Search results for: physiological sensors
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2174

Search results for: physiological sensors

224 Gene Expression and Staining Agents: Exploring the Factors That Influence the Electrophoretic Properties of Fluorescent Proteins

Authors: Elif Tugce Aksun Tumerkan, Chris Lowe, Hannah Krupa

Abstract:

Fluorescent proteins are self-sufficient in forming chromophores with a visible wavelength from 3 amino acids sequence within their own polypeptide structure. This chromophore – a molecule that absorbs a photon of light and exhibits an energy transition equal to the energy of the absorbed photon. Fluorescent proteins (FPs) consisted of a chain of 238 amino acid residues and composed of 11 beta strands shaped in a cylinder surrounding an alpha helix structure. A better understanding of the system of the chromospheres and the increasing advance in protein engineering in recent years, the properties of FPs offers the potential for new applications. They have used sensors and probes in molecular biology and cell-based research that giving a chance to observe these FPs tagged cell localization, structural variation and movement. For clarifying functional uses of fluorescent proteins, electrophoretic properties of these proteins are one of the most important parameters. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is used for determining electrophoretic properties commonly. While there are many techniques are used for determining the functionality of protein-based research, SDS-PAGE analysis can only provide a molecular level assessment of the proteolytic fragments. Before SDS-PAGE analysis, fluorescent proteins need to successfully purified. Due to directly purification of the target, FPs is difficult from the animal, gene expression is commonly used which must be done by transformation with the plasmid. Furthermore, used gel within electrophoresis and staining agents properties have a key role. In this review, the different factors that have the impact on the electrophoretic properties of fluorescent proteins explored. Fluorescent protein separation and purification are the essential steps before electrophoresis that should be done very carefully. For protein purification, gene expression process and following steps have a significant function. For successful gene expression, the properties of selected bacteria for expression, used plasmid are essential. Each bacteria has own characteristics which are very sensitive to gene expression, also used procedure is the important factor for fluorescent protein expression. Another important factors are gel formula and used staining agents. Gel formula has an effect on the specific proteins mobilization and staining with correct agents is a key step for visualization of electrophoretic bands of protein. Visuality of proteins can be changed depending on staining reagents. Apparently, this review has emphasized that gene expression and purification have a stronger effect than electrophoresis protocol and staining agents.

Keywords: cell biology, gene expression, staining agents, SDS-page

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223 Unmanned Aerial System Development for the Remote Reflectance Sensing Using Above-Water Radiometers

Authors: Sunghun Jung, Wonkook Kim

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Due to the difficulty of the utilization of satellite and an aircraft, conventional ocean color remote sensing has a disadvantage in that it is difficult to obtain images of desired places at desired times. These disadvantages make it difficult to capture the anomalies such as the occurrence of the red tide which requires immediate observation. It is also difficult to understand the phenomena such as the resuspension-precipitation process of suspended solids and the spread of low-salinity water originating in the coastal areas. For the remote sensing reflectance of seawater, above-water radiometers (AWR) have been used either by carrying portable AWRs on a ship or installing those at fixed observation points on the Ieodo ocean research station, Socheongcho base, and etc. In particular, however, it requires the high cost to measure the remote reflectance in various seawater environments at various times and it is even not possible to measure it at the desired frequency in the desired sea area at the desired time. Also, in case of the stationary observation, it is advantageous that observation data is continuously obtained, but there is the disadvantage that data of various sea areas cannot be obtained. It is possible to instantly capture various marine phenomena occurring on the coast using the unmanned aerial system (UAS) including vertical takeoff and landing (VTOL) type unmanned aerial vehicles (UAV) since it could move and hover at the one location and acquire data of the desired form at a high resolution. To remotely estimate seawater constituents, it is necessary to install an ultra-spectral sensor. Also, to calculate reflected light from the surface of the sea in consideration of the sun’s incident light, a total of three sensors need to be installed on the UAV. The remote sensing reflectance of seawater is the most basic optical property for remotely estimating color components in seawater and we could remotely estimate the chlorophyll concentration, the suspended solids concentration, and the dissolved organic amount. Estimating seawater physics from the remote sensing reflectance requires the algorithm development using the accumulation data of seawater reflectivity under various seawater and atmospheric conditions. The UAS with three AWRs is developed for the remote reflection sensing on the surface of the sea. Throughout the paper, we explain the details of each UAS component, system operation scenarios, and simulation and experiment results. The UAS consists of a UAV, a solar tracker, a transmitter, a ground control station (GCS), three AWRs, and two gimbals.

Keywords: above-water radiometers (AWR), ground control station (GCS), unmanned aerial system (UAS), unmanned aerial vehicle (UAV)

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222 Impact of Stress and Protein Malnutrition on the Potential Role of Epigallocatechin-3-Gallate in Providing Protection from Nephrotoxicity and Hepatotoxicity Induced by Aluminum in Rats

Authors: Azza A. Ali, Mona G. Khalil, Hemat A. Elariny, Shereen S. El Shaer

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Background: Aluminium (Al) is very abundant metal in the earth’s crust. It is a constituent of cooking utensils, medicines, cosmetics, some foods and food additives. Salts of Al are widely used in the treatment of drinking water for purification purposes. Excessive and prolonged exposure to Al causes oxidative stress and impairment of many physiological functions. Its accumulation in liver and kidney causes hepatotoxicity and nephrotoxicity. Social isolation (SI) or Protein malnutrition (PM) also increases oxidative stress and may enhance the toxicity of Al as well as the degeneration in liver and kidney. Epigallocatechin-3-gallate (EGCG) is the most abundant catechin in green tea and has strong antioxidant as well as anti-inflammatory activities and can protect against oxidative stress-induced degenerations. Objective: To study the influence of stress or PM on Al-induced nephrotoxicity and hepatotoxicity in rats, as well as on the potential role of EGCG in providing protection. Methods: Rats received daily AlCl3 (70 mg/kg, IP) for three weeks (Al-toxicity groups) except one normal control group received saline. Al-toxicity groups were divided into four treated and four untreated groups; treated rats received EGCG (10 mg/kg, IP) together with AlCl3. One group of both treated and untreated rats served as control for each of them, and the others were subjected to either stress (mild using isolation or high using electric shock) or to PM (10% casein diet). Specimens of liver and kidney were used for assessment of levels of inflammatory mediators as TNF-α, IL6β, nuclear factor kappa B (NF-κB), oxidative stress (MDA, SOD, TAC, NO), Caspase-3 and for DNA fragmentation as well as for histopathological examinations. Biochemical changes were also measured in the serum as total lipids, cholesterol, triglycerides, glucose, proteins, bilirubin, creatinine and urea as well as the level of Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and lactate deshydrogenase (LDH). Results: Nephrotoxicity and hepatotoxicity induced by Al were enhanced in rats exposed to stress and to PM. The influence of stress was more pronounced than PM. Al-toxicity was indicated by the increase in liver and kidney MDA, NO, TNF-α, IL-6β, NF-κB, caspase-3, DNA fragmentation and in ALT, AST, ALP, LDH and total lipids, cholesterol, triglycerides, glucose, proteins, bilirubin, creatinine and urea levels, together with the decrease in total proteins, SOD, TAC. EGCG provided protection against hazards of Al as indicated by the decrease in MDA, NO, TNF-α, IL-6β, NF-κB, caspase-3 and DNA fragmentation as well as in levels of ALT, AST, ALP, LDH and total lipids, cholesterol, triglycerides, glucose, proteins, bilirubin, creatinine and urea in liver and kidney, together with the increase in total proteins, SOD, TAC and confirmed by histopathological examinations. It provided more pronounced protection in high stressful conditions than in mild one than in PM. Conclusion: Stress have a bad impact on Al-induced nephrotoxicity and hepatotoxicity more than PM. Thus it can clarify and maximize the role of EGCG in providing protection. Consequently, administration of EGCG is advised with excessive Al-exposure to avoid nephrotoxicity and hepatotoxicity especially in populations more subjected to stress or PM.

