context of wind energy applications. For the generation of the turbulent box through KSEC method, firstly, the constrained data were collected from an auxiliary channel flow, and later processing was performed with the open-source tool PyconTurb, whereas for the precursor cyclic, only the data from the auxiliary channel were sufficient. The functionality of these methods was studied through various statistical properties such as variance, turbulent intensity, etc with respect to different Bulk Reynolds numbers, and a conclusion was drawn on the feasibility of KSEC method. Furthermore, it was found necessary to verify the obtained data with DNS case setup for its applicability to use it as a real field CFD simulation.

Keywords: Inlet Turbulence Generation, CFD, precursor cyclic, KSEC, large Eddy simulation, PyconTurb

Procedia PDF Downloads 74
867 Determining Cellular Biomarkers Sensitive to Low Damaging Exposure

Authors: Svetlana Guryeva, Inna Kornienko, Elena Petersen

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At present, translational medicine is a rapidly developing branch of biomedicine. The main idea of translational medicine is a practical application of fundamental research. One of the possible applications for translational medicine is researching therapies that improve human age-related organism condition. To fill the gap between experiments and clinical practice, it is necessary to create the standardized system for the investigation of different effects on cellular aging models. In this study, primary human fibroblasts derived from patients of different ages were used as a cellular aging model. The senescence-associated β-galactosidase activity, lipofuscin, γ-H2AX, the reactive oxygen species level, and cell death markers (annexin V/propidium iodide) were used as biomarkers of the cell functional state. The effects of damaging exposures (oxidative stress and heat shock), potential positive factors (metformin and acetaminophen), and their combinations were investigated using the described biomarkers. Oxidative stress and heat shock caused the increase in the levels of all biomarkers, and only the cells from young patients partly coped with stress 3 days after the exposures. Metformin improved the state of pretreatment cells from young and old patients. The acetaminophen did not show significant changes in the biomarker levels compare to the action of metformin. This study proved the opportunity to develop a standardized screening system based on biomarkers of the cell functional state to identify potential positive or negative effects of some physical and chemical exposures. Moreover, such a system can be useful for the aims of regenerative medicine to determine the effect of cell pretreatment before transplantation.

Keywords: biomarkers, primary fibroblasts, regenerative medicine, senescence, test system, translational medicine

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866 Liquid-Liquid Plug Flow Characteristics in Microchannel with T-Junction

Authors: Anna Yagodnitsyna, Alexander Kovalev, Artur Bilsky

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The efficiency of certain technological processes in two-phase microfluidics such as emulsion production, nanomaterial synthesis, nitration, extraction processes etc. depends on two-phase flow regimes in microchannels. For practical application in chemistry and biochemistry it is very important to predict the expected flow pattern for a large variety of fluids and channel geometries. In the case of immiscible liquids, the plug flow is a typical and optimal regime for chemical reactions and needs to be predicted by empirical data or correlations. In this work flow patterns of immiscible liquid-liquid flow in a rectangular microchannel with T-junction are investigated. Three liquid-liquid flow systems are considered, viz. kerosene – water, paraffin oil – water and castor oil – paraffin oil. Different flow patterns such as parallel flow, slug flow, plug flow, dispersed (droplet) flow, and rivulet flow are observed for different velocity ratios. New flow pattern of the parallel flow with steady wavy interface (serpentine flow) has been found. It is shown that flow pattern maps based on Weber numbers for different liquid-liquid systems do not match well. Weber number multiplied by Ohnesorge number is proposed as a parameter to generalize flow maps. Flow maps based on this parameter are superposed well for all liquid-liquid systems of this work and other experiments. Plug length and velocity are measured for the plug flow regime. When dispersed liquid wets channel walls plug length cannot be predicted by known empirical correlations. By means of particle tracking velocimetry technique instantaneous velocity fields in a plug flow regime were measured. Flow circulation inside plug was calculated using velocity data that can be useful for mass flux prediction in chemical reactions.

