Search results for: transport optimization

Authors: Doni Dermawan

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Modern sports competition as a consequence of the increase in the value of the business and entertainment in the field of sport has been demanding athletes to always have excellent physical endurance performance. Physical exercise is done in a long time, and intensive may pose a risk of muscle tissue damage caused by the increase of the enzyme creatine kinase. Branched Chain Amino Acids (BCAA) is an essential amino acid that is composed of leucine, isoleucine, and valine which serves to maintain muscle tissue, keeping the immune system, and prevent further loss of coordination and muscle pain. Pea (Pisum sativum L.) is a kind of leguminous plants that are rich in Branched Chain Amino Acids (BCAA) where every one gram of protein pea contains 82.7 mg of leucine; 56.3 mg isoleucine; and 56.0 mg of valine. This research aims to develop Branched Chain Amino Acids (BCAA) from pea extract is applied in dosage forms Gel PVP Kinesio Tape technology using Ultrasound-assisted Extraction. The method used in the writing of this paper is the Cochrane Collaboration Review that includes literature studies, testing the quality of the study, the characteristics of the data collection, analysis, interpretation of results, and clinical trials as well as recommendations for further research. Extraction of BCAA in pea done using ultrasound-assisted extraction technology with optimization variables includes the type of solvent extraction (NaOH 0.1%), temperature (20-250C), time (15-30 minutes) power (80 watt) and ultrasonic frequency (35 KHz). The advantages of this extraction method are the level of penetration of the solvent into the membrane of the cell is high and can increase the transfer period so that the BCAA substance separation process more efficient. BCAA extraction results are then applied to the polymer PVP (Polyvinylpyrrolidone) Gel powder composed of PVP K30 and K100 HPMC dissolved in 10 mL of water-methanol (1: 1) v / v. Preparations Kinesio Tape Gel PVP is the BCAA in the gel are absorbed into the muscle tissue, and joints through tensile force then provides stimulation to the muscle circulation with variable pressure so that the muscle can increase the biomechanical movement and prevent damage to the muscle enzyme creatine kinase. Analysis and evaluation of test preparation include interaction, thickness, weight uniformity, humidity, water vapor permeability, the levels of the active substance, content uniformity, percentage elongation, stability testing, release profile, permeation in vitro and in vivo skin irritation testing.

Keywords: branched chain amino acid, BCAA, Kinesio tape, pea, PVP gel, ultrasound-assisted extraction

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Authors: Shrabani Ghosh

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Automatic offensive language detection in social media has become a stirring task in today's NLP. Manual Offensive language detection is tedious and laborious work where automatic methods based on machine learning are only alternatives. Previous works have done sentiment analysis over social media in different ways such as supervised, semi-supervised, and unsupervised manner. Domain adaptation in a semi-supervised way has also been explored in NLP, where the source domain and the target domain are different. In domain adaptation, the source domain usually has a large amount of labeled data, while only a limited amount of labeled data is available in the target domain. Pretrained transformers like BERT, RoBERTa models are fine-tuned to perform text classification in an unsupervised manner to perform further pre-train masked language modeling (MLM) tasks. In previous work, hate speech detection has been explored in Gab.ai, which is a free speech platform described as a platform of extremist in varying degrees in online social media. In domain adaptation process, Twitter data is used as the source domain, and Gab data is used as the target domain. The performance of domain adaptation also depends on the cross-domain similarity. Different distance measure methods such as L2 distance, cosine distance, Maximum Mean Discrepancy (MMD), Fisher Linear Discriminant (FLD), and CORAL have been used to estimate domain similarity. Certainly, in-domain distances are small, and between-domain distances are expected to be large. The previous work finding shows that pretrain masked language model (MLM) fine-tuned with a mixture of posts of source and target domain gives higher accuracy. However, in-domain performance of the hate classifier on Twitter data accuracy is 71.78%, and out-of-domain performance of the hate classifier on Gab data goes down to 56.53%. Recently self-supervised learning got a lot of attention as it is more applicable when labeled data are scarce. Few works have already been explored to apply self-supervised learning on NLP tasks such as sentiment classification. Self-supervised language representation model ALBERTA focuses on modeling inter-sentence coherence and helps downstream tasks with multi-sentence inputs. Self-supervised attention learning approach shows better performance as it exploits extracted context word in the training process. In this work, a self-supervised attention mechanism has been proposed to detect hate speech on Gab.ai. This framework initially classifies the Gab dataset in an attention-based self-supervised manner. On the next step, a semi-supervised classifier trained on the combination of labeled data from the first step and unlabeled data. The performance of the proposed framework will be compared with the results described earlier and also with optimized outcomes obtained from different optimization techniques.

Keywords: attention learning, language model, offensive language detection, self-supervised learning

Procedia PDF Downloads 87

Authors: Hilal Turkoglu Sasmazel, Ozan Ozkan, Seda Surucu

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Tissue engineering is the field of treating defects caused by injuries, trauma or acute/chronic diseases by using artificial scaffolds that mimic the extracellular matrix (ECM), the natural biological support for the tissues and cells within the body. The main aspects of a successful artificial scaffold are (i) large surface area in order to provide multiple anchorage points for cells to attach, (ii) suitable porosity in order to achieve 3 dimensional growth of the cells within the scaffold as well as proper transport of nutrition, biosignals and waste and (iii) physical, chemical and biological compatibility of the material in order to obtain viability throughout the healing process. By hybrid scaffolds where two or more different materials were combined with advanced fabrication techniques into complex structures, it is possible to combine the advantages of individual materials into one single structure while eliminating the disadvantages of each. Adding this to the complex structure provided by advanced fabrication techniques enables obtaining the desired aspects of a successful artificial tissue scaffold. In this study, fibroblast compatibility of poly(ε-caprolactone) (PCL)/chitosan core-shell electrospun hybrid scaffolds with proper mechanical, chemical and physical properties successfully developed in our previous study was investigated. Standard 7-day cell culture was carried out with L929 fibroblast cell line. The viability of the cells cultured with the scaffolds was monitored with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay for every 48 h starting with 24 h after the initial seeding. In this assay, blank commercial tissue culture polystyrene (TCPS) Petri dishes, single electrospun PCL and single electrospun chitosan mats were used as control in order to compare and contrast the performance of the hybrid scaffolds. The adhesion, proliferation, spread and growth of the cells on/within the scaffolds were observed visually on the 3rd and the 7th days of the culture period with confocal laser scanning microscopy (CSLM) and scanning electron microscopy (SEM). The viability assay showed that the hybrid scaffolds caused no toxicity for fibroblast cells and provided a steady increase in cell viability, effectively doubling the cell density for every 48 h for the course of 7 days, as compared to TCPS, single electrospun PCL or chitosan mats. The cell viability on the hybrid scaffold was ~2 fold better compared to TCPS because of its 3D ECM-like structure compared to 2D flat surface of commercially cell compatible TCPS, and the performance was ~2 fold and ~10 fold better compared to single PCL and single chitosan mats, respectively, even though both fabricated similarly with electrospinning as non-woven fibrous structures, because single PCL and chitosan mats were either too hydrophobic or too hydrophilic to maintain cell attachment points. The viability results were verified with visual images obtained with CSLM and SEM, in which cells found to achieve characteristic spindle-like fibroblast shape and spread on the surface as well within the pores successfully at high densities.

Keywords: chitosan, core-shell, fibroblast, electrospinning, PCL

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Authors: M. Patterson, R. Bond, K. Cowan, M. Mulvenna, C. Reid, F. McMahon, P. McGowan, H. Cormican

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This paper outlines the design of a simulator to allow for the optimisation of clinical workflows through a pathology laboratory and to improve the laboratory’s efficiency in the processing, testing, and analysis of specimens. Often pathologists have difficulty in pinpointing and anticipating issues in the clinical workflow until tests are running late or in error. It can be difficult to pinpoint the cause and even more difficult to predict any issues which may arise. For example, they often have no indication of how many samples are going to be delivered to the laboratory that day or at a given hour. If we could model scenarios using past information and known variables, it would be possible for pathology laboratories to initiate resource preparations, e.g. the printing of specimen labels or to activate a sufficient number of technicians. This would expedite the clinical workload, clinical processes and improve the overall efficiency of the laboratory. The simulator design visualises the workflow of the laboratory, i.e. the clinical tests being ordered, the specimens arriving, current tests being performed, results being validated and reports being issued. The simulator depicts the movement of specimens through this process, as well as the number of specimens at each stage. This movement is visualised using an animated flow diagram that is updated in real time. A traffic light colour-coding system will be used to indicate the level of flow through each stage (green for normal flow, orange for slow flow, and red for critical flow). This would allow pathologists to clearly see where there are issues and bottlenecks in the process. Graphs would also be used to indicate the status of specimens at each stage of the process. For example, a graph could show the percentage of specimen tests that are on time, potentially late, running late and in error. Clicking on potentially late samples will display more detailed information about those samples, the tests that still need to be performed on them and their urgency level. This would allow any issues to be resolved quickly. In the case of potentially late samples, this could help to ensure that critically needed results are delivered on time. The simulator will be created as a single-page web application. Various web technologies will be used to create the flow diagram showing the workflow of the laboratory. JavaScript will be used to program the logic, animate the movement of samples through each of the stages and to generate the status graphs in real time. This live information will be extracted from an Oracle database. As well as being used in a real laboratory situation, the simulator could also be used for training purposes. ‘Bots’ would be used to control the flow of specimens through each step of the process. Like existing software agents technology, these bots would be configurable in order to simulate different situations, which may arise in a laboratory such as an emerging epidemic. The bots could then be turned on and off to allow trainees to complete the tasks required at that step of the process, for example validating test results.

