Search results for: batch injection

Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

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Authors: Jeremy Moloney

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A major oil and gas operator in western Canada producing approximately 50,000 BOE per day of sour fluids was experiencing increased water production along with decreased oil production over several years. The higher water volumes being produced meant an increase in the operator’s incumbent corrosion inhibitor (CI) chemical requirements but with reduced oil production revenues. Thus, a cost-effective corrosion inhibitor solution was sought to deliver enhanced corrosion mitigation of the carbon steel pipeline infrastructure but at reduced chemical injection dose rates. This paper presents the laboratory work conducted on the development of a corrosion inhibitor under the operator’s simulated sour operating conditions and then subsequent field testing of the product. The new CI not only provided extremely good levels of general and localized corrosion inhibition and outperformed the incumbent CI under the laboratory test conditions but did so at vastly lower concentrations. In turn, the novel CI product facilitated field chemical injection rates to be optimized and reduced by 40% compared with the incumbent whilst maintaining superior corrosion protection resulting in significant cost savings and associated sustainability benefits for the operator.

Keywords: carbon steel, sour gas, hydrogen sulphide, localized corrosion, pitting, corrosion inhibitor

Procedia PDF Downloads 51

Authors: Debabrata Adhikari, Mikhail Matveev, Louise Brown, Jan Kočí, Andy Long

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Resin transfer molding (RTM) is increasingly used for manufacturing high-quality composite structures due to its additional advantages over prepregs of low-cost out-of-autoclave processing. However, to retain the advantages, it is critical to reduce the void content during the injection. Reinforcements commonly used in RTM, such as woven fabrics, have dual-scale porosity with mesoscale pores between the yarns and the micro-scale pores within the yarns. Due to the fabric geometry and the nature of the dual-scale flow, the flow front during injection creates a complicated fingering formation which leads to void formation. Analytical modeling of void formation for woven fabrics has been widely studied elsewhere. However, there is scope for improvement to the reduction in void formation in 3D fabrics wherein the in-plane yarn layers are confined by additional through-thickness binder yarns. In the present study, the structural morphology of the tortuous pore spaces in the 3D fabric has been studied and implemented using open-source software TexGen. An analytical model for the void and the fingering formation has been implemented based on an idealized unit cell model of the 3D fabric. Since the pore spaces between the yarns are free domains, the region is treated as flow-through connected channels, whereas intra-yarn flow has been modeled using Darcy’s law with an additional term to account for capillary pressure. Later the void fraction has been characterised using the criterion of void formation by comparing the fill time for inter and intra yarn flow. Moreover, the dual-scale two-phase flow of resin with air has been simulated in the commercial CFD solver OpenFOAM/ANSYS to predict the probable location of voids and validate the analytical model. The use of an idealised unit cell model will give the insight to optimise the mesoscale geometry of the reinforcement and injection parameters to minimise the void content during the LCM process.

Keywords: 3D fiber, void formation, RTM, process modelling

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Authors: Junhai Liao, Yansong Shen, Aibing Yu

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A 3D numerical model is developed to simulate the complicated in-furnace combustion phenomena in the lower part of an ironmaking blast furnace (BF) while using pulverized coal injection (PCI) technology to reduce the consumption of relatively expensive coke. The computational domain covers blowpipe-tuyere-raceway-coke bed in the BF. The model is validated against experimental data in terms of gaseous compositions and coal burnout. Parameters, such as coal properties and some key operational variables, play an important role on the performance of coal combustion. Their diverse effects on different combustion characteristics are examined in the domain, in terms of gas compositions, temperature, and burnout. The heat generated by the combustion of upgraded Victorian brown coal is able to meet the heating requirement of a BF, hence making upgraded brown coal injected into BF possible. It is evidenced that the model is suitable to investigate the mechanism of the PCI operation in a BF. Prediction results provide scientific insights to optimize and control of the PCI operation. This model cuts the cost to investigate and understand the comprehensive combustion phenomena of upgraded Victorian brown coal in a full-scale BF.

Keywords: blast furnace, numerical study, pulverized coal injection, Victorian brown coal

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Authors: Ana Katrina C. Longos, Jarungchai Anton S. Vatanagul

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Objectives: To present a case of two Filipino male siblings initially seen with parkinsonism and eventually with dystonia and to present botulinum A toxin as part of the treatment for X-linked dystonia parkinsonism in Cebu City. Discussion: A 54 year old man presented initially with parkinsonian symptoms and later developed oromandibular and truncal dystonia. Further history revealed that he had an older brother who also presented with the same symptoms. Neuroimaging done on both patients revealed CVD infarcts in the pons and corona radiata respectively which where were not compatible with their symptoms. Family history revealed that their mother was originally from Panay and a diagnosis of X-linked dystonia parkinsonism (XDP) was made. Both patients were able to receive botulinum A toxin injections which provided temporary relief of symptoms. Conclusion: XDP was considered in 2 Filipino male siblings who presented with oromandibular dystonia, truncal dystonia, shuffling gait, resting tremors with ancestry from Panay on the maternal side. There is no cure for XDP, only symptomatic treatment. Until recently, only oral chemotherapy was available in Cebu. Botulinum A toxin injection done in both patients afforded temporary resolution of symptoms.

Keywords: XDP, dystonia of Panay, lubag, dystonia parkinsonism, botulinum a toxin

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Authors: Peter C. St. John, Michael F. Crowley, Yannick J. Bomble

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Production of chemicals from engineered organisms in a batch culture typically involves a trade-off between productivity, yield, and titer. However, strategies for strain design typically involve designing mutations to achieve the highest yield possible while maintaining growth viability. Such approaches tend to follow the principle of designing static networks with minimum metabolic functionality to achieve desired yields. While these methods are computationally tractable, optimum productivity is likely achieved by a dynamic strategy, in which intracellular fluxes change their distribution over time. One can use multi-stage fermentations to increase either productivity or yield. Such strategies would range from simple manipulations (aerobic growth phase, anaerobic production phase), to more complex genetic toggle switches. Additionally, some computational methods can also be developed to aid in optimizing two-stage fermentation systems. One can assume an initial control strategy (i.e., a single reaction target) in maximizing productivity - but it is unclear how close this productivity would come to a global optimum. The calculation of maximum theoretical yield in metabolic engineering can help guide strain and pathway selection for static strain design efforts. Here, we present a method for the calculation of a maximum theoretical productivity of a batch culture system. This method follows the traditional assumptions of dynamic flux balance analysis: that internal metabolite fluxes are governed by a pseudo-steady state and external metabolite fluxes are represented by dynamic system including Michealis-Menten or hill-type regulation. The productivity optimization is achieved via dynamic programming, and accounts explicitly for an arbitrary number of fermentation stages and flux variable changes. We have applied our method to succinate production in two common microbial hosts: E. coli and A. succinogenes. The method can be further extended to calculate the complete productivity versus yield Pareto surface. Our results demonstrate that nearly optimal yields and productivities can indeed be achieved with only two discrete flux stages.