Keywords: flow patterns, hydrodynamics, liquid-liquid flow, microchannel

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865 Comparative Proteomic Analysis of Rice bri1 Mutant Leaves at Jointing-Booting Stage

Authors: Jiang Xu, Daoping Wang, Yinghong Pan

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The jointing-booting stage is a critical period of both vegetative growth and reproductive growth in rice. Therefore, the proteomic analysis of the mutant Osbri1, whose corresponding gene OsBRI1 encodes the putative BRs receptor OsBRI1, at jointing-booting stage is very important for understanding the effects of BRs on vegetative and reproductive growth. In this study, the proteomes of leaves from an allelic mutant of the DWARF 61 (D61, OsBRI1) gene, Fn189 (dwarf54, d54) and its wild-type variety T65 (Taichung 65) at jointing-booting stage were analysed by using a Q Exactive plus orbitrap mass spectrometer, and more than 3,100 proteins were identified in each sample. Ontology analysis showed that these proteins distribute in various space of the cells, such as the chloroplast, mitochondrion, and nucleus, they functioned as structural components and/or catalytic enzymes and involved in many physiological processes. Moreover, quantitative analysis displayed that 266 proteins were differentially expressed in two samples, among them, 77 proteins decreased and 189 increased more than two times in Fn189 compared with T65, the proteins whose content decreased in Fn189 including b5-like Heme/Steroid binding domain containing protein, putative retrotransposon protein, putative glutaminyl-tRNA synthetase, and higher content proteins such as mTERF, putative Oligopeptidase homologue, zinc knuckle protein, and so on. A former study founded that the transcription level of a mTERF was up-regulated in the leaves of maize seedling after EBR treatment. In our experiments, it was interesting that one mTERF protein increased, but another mTERF decreased in leaves of Fn189 at jointing-booting stage, which suggested that BRs may have differential regulation mechanisms on the expression of various mTERF proteins. The relationship between other differential proteins with BRs is still unclear, and the effects of BRs on rice protein contents and its regulation mechanisms still need further research.

Keywords: bri1 mutant, jointing-booting stage, proteomic analysis, rice

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864 Modeling of Glycine Transporters in Mammalian Using the Probability Approach

Authors: K. S. Zaytsev, Y. R. Nartsissov

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Glycine is one of the key inhibitory neurotransmitters in Central nervous system (CNS) meanwhile glycinergic transmission is highly dependable on its appropriate reuptake from synaptic cleft. Glycine transporters (GlyT) of types 1 and 2 are the enzymes providing glycine transport back to neuronal and glial cells along with Na⁺ and Cl⁻ co-transport. The distribution and stoichiometry of GlyT1 and GlyT2 differ in details, and GlyT2 is more interesting for the research as it reuptakes glycine to neuron cells, whereas GlyT1 is located in glial cells. In the process of GlyT2 activity, the translocation of the amino acid is accompanied with binding of both one chloride and three sodium ions consequently (two sodium ions for GlyT1). In the present study, we developed a computer simulator of GlyT2 and GlyT1 activity based on known experimental data for quantitative estimation of membrane glycine transport. The trait of a single protein functioning was described using the probability approach where each enzyme state was considered separately. Created scheme of transporter functioning realized as a consequence of elemental steps allowed to take into account each event of substrate association and dissociation. Computer experiments using up-to-date kinetic parameters allowed receiving the number of translocated glycine molecules, Na⁺ and Cl⁻ ions per time period. Flexibility of developed software makes it possible to evaluate glycine reuptake pattern in time under different internal characteristics of enzyme conformational transitions. We investigated the behavior of the system in a wide range of equilibrium constant (from 0.2 to 100), which is not determined experimentally. The significant influence of equilibrium constant in the range from 0.2 to 10 on the glycine transfer process is shown. The environmental conditions such as ion and glycine concentrations are decisive if the values of the constant are outside the specified range.