Keywords: laboratory-process, optimization, pathology, computer simulation, workflow

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Authors: Huidan Yu, Anurag Deb, Rou Chen, I-Wen Wang

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The use of left ventricle assist devices (LVADs) in patients with heart failure has been a proven and effective therapy for patients with severe end-stage heart failure. Due to the limited availability of suitable donor hearts, LVADs will probably become the alternative solution for patient with heart failure in the near future. While the LVAD is being continuously improved toward enhanced performance, increased device durability, reduced size, a better understanding of implantation management becomes critical in order to achieve better long-term blood supplies and less post-surgical complications such as thrombi generation. Important issues related to the LVAD implantation include the location of outflow grafting (OG), the angle of the OG, the combination between LVAD and native heart pumping, uniform or pulsatile flow at OG, etc. We have hypothesized that an optimal implantation of LVAD is patient specific. To test this hypothesis, we employ a novel in-house computational modeling technique, named InVascular, to conduct a systematic evaluation of cardiac output at aortic arch together with other pertinent hemodynamic quantities for each patient under various implantation scenarios aiming to get an optimal implantation strategy. InVacular is a powerful computational modeling technique that integrates unified mesoscale modeling for both image segmentation and fluid dynamics with the cutting-edge GPU parallel computing. It first segments the aortic artery from patient’s CT image, then seamlessly feeds extracted morphology, together with the velocity wave from Echo Ultrasound image of the same patient, to the computation model to quantify 4-D (time+space) velocity and pressure fields. Using one NVIDIA Tesla K40 GPU card, InVascular completes a computation from CT image to 4-D hemodynamics within 30 minutes. Thus it has the great potential to conduct massive numerical simulation and analysis. The systematic evaluation for one patient includes three OG anastomosis (ascending aorta, descending thoracic aorta, and subclavian artery), three combinations of LVAD and native heart pumping (1:1, 1:2, and 1:3), three angles of OG anastomosis (inclined upward, perpendicular, and inclined downward), and two LVAD inflow conditions (uniform and pulsatile). The optimal LVAD implantation is suggested through a comprehensive analysis of the cardiac output and related hemodynamics from the simulations over the fifty-four scenarios. To confirm the hypothesis, 5 random patient cases will be evaluated.

Keywords: graphic processing unit (GPU) parallel computing, left ventricle assist device (LVAD), lumped-parameter model, patient-specific computational hemodynamics

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Authors: Michal Geca, Grzegorz Baranski, Ksenia Siadkowska

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The work is a part of the project which aims at developing innovative power and control systems for the high power aircraft piston engine ASz62IR. Developed electronically controlled ignition system will reduce emissions of toxic compounds as a result of lowered fuel consumption, optimized combustion and engine capability of efficient combustion of ecological fuels. The tested unit is an air-cooled four-stroke gasoline engine of 9 cylinders in a radial setup, mechanically charged by a radial compressor powered by the engine crankshaft. The total engine cubic capac-ity is 29.87 dm3, and the compression ratio is 6.4:1. The maximum take-off power is 1000 HP at 2200 rpm. The maximum fuel consumption is 280 kg/h. Engine powers aircrafts: An-2, M-18 „Dromader”, DHC-3 „OTTER”, DC-3 „Dakota”, GAF-125 „HAWK” i Y5. The main problems of the engine includes the imbalanced work of cylinders. The non-uniformity value in each cylinder results in non-uniformity of their work. In radial engine cylinders arrangement causes that the mixture movement that takes place in accordance (lower cylinder) or the opposite (upper cylinders) to the direction of gravity. Preliminary tests confirmed the presence of uneven workflow of individual cylinders. The phenomenon is most intense at low speed. The non-uniformity is visible on the waveform of cylinder pressure. Therefore two studies were conducted to determine the impact of this phenomenon on the engine performance: simulation and real tests. Simplified simulation was conducted on the element of the intake system coated with fuel film. The study shows that there is an effect of gravity on the movement of the fuel film inside the radial engine intake channels. Both in the lower and the upper inlet channels the film flows downwards. It follows from the fact that gravity assists the movement of the film in the lower cylinder channels and prevents the movement in the upper cylinder channels. Real tests on aircraft engine ASz62IR was conducted in transients condition (rapid change of the excess air in each cylinder were performed. Calculations were conducted for mass of fuel reaching the cylinders theoretically and really and on this basis, the factors of fuel evaporation “x” were determined. Therefore a simplified model of the fuel supply to cylinder was adopted. Model includes time constant of the fuel film τ, the number of engine transport cycles of non-evaporating fuel along the intake pipe γ and time between next cycles Δt. The calculation results of identification of the model parameters are presented in the form of radar graphs. The figures shows the averages declines and increases of the injection time and the average values for both types of stroke. These studies shown, that the change of the position of the cylinder will cause changes in the formation of fuel-air mixture and thus changes in the combustion process. Based on the results of the work of simulation and experiments was possible to develop individual algorithms for ignition control. This work has been financed by the Polish National Centre for Research and Development, INNOLOT, under Grant Agreement No. INNOLOT/I/1/NCBR/2013.

Keywords: radial engine, ignition system, non-uniformity, combustion process

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Authors: Melanie Redding

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Historic widespread nitrate contamination of the Lower Yakima Valley aquifer in Washington State initiated a community-based effort to reduce nitrate concentrations to below-drinking water standards. This group commissioned studies on characterizing local nitrogen sources, deep soil assessments, drinking water, and assessing nitrate concentrations at the water table. Nitrate is the most prevalent groundwater contaminant with common sources from animal and human waste, fertilizers, plants and precipitation. It is challenging to address groundwater contamination when common sources, such as agriculture, on-site sewage systems, and animal production, are widespread. Remediation is not possible, so mitigation is essential. The Lower Yakima Valley is located over 175,000 acres, with a population of 56,000 residents. Approximately 25% of the population do not have access to safe, clean drinking water, and 20% of the population is at or below the poverty level. Agriculture is the primary economic land-use activity. Irrigated agriculture and livestock production make up the largest percentage of acreage and nitrogen load. Commodities include apples, grapes, hops, dairy, silage corn, triticale, alfalfa and cherries. These commodities are important to the economic viability of the residents of the Lower Yakima Valley, as well as Washington State. Mitigation of nitrate in groundwater is challenging. The goal is to ensure everyone has safe drinking water. There are no easy remedies due to the extensive and pervasiveness of the contamination. Monitoring at the water table indicates that 45% of the 30 spatially distributed monitoring wells exceeded the drinking water standard. This indicates that there are multiple sources that are impacting water quality. Washington State has several areas which have extensive groundwater nitrate contamination. The groundwater in these areas continues to degrade over time. However, the Lower Yakima Valley is being successful in addressing this health issue because of the following reasons: the community is engaged and committed; there is one common goal; there has been extensive public education and outreach to citizens; and generating credible data using sound scientific methods. Work in this area is continuing as an ambient groundwater monitoring network is established to assess the condition of the aquifer over time. Nitrate samples are being collected from 170 wells, spatially distributed across the aquifer. This research entails quarterly sampling for two years to characterize seasonal variability and then continue annually afterward. This assessment will provide the data to statistically determine trends in nitrate concentrations across the aquifer, over time. Thirty-three of these wells are monitoring wells that are screened across the aquifer. The water quality from these wells are indicative of activities at the land surface. Additional work is being conducted to identify land use management practices that are effective in limiting nitrate migration through the soil column. Tracking nitrate in the soil column every season is an important component of bridging land-use practices with the fate and transport of nitrate through the subsurface. Patience, tenacity, and the ability to think outside the box are essential for dealing with widespread nitrate contamination of groundwater.

Keywords: community, groundwater, monitoring, nitrate

Procedia PDF Downloads 154

Authors: Yu-Mei Hsueh, Wei-Jen Chen, Yung-Kai Huang, Cheng-Shiuan Tsai, Kuo-Cheng Yeh

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Prostate cancer occurs in men over the age of 50, and rank sixth of the top ten cancers in Taiwan, and the incidence increased gradually over the past decade in Taiwan. Arsenic is confirmed as a carcinogen by International Agency for Research on (IARC). Arsenic induces oxidative stress may be a risk factor for prostate cancer, but the mechanism is not clear. Selenium is an important antioxidant element. Whether the association between plasma selenium levels and risk of prostate cancer are modified by different genotype of selenoprotein is still unknown. Glutathione peroxidase, selenoprotein P (SEPP1) and 15 kDa selenoprotein (SEP 15) are selenoprotein and regulates selenium transport and the oxidation and reduction reaction. However, the association between gene polymorphisms of selenoprotein and prostate cancer is not yet clear. The aim of this study is to determine the relationship between plasma selenium, polymorphism of selenoprotein, urinary total arsenic concentration and prostate cancer. This study is a hospital-based case-control study. Three hundred twenty-two cases of prostate cancer and age (±5 years) 1:1 matched 322 control group were recruited from National Taiwan University Hospital, Taipei Medical University Hospital, and Wan Fang Hospital. Well-trained personnel carried out standardized personal interviews based on a structured questionnaire. Information collected included demographic and socioeconomic characteristics, lifestyle and disease history. Blood and urine samples were also collected at the same time. The Research Ethics Committee of National Taiwan University Hospital, Taipei, Taiwan, approved the study. All patients provided informed consent forms before sample and data collection. Buffy coat was to extract DNA, and the polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) was used to measure the genotypes of SEPP1 rs3797310, SEP15 rs5859, GPX1 rs1050450, GPX2 rs4902346, GPX3 rs4958872, and GPX4 rs2075710. Plasma concentrations of selenium were determined by inductively coupled plasma mass spectrometry (ICP-MS).Urinary arsenic species concentrations were measured by high-performance liquid chromatography links hydride generator and atomic absorption spectrometer (HPLC-HG-AAS). Subject with high education level compared to those with low educational level had a lower prostate cancer odds ratio (OR) Mainland Chinese and aboriginal people had a lower OR of prostate cancer compared to Fukien Taiwanese. After adjustment for age, educational level, subjects with GPX1 rs1050450 CT and TT genotype compared to the CC genotype have lower, OR of prostate cancer, the OR and 95% confidence interval (Cl) was 0.53 (0.31-0.90). SEPP1 rs3797310 CT+TT genotype compared to those with CC genotype had a marginally significantly lower OR of PC. The low levels of plasma selenium and the high urinary total arsenic concentrations had the high OR of prostate cancer in a significant dose-response manner, and SEPP1 rs3797310 genotype modified this joint association.