Keywords: A. succinogenes, E. coli, metabolic engineering, metabolite fluxes, multi-stage fermentations, succinate

Procedia PDF Downloads 190

Authors: A. Threepanich, S. Youngme, P. Praipipat

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Although ground egg shells had the ability to eliminate lead in water, their efficiency may decrease in a case of contaminating of other cations such as Na⁺, Ca²⁺ in the water. The development of ground egg shells may solve this problem in which metal oxides are a good choice for this case since they have the ability to remove any heavy metals including lead in the water. Therefore, this study attempts to use this advantage for improving ground egg shells for the specific lead removal efficiency in the water. X-ray fluorescence (XRF) technique was used for the chemical element contents analysis of ground egg shells (GES) and ground egg shells with metal oxide (GESM), and Transmission electron microscope (TEM) technique was used to examine the material sizes. The batch test studies were designed to investigate the factor effects on dose (5, 10, 15 grams), pH (5, 7, 9), and settling time (1, 3, 5 hours) for the lead removal efficiency in the water. The XRF analysis results showed GES contained calcium (Ca) 91.41% and Silicon (Si) 4.03% and GESM contained calcium (Ca) 91.41%, Silicon (Si) 4.03%, and Iron (Fe) 3.05%. TEM results confirmed the sizes of GES and GESM in the range of 1-20 nm. The batch test studies showed the best optimum conditions for the lead removal in the water of GES and GESM in dose, pH, and settling time were 10 grams, pH 9, 5 hours and 5 grams, pH 9, 3 hours, respectively. The competing ions (Na⁺ and Ca²⁺) study reported GESM had the higher % lead removal efficiency than GES at 90% and 60%, respectively. Therefore, this result can confirm that adding of metal oxide to ground egg shells helps to improve the lead removal efficiency in the water.

Keywords: nano material, ground egg shells, metal oxide, lead

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Authors: Hemabushan K., Manikandan

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In a swirl atomizer, fluid undergoes a swirling motion as a result of centrifugal force created by opposed tangential inlets in the swirl chamber. The angular momentum of fluid continually increases as it reaches the exit orifice and forms a hollow sheet. Which disintegrates to form ligaments and droplets respectively as it flows downstream. This type of atomizers used in rocket injectors and oil burner furnaces. In this present investigation a swirl atomizer with two opposed tangential inlets has been designed. Water as working fluid, experiments had been conducted for the fluid injection pressures in regime of 0.033 bar to 0.519 bar. The fluid has been pressured by a 0.5hp pump and regulated by a pressure regulator valve. Injection pressure of fluid has been measured by a U-tube mercury manometer. The spray pattern and the droplets has been captured with a high resolution camera in black background with a high intensity flash highlighting the fluid. The unprocessed images were processed in ImageJ processing software for measuring the droplet diameters and its shape characteristics along the downstream. The parameters such as mean droplet diameter and distribution, wave pattern, rupture distance and spray angle were studied for this atomizer. The above results were compared with theoretical results and also analysed for deviation with design parameters.

Keywords: swirl atomizer, injector, spray, SWIRL

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Authors: Prasamsha Panta, Da Yeon Kim, Ja Yong Jang, Min Jae Kim, Jae Ho Kim, Moon Suk Kim

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Introduction: Among the many anticancer drugs used clinically, doxorubicin (Dox), was one of widely used drugs to treat many types of solid tumors such as liver, colon, breast, or lung. Intratumoral injection of chemotherapeutic agents is a potentially more effective alternative to systemic administration because direct delivery of the anticancer drug to the target may improve both the stability and efficacy of anticancer drugs. Injectable in situ-forming gels have attracted considerable attention because they can achieve site specific drug delivery, long term action periods, and improved patient compliance. Objective: Objective of present study is to confirm clinical benefit of intratumoral chemotherapy using injectable in situ-forming poly(ethylene glycol)-b-polycaprolactone diblock copolymer (MP) and Dox with increase in efficacy and reducing the toxicity in patients with cancer diseases. Methods and methodology: We prepared biodegradable MP hydrogel and measured viscosity for the evaluation of thermo-sensitive property. In vivo antitumor activity was performed with normal saline, MP only, single free Dox, repeat free Dox, and Dox-loaded MP gel. The remaining amount of Dox drug was measured using HPLC after the mouse was sacrified. For cytotoxicity studies WST-1 assay was performed. Histological analysis was done with H&E and TUNEL processes respectively. Results: The works in this experiment showed that Dox-loaded MP have biodegradable drug depot property. Dox-loaded MP gels showed remarkable in vitro cytotoxicity activities against cancer cells. Finally, this work indicates that injection of Dox-loaded MP allowed Dox to act effectively in the tumor and induced long-lasting supression of tumor growth. Conclusion: This work has examined the potential clinical utility of intratumorally injected Dox-loaded MP gel, which shows significant effect of higher local Dox retention compared with systemically administered Dox.