Keywords: aluminum, stress, protein malnutrition, nephrotoxicity, hepatotoxicity, epigallocatechin-3-gallate, rats

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221 Development of an Automatic Control System for ex vivo Heart Perfusion

Authors: Pengzhou Lu, Liming Xin, Payam Tavakoli, Zhonghua Lin, Roberto V. P. Ribeiro, Mitesh V. Badiwala

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Ex vivo Heart Perfusion (EVHP) has been developed as an alternative strategy to expand cardiac donation by enabling resuscitation and functional assessment of hearts donated from marginal donors, which were previously not accepted. EVHP parameters, such as perfusion flow (PF) and perfusion pressure (PP) are crucial for optimal organ preservation. However, with the heart’s constant physiological changes during EVHP, such as coronary vascular resistance, manual control of these parameters is rendered imprecise and cumbersome for the operator. Additionally, low control precision and the long adjusting time may lead to irreversible damage to the myocardial tissue. To solve this problem, an automatic heart perfusion system was developed by applying a Human-Machine Interface (HMI) and a Programmable-Logic-Controller (PLC)-based circuit to control PF and PP. The PLC-based control system collects the data of PF and PP through flow probes and pressure transducers. It has two control modes: the RPM-flow mode and the pressure mode. The RPM-flow control mode is an open-loop system. It influences PF through providing and maintaining the desired speed inputted through the HMI to the centrifugal pump with a maximum error of 20 rpm. The pressure control mode is a closed-loop system where the operator selects a target Mean Arterial Pressure (MAP) to control PP. The inputs of the pressure control mode are the target MAP, received through the HMI, and the real MAP, received from the pressure transducer. A PID algorithm is applied to maintain the real MAP at the target value with a maximum error of 1mmHg. The precision and control speed of the RPM-flow control mode were examined by comparing the PLC-based system to an experienced operator (EO) across seven RPM adjustment ranges (500, 1000, 2000 and random RPM changes; 8 trials per range) tested in a random order. System’s PID algorithm performance in pressure control was assessed during 10 EVHP experiments using porcine hearts. Precision was examined through monitoring the steady-state pressure error throughout perfusion period, and stabilizing speed was tested by performing two MAP adjustment changes (4 trials per change) of 15 and 20mmHg. A total of 56 trials were performed to validate the RPM-flow control mode. Overall, the PLC-based system demonstrated the significantly faster speed than the EO in all trials (PLC 1.21±0.03, EO 3.69±0.23 seconds; p < 0.001) and greater precision to reach the desired RPM (PLC 10±0.7, EO 33±2.7 mean RPM error; p < 0.001). Regarding pressure control, the PLC-based system has the median precision of ±1mmHg error and the median stabilizing times in changing 15 and 20mmHg of MAP are 15 and 19.5 seconds respectively. The novel PLC-based control system was 3 times faster with 60% less error than the EO for RPM-flow control. In pressure control mode, it demonstrates a high precision and fast stabilizing speed. In summary, this novel system successfully controlled perfusion flow and pressure with high precision, stability and a fast response time through a user-friendly interface. This design may provide a viable technique for future development of novel heart preservation and assessment strategies during EVHP.

Keywords: automatic control system, biomedical engineering, ex-vivo heart perfusion, human-machine interface, programmable logic controller

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220 Simulation and Characterization of Stretching and Folding in Microchannel Electrokinetic Flows

Authors: Justo Rodriguez, Daming Chen, Amador M. Guzman

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The detection, treatment, and control of rapidly propagating, deadly viruses such as COVID-19, require the development of inexpensive, fast, and accurate devices to address the urgent needs of the population. Microfluidics-based sensors are amongst the different methods and techniques for detection that are easy to use. A micro analyzer is defined as a microfluidics-based sensor, composed of a network of microchannels with varying functions. Given their size, portability, and accuracy, they are proving to be more effective and convenient than other solutions. A micro analyzer based on the concept of “Lab on a Chip” presents advantages concerning other non-micro devices due to its smaller size, and it is having a better ratio between useful area and volume. The integration of multiple processes in a single microdevice reduces both the number of necessary samples and the analysis time, leading the next generation of analyzers for the health-sciences. In some applications, the flow of solution within the microchannels is originated by a pressure gradient, which can produce adverse effects on biological samples. A more efficient and less dangerous way of controlling the flow in a microchannel-based analyzer is applying an electric field to induce the fluid motion and either enhance or suppress the mixing process. Electrokinetic flows are characterized by no less than two non-dimensional parameters: the electric Rayleigh number and its geometrical aspect ratio. In this research, stable and unstable flows have been studied numerically (and when possible, will be experimental) in a T-shaped microchannel. Additionally, unstable electrokinetic flows for Rayleigh numbers higher than critical have been characterized. The flow mixing enhancement was quantified in relation to the stretching and folding that fluid particles undergo when they are subjected to supercritical electrokinetic flows. Computational simulations were carried out using a finite element-based program while working with the flow mixing concepts developed by Gollub and collaborators. Hundreds of seeded massless particles were tracked along the microchannel from the entrance to exit for both stable and unstable flows. After post-processing, their trajectories, the folding and stretching values for the different flows were found. Numerical results show that for supercritical electrokinetic flows, the enhancement effects of the folding and stretching processes become more apparent. Consequently, there is an improvement in the mixing process, ultimately leading to a more homogenous mixture.

Keywords: microchannel, stretching and folding, electro kinetic flow mixing, micro-analyzer

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219 Analysis of Taxonomic Compositions, Metabolic Pathways and Antibiotic Resistance Genes in Fish Gut Microbiome by Shotgun Metagenomics

Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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218 Barbie in India: A Study of Effects of Barbie in Psychological and Social Health

Authors: Suhrita Saha

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Barbie is a fashion doll manufactured by the American toy company Mattel Inc and it made debut at the American International Toy Fair in New York in 9 March 1959. From being a fashion doll to a symbol of fetishistic commodification, Barbie has come a long way. A Barbie doll is sold every three seconds across the world, which makes the billion dollar brand the world’s most popular doll for the girls. The 11.5 inch moulded plastic doll has a height of 5 feet 9 inches at 1/6 scale. Her vital statistics have been estimated at 36 inches (chest), 18 inches (waist) and 33 inches (hips). Her weight is permanently set at 110 pounds which would be 35 pounds underweight. Ruth Handler, the creator of Barbie wanted a doll that represented adulthood and allowed children to imagine themselves as teenagers or adults. While Barbie might have been intended to be independent, imaginative and innovative, the physical uniqueness does not confine the doll to the status of a play thing. It is a cultural icon but with far reaching critical implications. The doll is a commodity bearing more social value than practical use value. The way Barbie is produced represents industrialization and commodification of the process of symbolic production. And this symbolic production and consumption is a standardized planned one that produce stereotypical ‘pseudo-individuality’ and suppresses cultural alternatives. Children are being subject to and also arise as subjects in this consumer context. A very gendered, physiologically dissected sexually charged symbolism is imposed upon children (both male and female), childhood, their social worlds, identity, and relationship formation. Barbie is also very popular among Indian children. While the doll is essentially an imaginative representation of the West, it is internalized by the Indian sensibilities. Through observation and questionnaire-based interview within a sample population of adolescent children (primarily female, a few male) and parents (primarily mothers) in Kolkata, an Indian metropolis, the paper puts forth findings of sociological relevance. 1. Barbie creates, recreates, and accentuates already existing divides between the binaries like male- female, fat- thin, sexy- nonsexy, beauty- brain and more. 2. The Indian girl child in her associative process with Barbie wants to be like her and commodifies her own self. The male child also readily accepts this standardized commodification. Definition of beauty is thus based on prejudice and stereotype. 3. Not being able to become Barbie creates health issues both psychological and physiological varying from anorexia to obesity as well as personality disorder. 4. From being a plaything Barbie becomes the game maker. Barbie along with many other forms of simulation further creates a consumer culture and market for all kind of fitness related hyper enchantment and subsequent disillusionment. The construct becomes the reality and the real gets lost in the play world. The paper would thus argue that Barbie from being an innocuous doll transports itself into becoming social construct with long term and irreversible adverse impact.

Keywords: barbie, commodification, personality disorder, sterotype

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217 The Use of Technology in Theatrical Performances as a Tool of Audience’S Engagement

Authors: Chrysoula Bousiouta

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Throughout the history of theatre, technology has played an important role both in influencing the relationship between performance and audience and offering different kinds of experiences. The use of technology dates back in ancient times, when the introduction of artifacts, such as “Deus ex machine” in ancient Greek theatre, started. Taking into account the key techniques and experiences used throughout history, this paper investigates how technology, through new media, influences contemporary theatre. In the context of this research, technology is defined as projections, audio environments, video-projections, sensors, tele-connections, all alongside with the performance, challenging audience’s participation. The theoretical framework of the research covers, except for the history of theatre, the theory of “experience economy” that took over the service and goods economy. The research is based on the qualitative and comparative analysis of two case studies, Contact Theatre in Manchester (United Kingdom) and Bios in Athens (Greece). The data selection includes desk research and is complemented with semi structured interviews. Building on the results of the research one could claim that the intended experience of modern/contemporary theatre is that of engagement. In this context, technology -as defined above- plays a leading role in creating it. This experience passes through and exists in the middle of the realms of entertainment, education, estheticism and escapism. Furthermore, it is observed that nowadays, theatre is not only about acting but also about performing; it is that one where the performances are unfinished without the participation of the audience. Both case studies try to achieve the experience of engagement through practices that promote the attraction of attention, the increase of imagination, the interaction, the intimacy and the true activity. These practices are achieved through the script, the scenery, the language and the environment of a performance. Contact and Bios consider technology as an intimate tool in order to accomplish the above, and they make an extended use of it. The research completes a notable record of technological techniques that modern theatres use. The use of technology, inside or outside the limits of film technique’s, helps to rivet the attention of the audience, to make performances enjoyable, to give the sense of the “unfinished” or to be used for things that take place around the spectators and force them to take action, being spect-actors. The advantage of technology is that it can be used as a hook for interaction in all stages of a performance. Further research on the field could involve exploring alternative ways of binding technology and theatre or analyzing how the performance is perceived through the use of technological artifacts.