Keywords: prostate cancer, plasma selenium concentration, urinary total arsenic concentrations, glutathione peroxidase, selenoprotein P, selenoprotein 15, gene polymorphism

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Authors: Iraklis Argyriou

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The transition to net-zero emission urban bus (ZEB) systems is receiving increased attention in research and policymaking throughout the globe. Most studies in this area tend to address techno-economic aspects and the perspectives of a narrow group of stakeholders, while they largely overlook analysis of current bus system dynamics. This offers limited insight into the types of ZEB governance challenges and opportunities that are encountered in real-world contexts, as well as into some of the immediate actions that need to be taken to set off the transition over the longer term. This research offers a multi-stakeholder perspective into both the technical and non-technical factors that influence ZEB transitions within a particular context, the UK. It does so by drawing from a recent transition visioning stakeholder workshop (June 2023) with key public, private and civic actors of the urban bus transportation system. Using NVivo software to qualitatively analyze the workshop discussions, the research examines the key technological and funding aspects, as well as the short-term actions (over the next five years), that need to be addressed for supporting the ZEB transition in UK cities. It finds that ZEB technology has reached a mature stage (i.e., high efficiency of batteries, motors and inverters), but important improvements can be pursued through greater control and integration of ZEB technological components and systems. In this regard, telemetry, predictive maintenance and adaptive control strategies pertinent to the performance and operation of ZEB vehicles have a key role to play in the techno-economic advancement of the transition. Yet, more pressing gaps were identified in the current ZEB funding regime. Whereas the UK central government supports greater ZEB adoption through a series of grants and subsidies, the scale of the funding and its fragmented nature do not match the needs for a UK-wide transition. Funding devolution arrangements (i.e., stable funding settlement deals between the central government and the devolved administrations/local authorities), as well as locally-driven schemes (i.e., congestion charging/workplace parking levy), could then enhance the financial prospects of the transition. As for short-term action, three areas were identified as critical: (1) the creation of whole value chains around the supply, use and recycling of ZEB components; (2) the ZEB retrofitting of existing fleets; and (3) integrated transportation that prioritizes buses as a first-choice, convenient and reliable mode while it simultaneously reduces car dependency in urban areas. Taken together, the findings point to the need for place-based transition approaches that create a viable techno-economic ecosystem for ZEB development but at the same time adopt a broader governance perspective beyond a ‘net-zero’ and ‘bus sectoral’ focus. As such, multi-actor collaborations and the coordination of wider resources and agency, both vertically across institutional scales and horizontally across transport, energy and urban planning, become fundamental features of comprehensive ZEB responses. The lessons from the UK case can inform a broader body of empirical contextual knowledge of ZEB transition governance within domestic political economies of public transportation.

Keywords: net-zero emission transition, stakeholders, transition governance, UK, urban bus transportation

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Authors: Feiyu Yang, Chunfang Ni, Rong Wang, Yun Zou, Wenbin Liu, Chenggong Zhang, Fenjin Sun, Chun Wang

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Cocaine is the most frequently used illegal drug globally, with the global annual prevalence of cocaine used ranging from 0.3% to 0.4 % of the adult population aged 15–64 years. Growing consumption trend of abused cocaine and drug crimes are a great concern, therefore urine sample testing has become an important noninvasive sampling whereas cocaine and its metabolites (COCs) are usually present in high concentrations and relatively long detection windows. However, direct analysis of urine samples is not feasible because urine complex medium often causes low sensitivity and selectivity of the determination. On the other hand, presence of low doses of analytes in urine makes an extraction and pretreatment step important before determination. Especially, in gathered taking drug cases, the pretreatment step becomes more tedious and time-consuming. So developing a sensitive, rapid and high-throughput method for detection of COCs in human body is indispensable for law enforcement officers, treatment specialists and health officials. In this work, a new automated magnetic dispersive solid-phase extraction (MDSPE) sampling method followed by high performance liquid chromatography-mass spectrometry (HPLC-MS) was developed for quantitative enrichment of COCs from human urine, using prepared magnetic nanoparticles as absorbants. The nanoparticles were prepared by silanizing magnetic Fe3O4 nanoparticles and modifying them with divinyl benzene and vinyl pyrrolidone, which possesses the ability for specific adsorption of COCs. And this kind of magnetic particle facilitated the pretreatment steps by electromagnetically controlled extraction to achieve full automation. The proposed device significantly improved the sampling preparation efficiency with 32 samples in one batch within 40mins. Optimization of the preparation procedure for the magnetic nanoparticles was explored and the performances of magnetic nanoparticles were characterized by scanning electron microscopy, vibrating sample magnetometer and infrared spectra measurements. Several analytical experimental parameters were studied, including amount of particles, adsorption time, elution solvent, extraction and desorption kinetics, and the verification of the proposed method was accomplished. The limits of detection for the cocaine and cocaine metabolites were 0.09-1.1 ng·mL-1 with recoveries ranging from 75.1 to 105.7%. Compared to traditional sampling method, this method is time-saving and environmentally friendly. It was confirmed that the proposed automated method was a kind of highly effective way for the trace cocaine and cocaine metabolites analyses in human urine.

Keywords: automatic magnetic dispersive solid-phase extraction, cocaine detection, magnetic nanoparticles, urine sample testing

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Authors: Huan Peng, Chenye Zeng, Zhao Wang

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Alzheimer’s disease (AD) is an age-related neurodegenerative disease that is a leading cause of dementia syndromes and has become a huge burden on society and families. The main pathological features of AD involve excessive deposition of β-amyloid (Aβ) and Tau proteins in the brain, resulting in loss of neurons, expansion of neuroinflammation, and cognitive dysfunction in patients. Researchers have found effective drugs to clear the brain of error-accumulating proteins or to slow the loss of neurons, but their direct administration has key bottlenecks such as single-drug limitation, rapid blood clearance rate, impenetrable blood-brain barrier (BBB), and poor ability to target tissues and cells. Therefore, we are committed to seeking a suitable and efficient delivery system. Inspired by the possibility that exosomes may be involved in the secretion and transport mechanism of many signaling molecules or proteins in the brain, exosomes have attracted extensive attention as natural nanoscale drug carriers. We selected exosomes derived from bone marrow mesenchymal stem cells (MSC-EXO) with low immunogenicity and exosomes derived from hippocampal neurons (HT22-EXO) that may have excellent homing ability to overcome the deficiencies of oral or injectable pathways and bypass the BBB through nasal administration and evaluated their delivery ability and effect on AD. First, MSC-EXO and HT22 cells were isolated and cultured, and MSCs were identified by microimaging and flow cytometry. Then MSC-EXO and HT22-EXO were obtained by gradient centrifugation and qEV SEC separation column, and a series of physicochemical characterization were performed by transmission electron microscope, western blot, nanoparticle tracking analysis and dynamic light scattering. Next, exosomes labeled with lipophilic fluorescent dye were administered to WT mice and APP/PS1 mice to obtain fluorescence images of various organs at different times. Finally, APP/PS1 mice were administered intranasally with two exosomes 20 times over 40 days and 20 μL each time. Behavioral analysis and pathological section analysis of the hippocampus were performed after the experiment. The results showed that MSC-EXO and HT22-EXO were successfully isolated and characterized, and they had good biocompatibility. MSC-EXO showed excellent brain enrichment in APP/PS1 mice after intranasal administration, could improve the neuronal damage and reduce inflammation levels in the hippocampus of APP/PS1 mice, and the improvement effect was significantly better than HT22-EXO. However, intranasal administration of the two exosomes did not cause depression and anxious-like phenotypes in APP/PS1 mice, nor significantly improved the short-term or spatial learning and memory ability of APP/PS1 mice, and had no significant effect on the content of Aβ plaques in the hippocampus, which also meant that MSC-EXO could use their own advantages in combination with other drugs to clear Aβ plaques. The possibility of realizing highly effective non-invasive synergistic treatment for AD provides new strategies and ideas for clinical research.

Keywords: Alzheimer’s disease, exosomes derived from mesenchymal stem cell, intranasal administration, therapy-carrier dual roles

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Authors: Renée M. Ripken, Johannes G. E. Gardeniers, Séverine Le Gac

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Controlling the multiphase behavior of aqueous biomass mixtures is essential when working in the biomass conversion industry. Here, the vapor/liquid equilibria (VLE) of ethylene glycol, glycerol, and xylitol were studied for temperatures between 25 and 200 °C and pressures of 1 to 10 bar. These experiments were performed in a microfluidic platform, which exhibits excellent heat transfer properties so that equilibrium is reached fast. Firstly, the saturated vapor pressure as a function of the temperature and the substrate mole fraction of the substrate was calculated using AspenPlus with a Redlich-Kwong-Soave Boston-Mathias (RKS-BM) model. Secondly, we developed a high-pressure and high-temperature microfluidic set-up for experimental validation. Furthermore, we have studied the multiphase flow pattern that occurs after the saturation temperature was achieved. A glass-silicon microfluidic device containing a 0.4 or 0.2 m long meandering channel with a depth of 250 μm and a width of 250 or 500 μm was fabricated using standard microfabrication techniques. This device was placed in a dedicated chip-holder, which includes a ceramic heater on the silicon side. The temperature was controlled and monitored by three K-type thermocouples: two were located between the heater and the silicon substrate, one to set the temperature and one to measure it, and the third one was placed in a 300 μm wide and 450 μm deep groove on the glass side to determine the heat loss over the silicon. An adjustable back pressure regulator and a pressure meter were added to control and evaluate the pressure during the experiment. Aqueous biomass solutions (10 wt%) were pumped at a flow rate of 10 μL/min using a syringe pump, and the temperature was slowly increased until the theoretical saturation temperature for the pre-set pressure was reached. First and surprisingly, a significant difference was observed between our theoretical saturation temperature and the experimental results. The experimental values were 10’s of degrees higher than the calculated ones and, in some cases, saturation could not be achieved. This discrepancy can be explained in different ways. Firstly, the pressure in the microchannel is locally higher due to both the thermal expansion of the liquid and the Laplace pressure that has to be overcome before a gas bubble can be formed. Secondly, superheating effects are likely to be present. Next, once saturation was reached, the flow pattern of the gas/liquid multiphase system was recorded. In our device, the point of nucleation can be controlled by taking advantage of the pressure drop across the channel and the accurate control of the temperature. Specifically, a higher temperature resulted in nucleation further upstream in the channel. As the void fraction increases downstream, the flow regime changes along the channel from bubbly flow to Taylor flow and later to annular flow. All three flow regimes were observed simultaneously. The findings of this study are key for the development and optimization of a microreactor for hydrogen production from biomass.