Keywords: injectable in-situ forming hydrogel, anticancer, doxorubicin, intratumoral injection

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Authors: Nomthandazo Precious Sibiya, Thembisile Patience Mahlangu, Sudesh Rathilal

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Heavy metal removal is critical in the wastewater treatment process due to its numerous harmful effects on human and aquatic life. There are several chemical and physical techniques for removing heavy metals from wastewater, including ion exchange, reverse osmosis, adsorption, electrodialysis, and ultrafiltration. However, adsorption technology has captivated researchers for years due to its low cost, high efficiency, and compatible with the environment. In this study, the adsorption effectiveness of three modified agro-waste materials was explored for the removal of lead from synthetic wastewater: banana peels (BP), orange peels (OP), and sugarcane bagasse (SB). The magnetite (Fe₃O₄) is incorporated with BP, OP, and SB at a ratio of 1:1 to create magnetic biosorbents. Characterization of biosorbents was carried out using and scanning electron microscopy (SEM) combined with energy-dispersive X-ray (EDX) to investigate surface morphology and elemental compositions, respectively. A series of batch experiments were carried out to investigate the effects of adsorbent mass, agitation time, and initial pH concentration on adsorption behaviour, as well as adsorption isotherms and kinetics. The removal efficiency of lead by the modified agro-waste materials proved to be superior to that of non-modified agro-waste materials. The proof of concept was achieved, and agro-waste materials can be paired with adsorption technology to effectively remove lead from aqueous media. The use of agricultural waste as biosorbents will aid in waste reduction and management.

Keywords: adsorption, isotherms, kinetics, agro waste, nanoparticles, batch

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Authors: Ma Yuzhe, Burra Venkata Durga Kumar

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The computer is a great invention. As people use computers more and more frequently, the demand for PCs is growing, and the performance of computer hardware is also rising to face more complex processing and operation. However, the operating system, which provides the soul for computers, has stopped developing at a stage. In the face of the high price of UNIX (Uniplexed Information and Computering System), batch after batch of personal computer owners can only give up. Disk Operating System is too simple and difficult to bring innovation into play, which is not a good choice. And MacOS is a special operating system for Apple computers, and it can not be widely used on personal computers. In this environment, Linux, based on the UNIX system, was born. Linux combines the advantages of the operating system and is composed of many microkernels, which is relatively powerful in the core architecture. Linux system supports all Internet protocols, so it has very good network functions. Linux supports multiple users. Each user has no influence on their own files. Linux can also multitask and run different programs independently at the same time. Linux is a completely open source operating system. Users can obtain and modify the source code for free. Because of these advantages of Linux, it has also attracted a large number of users and programmers. The Linux system is also constantly upgraded and improved. It has also issued many different versions, which are suitable for community use and commercial use. Linux system has good security because it relies on a file partition system. However, due to the constant updating of vulnerabilities and hazards, the using security of the operating system also needs to be paid more attention to. This article will focus on the analysis and discussion of Linux security issues.

Keywords: Linux, operating system, system management, security

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Authors: A. Drapeaux, S. Aviles, E. Garfoot

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Stem cell injections are a promising alternative treatment for hip osteoarthritis. Current literature has focused on short-term outcomes for both knee and hip osteoarthritis; however, there is a significant gap for longitudinal benefits for hip OA and limited firm conclusions due to small sample sizes. The purpose of this prospective study was to determine longitudinal changes in pain, function, and radiographs following bone marrow concentrate injection (BMAC) into the osteoarthritic hip joint. Methods: A prospective, cross-sectional study was conducted over the course of 12 months at an orthopedic practice. The study recruited 15 osteoarthritic pre-surgical hips with mild to moderate osteoarthritic severity who were scheduled to undergo hip arthroscopy. Data was collected at both pre-operative and post-operative time frames. Data collected included: hip radiographs, i-HOT-33 questionnaire data, BMAC autologous volume, and demographics. Questionnaire data was captured using Qualtrics XM software, and participants were sent an anonymous link at the following time frames: pre-operative, 2 weeks, 6 weeks, 12 weeks, 6 months, 12 months, and 24 months. Radiographic changes and BMAC volume were collected and reviewed by an orthopedic surgeon and sent to the primary investigator. Data was exported and analyzed in IBM-SPSS. Results: A total of 15 hips from 15 participants (mean age: 49, gender: 50% males, 50% females, BMI: 29.7) were used in the final analysis. Summative i-HOT 33 mean scores significantly changed between pre-operative status and 2-6 weeks post-operative status (p <.001) and pre-operative status and 3-6 months post-operative status (p <.001). There were no significant changes between other post-operative phases or between pre-operative status and 12 months post-operative. Significant improvements were found between summative i-HOT 33 mean (p<.001), daily pain (p<.001), daily sitting (p=.02), daily distance walked (p =.003), and daily limp (p=0.03) and post-operative status (2-6 weeks). No significant differences between demographic variables (gender, age, tobacco use, or diabetes) and i-HOT 33 summative mean scores. Discussion/Implications: The purpose of this study was to determine longitudinal changes in pain and function following a hip joint bone marrow concentrate injection. Results indicate that participants experience a significant improvement in pain and function between pre-operative and 2-6 weeks and 3-6 months post-injection. Participants also self-reported a significant change in average daily pain with sitting and walking between pre-operation and 2-6 weeks post-operative. This study includes a larger sample size of hip osteoarthritis cases; however, future research is warranted to include random controlled trials with a larger sample size.

Keywords: adult stem cell, orthopedics, osteoarthritis (hip), patient outcome assessment

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Authors: Georgios Koronis, Arlindo Silva

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This study seeks to understand the mechanical behavior of a natural fiber reinforced composite (epoxy/flax) in more depth, utilizing both experimental and numerical methods. It is attempted to identify relationships between the design parameters and the product performance, understand the effect of noise factors and reduce process variations. Optimization of the mechanical performance of manufactured goods has recently been implemented by numerous studies for green composites. However, these studies are limited and have explored in principal mass production processes. It is expected here to discover knowledge about composite’s manufacturing that can be used to design artifacts that are of low batch and tailored to niche markets. The goal is to reach greater consistency in the performance and further understand which factors play significant roles in obtaining the best mechanical performance. A prediction of response function (in various operating conditions) of the process is modeled by the DoE. Normally, a full factorial designed experiment is required and consists of all possible combinations of levels for all factors. An analytical assessment is possible though with just a fraction of the full factorial experiment. The outline of the research approach will comprise of evaluating the influence that these variables have and how they affect the composite mechanical behavior. The coupons will be fabricated by the vacuum infusion process defined by three process parameters: flow rate, injection point position and fiber treatment. Each process parameter is studied at 2-levels along with their interactions. Moreover, the tensile and flexural properties will be obtained through mechanical testing to discover the key process parameters. In this setting, an experimental phase will be followed in which a number of fabricated coupons will be tested to allow for a validation of the design of the experiment’s setup. Finally, the results are validated by performing the optimum set of in a final set of experiments as indicated by the DoE. It is expected that after a good agreement between the predicted and the verification experimental values, the optimal processing parameter of the biocomposite lamina will be effectively determined.