Keywords: glycine, inhibitory neurotransmitters, probability approach, single protein functioning

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863 Conversion of Sweet Sorghum Bagasse to Sugars for Succinic Acid Production

Authors: Enlin Lo, Ioannis Dogaris, George Philippidis

Abstract:

Succinic acid is a compound used for manufacturing lacquers, resins, and other coating chemicals. It is also used in the food and beverage industry as a flavor additive. It is predominantly manufactured from petrochemicals, but it can also be produced by fermentation of sugars from renewable feedstocks, such as plant biomass. Bio-based succinic acid has great potential in becoming a platform chemical (building block) for commodity and high-value chemicals. In this study, the production of bio-based succinic acid from sweet sorghum was investigated. Sweet sorghum has high fermentable sugar content and can be cultivated in a variety of climates. In order to avoid competition with food feedstocks, its non-edible ‘bagasse’ (the fiber part after extracting the juice) was targeted. Initially, various conditions of pretreating sweet sorghum bagasse (SSB) were studied in an effort to remove most of the non-fermentable components and expose the cellulosic fiber containing the fermentable sugars (glucose). Concentrated (83%) phosphoric acid was utilized at temperatures 50-80 oC for 30-60 min at various SSB loadings (10-15%), coupled with enzymatic hydrolysis using commercial cellulase (Ctec2, Novozymes) enzyme, to identify the conditions that lead to the highest glucose yields for subsequent fermentation to succinic acid. As the pretreatment temperature and duration increased, the bagasse color changed from light brown to dark brown-black, indicating decomposition, which ranged from 15% to 72%, while the theoretical glucose yield is 91%. With Minitab software statistical analysis, a model was built to identify the optimal pretreatment condition for maximum glucose released. The projected theoretical bio-based succinic acid production is 23g per 100g of SSB, which will be confirmed with fermentation experiments using the bacterium Actinobacillus succinogenes.

Keywords: biomass, cellulose, enzymatic hydrolysis, fermentation, pretreatment, succinic acid

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862 Feature Evaluation Based on Random Subspace and Multiple-K Ensemble

Authors: Jaehong Yu, Seoung Bum Kim

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Clustering analysis can facilitate the extraction of intrinsic patterns in a dataset and reveal its natural groupings without requiring class information. For effective clustering analysis in high dimensional datasets, unsupervised dimensionality reduction is an important task. Unsupervised dimensionality reduction can generally be achieved by feature extraction or feature selection. In many situations, feature selection methods are more appropriate than feature extraction methods because of their clear interpretation with respect to the original features. The unsupervised feature selection can be categorized as feature subset selection and feature ranking method, and we focused on unsupervised feature ranking methods which evaluate the features based on their importance scores. Recently, several unsupervised feature ranking methods were developed based on ensemble approaches to achieve their higher accuracy and stability. However, most of the ensemble-based feature ranking methods require the true number of clusters. Furthermore, these algorithms evaluate the feature importance depending on the ensemble clustering solution, and they produce undesirable evaluation results if the clustering solutions are inaccurate. To address these limitations, we proposed an ensemble-based feature ranking method with random subspace and multiple-k ensemble (FRRM). The proposed FRRM algorithm evaluates the importance of each feature with the random subspace ensemble, and all evaluation results are combined with the ensemble importance scores. Moreover, FRRM does not require the determination of the true number of clusters in advance through the use of the multiple-k ensemble idea. Experiments on various benchmark datasets were conducted to examine the properties of the proposed FRRM algorithm and to compare its performance with that of existing feature ranking methods. The experimental results demonstrated that the proposed FRRM outperformed the competitors.

