Search results for: biological sensor

Authors: Adel A. Ghobbar, Varun Raman

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The Corrective maintenance of components and systems is a problem plaguing almost every industry in the world today. Train operators’ and the maintenance repair and overhaul subsidiary of the Dutch railway company is also facing this problem. A considerable portion of the maintenance activities carried out by the company are unscheduled. This, in turn, severely stresses and stretches the workforce and resources available. One possible solution is to have a robust preventive maintenance plan. The other possible solution is to plan maintenance based on real-time data obtained from sensor-based ‘Health and Usage Monitoring Systems.’ The former has been investigated in this paper. The preventive maintenance model developed for train operator will subsequently be extended, to tackle the unscheduled maintenance problem also affecting the aerospace industry. The extension of the model to the aerospace sector will be dealt with in the second part of the research, and it would, in turn, validate the soundness of the model developed. Thus, there are distinct areas that will be addressed in this paper, including the mathematical modelling of preventive maintenance and optimization based on cost and system availability. The results of this research will help an organization to choose the right maintenance strategy, allowing it to save considerable sums of money as opposed to overspending under the guise of maintaining high asset availability. The concept of delay time modelling was used to address the practical problem of unscheduled maintenance in this paper. The delay time modelling can be used to help with support planning for a given asset. The model was run using MATLAB, and the results are shown that the ideal inspection intervals computed using the extended from a minimal cost perspective were 29 days, and from a minimum downtime, perspective was 14 days. Risk matrix integration was constructed to represent the risk in terms of the probability of a fault leading to breakdown maintenance and its consequences in terms of maintenance cost. Thus, the choice of an optimal inspection interval of 29 days, resulted in a cost of approximately 50 Euros and the corresponding value of b(T) was 0.011. These values ensure that the risk associated with component X being maintained at an inspection interval of 29 days is more than acceptable. Thus, a switch in maintenance frequency from 90 days to 29 days would be optimal from the point of view of cost, downtime and risk.

Keywords: delay time modelling, unscheduled maintenance, reliability, maintainability, availability

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Authors: G. C. Santini, C. Potrich, L. Lunelli, L. Vanzetti, S. Marasso, M. Cocuzza, C. Pederzolli

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The relevance of circulating miRNAs as non-invasive biomarkers for several pathologies is nowadays undoubtedly clear, as they have been found to have both diagnostic and prognostic value able to add fundamental information to patients’ clinical picture. The availability of these data, however, relies on a time-consuming process spanning from the sample collection and processing to the data analysis. In light of this, strategies which are able to ease this procedure are in high demand and considerable effort have been made in developing Lab-on-a-chip (LOC) devices able to speed up and standardise the bench work. In this context, a very promising polydimethylsiloxane (PDMS)-based microdevice which integrates the processing of the biological sample, i.e. purification of extracellular miRNAs, and reverse transcription was previously developed in our lab. In this study, we aimed at the improvement of the miRNA extraction performances of this micro device by increasing the ability of its surface to absorb extracellular miRNAs from biological samples. For this purpose, we focused on the modulation of two properties of the material: roughness and charge. PDMS surface roughness was modulated by casting with several templates (terminated with silicon oxide coated by a thin anti-adhesion aluminum layer), followed by a panel of curing conditions. Atomic force microscopy (AFM) was employed to estimate changes at the nanometric scale. To introduce modifications in surface charge we functionalized PDMS with different mixes of positively charged 3-aminopropyltrimethoxysilanes (APTMS) and neutral poly(ethylene glycol) silane (PEG). The surface chemical composition was characterized by X-ray photoelectron spectroscopy (XPS) and the number of exposed primary amines was quantified with the reagent sulfosuccinimidyl-4-o-(4,4-dimethoxytrityl) butyrate (s-SDTB). As our final end point, the adsorption rate of all these different conditions was assessed by fluorescence microscopy by incubating a synthetic fluorescently-labeled miRNA. Our preliminary analysis identified casting on thermally grown silicon oxide, followed by a curing step at 85°C for 1 hour, as the most efficient technique to obtain a PDMS surface roughness in the nanometric scaleable to trap miRNA. In addition, functionalisation with 0.1% APTMS and 0.9% PEG was found to be a necessary step to significantly increase the amount of microRNA adsorbed on the surface, therefore, available for further steps as on-chip reverse transcription. These findings show a substantial improvement in the extraction efficiency of our PDMS microdevice, ultimately leading to an important step forward in the development of an innovative, easy-to-use and integrated system for the direct purification of less abundant circulating microRNAs.

Keywords: circulating miRNAs, diagnostics, Lab-on-a-chip, polydimethylsiloxane (PDMS)

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Authors: Suchao Donpudsa, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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A crustin gene, LV-SWD1, previously found in the hemocyte cDNA library of Litopenaeus vannamei, contains the open reading frames of 288 bp encoding a putative protein of 96 amino acid residues. The putative signal peptides of the LV-SWD1 were identified using the online SignalP 3.0 with predicted cleavage sites between Ala24-Val25, resulting in 72 residue mature protein with calculated molecular mass of 7.4 kDa and predicted pI of 8.5. This crustin contains a Arg-Pro rich region at the amino-terminus and a single whey acidic protein (WAP) domain at the carboxyl-terminus. In order to characterize their properties and biological activities, the recombinant crustin protein was produced in the Escherichia coli expression system. Antimicrobial assays showed that the growth of Bacillus subtilis was inhibited by this recombinant crustin with MIC of about 25-50 µM.