Keywords: design of experiments, flax fabrics, mechanical performance, natural fiber reinforced composites

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Authors: M. Manohari, Himanshu Gupta, S. Priyadharshini, R.Santhanam, S.Chandrasekaran, B|.Venkatraman

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Injection is one of the potential routes of intake in a radioactive facility. The internal dose due to this intake is monitored at the radiation emergency medical centre, IGCAR using a portable planar HPGe detector. The contaminated wound may be having different shapes. In a reprocessing potential of wound contamination with actinide is more. Efficiency is one of the input parameters for estimation of internal dose. Estimating these efficiencies experimentally would be tedious and cumbersome. Numerical estimation can be a supplement to experiment. As an initial step in this study 241Am contamination of different shapes are studied. In this study portable planar HPGe detector was modeled using Monte Carlo code FLUKA and the effect of different parameters like distance of the contamination from the detector, radius of the circular contamination were studied. Efficiency values for point and surface contamination located at different distances were estimated. The effect of efficiency on the radius of the surface source was more predominant when the source is at 1 cm distance compared to when the source to detector distance is 10 cm. At 1 cm the efficiency decreased quadratically as the radius increased and at 10 cm it decreased linearly. The point source efficiency varied exponentially with source to detector distance.

Keywords: Planar HPGe, efficiency value, injection, surface source

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Authors: Tae Hyun Ahn, Gyo Woo Lee, Man Young Kim

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Because of high thermal efficiency and low CO2 emission, diesel engines are being used widely in many industrial fields although it makes many PM and NOx which give both human health and environment a negative effect. NOx regulations for diesel engines, however, are being strengthened and it is impossible to meet the emission standard without NOx reduction devices such as SCR (Selective Catalytic Reduction), LNC (Lean NOx Catalyst), and LNT (Lean NOx Trap). Among the NOx reduction devices, urea-SCR system is known as the most stable and efficient method to solve the problem of NOx emission. But this device has some issues associated with the ammonia slip phenomenon which is occurred by shortage of evaporation and thermolysis time, and that makes it difficult to achieve uniform distribution of the injected urea in front of monolith. Therefore, this study has focused on the mixing enhancement between urea and exhaust gases to enhance the efficiency of the SCR catalyst equipped in catalytic muffler by changing inlet gas temperature and spray conditions to improve the spray uniformity of the urea water solution. Finally, it can be found that various parameters such as inlet gas temperature and injector and injection angles significantly affect the evaporation and mixing of the urea water solution with exhaust gases, and therefore, optimization of these parameters are required.

Keywords: UWS (Urea-Water-Solution), selective catalytic reduction (SCR), evaporation, thermolysis, injection

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Authors: R. Gopakumar, G. Nagarajan

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Both DI and IDI systems possess inherent advantages as well as disadvantages. The objective of the present work is to obtain maximum advantages of both systems by implementing a hybrid design. A hybrid combustion chamber design consists of two combustion chambers viz., the main combustion chamber and an auxiliary combustion chamber. A fuel injector supplies major quantity of fuel to the auxiliary chamber. Due to the increased swirl motion in auxiliary chamber, mixing becomes more efficient which contributes to reduction in soot/particulate emissions. Also, by increasing the fuel injection pressure, NOx emissions can be reduced. The main objective of the hybrid combustion chamber design is to merge the positive features of both DI and IDI combustion chamber designs, which provides increased swirl motion and improved thermal efficiency. Due to the efficient utilization of fuel, low specific fuel consumption can be ensured. This system also aids in increasing the power output for same compression ratio and injection timing as compared with the conventional combustion chamber designs. The present system also reduces heat transfer and fluid dynamic losses which are encountered in IDI diesel engines. Since the losses are reduced, overall efficiency of the engine increases. It also minimizes the combustion noise and NOx emissions in conventional DI diesel engines.

Keywords: DI, IDI, hybrid combustion, diesel engines

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Authors: Shreya Poddar

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Legal drafting and revising are recognized as highly demanding tasks for legal professionals. This paper introduces an approach to automate and refine these processes through the use of advanced Artificial Intelligence (AI). The method employs Large Language Models (LLMs), with a specific focus on 'Chain of Thoughts' (CoT) and knowledge injection via prompt engineering. This approach differs from conventional methods that depend on comprehensive training or fine-tuning of models with extensive legal knowledge bases, which are often expensive and time-consuming. The proposed method incorporates knowledge injection directly into prompts, thereby enabling the AI to generate more accurate and contextually appropriate legal texts. This approach substantially decreases the necessity for thorough model training while preserving high accuracy and relevance in drafting. Additionally, the concept of guardrails is introduced. These are predefined parameters or rules established within the AI system to ensure that the generated content adheres to legal standards and ethical guidelines. The practical implications of this method for legal work are considerable. It has the potential to markedly lessen the time lawyers allocate to document drafting and revision, freeing them to concentrate on more intricate and strategic facets of legal work. Furthermore, this method makes high-quality legal drafting more accessible, possibly reducing costs and expanding the availability of legal services. This paper will elucidate the methodology, providing specific examples and case studies to demonstrate the effectiveness of 'Chain of Thoughts' and knowledge injection in legal drafting. The potential challenges and limitations of this approach will also be discussed, along with future prospects and enhancements that could further advance legal work. The impact of this research on the legal industry is substantial. The adoption of AI-driven methods by legal professionals can lead to enhanced efficiency, precision, and consistency in legal drafting, thereby altering the landscape of legal work. This research adds to the expanding field of AI in law, introducing a method that could significantly alter the nature of legal drafting and practice.