Keywords: clustering analysis, multiple-k ensemble, random subspace-based feature evaluation, unsupervised feature ranking

Procedia PDF Downloads 313
861 Fabrication of Superhydrophobic Galvanized Steel by Sintering Zinc Nanopowder

Authors: Francisco Javier Montes Ruiz-Cabello, Guillermo Guerrero-Vacas, Sara Bermudez-Romero, Miguel Cabrerizo Vilchez, Miguel Angel Rodriguez-Valverde

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Galvanized steel is one of the widespread metallic materials used in industry. It consists on a iron-based alloy (steel) coated with a layer of zinc with variable thickness. The zinc is aimed to prevent the inner steel from corrosion and staining. Its production is cheaper than the stainless steel and this is the reason why it is employed in the construction of materials with large dimensions in aeronautics, urban/ industrial edification or ski-resorts. In all these applications, turning the natural hydrophilicity of the metal surface into superhydrophobicity is particularly interesting and would open a wide variety of additional functionalities. However, producing a superhydrophobic surface on galvanized steel may be a very difficult task. Superhydrophobic surfaces are characterized by a specific surface texture which is reached either by coating the surface with a material that incorporates such texture, or by conducting several roughening methods. Since galvanized steel is already a coated material, the incorporation of a second coating may be undesired. On the other hand, the methods that are recurrently used to incorporate the surface texture leading to superhydrophobicity in metals are aggressive and may damage their surface. In this work, we used a novel strategy which goal is to produce superhydrophobic galvanized steel by a two-step non-aggressive process. The first process is aimed to create a hierarchical structure by incorporating zinc nanoparticles sintered on the surface at a temperature slightly lower than the zinc’s melting point. The second one is a hydrophobization by a thick fluoropolymer layer deposition. The wettability of the samples is characterized in terms of tilting plate and bouncing drop experiments, while the roughness is analyzed by confocal microscopy. The durability of the produced surfaces was also explored.

Keywords: galvanaized steel, superhydrophobic surfaces, sintering nanoparticles, zinc nanopowder

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860 Effect of Fiber Inclusion on the Geotechnical Parameters of Clayey Soil Subjected to Freeze-Thaw Cycles

Authors: Arun Prasad, P. B. Ramudu, Deep Shikha, Deep Jyoti Singh

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A number of studies have been conducted recently to investigate the influence of randomly oriented fibers on some engineering properties of cohesive soils.Freezing and thawing of soil affects the strength, durability and permeability of soil adversely. Experiments were carried out in order to investigate the effect of inclusion of randomly distributed polypropylene fibers on the strength, hydraulic conductivity and durability of local soil (CL) subjected to freeze–thaw cycles. For evaluating the change in strength of soil, a series of unconfined compression tests as well as tri-axial tests were carried out on reinforced and unreinforced soil samples. All the samples were subjected to seven cycles of freezing and thawing. Freezing was carried out at a temperature of - 15 to -18 °C; and thawing was carried out by keeping the samples at room temperature. The reinforcement of soil samples was done by mixing with polypropylene fibers, 12 mm long and with an aspect ratio of 240. The content of fibers was varied from 0.25 to 1% by dry weight of soil. The maximum strength of soil was found in samples having a fiber content of 0.75% for all the samples that were prepared at optimum moisture content (OMC), and if the OMC was increased (+2% OMC) or decreased (-2% OMC), the maximum strength observed at 0.5% fiber inclusion. The effect of fiber inclusion and freeze–thaw on the hydraulic conductivity was studied increased from around 25 times to 300 times that of the unreinforced soil, without subjected to any freeze-thaw cycles. For studying the increased durability of soil, mass loss after each freeze-thaw cycle was calculated and it was found that samples reinforced with polypropylene fibers show 50-60% less loss in weight than that of the unreinforced soil.

Keywords: fiber reinforcement, freezingand thawing, hydraulic conductivity, unconfined compressive strength

Procedia PDF Downloads 385
859 Performance Evaluation and Plugging Characteristics of Controllable Self-Aggregating Colloidal Particle Profile Control Agent

