Search results for: unreliable unpaced serial lines
968 Leveraging the HDAC Inhibitory Pharmacophore to Construct Deoxyvasicinone Based Tractable Anti-Lung Cancer Agent and pH-Responsive Nanocarrier
Authors: Ram Sharma, Esha Chatterjee, Santosh Kumar Guru, Kunal Nepali
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A tractable anti-lung cancer agent was identified via the installation of a Ring C expanded synthetic analogue of the alkaloid vasicinone [7,8,9,10-tetrahydroazepino[2,1-b] quinazolin-12(6H)-one (TAZQ)] as a surface recognition part in the HDAC inhibitory three-component model. Noteworthy to mention that the candidature of TAZQ was deemed suitable for accommodation in HDAC inhibitory pharmacophore as per the results of the fragment recruitment process conducted by our laboratory. TAZQ was pinpointed through the fragment screening program as a synthetically flexible fragment endowed with some moderate cell growth inhibitory activity against the lung cancer cell lines, and it was anticipated that the use of the aforementioned fragment to generate hydroxamic acid functionality (zinc-binding motif) bearing HDAC inhibitors would boost the antitumor efficacy of TAZQ. Consistent with our aim of applying epigenetic targets to the treatment of lung cancer, a strikingly potent anti-lung cancer scaffold (compound 6) was pinpointed through a series of in-vitro experiments. Notably, the compounds manifested a magnificent activity profile against KRAS and EGFR mutant lung cancer cell lines (IC50 = 0.80 - 0.96 µM), and the effects were found to be mediated through preferential HDAC6 inhibition (IC50 = 12.9 nM). In addition to HDAC6 inhibition, the compounds also elicited HDAC1 and HDAC3 inhibitory activity with an IC50 value of 49.9 nM and 68.5 nM, respectively. The HDAC inhibitory ability of compound 6 was also confirmed from the results of the western blot experiment that revealed its potential to decrease the expression levels of HDAC isoforms (HDAC1, HDAC3, and HDAC6). Noteworthy to mention that complete downregulation of the HDAC6 isoform was exerted by compound 6 at 0.5 and 1 µM. Moreover, in another western blot experiment, treatment with hydroxamic acid 6 led to upregulation of H3 acK9 and α-Tubulin acK40 levels, ascertaining its inhibitory activity toward both the class I HDACs and Class II B HDACs. The results of other assays were also encouraging as treatment with compound 6 led to the suppression of the colony formation ability of A549 cells, induction of apoptosis, and increase in autophagic flux. In silico studies led us to rationalize the results of the experimental assay, and some key interactions of compound 6 with the amino acid residues of HDAC isoforms were identified. In light of the impressive activity spectrum of compound 6, a pH-responsive nanocarrier (hyaluronic acid-compound 6 nanoparticles) was prepared. The dialysis bag approach was used for the assessment of the nanoparticles under both normal and acidic circumstances, and the pH-sensitive nature of hyaluronic acid-compound 6 nanoparticles was confirmed. Delightfully, the nanoformulation was devoid of cytotoxicity against the L929 mouse fibroblast cells (normal settings) and exhibited selective cytotoxicity towards the A549 lung cancer cell lines. In a nutshell, compound 6 appears to be a promising adduct, and a detailed investigation of this compound might yield a therapeutic for the treatment of lung cancer.Keywords: HDAC inhibitors, lung cancer, scaffold, hyaluronic acid, nanoparticles
Procedia PDF Downloads 90967 Genetic Diversity of Sugar Beet Pollinators
Authors: Ksenija Taški-Ajdukovic, Nevena Nagl, Živko Ćurčić, Dario Danojević
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Information about genetic diversity of sugar beet parental populations is of a great importance for hybrid breeding programs. The aim of this research was to evaluate genetic diversity among and within populations and lines of diploid sugar beet pollinators, by using SSR markers. As plant material were used eight pollinators originating from three USDA-ARS breeding programs and four pollinators from Institute of Field and Vegetable Crops, Novi Sad. Depending on the presence of self-fertility gene, the pollinators were divided into three groups: autofertile (inbred lines), autosterile (open-pollinating populations), and group with partial presence of autofertility gene. A total of 40 SSR primers were screened, out of which 34 were selected for the analysis of genetic diversity. A total of 129 different alleles were obtained with mean value 3.2 alleles per SSR primer. According to the results of genetic variability assessment the number and percentage of polymorphic loci was the maximal in pollinators NS1 and tester cms2 while effective number of alleles, expected heterozygosis and Shannon’s index was highest in pollinator EL0204. Analysis of molecular variance (AMOVA) showed that 77.34% of the total genetic variation was attributed to intra-varietal variance. Correspondence analysis results were very similar to grouping by neighbor-joining algorithm. Number of groups was smaller by one, because correspondence analysis merged IFVCNS pollinators with CZ25 into one group. Pollinators FC220, FC221 and C 51 were in the next group, while self-fertile pollinators CR10 and C930-35 from USDA-Salinas were separated. On another branch were self-sterile pollinators ЕL0204 and ЕL53 from USDA-East Lansing. Sterile testers cms1 and cms2 formed separate group. The presented results confirmed that SSR analysis can be successfully used in estimation of genetic diversity within and among sugar beet populations. Since the tested pollinator differed considering the presence of self-fertility gene, their heterozygosity differed as well. It was lower in genotypes with fixed self-fertility genes. Since the most of tested populations were open-pollinated, which rarely self-pollinate, high variability within the populations was expected. Cluster analysis grouped populations according to their origin.Keywords: auto fertility, genetic diversity, pollinator, SSR, sugar beet
Procedia PDF Downloads 456966 Transmission Line Inspection Using Drones
Authors: Jae Kyung Lee, Joon Young Park
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Maintenance on power transmission lines requires a lot of works. Sometimes they should be maintained on live-line environment with high altitude. Therefore, there always exist risks of falling from height and electric shock. To decline those risks, drones are recently applying on the electric power industry. This paper presents new operational technology while inspecting power transmission line. This paper also describes a technique for creating a flight path of a drone for transmission line inspection and a technique for controlling the drones of different types. Its technical and economical feasibilities have confirmed through experiments.Keywords: drones, transmission line, inspection, control system
Procedia PDF Downloads 346965 Application of Scoring Rubrics by Lecturers towards Objective Assessment of Essay Questions in the Department of Social Science Education, University of Calabar, Nigeria