Keywords: AI-driven legal drafting, legal automation, futureoflegalwork, largelanguagemodels

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Authors: M. Karzhauov, А. Mukhambetova, M. Sarsenova, E. Raimagambetov, V. Ogay

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Regeneration of injured articular cartilage remains one of the most difficult and unsolved problems in traumatology and orthopedics. Currently, for the treatment of cartilage defects surgical techniques for stimulation of the regeneration of cartilage in damaged joints such as multiple microperforation, mosaic chondroplasty, abrasion and microfractures is used. However, as shown by clinical practice, they can not provide a full and sustainable recovery of articular hyaline cartilage. In this regard, the current high hopes in the regeneration of cartilage defects reasonably are associated with the use of tissue engineering approaches to restore the structural and functional characteristics of damaged joints using stem cells, growth factors and biopolymers or scaffolds. The purpose of the present study was to investigate the effects of chondroinductive growth factors and synovium-derived mesenchymal stem cells (SD-MSCs) on the regeneration of cartilage defects in rabbits. SD-MSCs were isolated from the synovium membrane of Flemish giant rabbits, and expanded in complete culture medium α-MEM. Rabbit SD-MSCs were characterized by CFU-assay and by their ability to differentiate into osteoblasts, chondrocytes and adipocytes. The effects of growth factors (TGF-β1, BMP-2, BMP-4 and IGF-I) on MSC chondrogenesis were examined in micromass pellet cultures using histological and biochemical analysis. Articular cartilage defect (4mm in diameter) in the intercondylar groove of the patellofemoral joint was performed with a kit for the mosaic chondroplasty. The defect was made until subchondral bone plate. Delivery of SD-MSCs and growth factors was conducted in combination with hyaloronic acid (HA). SD-MSCs, growth factors and control groups were compared macroscopically and histologically at 10, 30, 60 and 90 days aftrer intra-articular injection. Our in vitro comparative study revealed that TGF-β1 and BMP-4 are key chondroinductive factors for both the growth and chondrogenesis of SD-MSCs. The highest effect on MSC chondrogenesis was observed with the synergistic interaction of TGF-β1 and BMP-4. In addition, biochemical analysis of the chondrogenic micromass pellets also revealed that the levels of glycosaminoglycans and DNA after combined treatment with TGF-β1 and BMP-4 was significantly higher in comparison to individual application of these factors. In vivo study showed that for complete regeneration of cartilage defects with intra-articular injection of SD-MSCs with HA takes time 90 days. However, single injection of SD-MSCs in combiantion with TGF-β1, BMP-4 and HA significantly promoted regeneration rate of the cartilage defects in rabbits. In this case, complete regeneration of cartilage defects was observed in 30 days after intra-articular injection. Thus, our in vitro and in vivo study demonstrated that combined application of rabbit SD-MSC with chondroinductive growth factors and HA results in strong synergistic effect on the chondrogenesis significantly enhancing regeneration of the damaged cartilage.

Keywords: Mesenchymal stem cells, synovium, chondroinductive factors, TGF-β1, BMP-2, BMP-4, IGF-I

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Authors: Y. Chiang, J. R. Wang, J. H. Yang, Y. S. Tseng, C. Shih, S. W. Chen

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Kuosheng nuclear power plant (NPP) is a BWR/6 plant in Taiwan. There is more concern for the safety of Spent Fuel Pools (SFPs) in Taiwan after Fukushima event. In order to estimate the safety of Kuosheng NPP SFP, by using MELCOR2.1 and SNAP, the safety analysis of Kuosheng NPP SFP was performed combined with the mitigation strategy of NEI 06-12 report. There were several steps in this research. First, the Kuosheng NPP SFP models were established by MELCOR2.1/SNAP. Second, the Station Blackout (SBO) analysis of Kuosheng SFP was done by TRACE and MELCOR under the cooling system failure condition. The results showed that the calculations of MELCOR and TRACE were very similar in this case. Second, the mitigation strategy analysis was done with the MELCOR model by following the NEI 06-12 report. The results showed the effectiveness of NEI 06-12 strategy in Kuosheng NPP SFP. Finally, a sensitivity study of SFP quenching was done to check the differences of different water injection time and the phenomena during the quenching. The results showed that if the cladding temperature was over 1600 K, the water injection may have chance to cause the accident more severe with more hydrogen generation. It was because of the oxidation heat and the “Breakaway” effect of the zirconium-water reaction. An animation model built by SNAP was also shown in this study.

Keywords: MELCOR, SNAP, spent fuel pool, quenching

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Authors: Suraya Mohamad Nadzir, Badrol Ahmad, Norlia Berahim

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T91 steel has been used as construction material for superheater tubes in sub-critical and super critical boiler. This steel was developed with higher creep strength property as compared to conventional low alloy steel. However, this steel is also susceptible to materials degradation due to its sensitivity to heat treatment especially Post Weld Heat Treatment (PWHT) after weld repair process. Review of PWHT process shows that the holding temperature may different from one batch to other batch of samples depending on the material composition. This issue was reviewed by many researchers and one of the potential solutions is the development of weld repair process without PWHT. This process is possible with the use of temper bead welding technique. However, study has shown the hardness value across the weld joint with exception of PWHT is much higher compare to recommended hardness value. Based on the above findings, a study to evaluate the microstructure and hardness changes of T91 weld joint after heating at 560°C at varying duration was carried out. This study was carried out to evaluate the possibility of self-tempering process during in-service period. In this study, the T91 weld joint was heat-up in air furnace at 560°C for duration of 50 and 150 hours. The heating process was controlled with heating rate of 200°C/hours, and cooling rate about 100°C/hours. Following this process, samples were prepared for the microstructure examination and hardness evaluation. Results have shown full tempered martensite structure and acceptance hardness value was achieved after 50 hours heating. This result shows that the thin component such as T91 superheater tubes is able to self-tempering during service hour.