Keywords: experience of engagement, interactive theatre, modern theatre, performance, technology

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216 Performance Assessment of the Gold Coast Desalination Plant Offshore Multiport Brine Diffuser during ‘Hot Standby’ Operation

Authors: M. J. Baum, B. Gibbes, A. Grinham, S. Albert, D. Gale, P. Fisher

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Alongside the rapid expansion of Seawater Reverse Osmosis technologies there is a concurrent increase in the production of hypersaline brine by-products. To minimize environmental impact, these by-products are commonly disposed into open-coastal environments via submerged diffuser systems as inclined dense jet outfalls. Despite the widespread implementation of this process, diffuser designs are typically based on small-scale laboratory experiments under idealistic quiescent conditions. Studies concerning diffuser performance in the field are limited. A set of experiments were conducted to assess the near field characteristics of brine disposal at the Gold Coast Desalination Plant offshore multiport diffuser. The aim of the field experiments was to determine the trajectory and dilution characteristics of the plume under various discharge configurations with production ranging 66 – 100% of plant operative capacity. The field monitoring system employed an unprecedented static array of temperature and electrical conductivity sensors in a three-dimensional grid surrounding a single diffuser port. Complimenting these measurements, Acoustic Doppler Current Profilers were also deployed to record current variability over the depth of the water column and wave characteristics. Recorded data suggested the open-coastal environment was highly active over the experimental duration with ambient velocities ranging 0.0 – 0.5 m∙s-1, with considerable variability over the depth of the water column observed. Variations in background electrical conductivity corresponding to salinity fluctuations of ± 1.7 g∙kg-1 were also observed. Increases in salinity were detected during plant operation and appeared to be most pronounced 10 – 30 m from the diffuser, consistent with trajectory predictions described by existing literature. Plume trajectories and respective dilutions extrapolated from salinity data are compared with empirical scaling arguments. Discharge properties were found to adequately correlate with modelling projections. Temporal and spatial variation of background processes and their subsequent influence upon discharge outcomes are discussed with a view to incorporating the influence of waves and ambient currents in the design of brine outfalls into the future.

Keywords: brine disposal, desalination, field study, negatively buoyant discharge

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215 Analysis and Modeling of Graphene-Based Percolative Strain Sensor

Authors: Heming Yao

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Graphene-based percolative strain gauges could find applications in many places such as touch panels, artificial skins or human motion detection because of its advantages over conventional strain gauges such as flexibility and transparency. These strain gauges rely on a novel sensing mechanism that depends on strain-induced morphology changes. Once a compression or tension strain is applied to Graphene-based percolative strain gauges, the overlap area between neighboring flakes becomes smaller or larger, which is reflected by the considerable change of resistance. Tiny strain change on graphene-based percolative strain sensor can act as an important leverage to tremendously increase resistance of strain sensor, which equipped graphene-based percolative strain gauges with higher gauge factor. Despite ongoing research in the underlying sensing mechanism and the limits of sensitivity, neither suitable understanding has been obtained of what intrinsic factors play the key role in adjust gauge factor, nor explanation on how the strain gauge sensitivity can be enhanced, which is undoubtedly considerably meaningful and provides guideline to design novel and easy-produced strain sensor with high gauge factor. We here simulated the strain process by modeling graphene flakes and its percolative networks. We constructed the 3D resistance network by simulating overlapping process of graphene flakes and interconnecting tremendous number of resistance elements which were obtained by fractionizing each piece of graphene. With strain increasing, the overlapping graphenes was dislocated on new stretched simulation graphene flake simulation film and a new simulation resistance network was formed with smaller flake number density. By solving the resistance network, we can get the resistance of simulation film under different strain. Furthermore, by simulation on possible variable parameters, such as out-of-plane resistance, in-plane resistance, flake size, we obtained the changing tendency of gauge factor with all these variable parameters. Compared with the experimental data, we verified the feasibility of our model and analysis. The increase of out-of-plane resistance of graphene flake and the initial resistance of sensor, based on flake network, both improved gauge factor of sensor, while the smaller graphene flake size gave greater gauge factor. This work can not only serve as a guideline to improve the sensitivity and applicability of graphene-based strain sensors in the future, but also provides method to find the limitation of gauge factor for strain sensor based on graphene flake. Besides, our method can be easily transferred to predict gauge factor of strain sensor based on other nano-structured transparent optical conductors, such as nanowire and carbon nanotube, or of their hybrid with graphene flakes.

Keywords: graphene, gauge factor, percolative transport, strain sensor

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214 Preparation of Biodegradable Methacrylic Nanoparticles by Semicontinuous Heterophase Polymerization for Drugs Loading: The Case of Acetylsalicylic Acid

Authors: J. Roberto Lopez, Hened Saade, Graciela Morales, Javier Enriquez, Raul G. Lopez

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Implementation of systems based on nanostructures for drug delivery applications have taken relevance in recent studies focused on biomedical applications. Although there are several nanostructures as drugs carriers, the use of polymeric nanoparticles (PNP) has been widely studied for this purpose, however, the main issue for these nanostructures is the size control below 50 nm with a narrow distribution size, due to they must go through different physiological barriers and avoid to be filtered by kidneys (< 10 nm) or the spleen (> 100 nm). Thus, considering these and other factors, it can be mentioned that drug-loaded nanostructures with sizes varying between 10 and 50 nm are preferred in the development and study of PNP/drugs systems. In this sense, the Semicontinuous Heterophase Polymerization (SHP) offers the possibility to obtain PNP in the desired size range. Considering the above explained, methacrylic copolymer nanoparticles were obtained under SHP. The reactions were carried out in a jacketed glass reactor with the required quantities of water, ammonium persulfate as initiator, sodium dodecyl sulfate/sodium dioctyl sulfosuccinate as surfactants, methyl methacrylate and methacrylic acid as monomers with molar ratio of 2/1, respectively. The monomer solution was dosed dropwise during reaction at 70 °C with a mechanical stirring of 650 rpm. Nanoparticles of poly(methyl methacrylate-co-methacrylic acid) were loaded with acetylsalicylic acid (ASA, aspirin) by a chemical adsorption technique. The purified latex was put in contact with a solution of ASA in dichloromethane (DCM) at 0.1, 0.2, 0.4 or 0.6 wt-%, at 35°C during 12 hours. According to the boiling point of DCM, as well as DCM and water densities, the loading process is completed when the whole DCM is evaporated. The hydrodynamic diameter was measured after polymerization by quasi-elastic light scattering and transmission electron microscopy, before and after loading procedures with ASA. The quantitative and qualitative analyses of PNP loaded with ASA were measured by infrared spectroscopy, differential scattering calorimetry and thermogravimetric analysis. Also, the molar mass distributions of polymers were determined in a gel permeation chromatograph apparatus. The load capacity and efficiency were determined by gravimetric analysis. The hydrodynamic diameter results for methacrylic PNP without ASA showed a narrow distribution with an average particle size around 10 nm and a composition methyl methacrylate/methacrylic acid molar ratio equal to 2/1, same composition of Eudragit S100, which is a commercial compound widely used as excipient. Moreover, the latex was stabilized in a relative high solids content (around 11 %), a monomer conversion almost 95 % and a number molecular weight around 400 Kg/mol. The average particle size in the PNP/aspirin systems fluctuated between 18 and 24 nm depending on the initial percentage of aspirin in the loading process, being the drug content as high as 24 % with an efficiency loading of 36 %. These average sizes results have not been reported in the literature, thus, the methacrylic nanoparticles here reported are capable to be loaded with a considerable amount of ASA and be used as a drug carrier.

Keywords: aspirin, biocompatibility, biodegradable, Eudragit S100, methacrylic nanoparticles

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213 Simscape Library for Large-Signal Physical Network Modeling of Inertial Microelectromechanical Devices

Authors: S. Srinivasan, E. Cretu

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The information flow (e.g. block-diagram or signal flow graph) paradigm for the design and simulation of Microelectromechanical (MEMS)-based systems allows to model MEMS devices using causal transfer functions easily, and interface them with electronic subsystems for fast system-level explorations of design alternatives and optimization. Nevertheless, the physical bi-directional coupling between different energy domains is not easily captured in causal signal flow modeling. Moreover, models of fundamental components acting as building blocks (e.g. gap-varying MEMS capacitor structures) depend not only on the component, but also on the specific excitation mode (e.g. voltage or charge-actuation). In contrast, the energy flow modeling paradigm in terms of generalized across-through variables offers an acausal perspective, separating clearly the physical model from the boundary conditions. This promotes reusability and the use of primitive physical models for assembling MEMS devices from primitive structures, based on the interconnection topology in generalized circuits. The physical modeling capabilities of Simscape have been used in the present work in order to develop a MEMS library containing parameterized fundamental building blocks (area and gap-varying MEMS capacitors, nonlinear springs, displacement stoppers, etc.) for the design, simulation and optimization of MEMS inertial sensors. The models capture both the nonlinear electromechanical interactions and geometrical nonlinearities and can be used for both small and large signal analyses, including the numerical computation of pull-in voltages (stability loss). Simscape behavioral modeling language was used for the implementation of reduced-order macro models, that present the advantage of a seamless interface with Simulink blocks, for creating hybrid information/energy flow system models. Test bench simulations of the library models compare favorably with both analytical results and with more in-depth finite element simulations performed in ANSYS. Separate MEMS-electronic integration tests were done on closed-loop MEMS accelerometers, where Simscape was used for modeling the MEMS device and Simulink for the electronic subsystem.