Authors: Donald B. Enu, Clement O. Ukpor, Abigail E. Okon
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Unreliable scoring of students’ performance by lecturers short-chains students’ assessment in terms of underequipping the school authority with facts as intended by society through the curriculum hence, the learners, the school and the society are cheated because the usefulness of testing is defeated. This study, therefore, examined lecturers’ scoring objectivity of essay items in the Department of Social Science Education, University of Calabar, Nigeria. Specifically, it assessed lecturers’ perception of the relevance of scoring rubrics and its level of application. Data were collected from all the 36 lecturers in the Department (28 members and 8 non-members adjourned to the department), through a 20-item questionnaire and checklist instruments. A case-study design was adopted. Descriptive statistics of frequency counts, weighted means, standard deviations, and percentages were used to analyze data gathered. A mean score of 2.5 and or 60 percent and above formed the acceptance or significant level in decision taking. It was found that lecturers perceived the use of scoring rubrics as a relevant practice to ensure fairness and reliable treatment of examiners scripts particularly in marking essay items and that there is a moderately high level of adherence to the application of scoring rubrics. It was also observed that some criteria necessary for the scoring objectivity of essay items were not fully put in place in the department. It was recommended strongly that students’ identities be hidden while marking and that pre-determined marking scheme should be prepared centrally and strictly adhered to during marking and recording of scores. Conference marking should be enforced in the department.Keywords: essay items, objective scoring, scorers reliability, scoring rubrics
Procedia PDF Downloads 173964 Closing the Gap: Efficient Voxelization with Equidistant Scanlines and Gap Detection
Authors: S. Delgado, C. Cerrada, R. S. Gómez
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This research introduces an approach to voxelizing the surfaces of triangular meshes with efficiency and accuracy. Our method leverages parallel equidistant scan-lines and introduces a Gap Detection technique to address the limitations of existing approaches. We present a comprehensive study showcasing the method's effectiveness, scalability, and versatility in different scenarios. Voxelization is a fundamental process in computer graphics and simulations, playing a pivotal role in applications ranging from scientific visualization to virtual reality. Our algorithm focuses on enhancing the voxelization process, especially for complex models and high resolutions. One of the major challenges in voxelization in the Graphics Processing Unit (GPU) is the high cost of discovering the same voxels multiple times. These repeated voxels incur in costly memory operations with no useful information. Our scan-line-based method ensures that each voxel is detected exactly once when processing the triangle, enhancing performance without compromising the quality of the voxelization. The heart of our approach lies in the use of parallel, equidistant scan-lines to traverse the interiors of triangles. This minimizes redundant memory operations and avoids revisiting the same voxels, resulting in a significant performance boost. Moreover, our method's computational efficiency is complemented by its simplicity and portability. Written as a single compute shader in Graphics Library Shader Language (GLSL), it is highly adaptable to various rendering pipelines and hardware configurations. To validate our method, we conducted extensive experiments on a diverse set of models from the Stanford repository. Our results demonstrate not only the algorithm's efficiency, but also its ability to produce 26 tunnel free accurate voxelizations. The Gap Detection technique successfully identifies and addresses gaps, ensuring consistent and visually pleasing voxelized surfaces. Furthermore, we introduce the Slope Consistency Value metric, quantifying the alignment of each triangle with its primary axis. This metric provides insights into the impact of triangle orientation on scan-line based voxelization methods. It also aids in understanding how the Gap Detection technique effectively improves results by targeting specific areas where simple scan-line-based methods might fail. Our research contributes to the field of voxelization by offering a robust and efficient approach that overcomes the limitations of existing methods. The Gap Detection technique fills a critical gap in the voxelization process. By addressing these gaps, our algorithm enhances the visual quality and accuracy of voxelized models, making it valuable for a wide range of applications. In conclusion, "Closing the Gap: Efficient Voxelization with Equidistant Scan-lines and Gap Detection" presents an effective solution to the challenges of voxelization. Our research combines computational efficiency, accuracy, and innovative techniques to elevate the quality of voxelized surfaces. With its adaptable nature and valuable innovations, this technique could have a positive influence on computer graphics and visualization.Keywords: voxelization, GPU acceleration, computer graphics, compute shaders
Procedia PDF Downloads 64963 An Explanatory Study Approach Using Artificial Intelligence to Forecast Solar Energy Outcome
Authors: Agada N. Ihuoma, Nagata Yasunori
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Artificial intelligence (AI) techniques play a crucial role in predicting the expected energy outcome and its performance, analysis, modeling, and control of renewable energy. Renewable energy is becoming more popular for economic and environmental reasons. In the face of global energy consumption and increased depletion of most fossil fuels, the world is faced with the challenges of meeting the ever-increasing energy demands. Therefore, incorporating artificial intelligence to predict solar radiation outcomes from the intermittent sunlight is crucial to enable a balance between supply and demand of energy on loads, predict the performance and outcome of solar energy, enhance production planning and energy management, and ensure proper sizing of parameters when generating clean energy. However, one of the major problems of forecasting is the algorithms used to control, model, and predict performances of the energy systems, which are complicated and involves large computer power, differential equations, and time series. Also, having unreliable data (poor quality) for solar radiation over a geographical location as well as insufficient long series can be a bottleneck to actualization. To overcome these problems, this study employs the anaconda Navigator (Jupyter Notebook) for machine learning which can combine larger amounts of data with fast, iterative processing and intelligent algorithms allowing the software to learn automatically from patterns or features to predict the performance and outcome of Solar Energy which in turns enables the balance of supply and demand on loads as well as enhance production planning and energy management.Keywords: artificial Intelligence, backward elimination, linear regression, solar energy