Authors: Zhiguo Yang, Xiangan Yue, Minglu Shao, Yue Yang, Rongjie Yan

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It is difficult to realize deep profile control because of the small pore-throats and easy water channeling in low-permeability heterogeneous reservoir, and the traditional polymer microspheres have the contradiction between injection and plugging. In order to solve this contradiction, the controllable self-aggregating colloidal particles (CSA) containing amide groups on the surface of microspheres was prepared based on emulsion polymerization of styrene and acrylamide. The dispersed solution of CSA colloidal particles, whose particle size is much smaller than the diameter of pore-throats, was injected into the reservoir. When the microspheres migrated to the deep part of reservoir, , these CSA colloidal particles could automatically self-aggregate into large particle clusters under the action of the shielding agent and the control agent, so as to realize the plugging of the water channels. In this paper, the morphology, temperature resistance and self-aggregation properties of CSA microspheres were studied by transmission electron microscopy (TEM) and bottle test. The results showed that CSA microspheres exhibited heterogeneous core-shell structure, good dispersion, and outstanding thermal stability. The microspheres remain regular and uniform spheres at 100℃ after aging for 35 days. With the increase of the concentration of the cations, the self-aggregation time of CSA was gradually shortened, and the influence of bivalent cations was greater than that of monovalent cations. Core flooding experiments showed that CSA polymer microspheres have good injection properties, CSA particle clusters can effective plug the water channels and migrate to the deep part of the reservoir for profile control.

Keywords: heterogeneous reservoir, deep profile control, emulsion polymerization, colloidal particles, plugging characteristic

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858 Combustion Characteristic of Propane/Acetylene Fuel Blends Pool Fire

Authors: Yubo Bi, Xiao Chen, Shouxiang Lu

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A kind of gas-fueled burner, named Burning Rate Emulator, was proposed for the purpose of the emulation of condensed fuel recently. The gaseous fuel can be pure combustible fuel gas or blends of gaseous fuel or inert gas. However, this concept was recently proposed without detailed study on the combustion characteristic of fuel blends. In this study, two kinds of common gaseous fuels were selected, propane and acetylene, to provide the combustion heat as well as a large amount of smoke, which widely exists in liquid and solid fuel burning process. A set of experiments were carried out using a gas-fueled burner with a diameter of 8 cm. The total volume flow rate of propane and acetylene was kept at 3 liters per minute. The volume fraction of propane varied from 0% to 100% at interval of 10%. It is found that the flame height increases with propane volume fraction, which may be caused by the increase of heat release rate, as the energy density of propane is larger than that of acetylene. The dimensionless flame height is correlated against dimensionless heat release rate, which shows a power function relationship. The radiation fraction of the flame does not show a monotonic relationship with propane volume fraction. With the increase of propane volume fraction from 0% to 100%, the value of radiation fraction increases first and reach a maximum value around 0.46 at a propane volume fraction of 10%, and then decreases continuously to a value of 0.25 at the propane volume fraction of 100%. The flame radiation is related to the soot in the flame. The trend of the radiation fraction reflects that there may be a synergistic effect of soot formation between propane and acetylene which can be guessed from the significantly high radiation fraction at a propane volume fraction of 10%. This work provides data for combustion of gaseous fuel blends pool fire and also give reference on the design of Burning Rate Emulator.

Keywords: Burning Rate Emulator, fuel blends pool fire, flame height, radiation fraction

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857 Solving LWE by Pregressive Pumps and Its Optimization

Authors: Leizhang Wang, Baocang Wang

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General Sieve Kernel (G6K) is considered as currently the fastest algorithm for the shortest vector problem (SVP) and record holder of open SVP challenge. We study the lattice basis quality improvement effects of the Workout proposed in G6K, which is composed of a series of pumps to solve SVP. Firstly, we use a low-dimensional pump output basis to propose a predictor to predict the quality of high-dimensional Pumps output basis. Both theoretical analysis and experimental tests are performed to illustrate that it is more computationally expensive to solve the LWE problems by using a G6K default SVP solving strategy (Workout) than these lattice reduction algorithms (e.g. BKZ 2.0, Progressive BKZ, Pump, and Jump BKZ) with sieving as their SVP oracle. Secondly, the default Workout in G6K is optimized to achieve a stronger reduction and lower computational cost. Thirdly, we combine the optimized Workout and the Pump output basis quality predictor to further reduce the computational cost by optimizing LWE instances selection strategy. In fact, we can solve the TU LWE challenge (n = 65, q = 4225, = 0:005) 13.6 times faster than the G6K default Workout. Fourthly, we consider a combined two-stage (Preprocessing by BKZ- and a big Pump) LWE solving strategy. Both stages use dimension for free technology to give new theoretical security estimations of several LWE-based cryptographic schemes. The security estimations show that the securities of these schemes with the conservative Newhope’s core-SVP model are somewhat overestimated. In addition, in the case of LAC scheme, LWE instances selection strategy can be optimized to further improve the LWE-solving efficiency even by 15% and 57%. Finally, some experiments are implemented to examine the effects of our strategies on the Normal Form LWE problems, and the results demonstrate that the combined strategy is four times faster than that of Newhope.