Keywords: crustin, single whey acidic protein, Litopenaeus vannamei, antimicrobial activity

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Authors: Ahmet Ozcan, Onder Alparslan, Anil Sezgin, Omer Cetin

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The use of unmanned systems for different purposes has become very popular over the past decade. Expectations from these systems have also shown an incredible increase in this parallel. But meeting the demands of the tasks are often not possible with the usage of a single unmanned vehicle in a mission, so it is necessary to use multiple autonomous vehicles with different abilities together in coordination. Therefore the usage of the same type of vehicles together as a swarm is helped especially to satisfy the time constraints of the missions effectively. In other words, it allows sharing the workload by the various numbers of homogenous platforms together. Besides, it is possible to say there are many kinds of problems that require the usage of the different capabilities of the heterogeneous platforms together cooperatively to achieve successful results. In this case, cooperative working brings additional problems beyond the homogeneous clusters. In the scenario presented as an example problem, it is expected that an autonomous ground vehicle, which is lack of its position information, manage to perform point-to-point navigation without losing its way in a previously unknown labyrinth. Furthermore, the ground vehicle is equipped with very limited sensors such as ultrasonic sensors that can detect obstacles. It is very hard to plan or complete the mission for the ground vehicle by self without lost its way in the unknown labyrinth. Thus, in order to assist the ground vehicle, the autonomous air drone is also used to solve the problem cooperatively. The autonomous drone also has limited sensors like downward looking camera and IMU, and it also lacks computing its global position. In this context, it is aimed to solve the problem effectively without taking additional support or input from the outside, just benefiting capabilities of two autonomous vehicles. To manage the point-to-point navigation in a previously unknown labyrinth, the platforms have to work together coordinated. In this paper, cooperative work of heterogeneous unmanned systems is handled in an applied sample scenario, and it is mentioned that how to work together with an autonomous ground vehicle and the autonomous flying platform together in a harmony to take advantage of different platform-specific capabilities. The difficulties of using heterogeneous multiple autonomous platforms in a mission are put forward, and the successful solutions are defined and implemented against the problems like spatially distributed tasks planning, simultaneous coordinated motion, effective communication, and sensor fusion.

Keywords: unmanned systems, heterogeneous autonomous vehicles, coordination, task planning

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Authors: R. Farhoud, G. Hermand, S. Delepine-lesoille

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Future underground facility of French radioactive waste disposal, named Cigeo, is designed to store intermediate and high level - long-lived French radioactive waste. Intermediate level waste cells are tunnel-like, about 400m length and 65 m² section, equipped with several concrete layers, which can be grouted in situ or composed of tunnel elements pre-grouted. The operating space into cells, to allow putting or removing waste containers, should be monitored for several decades without any maintenance. To provide the required information, design was performed and tested in situ in Andra’s underground laboratory (URL) at 500m under the surface. Based on distributed optic fiber sensors (OFS) and backscattered Brillouin for strain and Raman for temperature interrogation technics, the design consists of 2 loops of OFS, at 2 different radiuses, around the monitored section (Orthoradiale strains) and longitudinally. Strains measured by distributed OFS cables were compared to classical vibrating wire extensometers (VWE) and platinum probes (Pt). The OFS cables were composed of 2 cables sensitive to strains and temperatures and one only for temperatures. All cables were connected, between sensitive part and instruments, to hybrid cables to reduce cost. The connection has been made according to 2 technics: splicing fibers in situ after installation or preparing each fiber with a connector and only plugging them together in situ. Another challenge was installing OFS cables along a tunnel mad in several parts, without interruption along several parts. First success consists of the survival rate of sensors after installation and quality of measurements. Indeed, 100% of OFS cables, intended for long-term monitoring, survived installation. Few new configurations were tested with relative success. Measurements obtained were very promising. Indeed, after 3 years of data, no difference was observed between cables and connection methods of OFS and strains fit well with VWE and Pt placed at the same location. Data, from Brillouin instrument sensitive to strains and temperatures, were compensated with data provided by Raman instrument only sensitive to temperature and into a separated fiber. These results provide confidence in the next steps of the qualification processes which consists of testing several data treatment approach for direct analyses.

Keywords: monitoring, fiber optic, sensor, data treatment

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Authors: Danielle Sandler dos Passos, Helder Coelho, Flávia Mori Sarti

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Complexity science seeks the understanding of systems adopting diverse theories from various areas. Network analysis has been gaining space and credibility, namely with the biological, social and economic systems. Significant part of the literature focuses only monolayer representations of connections among agents considering one level of their relationships, and excludes other levels of interactions, leading to simplistic results in network analysis. Therefore, this work aims to demonstrate the advantages of the use of multilayer networks for the representation and analysis of networks. For this, we analyzed an interbank network, composed of 42 banks, comparing the centrality measures of the agents (degree and PageRank) resulting from each method (monolayer x multilayer). This proved to be the most reliable and efficient the multilayer analysis for the study of the current networks and highlighted JP Morgan and Deutsche Bank as the most important banks of the analyzed network.

Keywords: complexity, interbank networks, multilayer networks, network analysis

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Authors: Murray L. Ireland, Kevin J. Worrall, Rebecca Mackenzie, Thaleia Flessa, Euan McGookin, Douglas Thomson

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Robotic rovers which are designed to work in extra-terrestrial environments present a unique challenge in terms of the reliability and availability of systems throughout the mission. Should some fault occur, with the nearest human potentially millions of kilometres away, detection and identification of the fault must be performed solely by the robot and its subsystems. Faults in the system sensors are relatively straightforward to detect, through the residuals produced by comparison of the system output with that of a simple model. However, faults in the input, that is, the actuators of the system, are harder to detect. A step change in the input signal, caused potentially by the loss of an actuator, can propagate through the system, resulting in complex residuals in multiple outputs. These residuals can be difficult to isolate or distinguish from residuals caused by environmental disturbances. While a more complex fault detection method or additional sensors could be used to solve these issues, an alternative is presented here. Using inverse simulation (InvSim), the inputs and outputs of the mathematical model of the rover system are reversed. Thus, for a desired trajectory, the corresponding actuator inputs are obtained. A step fault near the input then manifests itself as a step change in the residual between the system inputs and the input trajectory obtained through inverse simulation. This approach avoids the need for additional hardware on a mass- and power-critical system such as the rover. The InvSim fault detection method is applied to a simple four-wheeled rover in simulation. Additive system faults and an external disturbance force and are applied to the vehicle in turn, such that the dynamic response and sensor output of the rover are impacted. Basic model-based fault detection is then employed to provide output residuals which may be analysed to provide information on the fault/disturbance. InvSim-based fault detection is then employed, similarly providing input residuals which provide further information on the fault/disturbance. The input residuals are shown to provide clearer information on the location and magnitude of an input fault than the output residuals. Additionally, they can allow faults to be more clearly discriminated from environmental disturbances.