Procedia PDF Downloads 154962 Assessing the Actual Status and Farmer’s Attitude towards Agroforestry in Chiniot, Pakistan
Authors: M. F. Nawaz, S. Gul, T. H. Farooq, M. T. Siddiqui, M. Asif, I. Ahmad, N. K. Niazi
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In Pakistan, major demands of fuel wood and timber wood are fulfilled by agroforestry. However, the information regarding economic significance of agroforestry and its productivity in Pakistan is still insufficient and unreliable. Survey of field conditions to examine the agroforestry status at local level helps us to know the future trends and to formulate the policies for sustainable wood supply. The objectives of this research were to examine the actual status and potential of agroforestry and to point out the barriers that are faced by farmers in the adoption of agroforestry. Research was carried out in Chiniot district, Pakistan because it is the famous city for furniture industry that is largely dependent on farm trees. A detailed survey of district Chiniot was carried out from 150 randomly selected farmer respondents using multi-objective oriented and pre-tested questionnaire. It was found that linear tree planting method was more adopted (45%) as compared to linear + interplanting (42%) and/or compact planting (12.6%). Chi-square values at P-value <0.5 showed that age (11.35) and education (17.09) were two more important factors in the quick adoption of agroforestry as compared to land holdings (P-value of 0.7). The major reason of agroforestry adoption was to obtain income, fodder and fuelwood. The most dominant species in farmlands was shisham (Dalbergia sissoo) but since last five years, mostly farmers were growing Sufeida (Eucalyptus camaldulensis), kikar (Acacia nilotica) and popular (Populus deltoides) on their fields due to “Shisham die-back” problem. It was found that agro-forestry can be increased by providing good quality planting material to farmers and improving wood markets.Keywords: agroforestry, trees, services, agriculture, farmers
Procedia PDF Downloads 446961 Performance Evaluation of Routing Protocol in Cognitive Radio with Multi Technological Environment
Authors: M. Yosra, A. Mohamed, T. Sami
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Over the past few years, mobile communication technologies have seen significant evolution. This fact promoted the implementation of many systems in a multi-technological setting. From one system to another, the Quality of Service (QoS) provided to mobile consumers gets better. The growing number of normalized standards extends the available services for each consumer, moreover, most of the available radio frequencies have already been allocated, such as 3G, Wifi, Wimax, and LTE. A study by the Federal Communications Commission (FCC) found that certain frequency bands are partially occupied in particular locations and times. So, the idea of Cognitive Radio (CR) is to share the spectrum between a primary user (PU) and a secondary user (SU). The main objective of this spectrum management is to achieve a maximum rate of exploitation of the radio spectrum. In general, the CR can greatly improve the quality of service (QoS) and improve the reliability of the link. The problem will reside in the possibility of proposing a technique to improve the reliability of the wireless link by using the CR with some routing protocols. However, users declared that the links were unreliable and that it was an incompatibility with QoS. In our case, we choose the QoS parameter "bandwidth" to perform a supervised classification. In this paper, we propose a comparative study between some routing protocols, taking into account the variation of different technologies on the existing spectral bandwidth like 3G, WIFI, WIMAX, and LTE. Due to the simulation results, we observe that LTE has significantly higher availability bandwidth compared with other technologies. The performance of the OLSR protocol is better than other on-demand routing protocols (DSR, AODV and DSDV), in LTE technology because of the proper receiving of packets, less packet drop and the throughput. Numerous simulations of routing protocols have been made using simulators such as NS3.Keywords: cognitive radio, multi technology, network simulator (NS3), routing protocol
Procedia PDF Downloads 54960 Implementing Effective Strategies to Improve Teaching and Learning in Higher Education: Balancing the Engagement Acts between Lecturers And Students
Authors: Jeffrey Siphiwe Mkhize
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Twelve years of schooling for most South African children, particularly those children from disadvantaged past, are confronted with numerous and diverse challenges. These challenges range from infrastructural limitations, language of teaching, poor resources and varying family backgrounds. Likewise, schools are categorized to signify schools’ geographic location, poverty lines, societal class and type of students that the school are likely to enroll. Such categorization perpetuates particular lines of identities that are indirectly reinforced by the same system that seeks to redress. South African universities prefer point systems to determine students’ suitability to gain access to their programmes. Once students are admitted based on the qualifying points there is an assumed equity in the manner in which they receive tuition. They are assumed as equal; noting the widened access to South African universities as means to redress past inequalities. Given the challenges, inequalities, it is necessary to view higher education as a site for knowledge construction that is accessible to all students. Epistemological access is key to all students irrespective of their socio-economic status. This paper seeks to contribute to the discourse of student engagement using lecturer-student relationship as a lens to understand this phenomenon. Data were generated using South African Survey of Student Engagement, focus group interviews, semi-structured one-on-one-interviews as well as document analysis. The focus was on students registered for the first year of a Bachelor of Education degree as well as lecturers that teach high risk modules in this qualification at the same level. The findings suggest that lecturers are challenged by overcrowded classrooms and over-enrolled modules; this challenge hampers their good intentions to become more efficient and innovative in their teaching. Students lack confidence in approaching lecturers for assistance. Collaborative learning has stronger results and students believe in self-support to deal with their challenges based on their individual strengths. Collaborative learning is key to student academic performance.Keywords: collaborative learning, consultations, student engagement, student performance