Keywords: T91, weld-joint, tempered martensite, self-tempering

Procedia PDF Downloads 351

Authors: Seyed Esmail Seyedi Bariran, Khairul Salleh Mohamed Sahari

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Apparent quality defects such as warpage, shrinkage, weld line, etc. are such an irresistible phenomenon in mass production of auto plastic appearance parts. These frequently occurred manufacturing defects should be satisfied concurrently so as to achieve a final product with acceptable quality standards. Determining the significant control factors that simultaneously affect multiple quality characteristics can significantly improve the optimization results by eliminating the deviating effect of the so-called ineffective outliers. Hence, a robust quantitative approach needs to be developed upon which major control factors and their level can be effectively determined to help improve the reliability of the optimal processing parameter design. Hence, the primary objective of current study was to develop a systematic methodology for selection of significant control factors (SCF) relevant to multiple quality optimization of auto plastic appearance part. Auto bumper was used as a specimen with the most identical quality and production characteristics to APAP group. A preliminary failure modes and effect analysis (FMEA) was conducted to nominate a database of pseudo significant significant control factors prior to the optimization phase. Later, CAE simulation Moldflow analysis was implemented to manipulate four rampant plastic injection quality defects concerned with APAP group including warpage deflection, volumetric shrinkage, sink mark and weld line. Furthermore, a step-backward elimination searching method (SESME) has been developed for systematic pre-optimization selection of SCF based on hierarchical orthogonal array design and priority-based one-way analysis of variance (ANOVA). The development of robust parameter design in the second phase was based on DOE module powered by Minitab v.16 statistical software. Based on the F-test (F 0.05, 2, 14) one-way ANOVA results, it was concluded that for warpage deflection, material mixture percentage was the most significant control factor yielding a 58.34% of contribution while for the other three quality defects, melt temperature was the most significant control factor with a 25.32%, 84.25%, and 34.57% contribution for sin mark, shrinkage and weld line strength control. Also, the results on the he least significant control factors meaningfully revealed injection fill time as the least significant factor for both warpage and sink mark with respective 1.69% and 6.12% contribution. On the other hand, for shrinkage and weld line defects, the least significant control factors were holding pressure and mold temperature with a 0.23% and 4.05% overall contribution accordingly.

Keywords: plastic injection moulding, quality optimization, FMEA, ANOVA, SESME, APAP

Procedia PDF Downloads 326

Authors: A. Zarenezhad, A. Esfandiari, E. Zarenezhad, M. Mardkhoshnood

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The aim of this study was to investigate the ultra-structure of the photoreceptor layer of male rats under the effect of methanol intoxication and protective effect of silymarin against the methanol toxicity. Fifteen adult male rats were divided into three groups: Control group, Experimental group I (received 4g/kg methanol by intraperitoneal injection for five days), Experimental group II (received 4 g/kg methanol by intraperitoneal injection for five days and received 250 mg/kg silymarin orally for three months). At the end of the experiment, the eyes were removed; retina was separated near the optic disc and studied by transmission electron microscope. Results showed that the retina in the experimental group I exhibited loss of outer segments and disorganization in inner segment. Increased extra cellular space, disappearance of outer limiting membrane and pyknotic nuclei were seen in this group. But normal outer segment, organized inner segment and normal outer limiting membrane were obvious after treatment with silymarin in experimental group II. These findings show that methanol causes damage in the photoreceptor layer of the rat retina and silymarin can protect the damage to retina against the methanol intoxication.

Keywords: ultra-structure, photoreceptor layer, methanol intoxication, silymarin, rat

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Authors: Yuri Boiko, Iluju Kiringa, Tet Yeap

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Current drainage attack suggested previously is further studied in regular settings of closed-loop controlled Brushless DC (BLDC) motor with Kalman filter in the feedback loop. Modeling and simulation experiments are conducted in a Matlab environment, implementing the closed-loop control model of BLDC motor operation in position sensorless mode under Kalman filter drive. The current increase in the motor windings is caused by the controller (p-controller in our case) affected by false data injection of substitution of the angular velocity estimates with distorted values. Operation of multiplication to distortion coefficient, values of which are taken from the distortion function synchronized in its periodicity with the rotor’s position change. A saw function with a triangular tooth shape is studied herewith for the purpose of carrying out the bias injection with current drainage consequences. The specific focus here is on how the asymmetry of the tooth in the saw function affects the flow of current drainage. The purpose is two-fold: (i) to produce and collect the signature of an asymmetric saw in the attack for further pattern recognition process, and (ii) to determine conditions of improving stealthiness of such attack via regulating asymmetry in saw function used. It is found that modification of the symmetry in the saw tooth affects the periodicity of current drainage modulation. Specifically, the modulation frequency of the drained current for a fully asymmetric tooth shape coincides with the saw function modulation frequency itself. Increasing the symmetry parameter for the triangle tooth shape leads to an increase in the modulation frequency for the drained current. Moreover, such frequency reaches the switching frequency of the motor windings for fully symmetric triangular shapes, thus becoming undetectable and improving the stealthiness of the attack. Therefore, the collected signatures of the attack can serve for attack parameter identification via the pattern recognition route.

Keywords: bias injection attack, Kalman filter, BLDC motor, control system, closed loop, P-controller, PID-controller, current drainage, saw-function, asymmetry

Procedia PDF Downloads 55

Authors: Natcha Ruamyat, Nichakorn Khondee

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The potential of an alkaliphilic bacteria isolated from soil in Thailand to utilized agro-industrial and agricultural wastes for the production of biosurfactants was evaluated in this study. Among five isolates, Pseudomonas mendocina NK41 used soapstock as substrate showing a high biosurfactant concentration of 7.10 g/L, oil displacement of 97.8 %, and surface tension reduction to 29.45 mN/m. Various agricultural residues were applied as mixed substrates with soapstock to enhance the synthesis of biosurfactants. The production of biosurfactant and bacterial growth was found to be the highest with coconut oil cake as compared to Sacha inchi shell, coconut kernel cake, and durian shell. The biodegradability of agro-industrial wastes was better than agricultural wastes, which allowed higher bacterial growth. The pretreatment of coconut oil cake by combined alkaline and hydrothermal method increased the production of biosurfactant from 12.69 g/L to 13.82 g/L. The higher microbial accessibility was improved by the swelling of the alkali-hydrothermal pretreated coconut oil cake, which enhanced its porosity and surface area. The pretreated coconut oil cake was reused twice in the repeated batch production, showing higher biosurfactant concentration up to 16.94 g/L from the second cycle. These results demonstrated the capability of using lignocellulosic wastes from agricultural and agro-industrial activities to produce a highly valuable biosurfactant. High biosurfactant yield with low-cost substrate reveals its potential towards further commercialization of biosurfactant on large-scale production.