Keywords: across-through variables, electromechanical coupling, energy flow, information flow, Matlab/Simulink, MEMS, nonlinear, pull-in instability, reduced order macro models, Simscape

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212 Tuning the Emission Colour of Phenothiazine by Introduction of Withdrawing Electron Groups

Authors: Andrei Bejan, Luminita Marin, Dalila Belei

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Phenothiazine with electron-rich nitrogen and sulfur heteroatoms has a high electron-donating ability which promotes a good conjugation and therefore low band-gap with consequences upon charge carrier mobility improving and shifting of light emission in visible domain. Moreover, its non-planar butterfly conformation inhibits molecular aggregation and thus preserves quite well the fluorescence quantum yield in solid state compared to solution. Therefore phenothiazine and its derivatives are promising hole transport materials for use in organic electronic and optoelectronic devices as light emitting diodes, photovoltaic cells, integrated circuit sensors or driving circuits for large area display devices. The objective of this paper was to obtain a series of new phenothiazine derivatives by introduction of different electron withdrawing substituents as formyl, carboxyl and cyanoacryl units in order to create a push pull system which has potential to improve the electronic and optical properties. Bromine atom was used as electrono-donor moiety to extend furthermore the existing conjugation. The understudy compounds were structural characterized by FTIR and 1H-NMR spectroscopy and single crystal X-ray diffraction. Besides, the single crystal X-ray diffraction brought information regarding the supramolecular architecture of the compounds. Photophysical properties were monitored by UV-vis and photoluminescence spectroscopy, while the electrochemical behavior was established by cyclic voltammetry. The absorption maxima of the studied compounds vary in a large range (322-455 nm), reflecting the different electronic delocalization degree, depending by the substituent nature. In a similar manner, the emission spectra reveal different color of emitted light, a red shift being evident for the groups with higher electron withdrawing ability. The emitted light is pure and saturated for the compounds containing strong withdrawing formyl or cyanoacryl units and reach the highest quantum yield of 71% for the compound containing bromine and cyanoacrilic units. Electrochemical study show reversible oxidative and reduction processes for all the compounds and a close correlation of the HOMO-LUMO band gap with substituent nature. All these findings suggest the obtained compounds as promising materials for optoelectronic devices.

Keywords: electrochemical properties, phenothiazine derivatives, photoluminescence, quantum yield

Procedia PDF Downloads 309
211 Kinematic Modelling and Task-Based Synthesis of a Passive Architecture for an Upper Limb Rehabilitation Exoskeleton

Authors: Sakshi Gupta, Anupam Agrawal, Ekta Singla

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An exoskeleton design for rehabilitation purpose encounters many challenges, including ergonomically acceptable wearing technology, architectural design human-motion compatibility, actuation type, human-robot interaction, etc. In this paper, a passive architecture for upper limb exoskeleton is proposed for assisting in rehabilitation tasks. Kinematic modelling is detailed for task-based kinematic synthesis of the wearable exoskeleton for self-feeding tasks. The exoskeleton architecture possesses expansion and torsional springs which are able to store and redistribute energy over the human arm joints. The elastic characteristics of the springs have been optimized to minimize the mechanical work of the human arm joints. The concept of hybrid combination of a 4-bar parallelogram linkage and a serial linkage were chosen, where the 4-bar parallelogram linkage with expansion spring acts as a rigid structure which is used to provide the rotational degree-of-freedom (DOF) required for lowering and raising of the arm. The single linkage with torsional spring allows for the rotational DOF required for elbow movement. The focus of the paper is kinematic modelling, analysis and task-based synthesis framework for the proposed architecture, keeping in considerations the essential tasks of self-feeding and self-exercising during rehabilitation of partially healthy person. Rehabilitation of primary functional movements (activities of daily life, i.e., ADL) is routine activities that people tend to every day such as cleaning, dressing, feeding. We are focusing on the feeding process to make people independent in respect of the feeding tasks. The tasks are focused to post-surgery patients under rehabilitation with less than 40% weakness. The challenges addressed in work are ensuring to emulate the natural movement of the human arm. Human motion data is extracted through motion-sensors for targeted tasks of feeding and specific exercises. Task-based synthesis procedure framework will be discussed for the proposed architecture. The results include the simulation of the architectural concept for tracking the human-arm movements while displaying the kinematic and static study parameters for standard human weight. D-H parameters are used for kinematic modelling of the hybrid-mechanism, and the model is used while performing task-based optimal synthesis utilizing evolutionary algorithm.

Keywords: passive mechanism, task-based synthesis, emulating human-motion, exoskeleton

Procedia PDF Downloads 118
210 Conceptual Methods of Mitigating Matured Urban Tree Roots Surviving in Conflicts Growth within Built Environment: A Review

Authors: Mohd Suhaizan Shamsuddin

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Urbanization exacerbates the environment quality and pressures of matured urban trees' growth and development in changing environment. The growth of struggled matured urban tree-roots by spreading within the existences of infrastructures, resulting in large damage to the structured and declined growth. Many physiological growths declined or damages by the present and installations of infrastructures within and nearby root zone. Afford to remain both matured urban tree and infrastructures as a service provider causes damage and death, respectively. Inasmuch, spending more expenditure on fixing both or removing matured urban trees as risky to the future environment as the mitigation methods to reduce the problems are unconcerned. This paper aims to explain mitigation method practices of reducing the encountered problems of matured urban tree-roots settling and infrastructures while modified urban soil to sustain at an optimum level. Three categories capturing encountered conflicts growth of matured urban tree-roots growth within and nearby infrastructures by mitigating the problems of limited soil spaces, poor soil structures and soil space barrier installations and maintenance. The limited soil space encountered many conflicts and identified six methods that mitigate the survival tree-roots, such as soil volume/mounding, soil replacement/amendment for the radial trench, soil spacing-root bridge, root tunneling, walkway/pavement rising/diverted, and suspended pavement. The limited soil spaces are mitigation affords of inadequate soil-roots and spreading root settling and modification of construction soil media since the barrier existed and installed in root trails or zones. This is the reason for enabling tree-roots spreading and finds adequate sources (nutrients, water uptake and oxygen), spaces and functioning to stability stand of root anchorage since the matured tree grows larger. The poor soil structures were identified as three methods to mitigate soil materials' problems, and fewer soil voids comprise skeletal soil, structural soil, and soil cell. Mitigation of poor soil structure is altering the existing and introducing new structures by modifying the quantities and materials ratio allowing more voids beneath for roots spreading by considering the above structure of foot and vehicle traffics functioning or load-bearing. The soil space barrier installations and maintenance recognized to sustain both infrastructures and tree-roots grown in limited spaces and its benefits, the root barrier installations and root pruning are recommended. In conclusion, these recommended methods attempt to mitigate the present problems encountered at a particular place and problems among tree-roots and infrastructures exist. The combined method is the best way to alleviates the conflicts since the recognized conflicts are between tree-roots and man-made while the urban soil is modified. These presenting methods are most considered to sustain the matured urban trees' lifespan growth in the urban environment.

Keywords: urban tree-roots, limited soil spaces, poor soil structures, soil space barrier and maintenance

Procedia PDF Downloads 168
209 Transport Properties of Alkali Nitrites

Authors: Y. Mateyshina, A.Ulihin, N.Uvarov

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Electrolytes with different type of charge carrier can find widely application in different using, e.g. sensors, electrochemical equipments, batteries and others. One of important components ensuring stable functioning of the equipment is electrolyte. Electrolyte has to be characterized by high conductivity, thermal stability, and wide electrochemical window. In addition to many advantageous characteristic for liquid electrolytes, the solid state electrolytes have good mechanical stability, wide working range of temperature range. Thus search of new system of solid electrolytes with high conductivity is an actual task of solid state chemistry. Families of alkali perchlorates and nitrates have been investigated by us earlier. In literature data about transport properties of alkali nitrites are absent. Nevertheless, alkali nitrites MeNO2 (Me= Li+, Na+, K+, Rb+ and Cs+), except for the lithium salt, have high-temperature phases with crystal structure of the NaCl-type. High-temperature phases of nitrites are orientationally disordered, i.e. non-spherical anions are reoriented over several equivalents directions in the crystal lattice. Pure lithium nitrite LiNO2 is characterized by ionic conductivity near 10-4 S/cm at 180°C and more stable as compared with lithium nitrate and can be used as a component for synthesis of composite electrolytes. In this work composite solid electrolytes in the binary system LiNO2 - A (A= MgO, -Al2O3, Fe2O3, CeO2, SnO2, SiO2) were synthesized and their structural, thermodynamic and electrical properties investigated. Alkali nitrite was obtained by exchange reaction from water solutions of barium nitrite and alkali sulfate. The synthesized salt was characterized by X-ray powder diffraction technique using D8 Advance X-Ray Diffractometer with Cu K radiation. Using thermal analysis, the temperatures of dehydration and thermal decomposition of salt were determined.. The conductivity was measured using a two electrode scheme in a forevacuum (6.7 Pa) with an HP 4284A (Precision LCR meter) in a frequency range 20 Hz < ν < 1 MHz. Solid composite electrolytes LiNO2 - A A (A= MgO, -Al2O3, Fe2O3, CeO2, SnO2, SiO2) have been synthesized by mixing of preliminary dehydrated components followed by sintering at 250°C. In the series of nitrite of alkaline metals Li+-Cs+, the conductivity varies not monotonically with increasing radius of cation. The minimum conductivity is observed for KNO2; however, with further increase in the radius of cation in the series, the conductivity tends to increase. The work was supported by the Russian Foundation for Basic research, grant #14-03-31442.