Procedia PDF Downloads 102959 A Biophysical Model of CRISPR/Cas9 on- and off-Target Binding for Rational Design of Guide RNAs
Authors: Iman Farasat, Howard M. Salis
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The CRISPR/Cas9 system has revolutionized genome engineering by enabling site-directed and high-throughput genome editing, genome insertion, and gene knockdowns in several species, including bacteria, yeast, flies, worms, and human cell lines. This technology has the potential to enable human gene therapy to treat genetic diseases and cancer at the molecular level; however, the current CRISPR/Cas9 system suffers from seemingly sporadic off-target genome mutagenesis that prevents its use in gene therapy. A comprehensive mechanistic model that explains how the CRISPR/Cas9 functions would enable the rational design of the guide-RNAs responsible for target site selection while minimizing unexpected genome mutagenesis. Here, we present the first quantitative model of the CRISPR/Cas9 genome mutagenesis system that predicts how guide-RNA sequences (crRNAs) control target site selection and cleavage activity. We used statistical thermodynamics and law of mass action to develop a five-step biophysical model of cas9 cleavage, and examined it in vivo and in vitro. To predict a crRNA's binding specificities and cleavage rates, we then compiled a nearest neighbor (NN) energy model that accounts for all possible base pairings and mismatches between the crRNA and the possible genomic DNA sites. These calculations correctly predicted crRNA specificity across 5518 sites. Our analysis reveals that cas9 activity and specificity are anti-correlated, and, the trade-off between them is the determining factor in performing an RNA-mediated cleavage with minimal off-targets. To find an optimal solution, we first created a scheme of safe-design criteria for Cas9 target selection by systematic analysis of available high throughput measurements. We then used our biophysical model to determine the optimal Cas9 expression levels and timing that maximizes on-target cleavage and minimizes off-target activity. We successfully applied this approach in bacterial and mammalian cell lines to reduce off-target activity to near background mutagenesis level while maintaining high on-target cleavage rate.Keywords: biophysical model, CRISPR, Cas9, genome editing
Procedia PDF Downloads 398958 Evaluating and Examining Pictures of Children of Five Years Old
Authors: Emine Yılmaz Bolat
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Early childhood is a very important period in terms of identifying and developing early skills and abilities. It is likely that the child's development will be in the same direction in the future. This study was conducted with 26 children for the purpose of examining pictures of children of five years old. In the survey, children were asked to draw a picture with pastel dyes. The drawings were collected and evaluated by the researcher. At the end of the research, it was found that the children used the yellow color (N = 17, 16,34%) and the least gray color (N = 1, 0,96%). When the features of children's pictures are examined, the children's paintings have been found to have hierarchy, transparency, completion, the use of vivid colors, and the presence of vertical and horizontal painting lines.Keywords: early childhood, kindergarten, pictures of children, features of pictures
Procedia PDF Downloads 298957 A Single Cell Omics Experiments as Tool for Benchmarking Bioinformatics Oncology Data Analysis Tools
Authors: Maddalena Arigoni, Maria Luisa Ratto, Raffaele A. Calogero, Luca Alessandri
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The presence of tumor heterogeneity, where distinct cancer cells exhibit diverse morphological and phenotypic profiles, including gene expression, metabolism, and proliferation, poses challenges for molecular prognostic markers and patient classification for targeted therapies. Understanding the causes and progression of cancer requires research efforts aimed at characterizing heterogeneity, which can be facilitated by evolving single-cell sequencing technologies. However, analyzing single-cell data necessitates computational methods that often lack objective validation. Therefore, the establishment of benchmarking datasets is necessary to provide a controlled environment for validating bioinformatics tools in the field of single-cell oncology. Benchmarking bioinformatics tools for single-cell experiments can be costly due to the high expense involved. Therefore, datasets used for benchmarking are typically sourced from publicly available experiments, which often lack a comprehensive cell annotation. This limitation can affect the accuracy and effectiveness of such experiments as benchmarking tools. To address this issue, we introduce omics benchmark experiments designed to evaluate bioinformatics tools to depict the heterogeneity in single-cell tumor experiments. We conducted single-cell RNA sequencing on six lung cancer tumor cell lines that display resistant clones upon treatment of EGFR mutated tumors and are characterized by driver genes, namely ROS1, ALK, HER2, MET, KRAS, and BRAF. These driver genes are associated with downstream networks controlled by EGFR mutations, such as JAK-STAT, PI3K-AKT-mTOR, and MEK-ERK. The experiment also featured an EGFR-mutated cell line. Using 10XGenomics platform with cellplex technology, we analyzed the seven cell lines together with a pseudo-immunological microenvironment consisting of PBMC cells labeled with the Biolegend TotalSeq™-B Human Universal Cocktail (CITEseq). This technology allowed for independent labeling of each cell line and single-cell analysis of the pooled seven cell lines and the pseudo-microenvironment. The data generated from the aforementioned experiments are available as part of an online tool, which allows users to define cell heterogeneity and generates count tables as an output. The tool provides the cell line derivation for each cell and cell annotations for the pseudo-microenvironment based on CITEseq data by an experienced immunologist. Additionally, we created a range of pseudo-tumor tissues using different ratios of the aforementioned cells embedded in matrigel. These tissues were analyzed using 10XGenomics (FFPE samples) and Curio Bioscience (fresh frozen samples) platforms for spatial transcriptomics, further expanding the scope of our benchmark experiments. The benchmark experiments we conducted provide a unique opportunity to evaluate the performance of bioinformatics tools for detecting and characterizing tumor heterogeneity at the single-cell level. Overall, our experiments provide a controlled and standardized environment for assessing the accuracy and robustness of bioinformatics tools for studying tumor heterogeneity at the single-cell level, which can ultimately lead to more precise and effective cancer diagnosis and treatment.Keywords: single cell omics, benchmark, spatial transcriptomics, CITEseq