Keywords: fault detection, ground robot, inverse simulation, rover

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Authors: Sudhir Kumar Sharma, Abiy D. Woldetsadik, Mazin Magzoub, Ramesh Jagannathan

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It is well known that bioactive ceramics exhibit specific biological affinities, especially in the area of tissue re-generation. In this context, we report the development of an eminently scalable, novel, supercritical CO₂ based process for the fabrication of hierarchically porous 'soft' CaCO₃ scaffolds. Porosity at the macro, micro, and nanoscales was obtained through process optimization of the so-called 'coffee-ring effect'. Exposure of these CaCO₃ scaffolds to monocytic THP-1 cells yielded negligible levels of tumor necrosis factor-alpha (TNF-α) thereby confirming the lack of immunogenicity of the scaffolds. ECM protein treatment of the scaffolds showed enhanced adsorption comparable to standard control such as glass. In vitro studies using osteoblast precursor cell line, MC3T3, also demonstrated the cytocompatibility of hierarchical porous CaCO₃ scaffolds.

Keywords: supercritical CO2, CaCO3 scaffolds, coffee-ring effect, ECM proteins

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Authors: Massimiliano Condotta, Paolo Ruggeri, Chiara Scanagatta, Giovanni Borga

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Architects and urban planners, when designing and renewing cities, have to face a complex set of problems, including the issues of noise and air pollution which are considered as hot topics (i.e., the Clean Air Act of London and the Soundscape definition). It is usually taken for granted that these problems go by together because the noise pollution present in cities is often linked to traffic and industries, and these produce air pollutants as well. Traffic congestion can create both noise pollution and air pollution, because NO₂ is mostly created from the oxidation of NO, and these two are notoriously produced by processes of combustion at high temperatures (i.e., car engines or thermal power stations). We can see the same process for industrial plants as well. What have to be investigated – and is the topic of this paper – is whether or not there really is a correlation between noise pollution and air pollution (taking into account NO₂) in urban areas. To evaluate if there is a correlation, some low-cost methodologies will be used. For noise measurements, the OpeNoise App will be installed on an Android phone. The smartphone will be positioned inside a waterproof box, to stay outdoor, with an external battery to allow it to collect data continuously. The box will have a small hole to install an external microphone, connected to the smartphone, which will be calibrated to collect the most accurate data. For air, pollution measurements will be used the AirMonitor device, an Arduino board to which the sensors, and all the other components, are plugged. After assembling the sensors, they will be coupled (one noise and one air sensor) and placed in different critical locations in the area of Mestre (Venice) to map the existing situation. The sensors will collect data for a fixed period of time to have an input for both week and weekend days, in this way it will be possible to see the changes of the situation during the week. The novelty is that data will be compared to check if there is a correlation between the two pollutants using graphs that should show the percentage of pollution instead of the values obtained with the sensors. To do so, the data will be converted to fit on a scale that goes up to 100% and will be shown thru a mapping of the measurement using GIS methods. Another relevant aspect is that this comparison can help to choose which are the right mitigation solutions to be applied in the area of the analysis because it will make it possible to solve both the noise and the air pollution problem making only one intervention. The mitigation solutions must consider not only the health aspect but also how to create a more livable space for citizens. The paper will describe in detail the methodology and the technical solution adopted for the realization of the sensors, the data collection, noise and pollution mapping and analysis.

Keywords: air quality, data analysis, data collection, NO₂, noise mapping, noise pollution, particulate matter

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Authors: Roaa Sait, Richard Cross

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The synthesis and characterization of one dimensional nanostructure such as nanowires has received considerable attention. Much effort has concentrated on TiN material especially in the biological field due to its useful and unique properties in this field. Therefore, for the purpose of this project, synthesis of Titanium Nitride (TiN) nanowires (NWs) will be presented. They will be synthesised by growing titanium dioxide (Ti) NWs in an aqueous solution at low temperatures under atmospheric pressure. Then the grown nanowires will undergo a 'Nitrodation process' in which results in the formation of TiN NWs. The structure, morphology and composition of the grown nanowires will be characterized using Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), X-ray Diffraction (XRD) and Cyclic Voltammetry (CV). Obtaining TiN NWs is a challenging task since it has not been formulated before, as far as we acknowledge. This might be due to the fact that nitriding Ti NWs can be difficult in terms of optimizing experimental parameters.

Keywords: nanowires, dissolution-growth, nucleation, PECVD, deposition, spin coating, scanning electron microscopic analysis, cyclic voltammetry analysis

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Authors: Branimir Jurun, Elza Jurun

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The focus of this paper is the description of an instrument called 'Quattuor 45' and defining of wave pressure measurement. Special attention is given to measurement of wave pressure created by the wind speed increasing obtained with the instrument 'Quattuor 45' in the investigated area. The study begins with respect to theoretical attitudes and numerous up to date investigations related to the waves approaching the coast. The detailed schematic view of the instrument is enriched with pictures from ground plan and side view. Horizontal stability of the instrument is achieved by mooring which relies on two concrete blocks. Vertical wave peak monitoring is ensured by one float above the instrument. The synthesis of horizontal stability and vertical wave peak monitoring allows to create a representative database for wave pressure measuring. Instrument ‘Quattuor 45' is named according to the way the database is received. Namely, the electronic part of the instrument consists of the main chip ‘Arduino', its memory, four load cells with the appropriate modules and the wind speed sensor 'Anemometers'. The 'Arduino' chip is programmed to store two data from each load cell and two data from the anemometer on SD card each second. The next part of the research is dedicated to data processing. All measured results are stored automatically in the database and after that detailed processing is carried out in the MS Excel. The result of the wave pressure measurement is synthesized by the unit of measurement kN/m². This paper also suggests a graphical presentation of the results by multi-line graph. The wave pressure is presented on the left vertical axis, while the wind speed is shown on the right vertical axis. The time of measurement is displayed on the horizontal axis. The paper proposes an algorithm for wind speed measurements showing the results for two characteristic winds in the Adriatic Sea, called 'Bura' and 'Jugo'. The first of them is the northern wind that reaches high speeds, causing low and extremely steep waves, where the pressure of the wave is relatively weak. On the other hand, the southern wind 'Jugo' has a lower speed than the northern wind, but due to its constant duration and constant speed maintenance, it causes extremely long and high waves that cause extremely high wave pressure.