Keywords: LWE, G6K, pump estimator, LWE instances selection strategy, dimension for free

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856 Optimization of Ultrasound Assisted Extraction of Polysaccharides from Plant Waste Materials: Selected Model Material is Hazelnut Skin

Authors: T. Yılmaz, Ş. Tavman

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In this study, optimization of ultrasound assisted extraction (UAE) of hemicellulose based polysaccharides from plant waste material has been studied. Selected material is hazelnut skin. Extraction variables for the operation are extraction time, amplitude and application temperature. Optimum conditions have been evaluated depending on responses such as amount of wet crude polysaccharide, total carbohydrate content and dried sample. Pretreated hazelnut skin powders were used for the experiments. 10 grams of samples were suspended in 100 ml water in a jacketed vessel with additional magnetic stirring. Mixture was sonicated by immersing ultrasonic probe processor. After the extraction procedures, ethanol soluble and insoluble sides were separated for further examinations. The obtained experimental data were analyzed by analysis of variance (ANOVA). Second order polynomial models were developed using multiple regression analysis. The individual and interactive effects of applied variables were evaluated by Box Behnken Design. The models developed from the experimental design were predictive and good fit with the experimental data with high correlation coefficient value (R2 more than 0.95). Extracted polysaccharides from hazelnut skin are assumed to be pectic polysaccharides according to the literature survey of Fourier Transform Spectrometry (FTIR) analysis results. No more change can be observed between spectrums of different sonication times. Application of UAE at optimized condition has an important effect on extraction of hemicellulose from plant material by satisfying partial hydrolysis to break the bounds with other components in plant cell wall material. This effect can be summarized by varied intensity of microjets and microstreaming at varied sonication conditions.

Keywords: hazelnut skin, optimization, polysaccharide, ultrasound assisted extraction

Procedia PDF Downloads 314
855 Investigation of the Effect of Anaerobic Digestate on Antifungal Activity and in Different Parameters of Maize

Authors: Nazia Zaffar, Alam Khan, Abdul Haq, Malik Badshah

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Pakistan is an agricultural country. The increasing population leads to an increase in demand for food. A large number of crops are infected by different microbes, and nutrient deficiency of soil adversely affects the yield of crops. Furthermore, the use of chemical fertilizers like Nitrogen, Phosphorus, Potassium (NPK) Urea, and Diammonium phosphate (DAP) and pesticides have environmental consequences. Therefore, there is an urgent need to explore alternative renewable and sustainable biofertilizers. Maize is one of the top growing crops in Pakistan, but it has low yield compared to other countries due to deficiency of organic matter, widespread nutrients deficiency (phosphorus and nitrogen), unbalanced use of fertilizers and various fungal diseases. In order to get rid of all these disadvantages, Digestate emerged as a win-win opportunity for the control of a few plant diseases and a replacement for the chemical fertilizers. The present study was designed to investigate the effect of Anerobic digestate on Antifungal Activity and in different parameters of Maize. The antifungal activity, minimum inhibitory concentration (MIC), and minimum fungicidal concentration (MFC) against selected phytopathogens (Colletotrichum coccodis, Pythium ultimum, Phytophthora capsci, Rhizoctonia solani, Bipolaris oryzae and Fusarium Fujikuroi) were determined by microtiter plate method. The effect of various fertilizers in different growth parameters height, diameter, chlorophyll, leaf area, biomass, and yield were studied in field experiments. The extracts from anaerobic digestate have shown antifungal activity against selected phytopathogens, the highest activity was noted against P. ultimum, the MIC activity was high in case of P. ultimum and B. oryzae. The present study concludes that anaerobic digestate have a positive effect on maize growth and yield as well as an antifungal activity which can be potentially a good biofertilizer.