Procedia PDF Downloads 106956 Small Scale Mobile Robot Auto-Parking Using Deep Learning, Image Processing, and Kinematics-Based Target Prediction
Authors: Mingxin Li, Liya Ni
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Autonomous parking is a valuable feature applicable to many robotics applications such as tour guide robots, UV sanitizing robots, food delivery robots, and warehouse robots. With auto-parking, the robot will be able to park at the charging zone and charge itself without human intervention. As compared to self-driving vehicles, auto-parking is more challenging for a small-scale mobile robot only equipped with a front camera due to the camera view limited by the robot’s height and the narrow Field of View (FOV) of the inexpensive camera. In this research, auto-parking of a small-scale mobile robot with a front camera only was achieved in a four-step process: Firstly, transfer learning was performed on the AlexNet, a popular pre-trained convolutional neural network (CNN). It was trained with 150 pictures of empty parking slots and 150 pictures of occupied parking slots from the view angle of a small-scale robot. The dataset of images was divided into a group of 70% images for training and the remaining 30% images for validation. An average success rate of 95% was achieved. Secondly, the image of detected empty parking space was processed with edge detection followed by the computation of parametric representations of the boundary lines using the Hough Transform algorithm. Thirdly, the positions of the entrance point and center of available parking space were predicted based on the robot kinematic model as the robot was driving closer to the parking space because the boundary lines disappeared partially or completely from its camera view due to the height and FOV limitations. The robot used its wheel speeds to compute the positions of the parking space with respect to its changing local frame as it moved along, based on its kinematic model. Lastly, the predicted entrance point of the parking space was used as the reference for the motion control of the robot until it was replaced by the actual center when it became visible again by the robot. The linear and angular velocities of the robot chassis center were computed based on the error between the current chassis center and the reference point. Then the left and right wheel speeds were obtained using inverse kinematics and sent to the motor driver. The above-mentioned four subtasks were all successfully accomplished, with the transformed learning, image processing, and target prediction performed in MATLAB, while the motion control and image capture conducted on a self-built small scale differential drive mobile robot. The small-scale robot employs a Raspberry Pi board, a Pi camera, an L298N dual H-bridge motor driver, a USB power module, a power bank, four wheels, and a chassis. Future research includes three areas: the integration of all four subsystems into one hardware/software platform with the upgrade to an Nvidia Jetson Nano board that provides superior performance for deep learning and image processing; more testing and validation on the identification of available parking space and its boundary lines; improvement of performance after the hardware/software integration is completed.Keywords: autonomous parking, convolutional neural network, image processing, kinematics-based prediction, transfer learning
Procedia PDF Downloads 129955 Effects of Lateness Gene on Yield and Related Traits in Indica Rice
Authors: B. B. Rana, M. Yokota, Y. Shimizu, Y. Koide, I. Takamure, T. Kawano, M. Murai
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Various genes which control or affect heading time have been found in rice. Out of them, Se1 and E1 loci play important roles in determining heading time by controlling photosensitivity. An isogenic-line pair of late and early lines were developed from progenies of the F1 from Suweon 258 × 36U. A lateness gene tentatively designated as “Ex” was found to control the difference in heading time between the early and late lines mentioned above. The present study was conducted to examine the effect of Ex on yield and related traits. Indica-type variety Suweon 258 was crossed with 36U, which is an Ur1 (Undulate rachis-1) isogenic line of IR36. In the F2 population, comparatively early-heading, late-heading and intermediate-heading plants were segregated. Segregation similar to that by the three types of heading was observed in the F3 and later generations. A late-heading plant and an early-heading plant were selected in the F8 population from an intermediate-heading F7 plant, for developing L and E of the isogenic-line pair, respectively. Experiments for L and E were conducted by randomized block design with three replications. Transplanting was conducted on May 3 at a planting distance of 30 cm × 15 cm with two seedlings per hill to an experimental field of the Faculty of Agriculture, Kochi University. Chemical fertilizers containing N, P2O5 and K2O were applied at the nitrogen levels of 4 g/m2, 9 g/m2 and 18 g/m2 in total being denoted by "N4", "N9" and "N18", respectively. Yield, yield components and other traits were measured. Ex delayed 80%-heading by 17 or 18 days in L as compared with E. In total brown rice yield (g/m2), L was 635, 606 and 590, and E was 577, 548 and 501, respectively, at N18, N9 and N4, indicating that Ex increased this trait by 10% to 18%. Ex increased yield-1.5 mm sieve (g/m2) b 9% to 15% at the three fertilizer levels. Ex increased the spikelet number per panicle by 16% to 22%. As a result, the spikelet number per m2 was increased by 11% to 18% at the three fertilizer levels. Ex decreased 1000-grain weight (g) by 2 to 4%. L was not significantly different from E in ripened-grain percentage, fertilized-spikelet percentage and percentage of ripened grains to fertilized spikelets. Hence, it is inferred that Ex increased yield by increasing spikelet number per panicle. Hence, Ex could be utilized to develop high yielding varieties for warmer districts.Keywords: heading time, lateness gene, photosensitivity, yield, yield components
Procedia PDF Downloads 193954 New Method for the Synthesis of Different Pyrroloquinazolinoquinolin Alkaloids
Authors: Abdulkareem M. Hamid, Yaseen Elhebshi, Adam Daïch
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Luotonins and its derivatives (Isoluotonins) are alkaloids from the aerial parts of Peganum nigellastrum Bunge that display three major skeleton types. Luotonins A, B, and E are pyrroloquinazolinoquinoline alkaloids. A few methods were known for the sysnthesis of Isoluotonin. All luotonins have shown promising cytotoxicities towards selected human cancer cell lines, especially against leukemia P-388 cells. Luotonin A is the most active one, with its activity stemming from topoisomerase I-dependent DNA-cleavage. Such intriguing biological activities and unique structures have led not only to the development of synthetic methods for the efficient synthesis of these compounds, but also to interest in structural modifications for improving the biological properties. Recent progress in the study of luotonins is covered.Keywords: luotonin A, isoluotonin, pyrroloquiolines, alkaloids