Keywords: alkaliphilic bacteria, agricultural/agro-industrial wastes, biosurfactant, combined alkaline-hydrothermal pretreatment

Procedia PDF Downloads 229

Authors: Oluwaseun Adeleke, Grace Amarachi Omenife, Jennifer Adebambo, Mopelola Raji, Anthony Nwala, Mogbonjubade Adesulure

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Introduction: The incorporation of digital technology in healthcare systems is instrumental in transforming the delivery, management, and overall experience of healthcare and holds the potential to scale up access through over 200 million active mobile phones used in Nigeria. Digital tools enable increased access to care, stronger client engagement, progress in research and data-driven insights, and more effective promotion of self-care and do-it-yourself practices. The Delivering Innovation in Self-Care (DISC) project 2021 has played a pivotal role in granting women greater autonomy over their sexual and reproductive health (SRH) through a variety of approaches, including information and training to self-inject contraception (DMPA-SC). To optimize its outcomes, the project also leverages digital technology platforms like social media: Facebook, Instagram, and Meet Tina (Chatbot) via WhatsApp, Customer Relationship Management (CRM) applications Freshworks, and Viamo. Methodology: The project has been successful at optimizing in-person digital cyberspace interaction to sensitize individuals effectively about self-injection and provide linkages to SI services. This platform employs the Freshworks CRM software application, along with specially trained personnel known as Cyber IPC Agents and DHIS calling centers. Integration of Freshworks CRM software with social media allows a direct connection with clients to address emerging issues, schedule follow-ups, send reminders to improve compliance with self-injection schedules, enhance the overall user experience for self-injection (SI) clients, and generate comprehensive reports and analytics on client interactions. Interaction covers a range of topics, including – How to use SI, learning more about SI, side-effects and its management, accessing services, fertility, ovulation, other family planning methods, inquiries related to Sexual Reproductive Health as well as uses an address log to connect them with nearby facilities or online pharmaceuticals. Results: Between the months of March to September, a total of 5,403 engagements were recorded. Among these, 4,685 were satisfactorily resolved. Since the program's inception, digital advertising has created 233,633,075 impressions, reached 12,715,582 persons, and resulted in 3,394,048 clicks. Conclusion: Leveraging digital technology has proven to be an invaluable tool in client management and improving client experience. The use of Cyber technology has enabled the successful development and maintenance of client relationships, which have been effective at providing support, facilitating delivery and compliance with DMPA-SC self-injection services, and ensuring overall client satisfaction. Concurrently, providing qualitative data, including user experience feedback, has enabled the derivation of crucial insights that inform the decision-making process and guide in normalizing self-care behavior.

Keywords: selfcare, DMPA-SC self-injection, digital technology, cyber technology, freshworks CRM software

Procedia PDF Downloads 42

Authors: Soraya Abtouche, Zeyneb Ghoualem, Syrine Daoudi, Lina Ouldmohamed, Xavier Assfeld

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This work reports density functional theory calculations of the optimized geometries, molecular reactivity, energy gap,and thermodynamic properties of the free base (H2P) and their Zn (II) metallated (ZnP), bearing one, two, or three carboxylic acid groups using the hybrid functional B3LYP, Cam-B3lYP, wb97xd with 6-31G(d,p) basis sets. When donating groups are attached to the molecular dye, the bond lengths are slightly decreased, which is important for the easy transfer of an electron from donating to the accepting group. For all dyes, the highest occupied molecular orbital/lowest occupied molecular orbital analysis results in positive outcomes upon electron injection to the semiconductor and subsequent dye regeneration by the electrolyte. The ionization potential increases with increasing conjugation; therefore, the compound dye attached to one carboxylic acid group has the highest ionization potential. The results show higher efficiencies of those sensitized with ZnP. These results have been explained, taking into account the electronic character of the metal ion, which acts as a mediator in the injection step, and, on the other hand, considering the number of anchoring groups to which it binds to the surface of TiO2.

Keywords: DSSC, porphyrin, TD-DFT, electronic properties, donor-acceptor groups

Procedia PDF Downloads 53

Authors: Catalina Iticescu, Lucian P. Georgescu, Mihaela Timofti, Gabriel Murariu

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Since the accumulation of large quantities of sewage sludge is producing serious environmental problems, numerous environmental specialists are looking for solutions to solve this problem. The sewage sludge obtained by treatment of municipal wastewater may be used as fertiliser on agricultural soils because such sludge contains large amounts of nitrogen, phosphorus and organic matter. In many countries, sewage sludge is used instead of chemical fertilizers in agriculture, this being the most feasible method to reduce the increasingly larger quantities of sludge. The use of sewage sludge on agricultural soils is allowed only with a strict monitoring of their physical and chemical parameters, because heavy metals exist in varying amounts in sewage sludge. Exceeding maximum permitted quantities of harmful substances may lead to pollution of agricultural soil and may cause their removal aside because the plants may take up the heavy metals existing in soil and these metals will most probably be found in humans and animals through food. The sewage sludge analyzed for the present paper was extracted from the Wastewater Treatment Station (WWTP) Galati, Romania. The physico-chemical parameters determined were: pH (upH), total organic carbon (TOC) (mg L⁻¹), N-total (mg L⁻¹), P-total (mg L⁻¹), N-NH₄ (mg L⁻¹), N-NO₂ (mg L⁻¹), N-NO₃ (mg L⁻¹), Fe-total (mg L⁻¹), Cr-total (mg L⁻¹), Cu (mg L⁻¹), Zn (mg L⁻¹), Cd (mg L⁻¹), Pb (mg L⁻¹), Ni (mg L⁻¹). The determination methods were electrometrical (pH, C, TSD) - with a portable HI 9828 HANNA electrodes committed multiparameter and spectrophotometric - with a Spectroquant NOVA 60 - Merck spectrophotometer and with specific Merck parameter kits. The tests made pointed out the fact that the sludge analysed is low heavy metal falling within the legal limits, the quantities of metals measured being much lower than the maximum allowed. The results of the tests made to determine the content of nutrients in the sewage sludge have shown that the existing nutrients may be used to increase the fertility of agricultural soils. Other tests were carried out on lands where sewage sludge was applied in order to establish the maximum quantity of sludge that may be used so as not to constitute a source of pollution. The tests were made on three plots: a first batch with no mud and no chemical fertilizers applied, a second batch on which only sewage sludge was applied, and a third batch on which small amounts of chemical fertilizers were applied in addition to sewage sludge. The results showed that the production increases when the soil is treated with sludge and small amounts of chemical fertilizers. Based on the results of the present research, a fertilization plan has been suggested. This plan should be reconsidered each year based on the crops planned, the yields proposed, the agrochemical indications, the sludge analysis, etc.