Keywords: instrument, measuring unit, waves pressure metering, wind seed measurement

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Authors: Shamim Mushtaq, Moazzam Shahid

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Breast cancer (BC) claims the lives of half a million women every year and is the most common cause of death in the developing world. In 2019, it was estimated that BC alone accounts for 15% of all cancer deaths in younger women (aged < 45 years old) with advanced-stage lung metastasis. According to the World Health Organization & International Union against Cancer, in Asia, a high number of cancer-related deaths will be observed in 2020, whereas the burden will be reduced in Western countries due to awareness about the disease, better health facilities and advanced treatments. In the last 15 years, it has been reported that the incidence of BC has increased by 1.1% among Asian compared to the US population from 2003 to 2012. To date, several BC biological subtypes have been reported so far, which are associated with different treatment responses. The heterogeneity and diversity of BC reflected these different subtypes, including Luminal A (23.7% prevalence) and B (38.8% prevalence) that have pathological estrogen receptor (ER+)-positive tumors, the human epidermal growth factor receptor 2 (HER2) (11.2% prevalence) and triple-negative breast cancer (TNBC) (25% prevalence). According to Shaukat Khanum Memorial Cancer Hospital and Research Centre – Pakistan, ten years of data showed that among 636 BC patients, 30.5% had TNBC who were <40 years of age, which is an extremely alarming situation. Therefore, there is a dire need to explore and develop therapeutic targets for the treatment of early TNBC. Since the last decade, unfortunately, there has been little success in understanding the complexity of TNBC and in discovering new biological therapeutic targets. However, conventional chemotherapy is the only choice of treatment for TNBC patients. Many investigators revealed advances in multi-omics (multiple "omes", e.g., genome, proteome, transcriptome, epigenome, and microbiome) which were later identified as actionable targets and increased prevalence in TNBC patients. However, various drugs have been identified so far which are related to a particular diagnostic and prognostic biomarker. For example, Epidermal growth factor receptor ( EGFR or ErbB-1), HER-2/neu (ErbB-2), HER-3 (ErbB-3), and HER-4 (ErbB-4). Protein Transglin-2 (TAGLN 2 ) and Profilins-1 (Pfn-1 ) are the ubiquitously expressed large family of proteins present in all eukaryotes, enabling actin cytoskeletal reorganization. It is known that the oncogenic transformation of cells is accompanied by alteration in the actin cytoskeleton. There are causal connections between altered expression of actin cytoskeletal regulators and cancer progression. Our case-control study identified TAGLN-2 and Pfn-1 proteins in TNBC blood by mass spectrometry. Both TAGLN-2 and Pfn-1 proteins are differentially expressed in early and advanced stages of TNBS patients, which could be potential predictors or therapeutic targets for TNBC.

Keywords: TNBC, blood biomarkers, mass spectrometry, qPCR, ELISA

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Authors: A. Y. Benabdellah, W. Lakhdari, A. Dahliz, Y. Bouchikh, A. Soud, R. M'lik, H. Hammi

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The direct comparison tests on the culture medium, between Trichoderma sp. and Fusarium oxysporum f. sp. radicis-lycopersici revealed that the latest one could inhibit the growth of F. oxysporum mycelial over than 40% compared to the control and that after four days of incubation at 26° C. Moreover, beyond this period and at the end of six days, Trichoderma sp. invading the colonies of F. oxysporum on what it sporule, thus revealing its power is highly myco-parasitic. Almost similar results were obtained against Alternaria solani is also a pathogen which is not causing a lot of damage, but we found it more sensitive to Trichoderma sp. with a percentage of inhibition more than 50%. So due to the in vitro test of Trichoderma sp. against these aggressive pathogens by direct contact has been found that can inhibit their mycelial growth with high speed and a high inhibition rate.

Keywords: Trichoderma sp., Fusarium oxysporum f. sp. radicis-lycopersici, Alternaria solani, biological control, antagonist

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Authors: May Fadheel Estephan, Richard Perks

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Context: Cancer is a serious health concern that affects millions of people worldwide. Early detection and treatment are essential for improving patient outcomes. However, current methods for cancer detection have limitations, such as low sensitivity and specificity. Research Aim: The aim of this study was to develop an optical sensor for cancer detection using elastic light scattering spectroscopy (ELSS). ELSS is a noninvasive optical technique that can be used to characterize the size and concentration of particles in a solution. Methodology: An optical probe was fabricated with a 100-μm-diameter core and a 132-μm centre-to-centre separation. The probe was used to measure the ELSS spectra of polystyrene spheres with diameters of 2, 0.8, and 0.413 μm. The spectra were then analysed to determine the size and concentration of the spheres. Findings: The results showed that the optical probe was able to differentiate between the three different sizes of polystyrene spheres. The probe was also able to detect the presence of polystyrene spheres in suspension concentrations as low as 0.01%. Theoretical Importance: The results of this study demonstrate the potential of ELSS for cancer detection. ELSS is a noninvasive technique that can be used to characterize the size and concentration of cells in a tissue sample. This information can be used to identify cancer cells and assess the stage of the disease. Data Collection: The data for this study were collected by measuring the ELSS spectra of polystyrene spheres with different diameters. The spectra were collected using a spectrometer and a computer. Analysis Procedures: The ELSS spectra were analysed using a software program to determine the size and concentration of the spheres. The software program used a mathematical algorithm to fit the spectra to a theoretical model. Question Addressed: The question addressed by this study was whether ELSS could be used to detect cancer cells. The results of the study showed that ELSS could be used to differentiate between different sizes of cells, suggesting that it could be used to detect cancer cells. Conclusion: The findings of this research show the utility of ELSS in the early identification of cancer. ELSS is a noninvasive method for characterizing the number and size of cells in a tissue sample. To determine cancer cells and determine the disease's stage, this information can be employed. Further research is needed to evaluate the clinical performance of ELSS for cancer detection.