Keywords: anaerobic digestate, antifungal activity, MIC, phytopathogens

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854 Analysis of Rock Cutting Progress with a New Axe-Shaped PDC Cutter to Improve PDC Bit Performance in Elastoplastic Formation

Authors: Fangyuan Shao, Wei Liu, Deli Gao

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Polycrystalline diamond compact (PDC) bits have occupied a large market of unconventional oil and gas drilling. The application of PDC bits benefits from the efficient rock breaking of PDC cutters. In response to increasingly complex formations, many shaped cutters have been invited, but many of them have not been solved by the mechanism of rock breaking. In this paper, two kinds of PDC cutters: a new axe-shaped (NAS) cutter and cylindrical cutter (benchmark) were studied by laboratory experiments. NAS cutter is obtained by optimizing two sides of axe-shaped cutter with curved surfaces. All the cutters were put on a vertical turret lathe (VTL) in the laboratory for cutting tests. According to the cutting distance, the VTL tests can be divided into two modes: single-turn rotary cutting and continuous cutting. The cutting depth of cutting (DOC) was set at 1.0 mm and 2.0 mm in the former mode. The later mode includes a dry VTL test for thermal stability and a wet VTL test for wear resistance. Load cell and 3D optical profiler were used to obtain the value of cutting forces and wear area, respectively. Based on the findings of the single-turn rotary cutting VTL tests, the performance of A NAS cutter was better than the benchmark cutter on elastoplastic material cutting. The cutting forces (normal forces, tangential force, and radial force) and special mechanical energy (MSE) of a NAS cutter were lower than that of the benchmark cutter under the same condition. It meant that a NAS cutter was more efficient on elastoplastic material breaking. However, the wear resistance of a new axe-shaped cutter was higher than that of a benchmark cutter. The results of the dry VTL test showed that the thermal stability of a NAS cutter was higher than that of a benchmark cutter. The cutting efficiency can be improved by optimizing the geometric structure of the PDC cutter. The change of thermal stability may be caused by the decrease of the contact area between cutter and rock at given DOC. The conclusions of this paper can be used as an important reference for PDC cutters designers.

Keywords: axe-shaped cutter, PDC cutter, rotary cutting test, vertical turret lathe

Procedia PDF Downloads 186
853 Design, Modeling, Fabrication, and Testing of a Scaled down Hybrid Rocket Engine

Authors: Pawthawala Nancy Manish, Syed Alay Hashim

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A hybrid rocket is a rocket engine which uses propellants in two different states of matter- one is in solid and the other either gas or liquid. A hybrid rocket exhibit advantages over both liquid rockets and solid rockets especially in terms of simplicity, stop-start-restart capabilities, safety and cost. This paper deals the design and development of a hybrid rocket having paraffin wax as solid fuel and liquid oxygen as oxidizer. Due to variation of pressure in combustion chamber there is significantly change in mass flow rate, burning rate and uneven regression along the length of the grain. This project describes the working model of a hybrid propellant rocket motor. We have designed a hybrid rocket thrust chamber based on the predetermined combustion chamber pressure and the properties of hybrid propellant. This project is all ready in working condition with normal oxygen injector. Now we have planned to modify the injector design to improve the combustion property. We will use spray type injector for injecting the oxidizer. This idea will increase the performance followed by the regression rate of the solid fuel. By employing mass conservation law, oxygen mass flux, oxidizer/fuel ratio and regression rate the thrust coefficient can be obtained for our current design. CATIA V5 R20 is our design software for the complete setup. This project is fully based on experimental evaluation and the collection of combustion and flow parameters. The thrust chamber is made of stainless steel and the duration of test is around 15-20 seconds (Maximum). These experiments indicates that paraffin based fuel provides the opportunity to satisfy a broad range of mission requirements for the next generation of the hybrid rocket system.