Keywords: biomass conversion, high pressure and high temperature microfluidics, multiphase, phase diagrams, superheating

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Authors: Utpal Das, Sona Das

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InAs/GaSb type-II superlattice (T2SL) Mid-wave infrared (MWIR) focal plane arrays (FPAs) have recently seen rapid development. However, in small pixel size large format FPAs, the occurrence of high mesa sidewall surface leakage current is a major constraint necessitating proper surface passivation. A simple pixel isolation technique in InAs/GaSb T2SL detector arrays without the conventional mesa etching has been proposed to isolate the pixels by forming a more resistive higher band gap material from the SL, in the inter-pixel region. Here, a single step femtosecond (fs) laser anneal of the T2SL structure of the inter-pixel T2SL regions, have been used to increase the band gap between the pixels by QW-intermixing and hence increase isolation between the pixels. The p-i-n photodiode structure used here consists of a 506nm, (10 monolayer {ML}) InAs:Si (1x10¹⁸cm⁻³)/(10ML) GaSb SL as the bottom n-contact layer grown on an n-type GaSb substrate. The undoped absorber layer consists of 1.3µm, (10ML)InAs/(10ML)GaSb SL. The top p-contact layer is a 63nm, (10ML)InAs:Be(1x10¹⁸cm⁻³)/(10ML)GaSb T2SL. In order to improve the carrier transport, a 126nm of graded doped (10ML)InAs/(10ML)GaSb SL layer was added between the absorber and each contact layers. A 775nm 150fs-laser at a fluence of ~6mJ/cm² is used to expose the array where the pixel regions are masked by a Ti(200nm)-Au(300nm) cap. Here, in the inter-pixel regions, the p+ layer have been reactive ion etched (RIE) using CH₄+H₂ chemistry and removed before fs-laser exposure. The fs-laser anneal isolation improvement in 200-400μm pixels due to spatially selective quantum well intermixing for a blue shift of ~70meV in the inter-pixel regions is confirmed by FTIR measurements. Dark currents are measured between two adjacent pixels with the Ti(200nm)-Au(300nm) caps used as contacts. The T2SL quality in the active photodiode regions masked by the Ti-Au cap is hardly affected and retains the original quality of the detector. Although, fs-laser anneal of p+ only etched p-i-n T2SL diodes show a reduction in the reverse dark current, no significant improvement in the full RIE-etched mesa structures is noticeable. Hence for a 128x128 array fabrication of 8μm square pixels and 10µm pitch, SU8 polymer isolation after RIE pixel delineation has been used. X-n+ row contacts and Y-p+ column contacts have been used to measure the optical response of the individual pixels. The photo-response of these 8μm and other 200μm pixels under a 2ns optical pulse excitation from an Optical-Parametric-Oscillator (OPO), shows a peak responsivity of ~0.03A/W and 0.2mA/W, respectively, at λ~3.7μm. Temporal response of this detector array is seen to have a fast response ~10ns followed typical slow decay with ringing, attributed to impedance mismatch of the connecting co-axial cables. In conclusion, response times of a few ns have been measured in 8µm pixels of a 128x128 array. Although fs-laser anneal has been found to be useful in increasing the inter-pixel isolation in InAs/GaSb T2SL arrays by QW inter-mixing, it has not been found to be suitable for passivation of full RIE etched mesa structures with vertical walls on InAs/GaSb T2SL.

Keywords: band-gap blue-shift, fs-laser-anneal, InAs/GaSb T2SL, Inter-pixel isolation, ns-Response, photodiode array

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Authors: Davide Cocchi, Andrea Zucchelli, Luca Raimondi, Maria Brugo Tommaso

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The increasing attention is given to the issues of environmental pollution and climate change is exponentially stimulating the development of electrically propelled vehicles powered by renewable energy, in particular, the solar one. Given the small amount of solar energy that can be stored and subsequently transformed into propulsive energy, it is necessary to develop vehicles with high mechanical, electrical and aerodynamic efficiencies along with reduced masses. The reduction of the masses is of fundamental relevance especially for the unsprung masses, that is the assembly of those elements that do not undergo a variation of their distance from the ground (wheel, suspension system, hub, upright, braking system). Therefore, the reduction of unsprung masses is fundamental in decreasing the rolling inertia and improving the drivability, comfort, and performance of the vehicle. This principle applies even more in solar propelled vehicles, equipped with an electric motor that is connected directly to the wheel hub. In this solution, the electric motor is integrated inside the wheel. Since the electric motor is part of the unsprung masses, the development of compact and lightweight solutions is of fundamental importance. The purpose of this research is the design development and optimization of a CFRP 16 wheel hub motor for solar propulsion vehicles that can carry up to four people. In addition to trying to maximize aspects of primary importance such as mass, strength, and stiffness, other innovative constructive aspects were explored. One of the main objectives has been to achieve a high geometric packing in order to ensure a reduced lateral dimension, without reducing the power exerted by the electric motor. In the final solution, it was possible to realize a wheel hub motor assembly completely comprised inside the rim width, for a total lateral overall dimension of less than 100 mm. This result was achieved by developing an innovative connection system between the wheel and the rotor with a double purpose: centering and transmission of the driving torque. This solution with appropriate interlocking noses allows the transfer of high torques and at the same time guarantees both the centering and the necessary stiffness of the transmission system. Moreover, to avoid delamination in critical areas, evaluated by means of FEM analysis using 3D Hashin damage criteria, electrospun nanofibrous mats have been interleaved between CFRP critical layers. In order to reduce rolling resistance, the rim has been designed to withstand high inflation pressure. Laboratory tests have been performed on the rim using the Digital Image Correlation technique (DIC). The wheel has been tested for fatigue bending according to E/ECE/324 R124e.

Keywords: composite laminate, delamination, DIC, lightweight vehicle, motor hub wheel, nanofiber

Procedia PDF Downloads 190

Authors: Kamolvara Sirisuksakulchai, Soamwadee Chaianansutcharit, Kazunori Sato

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Solid Oxide Fuel Cells (SOFCs) have been considered as one of the most efficient large unit power generators for household and industrial applications. The efficiency of an electronic cell depends mainly on the electrochemical reactions in the anode. The development of anode materials has been intensely studied to achieve higher kinetic rates of redox reactions and lower internal resistance. Recent studies have introduced an efficient cermet (ceramic-metallic) material for its ability in fuel oxidation and oxide conduction. This could expand the reactive site, also known as the triple-phase boundary (TPB), thus increasing the overall performance. In this study, a bimetallic catalyst Ni₀.₇₅Co₀.₂₅Oₓ was combined with Gd₀.₁Ce₀.₉O₁.₉₅ (GDC) to be used as a cermet anode (NiCo-GDC) for an anode-supported type SOFC. The synthesis of Ni₀.₇₅Co₀.₂₅Oₓ was carried out by ball milling NiO and Co3O4 powders in ethanol and calcined at 1000 °C. The Gd₀.₁Ce₀.₉O₁.₉₅ was prepared by a urea co-precipitation method. Precursors of Gd(NO₃)₃·6H₂O and Ce(NO₃)₃·6H₂O were dissolved in distilled water with the addition of urea and were heated subsequently. The heated mixture product was filtered and rinsed thoroughly, then dried and calcined at 800 °C and 1500 °C, respectively. The two powders were combined followed by pelletization and sintering at 1100 °C to form an anode support layer. The fabrications of an electrolyte layer and cathode layer were conducted. The electrochemical performance in H₂ was measured from 800 °C to 600 °C while for CH₄ was from 750 °C to 600 °C. The maximum power density at 750 °C in H₂ was 13% higher than in CH₄. The difference in performance was due to higher polarization resistances confirmed by the impedance spectra. According to the standard enthalpy, the dissociation energy of C-H bonds in CH₄ is slightly higher than the H-H bond H₂. The dissociation of CH₄ could be the cause of resistance within the anode material. The results from lower temperatures showed a descending trend of power density in relevance to the increased polarization resistance. This was due to lowering conductivity when the temperature decreases. The long-term stability was measured at 750 °C in CH₄ monitoring at 12-hour intervals. The maximum power density tends to increase gradually with time while the resistances were maintained. This suggests the enhanced stability from charge transfer activities in doped ceria due to the transition of Ce⁴⁺ ↔ Ce³⁺ at low oxygen partial pressure and high-temperature atmosphere. However, the power density started to drop after 60 h, and the cell potential also dropped from 0.3249 V to 0.2850 V. These phenomena was confirmed by a shifted impedance spectra indicating a higher ohmic resistance. The observation by FESEM and EDX-mapping suggests the degradation due to mass transport of ions in the electrolyte while the anode microstructure was still maintained. In summary, the electrochemical test and stability test for 60 h was achieved by NiCo-GDC cermet anode. Coke deposition was not detected after operation in CH₄, hence this confirms the superior properties of the bimetallic cermet anode over typical Ni-GDC.