Procedia PDF Downloads 411953 Phase Diagram Including a Negative Pressure Region for a Thermotropic Liquid Crystal in a Metal Berthelot Tube
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Thermodynamic properties of liquids under negative pressures are interesting and important in fields of scienceand technology. Here, phase transitions of a thermotropic liquid crystal are investigatedin a range from positive to negative pressures with a metal Berthelot tube using a commercial pressure transducer.Two co-existinglines, namely crystal (Kr) – nematic (N), and isotropic liquid (I) - nematic (N) lines, weredrawn in a pressure - temperature plane. The I-N line was drawn to ca. -5 (MPa).Keywords: Berthelot method, liquid crystal, negative pressure, phase transitions
Procedia PDF Downloads 399952 Augmented ADRC for Trajectory Tracking of a Novel Hydraulic Spherical Motion Mechanism
Authors: Bin Bian, Liang Wang
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A hydraulic spherical motion mechanism (HSMM) is proposed. Unlike traditional systems using serial or parallel mechanisms for multi-DOF rotations, the HSMM is capable of implementing continuous 2-DOF rotational motions in a single joint without the intermediate transmission mechanisms. It has some advantages of compact structure, low inertia and high stiffness. However, as HSMM is a nonlinear and multivariable system, it is very complicate to realize accuracy control. Therefore, an augmented active disturbance rejection controller (ADRC) is proposed in this paper. Compared with the traditional PD control method, three compensation items, i.e., dynamics compensation term, disturbance compensation term and nonlinear error elimination term, are added into the proposed algorithm to improve the control performance. The ADRC algorithm aims at offsetting the effects of external disturbance and realizing accurate control. Euler angles are applied to describe the orientation of rotor. Lagrange equations are utilized to establish the dynamic model of the HSMM. The stability of this algorithm is validated with detailed derivation. Simulation model is formulated in Matlab/Simulink. The results show that the proposed control algorithm has better competence of trajectory tracking in the presence of uncertainties.Keywords: hydraulic spherical motion mechanism, dynamic model, active disturbance rejection control, trajectory tracking
Procedia PDF Downloads 99951 Lean Commercialization: A New Dawn for Commercializing High Technologies
Authors: Saheed A. Gbadegeshin
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Lean Commercialization (LC) is a transformation of new technologies and knowledge to products and services through application of lean/agile principle. This principle focuses on how resources can be minimized on development, manufacturing, and marketing new products/services, which can be accepted by customers. To understand how the LC has been employed by the technology-based companies, a case study approach was employed by interviewing the founders, observing their high technologies, and interviewing the commercialization experts. Two serial entrepreneurs were interviewed in 2012, and their commercialized technologies were monitored from 2012 till 2016. Some results were collected, but to validate the commercialization strategies of these entrepreneurs, four commercialization experts were interviewed in 2017. Initial results, observation notes, and experts’ opinions were analyzed qualitatively. The final findings showed that the entrepreneurs applied the LC unknowingly, and the experts were aware of the LC. Similarly, the entrepreneurs used the LC due to the financial constraints, and their need for success. Additionally, their commercialization practices revealed that LC appeared to be one of their commercialization strategies. Thus, their practices were analyzed, and a framework was developed. Furthermore, the experts noted that LC is a new dawn, which technologists and scientists need to consider for their high technology commercialization. This article contributes to the theory and practice of commercialization. Theoretically, the framework adds value to the commercialization discussion. And, practically the framework can be used by the technology entrepreneurs (technologists and scientists), technology-based enterprises, and technology entrepreneurship educators as a guide in their commercialization adventures.Keywords: lean commercialization, high technologies, lean start-up, technology-based companies
Procedia PDF Downloads 157950 Caffeic Acid Methyl and Ethyl Esters Exhibit Beneficial Effect on Glucose and Lipid Metabolism in Cultured Murine Insulin-Sensitive Cells
Authors: Hoda M. Eid, Abir Nachar, Farah Thong, Gary Sweeney, Pierre S. Haddad
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Caffeic acid methyl ester (CAME) and caffeic ethyl esters (CAEE) were previously reported to potently stimulate glucose uptake in cultured C2C12 skeletal muscle cells via insulin-independent mechanisms involving the activation of adenosine monophosphate-activated protein kinase (AMPK). In the present study, we investigated the effect of the two compounds on the translocation of glucose transporter GLUT4 in L6 skeletal muscle cells. The cells were treated with the optimum non-toxic concentration (50 µM) of either CAME or CAEE for 18 h. Levels of GLUT4myc at the cell surface were measured by O-phenylenediamine dihydrochloride (OPD) assay. The effects of CAME and CAEE on GLUT1 and GLUT4 protein content were also measured by western immunoblot. Our results show that CAME and CAEE significantly increased glucose uptake, GLUT4 translocation and GLUT4 protein content. Furthermore, the effect of the two CA esters on two insulin-sensitive cell lines: H4IIE rat hepatoma and 3T3-L1 adipocytes were investigated. CAME and CAEE reduced the enzymatic activity of the key hepatic gluconeogenic enzyme glucose-6-phosphatase in a concentration-dependent manner. In addition, they exerted a concentration-dependent antiadipogenic effect on 3T3-L1 cells. Mitotic clonal expansion (MCE), a prerequisite for adipocytes differentiation was also concentration-dependently inhibited. The two compounds abrogated lipid droplet accumulation, blocked MCE and maintained cells in fibroblast-like state when applied at the maximum non-toxic concentration (100 µM). In addition, the expression of the early key adipogenic transcription factors CCAAT enhancer-binding protein beta (C/EBP-β) and the master regulator of adipogenesis peroxisome-proliferator-activated receptor gamma (PPAR-γ) were inhibited. We, therefore, conclude that CAME and CAEE exert pleiotropic benefits in several insulin-sensitive cell lines through insulin-independent mechanisms involving AMPK, hence they may treat obesity, diabetes and other metabolic diseases.Keywords: type 2 diabetes mellitus, insulin resistance, GLUT4, Akt, AMPK.