Keywords: burning rate, liquid oxygen, mass flow rate, paraffin wax and sugar

Procedia PDF Downloads 316
852 STD-NMR Based Protein Engineering of the Unique Arylpropionate-Racemase AMDase G74C

Authors: Sarah Gaßmeyer, Nadine Hülsemann, Raphael Stoll, Kenji Miyamoto, Robert Kourist

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Enzymatic racemization allows the smooth interconversion of stereocenters under very mild reaction conditions. Racemases find frequent applications in deracemization and dynamic kinetic resolutions. Arylmalonate decarboxylase (AMDase) from Bordetella Bronchiseptica has high structural similarity to amino acid racemases. These cofactor-free racemases are able to break chemically strong CH-bonds under mild conditions. The racemase-like catalytic machinery of mutant G74C conveys it a unique activity in the racemisation of pharmacologically relevant derivates of 2-phenylpropionic acid (profenes), which makes AMDase G74C an interesting object for the mechanistic investigation of cofactor-independent racemases. Structure-guided protein engineering achieved a variant of this unique racemase with 40-fold increased activity in the racemisation of several arylaliphatic carboxylic acids. By saturation–transfer–difference NMR spectroscopy (STD-NMR), substrate binding during catalysis was investigated. All atoms of the substrate showed interactions with the enzyme. STD-NMR measurements revealed distinct nuclear Overhauser effects in experiments with and without molecular conversion. The spectroscopic analysis led to the identification of several amino acid residues whose variation increased the activity of G74C. While single-amino acid exchanges increased the activity moderately, structure-guided saturation mutagenesis yielded a quadruple mutant with a 40 times higher reaction rate. This study presents STD-NMR as versatile tool for the analysis of enzyme-substrate interactions in catalytically competent systems and for the guidance of protein engineering.

Keywords: racemase, rational protein design, STD-NMR, structure guided saturation mutagenesis

Procedia PDF Downloads 286
851 The Dynamic of Nₘᵢₙ in Clay Loam Cambisol in Alternative Farming

Authors: Danute Jablonskyte-Rasce, Laura Masilionyte

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The field experiments of different farming systems were conducted at Joniškėlis Experimental Station of the Lithuanian Research Centre for Agriculture and Forestry in 2006–2016. The soil of the experimental site was Endocalcari-Endohypogleyic Cambisol (CMg-n-w-can). The research was designed to identify the effects of dry matter and nitrogen accumulated in the above-ground biomass of various catch crops grown after winter wheat on soil mineral nitrogen variation during the autumn and spring period in the presence of intensive leaching complex. Research was done in the soil differing in humus status in the organic and sustainable cropping systems by growing various plant mixtures as catch crops: narrow-leafed lupine (Lupinus angustifolius L.) and oil radish (Raphanus sativus var. Oleifera L.), white mustard (Sinapis alba L.) and buckwheat (Fagopyrum exculentum Moench.) and white mustard as a sole crop. All crop and soil management practices have shown optimal efficiency in late autumn – stubble breaking, catch crops and straw used during the post-harvest period of the main crops, reduced Nmin migration into deeper (40–80 cm) soil layer. The greatest Nmin reduction in the 0–40 cm soil layer during the period from late autumn to early spring was identified in the sustainable cropping system having applied N30 for the promotion of straw mineralization and with no catch crops cultivation. The sustainable cropping system, having applied N30 for straw mineralization and growing white mustard in combination with buckwheat as catch crops, Nmin difference in the spring compared with its status in the autumn in the soil low and moderate in humus was lower by 70.1% and 34.2%, respectively.

Keywords: soil nitrogen, catch crops, ecological and sustainable farming systems, Cambisol

Procedia PDF Downloads 244