Keywords: agricultural use, crops, physico–chemical parameters, sewage sludge

Procedia PDF Downloads 263

Authors: Guijie Sang, Geo Davis, Momchil Terziev

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Geological CO2 storage has been regarded as a promising strategy to mitigate the emission of greenhouse gas generated from traditional power stations and energy-intensive industry. Caprocks with very low permeability and ultra-fine pores create viscous and capillary barriers to guarantee CO2 sealing efficiency. However, caprock fractures, either naturally existing or artificially induced due to injection, could provide preferential paths for CO₂ escaping. Seeking an efficient technique to seal and strengthen caprock fractures is crucial. We apply microbial-induced-calcite-precipitation (MICP) technique for sealing and strengthening caprock fractures in the laboratory scale. The MICP bio-grouting technique has several advantages over conventional cement grouting methods, including its low viscosity, micron-size microbes (accessible to fine apertures), and low carbon footprint, among others. Different injection strategies are tested to achieve relatively homogenous calcite precipitation along the fractures, which is monitored dynamically based on laser ultrasonic technique. The MICP process in caprock fractures, which integrates the coupled flow and bio-chemical precipitation, is also modeled and validated through the experiment. The study could provide an effective bio-mediated grouting strategy for caprock sealing and thus ensuring a long-term safe geological CO2 storage.

Keywords: caprock sealing, geological CO2 storage, grouting strategy, microbial induced calcite precipitation

Procedia PDF Downloads 154

Authors: A. Alyami, S. Christmas, K. Neeltje, G. Pollakis, B. Paxton, Z. Al-Bayati

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The Human leukocyte antigen-G (HLA-G) molecule plays an important role in immunomodulation. To date, 16 untranslated regions (UTR) HLA-G haplotypes have been previously defined by sequenced SNPs in the coding region. From these, UTR-1, UTR-2, UTR-3, UTR-4, UTR-5, UTR-6 and UTR-7 are the most frequent 3’UTR haplotypes at the global level. UTR-1 is associated with higher levels of soluble HLA-G and HLA-G expression, whereas UTR-5 and UTR-7 are linked with low levels of soluble HLA-G and HLA-G expression. Human immunodeficiency virus type 1 (HIV-1) infection results in the progressive loss of immune function in infected individuals. The virus escape mechanism typically includes T lymphocytes and NK cell recognition and lyses by classical HLA-A and B down-regulation, which has been associated with non-classical HLA-G molecule up-regulation, respectively. We evaluated the haplotypes of the HLA-G 3′ untranslated region frequencies observed in three HIV-1 groups from the Netherlands and their susceptibility to develop infection. The three groups are made up of mainly men who have sex with men (MSM), injection drug users (IDU) and a high-risk-seronegative (HRSN) group. DNA samples were amplified with published primers prior sequencing. According to our results, the low expresser frequencies show higher in HRSN compared to other groups. This is indicating that 3’UTR polymorphisms may be identified as potential prognostic biomarkers to determine susceptibility to HIV.

Keywords: Human leukocyte antigen-G (HLA-G) , men who have sex with men (MSM), injection drug users (IDU), high-risk-seronegative (HRSN) group, high-untranslated region (UTR)

Procedia PDF Downloads 128

Authors: Mariam Dianat Sabet Gilani, Lars M. Blank, Birgitta E. Ebert

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Plant-derived lupane-type, pentacyclic triterpenoids are biologically active compounds that are highly interesting for applications in medical, pharmaceutical, and cosmetic industries. Due to the low abundance of these valuable compounds in their natural sources, and the environmentally harmful downstream process, alternative production methods, such as microbial cell factories, are investigated. Engineered Saccharomyces cerevisiae strains, harboring the heterologous genes for betulinic acid synthesis, can produce up to 2 g L-1 triterpenoids, showing high potential for large-scale production of triterpenoids. One limitation of the microbial synthesis is the intracellular product accumulation. It not only makes cell disruption a necessary step in the downstream processing but also limits productivity and product yield per cell. To overcome these restrictions, the aim of this study is to develop an in-situ extraction method, which extracts triterpenoids into a second organic phase. Such a continuous or sequential product removal from the biomass keeps the cells in an active state and enables extended production time or biomass recycling. After screening of twelve different solvents, selected based on product solubility, biocompatibility, as well as environmental and health impact, isopropyl myristate (IPM) was chosen as a suitable solvent for in-situ product removal from S. cerevisiae. Impedance-based single-cell analysis and off-gas measurement of carbon dioxide emission showed that cell viability and physiology were not affected by the presence of IPM. Initial experiments demonstrated that after the addition of 20 vol % IPM to cultures in the stationary phase, 40 % of the total produced triterpenoids were extracted from the cells into the organic phase. In future experiments, the application of IPM in a repeated batch process will be tested, where IPM is added at the end of each batch run to remove triterpenoids from the cells, allowing the same biocatalysts to be used in several sequential batch steps. Due to its high biocompatibility, the amount of IPM added to the culture can also be increased to more than 20 vol % to extract more than 40 % triterpenoids in the organic phase, allowing the cells to produce more triterpenoids. This highlights the potential for the development of a continuous large-scale process, which allows biocatalysts to produce intracellular products continuously without the necessity of cell disruption and without limitation of the cell capacity.

Keywords: betulinic acid, biocompatible solvent, in-situ extraction, isopropyl myristate, process development, secondary metabolites, triterpenoids, yeast

Procedia PDF Downloads 122