Procedia PDF Downloads 307949 Critical Velocities for Particle Transport from Experiments and CFD Simulations
Authors: Sajith Sajeev, Brenton McLaury, Siamack Shirazi
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In the petroleum industry, solid particles are often present along with the produced fluids. It is imperative to keep particles from accumulating in flow lines. In this study, various experiments are conducted to study sand particle transport, where critical velocity is defined as the average fluid velocity to keep particles continuously moving. Many parameters related to the fluid, particles and pipe affect the transport process. Experimental results are presented varying the particle concentration. Additionally, CFD simulations using a discrete element modeling (DEM) approach are presented to compare with experimental result.Keywords: particle transport, critical velocity, CFD, DEM
Procedia PDF Downloads 300948 Infusion Pump Historical Development, Measurement and Parts of Infusion Pump
Authors: Samuel Asrat
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Infusion pumps have become indispensable tools in modern healthcare, allowing for precise and controlled delivery of fluids, medications, and nutrients to patients. This paper provides an overview of the historical development, measurement, and parts of infusion pumps. The historical development of infusion pumps can be traced back to the early 1960s when the first rudimentary models were introduced. These early pumps were large, cumbersome, and often unreliable. However, advancements in technology and engineering over the years have led to the development of smaller, more accurate, and user-friendly infusion pumps. Measurement of infusion pumps involves assessing various parameters such as flow rate, volume delivered, and infusion duration. Flow rate, typically measured in milliliters per hour (mL/hr), is a critical parameter that determines the rate at which fluids or medications are delivered to the patient. Accurate measurement of flow rate is essential to ensure the proper administration of therapy and prevent adverse effects. Infusion pumps consist of several key parts, including the pump mechanism, fluid reservoir, tubing, and control interface. The pump mechanism is responsible for generating the necessary pressure to push fluids through the tubing and into the patient's bloodstream. The fluid reservoir holds the medication or solution to be infused, while the tubing serves as the conduit through which the fluid travels from the reservoir to the patient. The control interface allows healthcare providers to program and adjust the infusion parameters, such as flow rate and volume. In conclusion, infusion pumps have evolved significantly since their inception, offering healthcare providers unprecedented control and precision in delivering fluids and medications to patients. Understanding the historical development, measurement, and parts of infusion pumps is essential for ensuring their safe and effective use in clinical practice. Procedia PDF Downloads 53947 A Study on Design for Parallel Test Based on Embedded System
Authors: Zheng Sun, Weiwei Cui, Xiaodong Ma, Hongxin Jin, Dongpao Hong, Jinsong Yang, Jingyi Sun
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With the improvement of the performance and complexity of modern equipment, automatic test system (ATS) becomes widely used for condition monitoring and fault diagnosis. However, the conventional ATS mainly works in a serial mode, and lacks the ability of testing several equipments at the same time. That leads to low test efficiency and ATS redundancy. Especially for a large majority of equipment under test, the conventional ATS cannot meet the requirement of efficient testing. To reduce the support resource and increase test efficiency, we propose a method of design for the parallel test based on the embedded system in this paper. Firstly, we put forward the general framework of the parallel test system, and the system contains a central management system (CMS) and several distributed test subsystems (DTS). Then we give a detailed design of the system. For the hardware of the system, we use embedded architecture to design DTS. For the software of the system, we use test program set to improve the test adaption. By deploying the parallel test system, the time to test five devices is now equal to the time to test one device in the past. Compared with the conventional test system, the proposed test system reduces the size and improves testing efficiency. This is of great significance for equipment to be put into operation swiftly. Finally, we take an industrial control system as an example to verify the effectiveness of the proposed method. The result shows that the method is reasonable, and the efficiency is improved up to 500%.Keywords: parallel test, embedded system, automatic test system, automatic test system (ATS), central management system, central management system (CMS), distributed test subsystems, distributed test subsystems (DTS)
Procedia PDF Downloads 294946 Automatic Tuning for a Systemic Model of Banking Originated Losses (SYMBOL) Tool on Multicore
Authors: Ronal Muresano, Andrea Pagano
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Nowadays, the mathematical/statistical applications are developed with more complexity and accuracy. However, these precisions and complexities have brought as result that applications need more computational power in order to be executed faster. In this sense, the multicore environments are playing an important role to improve and to optimize the execution time of these applications. These environments allow us the inclusion of more parallelism inside the node. However, to take advantage of this parallelism is not an easy task, because we have to deal with some problems such as: cores communications, data locality, memory sizes (cache and RAM), synchronizations, data dependencies on the model, etc. These issues are becoming more important when we wish to improve the application’s performance and scalability. Hence, this paper describes an optimization method developed for Systemic Model of Banking Originated Losses (SYMBOL) tool developed by the European Commission, which is based on analyzing the application's weakness in order to exploit the advantages of the multicore. All these improvements are done in an automatic and transparent manner with the aim of improving the performance metrics of our tool. Finally, experimental evaluations show the effectiveness of our new optimized version, in which we have achieved a considerable improvement on the execution time. The time has been reduced around 96% for the best case tested, between the original serial version and the automatic parallel version.Keywords: algorithm optimization, bank failures, OpenMP, parallel techniques, statistical tool
Procedia PDF Downloads 363945 The Coalescence Process of Droplet Pairs in Different Junctions
Authors: Xiang Wang, Yan Pang, Zhaomiao Liu
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Droplet-based microfluidics have been studied extensively with the development of the Micro-Electro-Mechanical System (MEMS) which bears the advantages of high throughput, high efficiency, low cost and low polydispersity. Droplets, worked as versatile carriers, could provide isolated chambers as the internal dispersed phase is protected from the outside continuous phase. Droplets are used to add reagents to start or end bio-chemical reactions, to generate concentration gradients, to realize hydrate crystallization or protein analyses, while droplets coalescence acts as an important control technology. In this paper, deionized water is used as the dispersed phase, and several kinds of oil are used as the continuous phase to investigate the influence of the viscosity ratio of the two phases on the coalescence process. The microchannels are fabricated by coating a polydimethylsiloxane (PDMS) layer onto another PDMS flat plate after corona treatment. All newly made microchannels are rinsed with the continuous oil phase for hours before experiments to ensure the swelling fully developed. High-speed microscope system is used to document the serial videos with a maximum speed of 2000 frames per second. The critical capillary numbers (Ca*) of droplet pairs in various junctions are studied and compared. Ca* varies with different junctions or different liquids within the range of 0.002 to 0.01. However, droplets without extra control would have the problem of synchronism which reduces the coalescence efficiency.Keywords: coalescence, concentration, critical capillary number, droplet pair, split