Keywords: conductivity, alkali nitrites, composite electrolytes, transport properties

Procedia PDF Downloads 294
208 Feasibility of Small Autonomous Solar-Powered Water Desalination Units for Arid Regions

Authors: Mohamed Ahmed M. Azab

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The shortage of fresh water is a major problem in several areas of the world such as arid regions and coastal zones in several countries of Arabian Gulf. Fortunately, arid regions are exposed to high levels of solar irradiation most the year, which makes the utilization of solar energy a promising solution to such problem with zero harmful emission (Green System). The main objective of this work is to conduct a feasibility study of utilizing small autonomous water desalination units powered by photovoltaic modules as a green renewable energy resource to be employed in different isolated zones as a source of drinking water for some scattered societies where the installation of huge desalination stations are discarded owing to the unavailability of electric grid. Yanbu City is chosen as a case study where the Renewable Energy Center exists and equipped with all sensors to assess the availability of solar energy all over the year. The study included two types of available water: the first type is brackish well water and the second type is seawater of coastal regions. In the case of well water, two versions of desalination units are involved in the study: the first version is based on day operation only. While the second version takes into consideration night operation also, which requires energy storage system as batteries to provide the necessary electric power at night. According to the feasibility study results, it is found that utilization of small autonomous desalinations unit is applicable and economically accepted in the case of brackish well water. While in the case of seawater the capital costs are extremely high and the cost of desalinated water will not be economically feasible unless governmental subsidies are provided. In addition, the study indicated that, for the same water production, the utilization of energy storage version (day-night) adds additional capital cost for batteries, and extra running cost for their replacement, which makes the unit price not only incompetent with day-only unit but also with conventional units powered by diesel generator (fossil fuel) owing to the low prices of fuel in the kingdom. However, the cost analysis shows that the price of the produced water per cubic meter of day-night unit is similar to that produced from the day-only unit provided that the day-night unit operates theoretically for a longer period of 50%.

Keywords: solar energy, water desalination, reverse osmosis, arid regions

Procedia PDF Downloads 421
207 Identification of Damage Mechanisms in Interlock Reinforced Composites Using a Pattern Recognition Approach of Acoustic Emission Data

Authors: M. Kharrat, G. Moreau, Z. Aboura

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The latest advances in the weaving industry, combined with increasingly sophisticated means of materials processing, have made it possible to produce complex 3D composite structures. Mainly used in aeronautics, composite materials with 3D architecture offer better mechanical properties than 2D reinforced composites. Nevertheless, these materials require a good understanding of their behavior. Because of the complexity of such materials, the damage mechanisms are multiple, and the scenario of their appearance and evolution depends on the nature of the exerted solicitations. The AE technique is a well-established tool for discriminating between the damage mechanisms. Suitable sensors are used during the mechanical test to monitor the structural health of the material. Relevant AE-features are then extracted from the recorded signals, followed by a data analysis using pattern recognition techniques. In order to better understand the damage scenarios of interlock composite materials, a multi-instrumentation was set-up in this work for tracking damage initiation and development, especially in the vicinity of the first significant damage, called macro-damage. The deployed instrumentation includes video-microscopy, Digital Image Correlation, Acoustic Emission (AE) and micro-tomography. In this study, a multi-variable AE data analysis approach was developed for the discrimination between the different signal classes representing the different emission sources during testing. An unsupervised classification technique was adopted to perform AE data clustering without a priori knowledge. The multi-instrumentation and the clustered data served to label the different signal families and to build a learning database. This latter is useful to construct a supervised classifier that can be used for automatic recognition of the AE signals. Several materials with different ingredients were tested under various solicitations in order to feed and enrich the learning database. The methodology presented in this work was useful to refine the damage threshold for the new generation materials. The damage mechanisms around this threshold were highlighted. The obtained signal classes were assigned to the different mechanisms. The isolation of a 'noise' class makes it possible to discriminate between the signals emitted by damages without resorting to spatial filtering or increasing the AE detection threshold. The approach was validated on different material configurations. For the same material and the same type of solicitation, the identified classes are reproducible and little disturbed. The supervised classifier constructed based on the learning database was able to predict the labels of the classified signals.

Keywords: acoustic emission, classifier, damage mechanisms, first damage threshold, interlock composite materials, pattern recognition

Procedia PDF Downloads 135
206 Exploring Fluoroquinolone-Resistance Dynamics Using a Distinct in Vitro Fermentation Chicken Caeca Model

Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

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Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

Procedia PDF Downloads 34
205 Neonatology Clinical Routine in Cats and Dogs: Cases, Main Conditions and Mortality

Authors: Maria L. G. Lourenço, Keylla H. N. P. Pereira, Viviane Y. Hibaru, Fabiana F. Souza, João C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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The neonatal care of cats and dogs represents a challenge to veterinarians due to the small size of the newborns and their physiological particularities. In addition, many Veterinary Medicine colleges around the world do not include neonatology in the curriculum, which makes it less likely for the veterinarian to have basic knowledge regarding neonatal care and worsens the clinical care these patients receive. Therefore, lack of assistance and negligence have become frequent in the field, which contributes towards the high mortality rates. This study aims at describing cases and the main conditions pertaining to the neonatology clinical routine in cats and dogs, highlighting the importance of specialized care in this field of Veterinary Medicine. The study included 808 neonates admitted to the São Paulo State University (UNESP) Veterinary Hospital, Botucatu, São Paulo, Brazil, between January 2018 and November 2019. Of these, 87.3% (705/808) were dogs and 12.7% (103/808) were cats. Among the neonates admitted, 57.3% (463/808) came from emergency c-sections due to dystocia, 8.7% (71/808) cane from vaginal deliveries with obstetric maneuvers due to dystocia, and 34% (274/808) were admitted for clinical care due to neonatal conditions. Among the neonates that came from emergency c-sections and vaginal deliveries, 47.3% (253/534) was born in respiratory distress due to severe hypoxia or persistent apnea and required resuscitation procedure, such as the Jen Chung acupuncture point (VG26), oxygen therapy with mask, pulmonary expansion with resuscitator, heart massages and administration of emergency medication, such as epinephrine. On the other hand, in the neonatal clinical care, the main conditions and alterations observed in the newborns were omphalophlebitis, toxic milk syndrome, neonatal conjunctivitis, swimmer puppy syndrome, neonatal hemorrhagic syndrome, pneumonia, trauma, low weight at birth, prematurity, congenital malformations (cleft palate, cleft lip, hydrocephaly, anasarca, vascular anomalies in the heart, anal atresia, gastroschisis, omphalocele, among others), neonatal sepsis and other local and systemic bacterial infections, viral infections (feline respiratory complex, parvovirus, canine distemper, canine infectious traqueobronchitis), parasitical infections (Toxocara spp., Ancylostoma spp., Strongyloides spp., Cystoisospora spp., Babesia spp. and Giardia spp.) and fungal infections (dermatophytosis by Microsporum canis). The most common clinical presentation observed was the neonatal triad (hypothermia, hypoglycemia and dehydration), affecting 74.6% (603/808) of the patients. The mortality rate among the neonates was 10.5% (85/808). Being knowledgeable about neonatology is essential for veterinarians to provide adequate care for these patients in the clinical routine. Adding neonatology to college curriculums, improving the dissemination of information on the subject, and providing annual training in neonatology for veterinarians and employees are important to improve immediate care and reduce the mortality rates.