Keywords: bimetallic catalyst, ceria-based SOFCs, methane oxidation, solid oxide fuel cell

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Authors: Keda Li, Hong Hu

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Auxetic materials, as an emerging artificial designed metamaterial has attracted growing attention due to their promising negative Poisson’s ratio behaviors and tunable properties. The conventional auxetic lattice structures for which the deformation process is governed by a bending-dominated mechanism have faced the limitation of poor mechanical performance for many potential engineering applications. Recently, both load-bearing and energy absorption capabilities have become a crucial consideration in auxetic structure design. This study reports the finite element analysis of a class of hexagonal double-arrowhead auxetic structures with enhanced stiffness and energy absorption performance. The structure design was developed by extending the traditional double-arrowhead honeycomb to a hexagon frame, the stretching-dominated deformation mechanism was determined according to Maxwell’s stability criterion. The finite element (FE) models of 2D lattice structures established with stainless steel material were analyzed in ABAQUS/Standard for predicting in-plane structural deformation mechanism, failure process, and compressive elastic properties. Based on the computational simulation, the parametric analysis was studied to investigate the effect of the structural parameters on Poisson’s ratio and mechanical properties. The geometrical optimization was then implemented to achieve the optimal Poisson’s ratio for the maximum specific energy absorption. In addition, the optimized 2D lattice structure was correspondingly converted into a 3D geometry configuration by using the orthogonally splicing method. The numerical results of 2D and 3D structures under compressive quasi-static loading conditions were compared separately with the traditional double-arrowhead re-entrant honeycomb in terms of specific Young's moduli, Poisson's ratios, and specified energy absorption. As a result, the energy absorption capability and stiffness are significantly reinforced with a wide range of Poisson’s ratio compared to traditional double-arrowhead re-entrant honeycomb. The auxetic behaviors, energy absorption capability, and yield strength of the proposed structure are adjustable with different combinations of joint angle, struts thickness, and the length-width ratio of the representative unit cell. The numerical prediction in this study suggests the proposed concept of hexagonal double-arrowhead structure could be a suitable candidate for the energy absorption applications with a constant request of load-bearing capacity. For future research, experimental analysis is required for the validation of the numerical simulation.

Keywords: auxetic, energy absorption capacity, finite element analysis, negative Poisson's ratio, re-entrant hexagonal honeycomb

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Authors: Sergey Nikitenko, Vladimir Klishin, Yury Malakhov, Elena Goosen

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Transition to renewable energy sources (solar, wind, bioenergy, etc.) and launching of alternative energy generation has weakened the role of coal as a source of energy. The Paris Agreement and assumption of obligations by many nations to orderly reduce CO₂ emissions by means of technological modernization and climate change adaptation has abridged coal demand yet more. This paper aims to assess current resilience of the coal industry to stress and to define prospects for coal production optimization using high technologies pursuant to global challenges and requirements of energy transition. Our research is based on the resilience concept adapted to the coal industry. It is proposed to divide the coal sector into segments depending on the prevailing value chains (VC). Four representative models of VC are identified in the coal sector. The most promising lines of upgrading VC in the coal industry include: •Elongation of VC owing to introduction of clean technologies of coal conversion and utilization; •Creation of parallel VC by means of waste management; •Branching of VC (conversion of a company’s VC into a production network). The upgrade effectiveness is governed in many ways by applicability of advanced coal processing technologies, usability of waste, expandability of production, entrance to non-rival markets and localization of new segments of VC in receiving regions. It is also important that upgrade of VC by means of formation of agile high-tech inter-industry production networks within the framework of operating surface and underground mines can reduce social, economic and ecological risks associated with closure of coal mines. Such promising route of VC upgrade is application of methanotrophic bacteria to produce protein to be used as feed-stuff in fish, poultry and cattle breeding, or in production of ferments, lipoids, sterols, antioxidants, pigments and polysaccharides. Closed mines can use recovered methane as a clean energy source. There exist methods of methane utilization from uncontrollable sources, including preliminary treatment and recovery of methane from air-and-methane mixture, or decomposition of methane to hydrogen and acetylene. Separated hydrogen is used in hydrogen fuel cells to generate power to feed the process of methane utilization and to supply external consumers. Despite the recent paradigm of carbon-free energy generation, it is possible to preserve the coal mining industry using the differentiated approach to upgrade of value chains based on flexible technologies with regard to specificity of mining companies.

Keywords: resilience, resilience concept, resilience indicator, resilience in the Russian coal industry, value chains

Procedia PDF Downloads 82

Authors: David A. Ullisch, Yantree D. Sankar-Thomas, Stefan Wilke, Thomas Selge, Matthias Pump, Thomas Leibold, Kai Schütte, Gilbert Gorr

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Plants have been considered as a source of natural substances for ages. Secondary metabolites from plants are utilized especially in medical applications but are more and more interesting as cosmetical ingredients and in the field of nutraceuticals. However, supply of compounds from natural harvest can be limited by numerous factors i.e. endangered species, low product content, climate impacts and cost intensive extraction. Especially in the pharmaceutical industry the ability to provide sufficient amounts of product and high quality are additional requirements which in some cases are difficult to fulfill by plant harvest. Whereas in many cases the complexity of secondary metabolites precludes chemical synthesis on a reasonable commercial basis, plant cells contain the biosynthetic pathway – a natural chemical factory – for a given compound. A promising approach for the sustainable production of natural products can be plant cell fermentation (PCF®). A thoroughly accomplished development process comprises the identification of a high producing cell line, optimization of growth and production conditions, the development of a robust and reliable production process and its scale-up. In order to address persistent, long lasting production, development of cryopreservation protocols and generation of working cell banks is another important requirement to be considered. So far the most prominent example using a PCF® process is the production of the anticancer compound paclitaxel. To demonstrate the power of plant suspension cultures here we present three case studies: 1) For more than 17 years Phyton produces paclitaxel at industrial scale i.e. up to 75,000 L in scale. With 60 g/kg dw this fully controlled process which is applied according to GMP results in outstanding high yields. 2) Thapsigargin is another anticancer compound which is currently isolated from seeds of Thapsia garganica. Thapsigargin is a powerful cytotoxin – a SERCA inhibitor – and the precursor for the derivative ADT, the key ingredient of the investigational prodrug Mipsagargin (G-202) which is in several clinical trials. Phyton successfully generated plant cell lines capable to express this compound. Here we present data about the screening for high producing cell lines. 3) The third case study covers ingenol-3-mebutate. This compound is found in the milky sap of the intact plants of the Euphorbiacae family at very low concentrations. Ingenol-3-mebutate is used in Picato® which is approved against actinic keratosis. Generation of cell lines expressing significant amounts of ingenol-3-mebutate is another example underlining the strength of plant cell culture. The authors gratefully acknowledge Inspyr Therapeutics for funding.

Keywords: Ingenol-3-mebutate, plant cell culture, sustainability, thapsigargin

Procedia PDF Downloads 222

Authors: A. G. Back

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Urban planning is a relevant tool to address, in a systemic way, several sectoral policies capable of linking the urban agenda with the reduction of socio-environmental risks. The Sao Paulo’s master plan (2014) presents innovations capable of promoting the transition to sustainability in the urban space, with a view to its regulatory instruments related to i) promotion of density in the axes of mass transport involving the mixture of commercial, residential, services, and leisure uses (principles related to the compact city); ii) vulnerabilities reduction based on housing policies including regular sources of funds for social housing and land reservation in urbanized areas; iii) reserve of green areas in the city to create parks and environmental regulations for new buildings focused on reducing the effects of heat island and improving urban drainage. However, its long-term implementation involves distributive conflicts and can undergo changes in different political, economic, and social contexts over time. Thus, the main objective of this paper is to identify and analyze the dynamics of conflicts of interest between social groups in the implementation of Sao Paulo’s urban development policy, particularly in relation to recent attempts at a (re) interpretation of the Master Plan guidelines, in view of the proposals for revision of the urban zoning law. In this sense, we seek to identify the demands, narratives of urban actors, including the real estate market, middle-class neighborhood associations ('not in my backyard' movements), and social housing rights movements. And we seek to analyze the power resources that these actors mobilize to influence the decision-making process, involving five categories: social capital, political access; discursive resource; media, juridical resource. The major findings of this research suggest that the interests and demands of the real estate market do not always prevail in urban regulation. After all, other actors also press for the definition of urban law with interests opposite to those of the real estate market. This is the case of associations of middle-class neighborhoods, which work to protect the characteristics of the locality, acting, in general, to prevent constructive and population densification in neighborhoods well located near the center, in São Paulo. One of the main demands of these “not in my backyard” movements is the delimitation of exclusively residential areas in the central region of the city, which is not only contrary to the interests of the real state market but also contrary to the principles of the compact city. On the other hand, social housing rights movements have also made progress in delimiting special areas of social interest in well-located and valued areas in the city dedicated to building social housing, also contrary to the interests of the real estate market. An urban development that follows the principles of the compact city must take into account the insertion of low-income populations in well-located regions; otherwise, such a development model may continue to push the less favored to the peripheries towards the preservation areas and/or risk areas.