Keywords: elastic light scattering spectroscopy, polystyrene spheres in suspension, optical probe, fibre optics

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Authors: Marie-Elise Beydon, Daniel Sacristan, Isabel Ruppen

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MB02 (Alymsys®) is a candidate biosimilar to bevacizumab, which was developed against the reference product (RP) Avastin® sourced from both the European Union (EU) and United States (US). MB02 has been extensively characterized comparatively to Avastin® at a physicochemical and biological level using sensitive orthogonal state-of-the-art analytical methods. MB02 has been demonstrated similar to the RP with regard to its primary and higher-order structure, post- and co-translational profiles such as glycosylation, charge, and size variants. Specific focus has been put on the characterization of Fab-related activities, such as binding to VEGF A 165, which directly reflect the bevacizumab mechanism of action. Fc-related functionality was also investigated, including binding to FcRn, which is indicative of antibodies' half-life. The data generated during the analytical similarity assessment demonstrate the high analytical similarity of MB02 to its RP.

Keywords: analytical similarity, bevacizumab, biosimilar, MB02

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Authors: Igor V. Grigoriev

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Understanding of biological processes encoded in fungi is instrumental in addressing future food, feed, and energy demands of the growing human population. Genomics is a powerful and quickly evolving tool to understand these processes. The Fungal Genomics Program of the US Department of Energy Joint Genome Institute (JGI) partners with researchers around the world to explore fungi in several large scale genomics projects, changing the fungal genomics landscape. The key trends of these changes include: (i) rapidly increasing scale of sequencing and analysis, (ii) developing approaches to go beyond culturable fungi and explore fungal ‘dark matter,’ or unculturables, and (iii) functional genomics and multi-omics data integration. Power of comparative genomics has been recently demonstrated in several JGI projects targeting mycorrhizae, plant pathogens, wood decay fungi, and sugar fermenting yeasts. The largest JGI project ‘1000 Fungal Genomes’ aims at exploring the diversity across the Fungal Tree of Life in order to better understand fungal evolution and to build a catalogue of genes, enzymes, and pathways for biotechnological applications. At this point, at least 65% of over 700 known families have one or more reference genomes sequenced, enabling metagenomics studies of microbial communities and their interactions with plants. For many of the remaining families no representative species are available from culture collections. To sequence genomes of unculturable fungi two approaches have been developed: (a) sequencing DNA from fruiting bodies of ‘macro’ and (b) single cell genomics using fungal spores. The latter has been tested using zoospores from the early diverging fungi and resulted in several near-complete genomes from underexplored branches of the Fungal Tree, including the first genomes of Zoopagomycotina. Genome sequence serves as a reference for transcriptomics studies, the first step towards functional genomics. In the JGI fungal mini-ENCODE project transcriptomes of the model fungus Neurospora crassa grown on a spectrum of carbon sources have been collected to build regulatory gene networks. Epigenomics is another tool to understand gene regulation and recently introduced single molecule sequencing platforms not only provide better genome assemblies but can also detect DNA modifications. For example, 6mC methylome was surveyed across many diverse fungi and the highest among Eukaryota levels of 6mC methylation has been reported. Finally, data production at such scale requires data integration to enable efficient data analysis. Over 700 fungal genomes and other -omes have been integrated in JGI MycoCosm portal and equipped with comparative genomics tools to enable researchers addressing a broad spectrum of biological questions and applications for bioenergy and biotechnology.

Keywords: fungal genomics, single cell genomics, DNA methylation, comparative genomics

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Authors: Ahmed Elaksher, Islam Omar

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For many years, Mars surface has been a mystery for scientists. Lately with the help of geospatial data and photogrammetric procedures researchers were able to capture some insights about this planet. Two of the most imperative data sources to explore Mars are the The High Resolution Imaging Science Experiment (HiRISE) and the Mars Orbiter Laser Altimeter (MOLA). HiRISE is one of six science instruments carried by the Mars Reconnaissance Orbiter, launched August 12, 2005, and managed by NASA. The MOLA sensor is a laser altimeter carried by the Mars Global Surveyor (MGS) and launched on November 7, 1996. In this project, we used MOLA-based DEMs to orthorectify HiRISE optical images for generating a more accurate and trustful surface of Mars. The MOLA data was interpolated using the kriging interpolation technique. Corresponding tie points were digitized from both datasets. These points were employed in co-registering both datasets using GIS analysis tools. In this project, we employed three different 3D to 2D transformation models. These are the parallel projection (3D affine) transformation model; the extended parallel projection transformation model; the Direct Linear Transformation (DLT) model. A set of tie-points was digitized from both datasets. These points were split into two sets: Ground Control Points (GCPs), used to evaluate the transformation parameters using least squares adjustment techniques, and check points (ChkPs) to evaluate the computed transformation parameters. Results were evaluated using the RMSEs between the precise horizontal coordinates of the digitized check points and those estimated through the transformation models using the computed transformation parameters. For each set of GCPs, three different configurations of GCPs and check points were tested, and average RMSEs are reported. It was found that for the 2D transformation models, average RMSEs were in the range of five meters. Increasing the number of GCPs from six to ten points improve the accuracy of the results with about two and half meters. Further increasing the number of GCPs didn’t improve the results significantly. Using the 3D to 2D transformation parameters provided three to two meters accuracy. Best results were reported using the DLT transformation model. However, increasing the number of GCPS didn’t have substantial effect. The results support the use of the DLT model as it provides the required accuracy for ASPRS large scale mapping standards. However, well distributed sets of GCPs is a key to provide such accuracy. The model is simple to apply and doesn’t need substantial computations.

Keywords: mars, photogrammetry, MOLA, HiRISE

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Authors: Tuisem Shimrah

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One general agreement on climate change is that its causes may be local but the effects are global. Indigenous people are subscribed to “low-carbon” traditional ways of life and as such they have contributed little to causes of climate change. On the contrary they are the most adversely affected by climate change due to their dependence on surrounding rich biological wealth as a source of their livelihood, health care, entertainment and cultural activities This paper deals with the results of the investigation of various adaptation strategies adopted to combat climate change by traditional community. The result shows effective ways of application of traditional knowledge and wisdom applied by Tangkhul traditional community at local and community level in remote areas in Northeast India. Four adaptation measures are being presented in this paper.