Keywords: neonatal care, puppies, neonatal, conditions

Procedia PDF Downloads 200
204 The Role of Two Macrophyte Species in Mineral Nutrient Cycling in Human-Impacted Water Reservoirs

Authors: Ludmila Polechonska, Agnieszka Klink

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The biogeochemical studies of macrophytes shed light on elements bioavailability, transfer through the food webs and their possible effects on the biota, and provide a basis for their practical application in aquatic monitoring and remediation. Measuring the accumulation of elements in plants can provide time-integrated information about the presence of chemicals in aquatic ecosystems. The aim of the study was to determine and compare the contents of micro- and macroelements in two cosmopolitan macrophytes, submerged Ceratophyllum demersum (hornworth) and free-floating Hydrocharis morsus-ranae (European frog-bit), in order to assess their bioaccumulation potential, elements stock accumulated in each plant and their role in nutrients cycling in small water reservoirs. Sampling sites were designated in 25 oxbow lakes in urban areas in Lower Silesia (SW Poland). In each sampling site, fresh whole plants of C. demersum and H. morsus-ranae were collected from squares of 1x1 meters each where the species coexisted. European frog-bit was separated into leaves, stems and roots. For biomass measurement all plants growing on 1 square meter were collected, dried and weighed. At the same time, water samples were collected from each reservoir and their pH and EC were determined. Water samples were filtered and acidified and plant samples were digested in concentrated nitric acid. Next, the content of Ca, Cu, Fe, K, Mg, Mn, Ni and Zn was determined using atomic absorption method (AAS). Statistical analysis showed that C. demersum and organs of H. morsus-ranae differed significantly in respect of metals content (Kruskal-Wallis Anova, p<0.05). Contents of Cu, Mn, Ni and Zn were higher in hornwort, while European frog-bit contained more Ca, Fe, K, Mg. Bioaccumulation Factors (BCF=content in plant/concentration in water) showed similar pattern of metal bioaccumulation – microelements were more intensively accumulated by hornwort and macroelements by frog-bit. Based on BCF values both species may be positively evaluated as good accumulators of Cu, Fe, Mn, Ni and Zn. However, the distribution of metals in H. morsus-ranae was uneven – the majority of studied elements were retained in roots, which may indicate to existence of physiological barriers developed for dealing with toxicity. Some percent of Ca and K was actively transported to stems, but to leaves Mg only. Although the biomass of C. demersum was two times greater than biomass of H. morsus-ranae, the element off-take was greater only for Cu, Mn, Ni and Zn. Nevertheless, it can be stated that despite a relatively small biomass, compared to other macrophytes, both species may have an influence on the removal of trace elements from aquatic ecosystems and, as they serve as food for some animals, also on the incorporation of toxic elements into food chains. There was a significant positive correlation between content of Mn and Fe in water and roots of H. morus-ranae (R=0.51 and R=0.60, respectively) as well as between Cu concentration in water and in C. demersum (R=0.41) (Spearman rank correlation, p<0.05). High bioaccumulation rates and correlation between plants and water elements concentrations point to their possible use as passive biomonitors of aquatic pollution.

Keywords: aquatic plants, bioaccumulation, biomonitoring, macroelements, phytoremediation, trace metals

Procedia PDF Downloads 157
203 Computational and Experimental Determination of Acoustic Impedance of Internal Combustion Engine Exhaust

Authors: A. O. Glazkov, A. S. Krylova, G. G. Nadareishvili, A. S. Terenchenko, S. I. Yudin

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The topic of the presented materials concerns the design of the exhaust system for a certain internal combustion engine. The exhaust system can be divided into two parts. The first is the engine exhaust manifold, turbocharger, and catalytic converters, which are called “hot part.” The second part is the gas exhaust system, which contains elements exclusively for reducing exhaust noise (mufflers, resonators), the accepted designation of which is the "cold part." The design of the exhaust system from the point of view of acoustics, that is, reducing the exhaust noise to a predetermined level, consists of working on the second part. Modern computer technology and software make it possible to design "cold part" with high accuracy in a given frequency range but with the condition of accurately specifying the input parameters, namely, the amplitude spectrum of the input noise and the acoustic impedance of the noise source in the form of an engine with a "hot part". Getting this data is a difficult problem: high temperatures, high exhaust gas velocities (turbulent flows), and high sound pressure levels (non-linearity mode) do not allow the calculated results to be applied with sufficient accuracy. The aim of this work is to obtain the most reliable acoustic output parameters of an engine with a "hot part" based on a complex of computational and experimental studies. The presented methodology includes several parts. The first part is a finite element simulation of the "cold part" of the exhaust system (taking into account the acoustic impedance of radiation of outlet pipe into open space) with the result in the form of the input impedance of "cold part". The second part is a finite element simulation of the "hot part" of the exhaust system (taking into account acoustic characteristics of catalytic units and geometry of turbocharger) with the result in the form of the input impedance of the "hot part". The next third part of the technique consists of the mathematical processing of the results according to the proposed formula for the convergence of the mathematical series of summation of multiple reflections of the acoustic signal "cold part" - "hot part". This is followed by conducting a set of tests on an engine stand with two high-temperature pressure sensors measuring pulsations in the nozzle between "hot part" and "cold part" of the exhaust system and subsequent processing of test results according to a well-known technique in order to separate the "incident" and "reflected" waves. The final stage consists of the mathematical processing of all calculated and experimental data to obtain a result in the form of a spectrum of the amplitude of the engine noise and its acoustic impedance.

Keywords: acoustic impedance, engine exhaust system, FEM model, test stand

Procedia PDF Downloads 29
202 Influence of the Use of Fruits Byproducts on the Lipid Profile of Hermetia illucens, Tenebrio molitor and Zophoba morio Larvae

Authors: Rebeca P Ramos-Bueno, Maria Jose Gonzalez-Fernandez, Rosa M. Moreno-Zamora, Antonia Barros Heras, Yolanda Serrano Alonso, Carolina Sanchez Barranco

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Insects are a new source of fatty acids (FA), so they are considered a sustainable and environmentally friendly alternative for both animal feed and the human diet, and furthermore, their harvesting/rearing require a low-tech and low capital investment. For that reason, lipids obtained by insect breeding open interesting possibilities with alimentary and industrial purposes, i.e., the production of biodiesel. Particularly, certain insect species, especially during the larval stage, contain high proportions of fat which is highly dependent on their feed and stage of development. Among them, Hermetia illucens larvae can be bred on food wastes to produce fat- and protein-rich raw materials for food by-product management. So, insects can act as excellent bioconverters of organic waste to nutrient-rich materials. In this regard, the aim of the study was to evaluate the effects of fruit byproducts on the FA compositions of Tenebrio molitor, Zophoba morio, and H. illucens larvae. Firstly, oil was extracted with the green solvent ethyl acetate, and FA methyl ester was obtained and analyzed by GC to show the FA profile. In addition, the triacylglycerol (TAG) profile was obtained by HPLC. Dehydrated watermelon, tomato, and papaya by-products, as well as wheat-based control feed, were assayed. High FA content was reached by Z. morio larvae fed with all fruits; however, no differences were shown in lipid profile with any change. It is worth highlighting that both Z. morio and H. illucens could be selected as the best candidates for biodiesel production due to their high content of saturated FA. On the other hand, T. molitor larvae showed a higher content of monounsaturated FA than control larvae, whereas the n-6 polyunsaturated FA content decreased in larvae fed with fruits. This result indicates that the improvement of the FA profile of Tenebrio can depend on both the type of feeding and the intended use. The lipid profile of H. illucens larvae fed with papaya and tomato showed a slight increase in the content of α-linoleic acid (ALA, 18:3n3). This FA is the precursor of docosahexaenoic acid (DHA, 22:6n3), which plays an important role as a component of structural lipids in cell membranes as well as in the synthesis of eicosanoids, protecting and resolving. Also, it was evaluated the TAG profile of Z. morio larvae due to their highest oil content. The results showed a high oleic acid (OA, 18:1n9) content, which displays modulatory effects in a wide range of physiological functions, having anti-inflammatory and anti-atherogenic properties. In conclusion, this study clearly shows that Z. morio and H. illucens larvae constitute an alternative source of OA- and ALA-rich oils, respectively, which can be devoted for food use, as well as for using in the food and pharmaceutical industries, with agronomic implications. Finally, although the profile of Z. morio was not improved with fruit feeding, this kind of feeding could be used due to its low environmental impact.

Keywords: fatty acids, fruit byproducts, Hermetia illucens, Zophoba morio, Tenebrio molitor, insect rearing

Procedia PDF Downloads 121
201 Robust Processing of Antenna Array Signals under Local Scattering Environments

Authors: Ju-Hong Lee, Ching-Wei Liao

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An adaptive array beamformer is designed for automatically preserving the desired signals while cancelling interference and noise. Providing robustness against model mismatches and tracking possible environment changes calls for robust adaptive beamforming techniques. The design criterion yields the well-known generalized sidelobe canceller (GSC) beamformer. In practice, the knowledge of the desired steering vector can be imprecise, which often occurs due to estimation errors in the DOA of the desired signal or imperfect array calibration. In these situations, the SOI is considered as interference, and the performance of the GSC beamformer is known to degrade. This undesired behavior results in a reduction of the array output signal-to-interference plus-noise-ratio (SINR). Therefore, it is worth developing robust techniques to deal with the problem due to local scattering environments. As to the implementation of adaptive beamforming, the required computational complexity is enormous when the array beamformer is equipped with massive antenna array sensors. To alleviate this difficulty, a generalized sidelobe canceller (GSC) with partially adaptivity for less adaptive degrees of freedom and faster adaptive response has been proposed in the literature. Unfortunately, it has been shown that the conventional GSC-based adaptive beamformers are usually very sensitive to the mismatch problems due to local scattering situations. In this paper, we present an effective GSC-based beamformer against the mismatch problems mentioned above. The proposed GSC-based array beamformer adaptively estimates the actual direction of the desired signal by using the presumed steering vector and the received array data snapshots. We utilize the predefined steering vector and a presumed angle tolerance range to carry out the required estimation for obtaining an appropriate steering vector. A matrix associated with the direction vector of signal sources is first created. Then projection matrices related to the matrix are generated and are utilized to iteratively estimate the actual direction vector of the desired signal. As a result, the quiescent weight vector and the required signal blocking matrix required for performing adaptive beamforming can be easily found. By utilizing the proposed GSC-based beamformer, we find that the performance degradation due to the considered local scattering environments can be effectively mitigated. To further enhance the beamforming performance, a signal subspace projection matrix is also introduced into the proposed GSC-based beamformer. Several computer simulation examples show that the proposed GSC-based beamformer outperforms the existing robust techniques.