Keywords: interest groups, Sao Paulo, sustainable urban development, urban policies implementation

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Authors: Alex B. Cusick

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The objective of this study is to improve upon the current ²³⁸PuO₂ fuel technology for space and defense applications. Modern RTGs (radioisotope thermoelectric generators) utilize the heat generated from the radioactive decay of ²³⁸Pu to create heat and electricity for long term and remote missions. Application of RTG technology is limited by the scarcity and expense of producing the isotope, as well as the power output which is limited to only a few hundred watts. The scarcity and expense make the efficient use of ²³⁸Pu absolutely necessary. By utilizing the decay of ²³⁸Pu, not only to produce heat directly but to also indirectly induce fission in ²³⁹Pu (which is already present within currently used fuel), it is possible to see large increases in temperature which allows for a more efficient conversion to electricity and a higher power-to-weight ratio. This concept can reduce the quantity of ²³⁸Pu necessary for these missions, potentially saving millions on investment, while yielding higher power output. Current work investigating radioisotope power systems have focused on improving efficiency of the thermoelectric components and replacing systems which produce heat by virtue of natural decay with fission reactors. The technical feasibility of utilizing (α,n) reactions to induce fission within current radioisotopic fuels has not been investigated in any appreciable detail, and our study aims to thoroughly investigate the performance of many such designs, develop those with highest capabilities, and facilitate experimental testing of these designs. In order to determine the specific design parameters that maximize power output and the efficient use of ²³⁸Pu for future RTG units, MCNP6 simulations have been used to characterize the effects of modifying fuel composition, geometry, and porosity, as well as introducing neutron moderating, reflecting, and shielding materials to the system. Although this project is currently in the preliminary stages, the final deliverables will include sophisticated designs and simulation models that define all characteristics of multiple novel RTG fuels, detailed enough to allow immediate fabrication and testing. Preliminary work has consisted of developing a benchmark model to accurately represent the ²³⁸PuO₂ pellets currently in use by NASA; this model utilizes the alpha transport capabilities of MCNP6 and agrees well with experimental data. In addition, several models have been developed by varying specific parameters to investigate their effect on (α,n) and (n,fi ssion) reaction rates. Current practices in fuel processing are to exchange out the small portion of naturally occurring ¹⁸O and ¹⁷O to limit (α,n) reactions and avoid unnecessary neutron production. However, we have shown that enriching the oxide in ¹⁸O introduces a sufficient (α,n) reaction rate to support significant fission rates. For example, subcritical fission rates above 10⁸ f/cm³-s are easily achievable in cylindrical ²³⁸PuO₂ fuel pellets with a ¹⁸O enrichment of 100%, given an increase in size and a ⁹Be clad. Many viable designs exist and our intent is to discuss current results and future endeavors on this project.

Keywords: radioisotope thermoelectric generators (RTG), Pu-238, subcritical reactors, (alpha, n) reactions

Procedia PDF Downloads 155

Authors: Toufic Sarieddine

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The current debate on the hegemonic impact of China’s Belt and Road Initiative (BRI) is of two opposing strands: Resilient and absolute US hegemony on the one hand and various models of multipolar hegemony such as bifurcation on the other. Bifurcation theories illustrate an unprecedented division of hegemonic functions between China and the US, whereby Beijing becomes the world’s economic hegemon, leaving Washington the world’s military hegemon and security guarantor. While consensus points to China being the main driver of unipolarity’s rupturing, the debate among bifurcationists is on the location of the first rupture. In this regard, the Middle East and North Africa (MENA) region has seen increasing Chinese foreign direct investment in recent years while that to other regions has declined, ranking it second in 2018 as part of the financing for the Maritime Silk Road Initiative (MSRI). China has also become the top trade partner of 11 states in the MENA region, as well as its top source of machine imports, surpassing the US and achieving an overall trade surplus almost double that of Washington’s. These are among other features outlined in world-systems analysis (WSA) literature which correspond with the emergence of a new hegemon. WSA is further utilized to gauge other facets of China’s increasing involvement in MENA and assess whether bifurcation is unfolding therein. These features of hegemony include the adoption of China’s modi operandi, economic dominance in production, trade, and finance, military capacity, cultural hegemony in ideology, education, and language, and the promotion of a general interest around which to rally potential peripheries (MENA states in this case). China’s modi operandi has seen some adoption with regards to support against the United Nations Convention on the Law of the Sea, oil bonds denominated in the yuan, and financial institutions such as the Shanghai Gold Exchange enjoying increasing Arab patronage. However, recent elections in Qatar, as well as liberal reforms in Saudi Arabia, demonstrate Washington’s stronger normative influence. Meanwhile, Washington’s economic dominance is challenged by China’s sizable machine exports, increasing overall imports, and widening trade surplus, but retains some clout via dominant arms and transport exports, as well as free-trade deals across the region. Militarily, Washington bests Beijing’s arms exports, has a dominant and well-established presence in the region, and successfully blocked Beijing’s attempt to penetrate through the UAE. Culturally, Beijing enjoys higher favorability in Arab public opinion, and its broadcast networks have found some resonance with Arab audiences. In education, the West remains MENA students’ preferred destination. Further, while Mandarin has become increasingly available in schools across MENA, its usage and availability still lag far behind English. Finally, Beijing’s general interest in infrastructure provision and prioritizing economic development over social justice and democracy provides an avenue for increased incorporation between Beijing and the MENA region. The overall analysis shows solid progress towards bifurcation in MENA.

Keywords: belt and road initiative, hegemony, Middle East and North Africa, world-systems analysis

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Authors: komal verma, V. S. Moholkar

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In this current study, the challenge of effectively treating and mineralizing industrial wastewater prior to its discharge into natural water bodies, such as rivers and lakes, is being addressed. Particularly, the focus is on the wastewater produced by chemical process industries, including refineries, petrochemicals, fertilizer, pharmaceuticals, pesticides, and dyestuff industries. These wastewaters often contain stubborn organic pollutants that conventional techniques, such as microbial processes cannot efficiently degrade. To tackle this issue, a ternary hybrid technique comprising of adsorption, heterogeneous Fenton process, and sonication has been employed. The study aims to evaluate the effectiveness of this approach for treating and mineralizing wastewater from a fertilizer industry located in Northeast India. The study comprises several key components, starting with the synthesis of the Fe3O4@AC nanocomposite using the co-precipitation method. The nanocomposite is then subjected to comprehensive characterization through various standard techniques, including FTIR, FE-SEM, EDX, TEM, BET surface area analysis, XRD, and magnetic property determination using VSM. Next, the process parameters of wastewater treatment are statistically optimized, focusing on achieving a high level of COD (Chemical Oxygen Demand) removal as the response variable. The Fe3O4@AC nanocomposite's adsorption characteristics and kinetics are also assessed in detail. The remarkable outcome of this study is the successful application of the ternary hybrid technique, combining adsorption, Fenton process, and sonication. This approach proves highly effective, leading to nearly complete mineralization (or TOC removal) of the fertilizer industry wastewater. The results highlight the potential of the Fe3O4@AC nanocomposite and the ternary hybrid technique as a promising solution for tackling challenging wastewater pollutants from various chemical process industries. This paper reports investigations in the mineralization of industrial wastewater (COD = 3246 mg/L, TOC = 2500 mg/L) using a ternary (ultrasound + Fenton + adsorption) hybrid advanced oxidation process. Fe3O4 decorated activated charcoal (Fe3O4@AC) nanocomposites (surface area = 538.88 m2/g; adsorption capacity = 294.31 mg/g) were synthesized using co-precipitation. The wastewater treatment process was optimized using central composite statistical design. At optimum conditions, viz. pH = 4.2, H2O2 loading = 0.71 M, adsorbent dose = 0.34 g/L, reduction in COD and TOC of wastewater were 94.75% and 89%, respectively. This result results from synergistic interactions among the adsorption of pollutants onto activated charcoal and surface Fenton reactions induced due to the leaching of Fe2+/Fe3+ ions from the Fe3O4 nanoparticles. Micro-convection generated due to sonication assisted faster mass transport (adsorption/desorption) of pollutants between Fe3O4@AC nanocomposite and the solution. The net result of this synergism was high interactions and reactions among and radicals and pollutants that resulted in the effective mineralization of wastewater. The Fe3O4@AC showed excellent recovery (> 90 wt%) and reusability (> 90% COD removal) in 5 successive cycles of treatment. LC-MS analysis revealed effective (> 50%) degradation of more than 25 significant contaminants (in the form of herbicides and pesticides) after the treatment with ternary hybrid AOP. Similarly, the toxicity analysis test using the seed germination technique revealed ~ 60% reduction in the toxicity of the wastewater after treatment.

Keywords: chemical oxygen demand (cod), fe3o4@ac nanocomposite, kinetics, lc-ms, rsm, toxicity

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Authors: Dian Mayasari, Yosi Bayu Murti, Sylvia Utami Tunjung Pratiwi, Sudarsono

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Melastoma malabathricum L. is an Indo-Pacific herb that has been traditionally used to treat several ailments such as wounds, dysentery, diarrhea, toothache, and diabetes. This plant is common across tropical Indo-Pacific archipelagos and is tolerant of a range of soils, from low-lying areas subject to saltwater inundation to the salt-free conditions of mountain slopes. How the soil and environmental variation influences secondary metabolite production in the herb, and an understanding of the plant’s utility as traditional medicine, remain largely unknown and unexplored. The objective of this study is to evaluate the variability of the metabolic profiles of M. malabathricum L. across its geographic distribution. By employing high-performance liquid chromatography-diode array detector (HPLC-DAD), a highly established, simple, sensitive, and reliable method was employed for establishing the chemical fingerprints of 72 samples of M. malabathricum L. leaves from various geographical locations in Indonesia. Specimens collected from six terrestrial and archipelago regions of Indonesia were analyzed by HPLC to generate chromatogram peak profiles that could be compared across each region. Data corresponding to the common peak areas of HPLC chromatographic fingerprint were analyzed by hierarchical component analysis (HCA) and principal component analysis (PCA) to extract information on the most significant variables contributing to characterization and classification of analyzed samples data. Principal component values were identified as PC1 and PC2 with 41.14% and 19.32%, respectively. Based on variety and origin, the high-performance liquid chromatography method validated the chemical fingerprint results used to screen the in vitro antioxidant activity of M. malabathricum L. The result shows that the developed method has potential values for the quality of similar M. malabathrium L. samples. These findings provide a pathway for the development and utilization of references for the identification of M. malabathricum L. Our results indicate the importance of considering geographic distribution during field-collection efforts as they demonstrate regional metabolic variation in secondary metabolites of M. malabathricum L., as illustrated by HPLC chromatogram peaks and their antioxidant activities. The results also confirm the utility of this simple approach to a rapid evaluation of metabolic variation between plants and their potential ethnobotanical properties, potentially due to the environments from whence they were collected. This information will facilitate the optimization of growth conditions to suit particular medicinal qualities.