Procedia PDF Downloads 240944 Autism Disease Detection Using Transfer Learning Techniques: Performance Comparison between Central Processing Unit vs. Graphics Processing Unit Functions for Neural Networks
Authors: Mst Shapna Akter, Hossain Shahriar
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Neural network approaches are machine learning methods used in many domains, such as healthcare and cyber security. Neural networks are mostly known for dealing with image datasets. While training with the images, several fundamental mathematical operations are carried out in the Neural Network. The operation includes a number of algebraic and mathematical functions, including derivative, convolution, and matrix inversion and transposition. Such operations require higher processing power than is typically needed for computer usage. Central Processing Unit (CPU) is not appropriate for a large image size of the dataset as it is built with serial processing. While Graphics Processing Unit (GPU) has parallel processing capabilities and, therefore, has higher speed. This paper uses advanced Neural Network techniques such as VGG16, Resnet50, Densenet, Inceptionv3, Xception, Mobilenet, XGBOOST-VGG16, and our proposed models to compare CPU and GPU resources. A system for classifying autism disease using face images of an autistic and non-autistic child was used to compare performance during testing. We used evaluation matrices such as Accuracy, F1 score, Precision, Recall, and Execution time. It has been observed that GPU runs faster than the CPU in all tests performed. Moreover, the performance of the Neural Network models in terms of accuracy increases on GPU compared to CPU.Keywords: autism disease, neural network, CPU, GPU, transfer learning
Procedia PDF Downloads 107943 Climate Adaptations to Traditional Milpa Farming Practices in Mayan Communities of Southern Belize: A Socio-Ecological Systems Approach
Authors: Kristin Drexler
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Climate change has exacerbated food and livelihood insecurity for Mayan milpa farmers in Central America. For centuries, milpa farming has been sustainable for subsistence; however, in the last 50 years, milpas have become less reliable due to accelerating climate change, resource degradation, declining markets, poverty, and other factors. Using interviews with extension leaders and milpa farmers in Belize, this qualitative study examines the capacity for increasing climate-smart agriculture (CSA) aspects of existing traditional milpa practices, specifically no-burn mulching, soil enrichment, and the use of cover plants. Applying community capitals and socio-ecological systems frameworks, this study finds four key capitals were perceived by farmers and agriculture extension leaders as barriers for increasing CSA practices: (1) human-capacity, (2) financial, (3) infrastructure, and (4) governance-justice capitals. The key barriers include a lack of CSA technology and pest management knowledge-sharing (human-capacity), unreliable roads and utility services (infrastructure), the closure of small markets and crop-buying programs in Belize (financial), and constraints on extension services and exacerbating a sense of marginalization in Maya communities (governance-justice). Recommendations are presented for government action to reduce barriers and facilitate an increase in milpa crop productivity, promote food and livelihood security, and enable climate resilience of Mayan milpa communities in Belize.Keywords: socio-ecological systems, community capitals, climate-smart agriculture, food security, milpa, Belize
Procedia PDF Downloads 88942 Counting People Utilizing Space-Time Imagery
Authors: Ahmed Elmarhomy, K. Terada
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An automated method for counting passerby has been proposed using virtual-vertical measurement lines. Space-time image is representing the human regions which are treated using the segmentation process. Different color space has been used to perform the template matching. A proper template matching has been achieved to determine direction and speed of passing people. Distinguish one or two passersby has been investigated using a correlation between passerby speed and the human-pixel area. Finally, the effectiveness of the presented method has been experimentally verified.Keywords: counting people, measurement line, space-time image, segmentation, template matching
Procedia PDF Downloads 446941 Analysis of Brake System for Vehicle Off-Road
Authors: Elmo Thiago Lins Cöuras Ford, Valentina Alessandra Carvalho do Vale, José Ubiragi de Lima Mendes
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In elapsing of the years it elaborates automobile it is developing automobiles more and more modern that, every year, the vehicles recently of the assembly lines, practically they push for the past produced models there is very little time. Those innovations didn't also pass unperceived in 0respect the safety of the vehicles. It is in this development apprenticeship the brakes systems equipped more and more with resources sophisticated. In that way, before of that context, this research tried to project a brake system for a vehicle off-road and to analyze your acting as the brakes efficiency: distances traveled and time, concluding with possible improvements in the system.Keywords: brakes system, off-road, vehicle acting, automotive and mechanical engineering
Procedia PDF Downloads 479940 Biflavonoids from Selaginellaceae as Epidermal Growth Factor Receptor Inhibitors and Their Anticancer Properties
Authors: Adebisi Adunola Demehin, Wanlaya Thamnarak, Jaruwan Chatwichien, Chatchakorn Eurtivong, Kiattawee Choowongkomon, Somsak Ruchirawat, Nopporn Thasana
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The epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein involved in cellular signalling processes and, its aberrant activity is crucial in the development of many cancers such as lung cancer. Selaginellaceae are fern allies that have long been used in Chinese traditional medicine to treat various cancer types, especially lung cancer. Biflavonoids, the major secondary metabolites in Selaginellaceae, have numerous pharmacological activities, including anti-cancer and anti-inflammatory. For instance, amentoflavone induces a cytotoxic effect in the human NSCLC cell line via the inhibition of PARP-1. However, to the best of our knowledge, there are no studies on biflavonoids as EGFR inhibitors. Thus, this study aims to investigate the EGFR inhibitory activities of biflavonoids isolated from Selaginella siamensis and Selaginella bryopteris. Amentoflavone, tetrahydroamentoflavone, sciadopitysin, robustaflavone, robustaflavone-4-methylether, delicaflavone, and chrysocauloflavone were isolated from the ethyl-acetate extract of the whole plants. The structures were determined using NMR spectroscopy and mass spectrometry. In vitro study was conducted to evaluate their cytotoxicity against A549, HEPG2, and T47D human cancer cell lines using the MTT assay. In addition, a target-based assay was performed to investigate their EGFR inhibitory activity using the kinase inhibition assay. Finally, a molecular docking study was conducted to predict the binding modes of the compounds. Robustaflavone-4-methylether and delicaflavone showed the best cytotoxic activity on all the cell lines with IC50 (µM) values of 18.9 ± 2.1 and 22.7 ± 3.3 on A549, respectively. Of these biflavonoids, delicaflavone showed the most potent EGFR inhibitory activity with an 84% relative inhibition at 0.02 nM using erlotinib as a positive control. Robustaflavone-4-methylether showed a 78% inhibition at 0.15 nM. The docking scores obtained from the molecular docking study correlated with the kinase inhibition assay. Robustaflavone-4-methylether and delicaflavone had a docking score of 72.0 and 86.5, respectively. The inhibitory activity of delicaflavone seemed to be linked with the C2”=C3” and 3-O-4”’ linkage pattern. Thus, this study suggests that the structural features of these compounds could serve as a basis for developing new EGFR-TK inhibitors.Keywords: anticancer, biflavonoids, EGFR, molecular docking, Selaginellaceae
Procedia PDF Downloads 192939 Oncolytic Efficacy of Thymidine Kinase-Deleted Vaccinia Virus Strain Tiantan (oncoVV-TT) in Glioma
Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi
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Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus
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