Keywords: adaptation, climate change, Northeast India, Tangkhul, traditional community

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Authors: Md. Khayrul Bashar, Md. Saiful Islam, Kimiko Yamashita, Yano Midori

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Despite having some progress in human authentication, conventional biometrics (e.g., facial features, fingerprints, retinal scans, gait, voice patterns) are not robust against falsification because they are neither confidential nor secret to an individual. As a non-invasive tool, electrocardiogram (ECG) has recently shown a great potential in human recognition due to its unique rhythms characterizing the variability of human heart structures (chest geometry, sizes, and positions). Moreover, ECG has a real-time vitality characteristic that signifies the live signs, which ensure legitimate individual to be identified. However, the detection accuracy of the current ECG-based methods is not sufficient due to a high variability of the individual’s heartbeats at a different instance of time. These variations may occur due to muscle flexure, the change of mental or emotional states, and the change of sensor positions or long-term baseline shift during the recording of ECG signal. In this study, a new method is proposed for human identification, which is based on the extraction of the local roughness of ECG heartbeat signals. First ECG signal is preprocessed using a second order band-pass Butterworth filter having cut-off frequencies of 0.00025 and 0.04. A number of local binary patterns are then extracted by applying a moving neighborhood window along the ECG signal. At each instant of the ECG signal, the pattern is formed by comparing the ECG intensities at neighboring time points with the central intensity in the moving window. Then, binary weights are multiplied with the pattern to come up with the local roughness description of the signal. Finally, histograms are constructed that describe the heartbeat signals of individual subjects in the database. One advantage of the proposed feature is that it does not depend on the accuracy of detecting QRS complex, unlike the conventional methods. Supervised recognition methods are then designed using minimum distance to mean and Bayesian classifiers to identify authentic human subjects. An experiment with sixty (60) ECG signals from sixty adult subjects from National Metrology Institute of Germany (NMIG) - PTB database, showed that the proposed new method is promising compared to a conventional interval and amplitude feature-based method.

Keywords: human identification, ECG biometrics, local roughness patterns, supervised classification

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Authors: Jesús Horacio, Alfredo Ollero, Víctor Bouzas-Blanco, Augusto Pérez-Alberti

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Rivers have traditionally been classified, assessed and managed in terms of hydrological, chemical and / or biological criteria. Geomorphological classifications had in the past a secondary role, although proposals like River Styles Framework, Catchment Baseline Survey or Stroud Rural Sustainable Drainage Project did incorporate geomorphology for management decision-making. In recent years many studies have been attracted to the geomorphological component. The geomorphological processes and their associated forms determine the structure of a river system. Understanding these processes and forms is a critical component of the sustainable rehabilitation of aquatic ecosystems. The fluvial auto-classification approach suggests that a river is a self-built natural system, with processes and forms designed to effectively preserve their ecological function (hydrologic, sedimentological and biological regime). Fluvial systems are formed by a wide range of elements with multiple non-linear interactions on different spatial and temporal scales. Besides, the fluvial auto-classification concept is built using data from the river itself, so that each classification developed is peculiar to the river studied. The variables used in the classification are specific stream power and mean grain size. A discriminant analysis showed that these variables are the best characterized processes and forms. The statistical technique applied allows to get an individual discriminant equation for each geomorphological type. The geomorphological classification was developed using sites with high naturalness. Each site is a control point of high ecological and geomorphological quality. The changes in the conditions of the control points will be quickly recognizable, and easy to apply a right management measures to recover the geomorphological type. The study focused on Galicia (NW Spain) and the mapping was made analyzing 122 control points (sites) distributed over eight river basins. In sum, this study provides a method for fluvial geomorphological classification that works as an open and flexible tool underlying the fluvial auto-classification concept. The hydrographic mapping is the visual expression of the results, such that each river has a particular map according to its geomorphological characteristics. Each geomorphological type is represented by a particular type of hydraulic geometry (channel width, width-depth ratio, hydraulic radius, etc.). An alteration of this geometry is indicative of a geomorphological disturbance (whether natural or anthropogenic). Hydrographic mapping is also dynamic because its meaning changes if there is a modification in the specific stream power and/or the mean grain size, that is, in the value of their equations. The researcher has to check annually some of the control points. This procedure allows to monitor the geomorphology quality of the rivers and to see if there are any alterations. The maps are useful to researchers and managers, especially for conservation work and river restoration.

Keywords: fluvial auto-classification concept, mapping, geomorphology, river

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Authors: L. Mallesha, C. S. Karthik, P. Mallu

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A series of new [1-(substituted-benzoyl)-piperidin-4-yl]-(2,4-difluoro-phenyl)-methanone oxime derivatives, 3(a-f) were synthesized and characterized by different spectral studies. All compounds were evaluated for their in vitro antibacterial activity against bacterial strains. These compounds were screened for their antioxidant activity by DPPH• and Fe2+ chelating assay. Antiproliferative effects were evaluated using the MTT assay method against two human cancer cell lines and one astrocytoma brain tumor cell line. Compound 3b exhibited moderate antibacterial activity when compared with other compounds. All the compounds showed antioxidant activity, where compound 3f was the best radical scavenger and Fe2+ ion scavenger. Compounds, 3b, and 3d showed good activity on all cell lines, whereas the other compounds in the series exhibited moderate activity.

Keywords: Piperidine, antibacterial, antioxidant, antiproliferative

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Authors: Mahsa Mafi

Abstract:

Encapsulation of Micrococcus Luteus (M. Luteus) in polymeric composites has been employed for the bioremediation, sequestration of metals and for the biodegradation of chemical pollutants and toxic components in waste water. Polymer composites in the form of nonwovens of nanofibers, or core/shell particles can provide a bacterial friendly environment for transfer of nutrients and metabolisms, with the least leakage of bacteria. M. Luteus is encapsulated in a hydrophilic core of poly (vinyl alcohol), following by synthesis or coating of a proper shell as a support to maintain the chemical and mechanical strength. The biological activity of bacteria is confirmed by Live/Dead analysis and agar plate tests. SEM and TEM analysis were utilized for morphological studies of polymer composites. As a result of the successful encapsulation of the alive bacteria in polymers, longer storage time in their functional state were achieved.