Keywords: fingerprint, high performance liquid chromatography, Melastoma malabathricum l., metabolic profiles, principal component analysis

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Authors: E. Nakamachi, S. Tanaka, K. Yamamoto, Y. Morita

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In this study, we developed a three-dimensional (3D) direct current electric field (DCEF) stimulation bio-reactor for axonal outgrowth enhancement to generate the neural network of the central nervous system (CNS). By using our newly developed 3D DCEF stimulation bio-reactor, we cultured the rat pheochromocytoma cells (PC12) and investigated the effects on the axonal extension enhancement and network generation. Firstly, we designed and fabricated a 3D bio-reactor, which can load DCEF stimulation on PC12 cells embedded in the collagen gel as extracellular environment. The connection between the electrolyte and the medium using salt bridges for DCEF stimulation was introduced to avoid the cell death by the toxicity of metal ion. The distance between the salt bridges was adopted as the design variable to optimize a structure for uniform DCEF stimulation, where the finite element (FE) analyses results were used. Uniform DCEF strength and electric flux vector direction in the PC12 cells embedded in collagen gel were examined through measurements of the fabricated 3D bio-reactor chamber. Measurement results of DCEF strength in the bio-reactor showed a good agreement with FE results. In addition, the perfusion system was attached to maintain pH 7.2 ~ 7.6 of the medium because pH change was caused by DCEF stimulation loading. Secondly, we disseminated PC12 cells in collagen gel and carried out 3D culture. Finally, we measured the morphology of PC12 cell bodies and neurites by the multiphoton excitation fluorescence microscope (MPM). The effectiveness of DCEF stimulation to enhance the axonal outgrowth and the neural network generation was investigated. We confirmed that both an increase of mean axonal length and axogenesis rate of PC12, which have been exposed 5 mV/mm for 6 hours a day for 4 days in the bioreactor. We found following conclusions in our study. 1) Design and fabrication of DCEF stimulation bio-reactor capable of 3D culture nerve cell were completed. A uniform electric field strength of average value of 17 mV/mm within the 1.2% error range was confirmed by using FE analyses, after the structure determination through the optimization process. In addition, we attached a perfusion system capable of suppressing the pH change of the culture solution due to DCEF stimulation loading. 2) Evaluation of DCEF stimulation effects on PC12 cell activity was executed. The 3D culture of PC 12 was carried out adopting the embedding culture method using collagen gel as a scaffold for four days under the condition of 5.0 mV/mm and 10mV/mm. There was a significant effect on the enhancement of axonal extension, as 11.3% increase in an average length, and the increase of axogenesis rate. On the other hand, no effects on the orientation of axon against the DCEF flux direction was observed. Further, the network generation was enhanced to connect longer distance between the target neighbor cells by DCEF stimulation.

Keywords: PC12, DCEF stimulation, 3D bio-reactor, axonal extension, neural network generation

Procedia PDF Downloads 168

Authors: Jeanne Leblond, Warren Viricel, Amira Mbarek

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Although several products have reached the market, gene therapeutics are still in their first stages and require optimization. It is possible to improve their lacking efficiency by the use of carefully engineered vectors, able to carry the genetic material through each of the biological barriers they need to cross. In particular, getting inside the cell is a major challenge, because these hydrophilic nucleic acids have to cross the lipid-rich plasmatic and/or endosomal membrane, before being degraded into lysosomes. It takes less than one hour for newly endocytosed liposomes to reach highly acidic lysosomes, meaning that the degradation of the carried gene occurs rapidly, thus limiting the transfection efficiency. We propose to use a new pH-sensitive lipid able to change its conformation upon protonation at endosomal pH values, leading to the disruption of the lipidic bilayer and thus to the fast release of the nucleic acids into the cytosol. It is expected that this new pH-sensitive mechanism promote endosomal escape of the gene, thereby its transfection efficiency. The main challenge of this work was to design a preparation presenting fast-responding lipidic bilayer destabilization properties at endosomal pH 5 while remaining stable at blood pH value and during storage. A series of pH-sensitive lipids able to perform a conformational switch upon acidification were designed and synthesized. Liposomes containing these switchable lipids, as well as co-lipids were prepared and characterized. The liposomes were stable at 4°C and pH 7.4 for several months. Incubation with siRNA led to the full entrapment of nucleic acids as soon as the positive/negative charge ratio was superior to 2. The best liposomal formulation demonstrated a silencing efficiency up to 10% on HeLa cells, very similar to a commercial agent, with a lowest toxicity than the commercial agent. Using flow cytometry and microscopy assays, we demonstrated that drop of pH was required for the transfection efficiency, since bafilomycin blocked the transfection efficiency. Additional evidence was brought by the synthesis of a negative control lipid, which was unable to switch its conformation, and consequently exhibited no transfection ability. Mechanistic studies revealed that the uptake was mediated through endocytosis, by clathrin and caveolae pathways, as reported for previous lipid nanoparticle systems. This potent system was used for the treatment of hypercholesterolemia. The switchable lipids were able to knockdown PCSK9 expression on human hepatocytes (Huh-7). Its efficiency is currently evaluated on in vivo mice model of PCSK9 KO mice. In summary, we designed and optimized a new cationic pH-sensitive lipid for gene delivery. Its transfection efficiency is similar to the best available commercial agent, without the usually associated toxicity. The promising results lead to its use for the treatment of hypercholesterolemia on a mice model. Anticancer applications and pulmonary chronic disease are also currently investigated.

Keywords: liposomes, siRNA, pH-sensitive, molecular switch

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Authors: V. Jayanthi

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Urbanization and pace of urbanization have increased tremendously in last few decades. More towns are now getting converted into cities. Urbanization trend is seen all over the world but is becoming most dominant in Asia. Today, the scale of urbanization in India is so huge that Indian cities are among the fastest-growing in the world, including Bangalore, Hyderabad, Pune, Chennai, Delhi, and Mumbai. Urbanization remains a single predominant factor that is continuously linked to the destruction of urban green spaces. With reference to Chennai as a case study, which is suffering from rapid deterioration of its green spaces, this paper sought to fill this gap by exploring key factors aside urbanization that is responsible for the destruction of green spaces. The paper relied on a research approach and triangulated data collection techniques such as interviews, focus group discussion, personal observation and retrieval of archival data. It was observed that apart from urbanization, problem of ownership of green space lands, low priority to green spaces, poor maintenance, enforcement of development controls, wastage of underpass spaces, and uncooperative attitudes of the general public, play a critical role in the destruction of urban green spaces. Therefore the paper narrows down to a point, that for a city to have a proper sustainable urban green space, broader city development plans are essential. Though rapid urbanization is an indicator of positive development, it is also accompanied by a host of challenges. Chennai lost a lot of greenery, as the city urbanized rapidly that led to a steep fall in vegetation cover. Environmental deterioration will be the big price we pay if Chennai continues to grow at the expense of greenery. Soaring skyscrapers, multistoried complexes, gated communities, and villas, frame the iconic skyline of today’s Chennai city which reveals that we overlook the importance of our green cover, which is important to balance our urban and lung spaces. Chennai, with a clumped landscape at the center of the city, is predicted to convert 36% of its total area into urban areas by 2026. One major issue is that a city designed and planned in isolation creates underused spaces all around the cities which are of negligence. These urban voids are dead, underused, unused spaces in the cities that are formed due to inefficient decision making, poor land management, and poor coordination. Urban voids have huge potential of creating a stronger urban fabric, exploited as public gathering spaces, pocket parks or plazas or just enhance public realm, rather than dumping of debris and encroachments. Flyovers need to justify their existence themselves by being more than just traffic and transport solutions. The vast, unused space below the Kathipara flyover is a case in point. This flyover connects three major routes: Tambaram, Koyambedu, and Adyar. This research will focus on the concept of urban voids, how these voids under the flyovers, can be used for place making process, how this space beneath flyovers which are neglected, can be a part of the urban realm through urban design and landscaping.

Keywords: landscape design, flyovers, public spaces, reclaiming lost spaces, urban voids

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Authors: Anubhav Kumar Dwivedi, Arshad Khan, S. N. Tripathi, Manish Joshi, Gaurav Mishra, Dinesh Nath, Naveen Tiwari, B. K. Sapra

Abstract:

In a nuclear reactor accident scenario, a large number of fission products may release to the piping system of the primary heat transport. The released fission products, mostly in the form of the aerosol, get deposited on the inner surface of the piping system mainly due to gravitational settling and thermophoretic deposition. The removal processes in the complex piping system are controlled to a large extent by the thermal-hydraulic conditions like temperature, pressure, and flow rates. These parameters generally vary with time and therefore must be carefully monitored to predict the aerosol behavior in the piping system. The removal process of aerosol depends on the size of particles that determines how many particles get deposit or travel across the bends and reach to the other end of the piping system. The released aerosol gets deposited onto the inner surface of the piping system by various mechanisms like gravitational settling, Brownian diffusion, thermophoretic deposition, and by other deposition mechanisms. To quantify the correct estimate of deposition, the identification and understanding of the aforementioned deposition mechanisms are of great importance. These mechanisms are significantly affected by different flow and thermodynamic conditions. Thermophoresis also plays a significant role in particle deposition. In the present study, a series of experiments were performed in the piping system of the National Aerosol Test Facility (NATF), BARC using metal aerosols (zinc) in dry environments to study the spatial distribution of particles mass and number concentration, and their depletion due to various removal mechanisms in the piping system. The experiments were performed at two different carrier gas flow rates. The commercial CFD software FLUENT is used to determine the distribution of temperature, velocity, pressure, and turbulence quantities in the piping system. In addition to the in-built models for turbulence, heat transfer and flow in the commercial CFD code (FLUENT), a new sub-model PBM (population balance model) is used to describe the coagulation process and to compute the number concentration along with the size distribution at different sections of the piping. In the sub-model coagulation kernels are incorporated through user-defined function (UDF). The experimental results are compared with the CFD modeled results. It is found that most of the Zn particles (more than 35 %) deposit near the inlet of the plenum chamber and a low deposition is obtained in piping sections. The MMAD decreases along the length of the test assembly, which shows that large particles get deposited or removed in the course of flow, and only fine particles travel to the end of the piping system. The effect of a bend is also observed, and it is found that the relative loss in mass concentration at bends is more in case of a high flow rate. The simulation results show that the thermophoresis and depositional effects are more dominating for the small and larger sizes as compared to the intermediate particles size. Both SEM and XRD analysis of the collected samples show the samples are highly agglomerated non-spherical and composed mainly of ZnO. The coupled model framed in this work could be used as an important tool for predicting size distribution and concentration of some other aerosol released during a reactor accident scenario.