Keywords: adaptive antenna beamforming, local scattering, signal blocking, steering mismatch

Procedia PDF Downloads 86
200 Productivity of Grain Sorghum-Cowpea Intercropping System: Climate-Smart Approach

Authors: Mogale T. E., Ayisi K. K., Munjonji L., Kifle Y. G.

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Grain sorghum and cowpea are important staple crops in many areas of South Africa, particularly the Limpopo Province. The two crops are produced under a wide range of unsustainable conventional methods, which reduces productivity in the long run. Climate-smart traditional methods such as intercropping can be adopted to ensure sustainable production of these important two crops in the province. A no-tillage field experiment was laid out in a randomised complete block design (RCBD) with four replications over two seasons in two distinct agro-ecological zones, Syferkuil and Ofcolacoin, the province to assess the productivity of sorghum-cowpea intercropped under two cowpea densities.LCi Ultra compact photosynthesis machine was used to collect photosynthetic rate data biweekly between 11h00 and 13h00 until physiological maturity. Biomass and grain yield of the component crops in binary and sole cultures were determined at harvest maturity from middle rows of 2.7 m2 area. The biomass was oven dried in the laboratory at 65oC till constant weight. To obtain grain yield, harvested sorghum heads and cowpea pods were threshed, cleaned, and weighed. Harvest index (HI) and land equivalent ratio (LER) of the two crops were calculated to assess intercrop productivity relative to sole cultures. Data was analysed using the statistical analysis software system (SAS) 9.4 version, followed by mean separation using the least significant difference method. The photosyntheticrate of sorghum-cowpea intercrop was influenced by cowpea density and sorghum cultivar. Photosynthetic rate under low density was higher compared to high density, but this was dependent on the growing conditions. Dry biomass accumulation, grain yield, and harvest index differed among the sorghum cultivars and cowpea in both binary and sole cultures at the two test locations during the 2018/19 and 2020/21 growing seasons. Cowpea grain and dry biomass yields werein excess of 60% under high density compared to low density in both binary and sole cultures. The results revealed that grain yield accumulation of sorghum cultivars was influenced by the density of the companion cowpea crop as well as the production season. For instant, at Syferkuil, Enforcer and Ns5511 accumulated high yield under low density, whereas, at Ofcolaco, the higher yield was recorded under high density. Generally, under low cowpea density, cultivar Enforcer produced relatively higher grain yield whereas, under higher density, Titan yield was superior. The partial and total LER varied with growing season and the treatments studied. The total LERs exceeded 1.0 at the two locations across seasons, ranging from 1.3 to 1.8. From the results, it can be concluded that resources were used more efficiently in sorghum-cowpea intercrop at both Syferkuil and Ofcolaco. Furthermore, intercropping system improved photosynthetic rate, grain yield, and dry matter accumulation of sorghum and cowpea depending on growing conditions and density of cowpea. Hence, the sorghum-cowpea intercropping system can be adopted as a climate-smart practice for sustainable production in the Limpopo province.

Keywords: cowpea, climate-smart, grain sorghum, intercropping

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199 Magnetic Navigation in Underwater Networks

Authors: Kumar Divyendra

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Underwater Sensor Networks (UWSNs) have wide applications in areas such as water quality monitoring, marine wildlife management etc. A typical UWSN system consists of a set of sensors deployed randomly underwater which communicate with each other using acoustic links. RF communication doesn't work underwater, and GPS too isn't available underwater. Additionally Automated Underwater Vehicles (AUVs) are deployed to collect data from some special nodes called Cluster Heads (CHs). These CHs aggregate data from their neighboring nodes and forward them to the AUVs using optical links when an AUV is in range. This helps reduce the number of hops covered by data packets and helps conserve energy. We consider the three-dimensional model of the UWSN. Nodes are initially deployed randomly underwater. They attach themselves to the surface using a rod and can only move upwards or downwards using a pump and bladder mechanism. We use graph theory concepts to maximize the coverage volume while every node maintaining connectivity with at least one surface node. We treat the surface nodes as landmarks and each node finds out its hop distance from every surface node. We treat these hop-distances as coordinates and use them for AUV navigation. An AUV intending to move closer to a node with given coordinates moves hop by hop through nodes that are closest to it in terms of these coordinates. In absence of GPS, multiple different approaches like Inertial Navigation System (INS), Doppler Velocity Log (DVL), computer vision-based navigation, etc., have been proposed. These systems have their own drawbacks. INS accumulates error with time, vision techniques require prior information about the environment. We propose a method that makes use of the earth's magnetic field values for navigation and combines it with other methods that simultaneously increase the coverage volume under the UWSN. The AUVs are fitted with magnetometers that measure the magnetic intensity (I), horizontal inclination (H), and Declination (D). The International Geomagnetic Reference Field (IGRF) is a mathematical model of the earth's magnetic field, which provides the field values for the geographical coordinateson earth. Researchers have developed an inverse deep learning model that takes the magnetic field values and predicts the location coordinates. We make use of this model within our work. We combine this with with the hop-by-hop movement described earlier so that the AUVs move in such a sequence that the deep learning predictor gets trained as quickly and precisely as possible We run simulations in MATLAB to prove the effectiveness of our model with respect to other methods described in the literature.

Keywords: clustering, deep learning, network backbone, parallel computing

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198 Mathematical Modeling of Avascular Tumor Growth and Invasion

Authors: Meitham Amereh, Mohsen Akbari, Ben Nadler

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Cancer has been recognized as one of the most challenging problems in biology and medicine. Aggressive tumors are a lethal type of cancers characterized by high genomic instability, rapid progression, invasiveness, and therapeutic resistance. Their behavior involves complicated molecular biology and consequential dynamics. Although tremendous effort has been devoted to developing therapeutic approaches, there is still a huge need for new insights into the dark aspects of tumors. As one of the key requirements in better understanding the complex behavior of tumors, mathematical modeling and continuum physics, in particular, play a pivotal role. Mathematical modeling can provide a quantitative prediction on biological processes and help interpret complicated physiological interactions in tumors microenvironment. The pathophysiology of aggressive tumors is strongly affected by the extracellular cues such as stresses produced by mechanical forces between the tumor and the host tissue. During the tumor progression, the growing mass displaces the surrounding extracellular matrix (ECM), and due to the level of tissue stiffness, stress accumulates inside the tumor. The produced stress can influence the tumor by breaking adherent junctions. During this process, the tumor stops the rapid proliferation and begins to remodel its shape to preserve the homeostatic equilibrium state. To reach this, the tumor, in turn, upregulates epithelial to mesenchymal transit-inducing transcription factors (EMT-TFs). These EMT-TFs are involved in various signaling cascades, which are often associated with tumor invasiveness and malignancy. In this work, we modeled the tumor as a growing hyperplastic mass and investigated the effects of mechanical stress from surrounding ECM on tumor invasion. The invasion is modeled as volume-preserving inelastic evolution. In this framework, principal balance laws are considered for tumor mass, linear momentum, and diffusion of nutrients. Also, mechanical interactions between the tumor and ECM is modeled using Ciarlet constitutive strain energy function, and dissipation inequality is utilized to model the volumetric growth rate. System parameters, such as rate of nutrient uptake and cell proliferation, are obtained experimentally. To validate the model, human Glioblastoma multiforme (hGBM) tumor spheroids were incorporated inside Matrigel/Alginate composite hydrogel and was injected into a microfluidic chip to mimic the tumor’s natural microenvironment. The invasion structure was analyzed by imaging the spheroid over time. Also, the expression of transcriptional factors involved in invasion was measured by immune-staining the tumor. The volumetric growth, stress distribution, and inelastic evolution of tumors were predicted by the model. Results showed that the level of invasion is in direct correlation with the level of predicted stress within the tumor. Moreover, the invasion length measured by fluorescent imaging was shown to be related to the inelastic evolution of tumors obtained by the model.