Keywords: Polymer composites, Bacteria encapsulation, Bioremediation, Waste water treatment

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Authors: Adesiji Adeolu Richard, Hassa Musa, Osita Evaritus Asogwa, Mary Oluwatobi Odekunle, Mangey Jarumi Akila

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In this paper, extract from raw Moringa Oleifera seeds for the treatment of 40 liters of abattoir wastewater was studied for a period of ten (10) weeks. A completely randomized design with loading dosages of 10, 12, 14, 16, 18, and 20g of processed Moringa Oleifera seed was used in the treatment. A control sample (with no Moringa Oleifera treatment) was also included. The physical and chemical properties of abattoir wastewater were investigated before and after treatment. The turbidity value was reduced drastically after the treatments from 15.40 to 7.63 mg/l for 16g dosage in week 7. Total alkalinity, Total hardness, Conductivity, Calcium, and Biological Oxygen Demand were all found to be reduced in concentration within the second and fourth weeks of the experiment with 14 to 16g of Moringa Oleifera dosage. The results generally showed that 16g/500ml of Moringa Oleifera was able to treat abattoir wastewater after weeks of the experiment.

Keywords: Moringa Oleifera, abattoir wastewater, turbidity, conductivity, pH

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Authors: Yogesh D. Bansod, Jiri Bursa

Abstract:

The quantitative study of cell mechanics is of paramount interest since it regulates the behavior of the living cells in response to the myriad of extracellular and intracellular mechanical stimuli. The novel experimental techniques together with robust computational approaches have given rise to new theories and models, which describe cell mechanics as a combination of biomechanical and biochemical processes. This review paper encapsulates the existing continuum-based computational approaches that have been developed for interpreting the mechanical responses of living cells under different loading and boundary conditions. The salient features and drawbacks of each model are discussed from both structural and biological points of view. This discussion can contribute to the development of even more precise and realistic computational models of cell mechanics based on continuum approaches or on their combination with microstructural approaches, which in turn may provide a better understanding of mechanotransduction in living cells.

Keywords: cell mechanics, computational models, continuum approach, mechanical models

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Authors: Aravind Ravichandran, Marc Ramuz, Sylvain Blayac

Abstract:

With the rapid development of wearable electronics and sensor networks, batteries cannot meet the sustainable energy requirement due to their limited lifetime, size and degradation. Ambient energies such as wind have been considered as an attractive energy source due to its copious, ubiquity, and feasibility in nature. With miniaturization leading to high-power and robustness, triboelectric nanogenerator (TENG) have been conceived as a promising technology by harvesting mechanical energy for powering small electronics. TENG integration in large-scale applications is still unexplored considering its attractive properties. In this work, a state of the art design TENG based on wind venturi system is demonstrated for use in any complex environment. When wind introduces into the air gap of the homemade TENG venturi system, a thin flexible polymer repeatedly contacts with and separates from electrodes. This device structure makes the TENG suitable for large scale harvesting without massive volume. Multiple stacking not only amplifies the output power but also enables multi-directional wind utilization. The system converts ambient mechanical energy to electricity with 400V peak voltage by charging of a 1000mF super capacitor super rapidly. Its future implementation in an array of applications aids in environment friendly clean energy production in large scale medium and the proposed design performs with an exhaustive material testing. The relation between the interfacial micro-and nano structures and the electrical performance enhancement is comparatively studied. Nanostructures are more beneficial for the effective contact area, but they are not suitable for the anti-adhesion property due to the smaller restoring force. Considering these issues, the nano-patterning is proposed for further enhancement of the effective contact area. By considering these merits of simple fabrication, outstanding performance, robust characteristic and low-cost technology, we believe that TENG can open up great opportunities not only for powering small electronics, but can contribute to large-scale energy harvesting through engineering design being complementary to solar energy in remote areas.

Keywords: triboelectric nanogenerator, wind energy, vortex design, large scale energy

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Authors: Malkhaz Jokhadze, Vakhtang Mshvildadze, Levan Makaradze, Ekaterine Mosidze, Salome Barbaqadze, Mariam Murtazashvili, Dali Berashvili, Koba sivsivadze, Lasha Bakuridze, Aliosha Bakuridze

Abstract:

Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.

Keywords: clary, essential oils, monarda, poultry, phytobiotics, thyme

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Authors: Selma Hamimed, Abdelwaheb Chatti

Abstract:

The current study deals with the biological treatment of saline wastewaters generated by various agro-food industries using Yarrowia lipolytica. The ability of this yeast was studied on the mixture of olive mill wastewater and tuna wash processing wastewater. Results showed that the high proportion of olive mill wastewater in the mixture about (75:25) is the suitable one for the highest Y. lipolytica biomass production, reaching 11.3 g L⁻¹ after seven days. In addition, results showed significant removal of chemical oxygen demand (COD) and phosphorous of 97.49 % and 98.90 %, respectively. On the other hand, Y. lipolytica was found to be effective to desalinate all mixtures reaching a removal of 92.21 %. Moreover, the analytical results using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM) confirmed the biosorption of NaCl on the surface of the yeast as nanocrystals form with a size of 47.3 nm.

Keywords: nanocrystallization of NaCl, desalination, wastewater treatment, yarrowia lipolytica

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Authors: Raquel C. A. G. Mota, Livia R. Menezes, Emerson O. da Silva

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Both silver nanoparticles and titanium dioxide have been extensively used in tissue engineering since they’ve been approved by the Food and Drug Administration (FDA), and present a bactericide effect when added to a polymeric matrix. In this work, the focus is on fabricating binary systems with both nanoparticles so that the synergistic effect can be investigated. The systems were tested by Nuclear Magnetic Resonance (NMR), Thermogravimetric Analysis (TGA), Fourier-Transformed Infrared (FTIR), and Differential Scanning Calorimetry (DSC), and X-ray Diffraction (XRD), and had both their bioactivity and bactericide effect tested. The binary systems presented different properties than the individual systems, enhancing both the thermal and biological properties as was to be expected. The crystallinity was also affected, as indicated by the finding of the DSC and XDR techniques, and the NMR showed a good dispersion of both nanoparticles in the polymer matrix. These findings indicate the potential of combining TiO₂ and silver nanoparticles in biomedicine.