Keywords: aerosol, CFD, deposition, coagulation

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Authors: K. Verma, v. S. Moholkar

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In developed countries, water pollution caused by industrial discharge has emerged as a significant environmental concern over the past decades. However, despite ongoing efforts, a fully effective and sustainable remediation strategy has yet to be identified. This paper describes how enzymatic and sonochemical treatments have demonstrated great promise in degrading bio-refractory pollutants. Mainly, a compelling area of interest lies in the combined technique of sono-enzymatic treatment, which has exhibited a synergistic enhancement effect surpassing that of the individual techniques. This study employed the covalent attachment method to immobilize Laccase from Trametes versicolor onto amino-functionalized magnetic Fe₃O₄ nanoparticles. To comprehensively characterize the synthesized free nanoparticles and the laccase-immobilized nanoparticles, various techniques such as X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM), vibrating sample magnetometer (VSM), and surface area through Brunauer-Emmett-Teller (BET) were employed. The size of immobilized Fe₃O₄@Laccase was found to be 60 nm, and the maximum loading of laccase was found to be 24 mg/g of nanoparticle. An investigation was conducted to study the effect of various process parameters, such as immobilized Fe₃O₄ Laccase dose, temperature, and pH, on the % Chemical oxygen demand (COD) removal as a response. The statistical design pinpointed the optimum conditions (immobilized Fe₃O₄ Laccase dose = 1.46 g/L, pH = 4.5, and temperature = 66 oC), resulting in a remarkable 65.58% COD removal within 60 minutes. An even more significant improvement (90.31% COD removal) was achieved with ultrasound-assisted enzymatic reaction utilizing a 10% duty cycle. The investigation of various kinetic models for free and immobilized laccase, such as the Haldane, Yano, and Koga, and Michaelis-Menten, showed that ultrasound application impacted the kinetic parameters Vmax and Km. Specifically, Vmax values for free and immobilized laccase were found to be 0.021 mg/L min and 0.045 mg/L min, respectively, while Km values were 147.2 mg/L for free laccase and 136.46 mg/L for immobilized laccase. The lower Km and higher Vmax for immobilized laccase indicate its enhanced affinity towards the substrate, likely due to ultrasound-induced alterations in the enzyme's confirmation and increased exposure of active sites, leading to more efficient degradation. Furthermore, the toxicity and Liquid chromatography-mass spectrometry (LC-MS) analysis revealed that after the treatment process, the wastewater exhibited 70% less toxicity than before treatment, with over 25 compounds degrading by more than 75%. At last, the prepared immobilized laccase had excellent recyclability retaining 70% activity up to 6 consecutive cycles. A straightforward manufacturing strategy and outstanding performance make the recyclable magnetic immobilized Laccase (Fe₃O₄ Laccase) an up-and-coming option for various environmental applications, particularly in water pollution control and treatment.

Keywords: kinetic, laccase enzyme, sonoenzymatic, ultrasound irradiation

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Authors: Aleksander Ejsmont, Stefan Wuttke, Joanna Goscianska

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Gaining control over particle shape, size and crystallinity is an ongoing challenge for many materials. Especially metalorganic frameworks (MOFs) are recently widely studied. Besides their remarkable porosity and interesting topologies, morphology has proven to be a significant feature. It can affect the further material application. Thus seeking new approaches that enable MOF morphology modulation is important. MOFs are reticular structures, where building blocks are made up of organic linkers and metallic nodes. The most common strategy of ensuring metal source is using salts, which usually exhibit high solubility and hinder morphology control. However, there has been a growing interest in using metal oxides as structure-directing agents towards MOFs due to their very low solubility and shape preservation. Metal oxides can be treated as a metal reservoir during MOF synthesis. Up to now, reports in which receiving MOFs from metal oxides mostly present ZnO conversion to ZIF-8. However, there are other oxides, for instance, Co₃O₄, which often is overlooked due to their structural stability and insolubility in aqueous solutions. Cobalt-based materials are famed for catalytic activity. Therefore the development of their efficient synthesis is worth attention. In the presented work, an optimized Co₃O₄transition to Co-MOFviaa solvothermal approach was proposed. The starting point of the research was the synthesis of Co₃O₄ flower petals and needles under hydrothermal conditions using different cobalt salts (e.g., cobalt(II) chloride and cobalt(II) nitrate), in the presence of urea, and hexadecyltrimethylammonium bromide (CTAB) surfactant as a capping agent. After receiving cobalt hydroxide, the calcination process was performed at various temperatures (300–500 °C). Then cobalt oxides as a source of cobalt cations were subjected to reaction with trimesic acid in solvothermal environment and temperature of 120 °C leading to Co-MOF fabrication. The solution maintained in the system was a mixture of water, dimethylformamide, and ethanol, with the addition of strong acids (HF and HNO₃). To establish how solvents affect metal oxide conversion, several different solvent ratios were also applied. The materials received were characterized with analytical techniques, including X-ray powder diffraction, energy dispersive spectroscopy,low-temperature nitrogen adsorption/desorption, scanning, and transmission electron microscopy. It was confirmed that the synthetic routes have led to the formation of Co₃O₄ and Co-based MOF varied in shape and size of particles. The diffractograms showed receiving crystalline phase for Co₃O₄, and also for Co-MOF. The Co₃O₄ obtained from nitrates and with using low-temperature calcination resulted in smaller particles. The study indicated that cobalt oxide particles of different size influence the efficiency of conversion and morphology of Co-MOF. The highest conversion was achieved using metal oxides with small crystallites.

Keywords: Co-MOF, solvothermal synthesis, morphology control, core-shell

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Authors: Yi-Hao Wong, Li-Li Chan, Chee-Onn Leong, Stephen Ambu, Joon-Wah Mak, Priyasashi Sahu

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Background: Acanthamoeba is a genus of amoebae which lives as a free-living in nature or as a human pathogen that causes severe brain and eye infections. Virulence potential of Acanthamoeba is not constant and can change with growth conditions. DNA methylation, an epigenetic process which adds methyl groups to DNA, is used by eukaryotic cells, including several human parasites to control their gene expression. We used qPCR, siRNA gene silencing, and RNA sequencing (RNA-Seq) to study DNA-methyltransferase gene family (DNMT) in order to indicate the possibility of its involvement in programming Acanthamoeba virulence potential. Methods: A virulence-attenuated Acanthamoeba isolate (designation: ATCC; original isolate: ATCC 50492) was subjected to mouse passages to restore its pathogenicity; a virulence-reactivated isolate (designation: AC/5) was generated. Several established factors associated with Acanthamoeba virulence phenotype were examined to confirm the succession of reactivation process. Differential gene expression of DNMT between ATCC and AC/5 isolates was performed by qPCR. Silencing on DNMT gene expression in AC/5 isolate was achieved by siRNA duplex. Total RNAs extracted from ATCC, AC/5, and siRNA-treated (designation: si-146) were subjected to RNA-Seq for comparative transcriptomic analysis in order to identify the genome-wide effect of DNMT in regulating Acanthamoeba gene expression. qPCR was performed to validate the RNA-Seq results. Results: Physiological and cytophatic assays demonstrated an increased in virulence potential of AC/5 isolate after mouse passages. DNMT gene expression was significantly higher in AC/5 compared to ATCC isolate (p ≤ 0.01) by qPCR. si-146 duplex reduced DNMT gene expression in AC/5 isolate by 30%. Comparative transcriptome analysis identified the differentially expressed genes, with 3768 genes in AC/5 vs ATCC isolate; 2102 genes in si-146 vs AC/5 isolate and 3422 genes in si-146 vs ATCC isolate, respectively (fold-change of ≥ 2 or ≤ 0.5, p-value adjusted (padj) < 0.05). Of these, 840 and 1262 genes were upregulated and downregulated, respectively, in si-146 vs AC/5 isolate. Eukaryotic orthologous group (KOG) assignments revealed a higher percentage of downregulated gene expression in si-146 compared to AC/5 isolate, were related to posttranslational modification, signal transduction and energy production. Gene Ontology (GO) terms for those downregulated genes shown were associated with transport activity, oxidation-reduction process, and metabolic process. Among these downregulated genes were putative genes encoded for heat shock proteins, transporters, ubiquitin-related proteins, proteins for vesicular trafficking (small GTPases), and oxidoreductases. Functional analysis of similar predicted proteins had been described in other parasitic protozoa for their survival and pathogenicity. Decreased expression of these genes in si146-treated isolate may account in part for Acanthamoeba reduced pathogenicity. qPCR on 6 selected genes upregulated in AC/5 compared to ATCC isolate corroborated the RNA sequencing findings, indicating a good concordance between these two analyses. Conclusion: To the best of our knowledge, this study represents the first genome-wide analysis of DNA methylation and its effects on gene expression in Acanthamoeba spp. The present data indicate that DNA methylation has substantial effect on global gene expression, allowing further dissection of the genome-wide effects of DNA-methyltransferase gene in regulating Acanthamoeba pathogenicity.

Keywords: Acanthamoeba, DNA methylation, RNA sequencing, virulence

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