Keywords: cancer, invasion, mathematical modeling, microfluidic chip, tumor spheroids

Procedia PDF Downloads 91
197 Biomimetic Dinitrosyl Iron Complexes: A Synthetic, Structural, and Spectroscopic Study

Authors: Lijuan Li

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Nitric oxide (NO) has become a fascinating entity in biological chemistry over the past few years. It is a gaseous lipophilic radical molecule that plays important roles in several physiological and pathophysiological processes in mammals, including activating the immune response, serving as a neurotransmitter, regulating the cardiovascular system, and acting as an endothelium-derived relaxing factor. NO functions in eukaryotes both as a signal molecule at nanomolar concentrations and as a cytotoxic agent at micromolar concentrations. The latter arises from the ability of NO to react readily with a variety of cellular targets leading to thiol S-nitrosation, amino acid N-nitrosation, and nitrosative DNA damage. Nitric oxide can readily bind to metals to give metal-nitrosyl (M-NO) complexes. Some of these species are known to play roles in biological NO storage and transport. These complexes have different biological, photochemical, or spectroscopic properties due to distinctive structural features. These recent discoveries have spawned a great interest in the development of transition metal complexes containing NO, particularly its iron complexes that are central to the role of nitric oxide in the body. Spectroscopic evidence would appear to implicate species of “Fe(NO)2+” type in a variety of processes ranging from polymerization, carcinogenesis, to nitric oxide stores. Our research focuses on isolation and structural studies of non-heme iron nitrosyls that mimic biologically active compounds and can potentially be used for anticancer drug therapy. We have shown that reactions between Fe(NO)2(CO)2 and a series of imidazoles generated new non-heme iron nitrosyls of the form Fe(NO)2(L)2 [L = imidazole, 1-methylimidazole, 4-methylimidazole, benzimidazole, 5,6-dimethylbenzimidazole, and L-histidine] and a tetrameric cluster of [Fe(NO)2(L)]4 (L=Im, 4-MeIm, BzIm, and Me2BzIm), resulted from the interactions of Fe(NO)2 with a series of substituted imidazoles was prepared. Recently, a series of sulfur bridged iron di nitrosyl complexes with the general formula of [Fe(µ-RS)(NO)2]2 (R = n-Pr, t-Bu, 6-methyl-2-pyridyl, and 4,6-dimethyl-2-pyrimidyl), were synthesized by the reaction of Fe(NO)2(CO)2 with thiols or thiolates. Their structures and properties were studied by IR, UV-vis, 1H-NMR, EPR, electrochemistry, X-ray diffraction analysis and DFT calculations. IR spectra of these complexes display one weak and two strong NO stretching frequencies (νNO) in solution, but only two strong νNO in solid. DFT calculations suggest that two spatial isomers of these complexes bear 3 Kcal energy difference in solution. The paramagnetic complexes [Fe2(µ-RS)2(NO)4]-, have also been investigated by EPR spectroscopy. Interestingly, the EPR spectra of complexes exhibit an isotropic signal of g = 1.998 - 2.004 without hyperfine splitting. The observations are consistent with the results of calculations, which reveal that the unpaired electron dominantly delocalize over the two sulfur and two iron atoms. The difference of the g values between the reduced form of iron-sulfur clusters and the typical monomeric di nitrosyl iron complexes is explained, for the first time, by of the difference in unpaired electron distributions between the two types of complexes, which provides the theoretical basis for the use of g value as a spectroscopic tool to differentiate these biologically active complexes.

Keywords: di nitrosyl iron complex, metal nitrosyl, non-heme iron, nitric oxide

Procedia PDF Downloads 282
196 New Derivatives 7-(diethylamino)quinolin-2-(1H)-one Based Chalcone Colorimetric Probes for Detection of Bisulfite Anion in Cationic Micellar Media

Authors: Guillermo E. Quintero, Edwin G. Perez, Oriel Sanchez, Christian Espinosa-Bustos, Denis Fuentealba, Margarita E. Aliaga

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Bisulfite ion (HSO3-) has been used as a preservative in food, drinks, and medication. However, it is well-known that HSO3- can cause health problems like asthma and allergic reactions in people. Due to the above, the development of analytical methods for detecting this ion has gained great interest. In line with the above, the current use of colorimetric and/or fluorescent probes as a detection technique has acquired great relevance due to their high sensitivity and accuracy. In this context, 2-quinolinone derivatives have been found to possess promising activity as antiviral agents, sensitizers in solar cells, antifungals, antioxidants, and sensors. In particular, 7-(diethylamino)-2-quinolinone derivatives have attracted attention in recent years since their suitable photophysical properties become promising fluorescent probes. In Addition, there is evidence that photophysical properties and reactivity can be affected by the study medium, such as micellar media. Based on the above background, 7-(diethylamino)-2-quinolinone derivatives based chalcone will be able to be incorporated into a cationic micellar environment (Cetyltrimethylammonium bromide, CTAB). Furthermore, the supramolecular control induced by the micellar environment will increase the reactivity of these derivatives towards nucleophilic analytes such as HSO3- (Michael-type addition reaction), leading to the generation of new colorimetric and/or fluorescent probes. In the present study, two derivatives of 7-(diethylamino)-2-quinolinone based chalcone DQD1-2 were synthesized according to the method reported by the literature. These derivatives were structurally characterized by 1H, 13C NMR, and HRMS-ESI. In addition, UV-VIS and fluorescence studies determined absorption bands near 450 nm, emission bands near 600 nm, fluorescence quantum yields near 0.01, and fluorescence lifetimes of 5 ps. In line with the foregoing, these photophysical properties aforementioned were improved in the presence of a cationic micellar medium using CTAB thanks to the formation of adducts presenting association constants of the order of 2,5x105 M-1, increasing the quantum yields to 0.12 and the fluorescence lifetimes corresponding to two lifetimes near to 120 and 400 ps for DQD1 and DQD2. Besides, thanks to the presence of the micellar medium, the reactivity of these derivatives with nucleophilic analytes, such as HSO3-, was increased. This was achieved through kinetic studies, which demonstrated an increase in the bimolecular rate constants in the presence of a micellar medium. Finally, probe DQD1 was chosen as the best sensor since it was assessed to detect HSO3- with excellent results.

Keywords: bisulfite detection, cationic micelle, colorimetric probes, quinolinone derivatives

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195 Effects of Hydrogen Bonding and Vinylcarbazole Derivatives on 3-Cyanovinylcarbazole Mediated Photo-Cross-Linking Induced Cytosine Deamination

Authors: Siddhant Sethi, Yasuharu Takashima, Shigetaka Nakamura, Kenzo Fujimoto

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Site-directed mutagenesis is a renowned technique to introduce specific mutations in the genome. To achieve site-directed mutagenesis, many chemical and enzymatic approaches have been reported in the past like disulphite induced genome editing, CRISPR-Cas9, TALEN etc. The chemical methods are invasive whereas the enzymatic approaches are time-consuming and expensive. Most of these techniques are unusable in the cellular application due to their toxicity and other limitations. Photo-chemical cytosine deamination, introduced in 2010, is one of the major technique for enzyme-free single-point mutation of cytosine to uracil in DNA and RNA, wherein, 3-cyanovinylcarbazole nucleoside (CNVK) containing oligodeoxyribonucleotide (ODN) having CNVK at -1 position to that of target cytosine is reversibly crosslinked to target DNA strand using 366 nm and then incubated at 90ºC to accommodate deamination. This technique is superior to enzymatic methods of site-directed mutagenesis but has a disadvantage that it requires the use of high temperature for the deamination step which restricts its applicability in the in vivo applications. This study has been focused on improving the technique by reducing the temperature required for deamination. Firstly, the photo-cross-linker, CNVK has been modified by replacing cyano group attached to vinyl group with methyl ester (OMeVK), amide (NH2VK), and carboxylic acid (OHVK) to observe the acceleration in the deamination of target cytosine cross-linked to vinylcarbazole derivative. Among the derivatives, OHVK has shown 2 times acceleration in deamination reaction as compared to CNVK, while the other two derivatives have shown deceleration towards deamination reaction. The trend of rate of deamination reaction follows the same order as that of hydrophilicity of the vinylcarbazole derivatives. OHVK being most hydrophilic has shown highest acceleration while OMeVK is least hydrophilic has proven to be least active for deamination. Secondly, in the related study, the counter-base of the target cytosine, guanine has been replaced by inosine, 2-aminopurine, nebularine, and 5-nitroindole having distinct hydrogen bonding patterns with target cytosine. Among the ODNs with these counter bases, ODN with inosine has shown 12 fold acceleration towards deamination of cytosine cross-linked to CNVK at physiological conditions as compared to guanosine. Whereas, when 2-aminopurine, nebularine, and 5-nitroindole were used, no deamination reaction took place. It can be concluded that inosine has potential to be used as the counter base of target cytosine for the CNVK mediated photo-cross-linking induced deamination of cytosine. The increase in rate of deamination reaction has been attributed to pattern and number of hydrogen bonding between the cytosine and counter base. One of the important factor is presence of hydrogen bond between exo-cyclic amino group of cytosine and the counter base. These results will be useful for development of more efficient technique for site-directed mutagenesis for C → U transformations in the DNA/RNA which might be used in the living system for treatment of various genetic disorders and genome engineering for making designer and non-native proteins.

Keywords: C to U transformation, DNA editing, genome engineering, ultra-fast photo-cross-linking

Procedia PDF Downloads 212