Keywords: metallic nanoparticles, nanotechnology, polymer nanocomposites, polymer science

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Authors: Ali Mirtaghavi, Andy Baldwin, Rajendarn Muthuraj, Jack Luo

Abstract:

Recent advances in biomaterials have led to utilizing biopolymers to develop 3D scaffolds in tissue regeneration. One of the major challenges of designing biomaterials for 3D scaffolds is to mimic the building blocks similar to the extracellular matrix (ECM) of the native tissues. Biopolymer based aerogels obtained by freeze-drying have shown to provide structural similarities to the ECM owing to their 3D format and a highly porous structure with interconnected pores, similar to the ECM. Gelatin (GEL) is known to be a promising biomaterial with inherent regenerative characteristics owing to its chemical similarities to the ECM in native tissue, biocompatibility abundance, cost-effectiveness and accessible functional groups, which makes it facile for chemical modifications with other biomaterials to form biocomposites. Despite such advantages, gelatin offers poor mechanical properties, sensitive enzymatic degradation and high viscosity at room temperature which limits its application and encourages its use to develop biocomposites. Hydrophilic biomass-based cellulose nanofibrous (CNF) has been explored to use as suspension for biocomposite aerogels for the development of 3D porous structures with excellent mechanical properties, biocompatibility and slow enzymatic degradation. In this work, CNF biocomposite aerogels with various ratios of CNF:GEL) (90:10, 70:30 and 50:50) were prepared by freeze-drying technique, and their properties were investigated in terms of physicochemical, mechanical and biological characteristics. Epichlorohydrin (EPH) was used to investigate the effect of chemical crosslinking on the molecular interaction of CNF: GEL, and its effects on physicochemical, mechanical and biological properties of the biocomposite aerogels. Ultimately, chemical crosslinking helped to improve the mechanical resilience of the resulting aerogels. Amongst all the CNF-GEL composites, the crosslinked CNF: GEL (70:30) biocomposite was found to be favourable for cell attachment and viability. It possessed highly porous structure (porosity of ~93%) with pore sizes ranging from 16-110 µm, adequate mechanical properties (compression modulus of ~47 kPa) and optimal biocompatibility both in-vitro and in-vivo, as well as controlled enzymatic biodegradation, high water penetration, which could be considered a suitable option for wound healing application. In-vivo experiments showed improvement on inflammation and foreign giant body cell reaction for the crosslinked CNF: GEL (70:30) compared to the other samples. This could be due to the superior interaction of CNF with gelatin through chemical crosslinking, resulting in more optimal in-vivo improvement. In-vitro cell culture investigation on human dermal fibroblasts showed satisfactory 3D cell attachment over time. Overall, it has been observed that the developed CNF: GEL aerogel can be considered as a potential scaffold for soft tissue regeneration application.

Keywords: 3D scaffolds, aerogels, Biocomposites , tissue engineering

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Authors: Precious Barnes, Abraham Mensah, Leonard Derkyi-Kwarteng, Benjamin Amoani, George Adjei, Ernest Adankwah, Faustina Pappoe, Kwabena Dankwah, Daniel Amoako-Sakyi, Samuel Victor Nuvor, Dorcas Obiri-Yeboah, Ewura Seidu Yahaya, Patrick Kafui Akakpo, Roland Osei Saahene

Abstract:

Context: Breast cancer is a prevalent and aggressive type of cancer among African women, with high mortality rates in Ghana. Nuclear factor kappa B (NF-kB) is a transcription factor that has been associated with tumor progression in breast cancer. However, there is a lack of published data on NF-kB in breast cancer patients in Ghana or other African countries. Research Aim: The aim of this study was to assess the prognostic significance of NF-kB (p65) expression and its association with various clinicopathological features in breast cancer patients at the Cape Coast Teaching Hospital in Ghana. Methodology: A total of 90 formalin-fixed breast cancer tissues and 15 normal breast tissues were used in this study. The expression level of NF-kB (p65) was examined using immunohistochemical techniques. Correlation analysis between NF-kB (p65) expression and clinicopathological features was performed using SPSS version 25. Findings: The study found that NF-kB (p65) was expressed in 86.7% of breast cancer tissues. There was a significant relationship between NF-kB (p65) expression and tumor grade, proliferation index (Ki67), and molecular subtype. High-level expression of NF-kB (p65) was more common in tumor grade 3 compared to grade 1, and Ki67 > 20 had higher expression of NF-kB (p65) compared to Ki67 ≤ 20. Triple-negative breast cancer patients had the highest overexpression of NF-kB (p65) compared to other molecular subtypes. There was no significant association between NF-kB (p65) expression and other clinicopathological parameters. Theoretical Importance: This study provides important insights into the expression of NF-kB (p65) in breast cancer patients in Ghana, particularly in relation to tumor grade and proliferation index. The findings suggest that NF-kB (p65) could serve as a potential biological marker for cancer stage, progression, prognosis and as a therapeutic target. Data Collection and Analysis Procedures: Formalin-fixed breast cancer tissues and normal breast tissues were collected and analyzed using immunohistochemical techniques. Correlation analysis between NF-kB (p65) expression and clinicopathological features was performed using SPSS version 25. Question Addressed: This study addressed the question of the prognostic significance of NF-kB (p65) expression and its association with clinicopathological features in breast cancer patients in Ghana. Conclusion: This study, the first of its kind in Ghana, demonstrates that NF-kB (p65) is highly expressed among breast cancer patients at the Cape Coast Teaching Hospital, especially in triple-negative breast cancer patients. The expression of NF-kB (p65) is associated with tumor grade and proliferation index. NF-kB (p65) could potentially serve as a biological marker for cancer stage, progression, prognosis, and as a therapeutic target.

Keywords: breast cancer, Ki67, NF-kB (p65), tumor grade

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