Search results for: protein structure classification
11469 Alterations of Malondialdehyde and Heat Shock Protein-27 in Sheep with Naturally Infected Liver Cystic Echinococcosis
Authors: K. Azimzadeh, S. Rasouli
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The present study investigates whether malondialdehyde (MDA) and heat shock protein-27 (HSP-27) are altered in sheep with cystic echinococcosis (CE). For this purpose, forty parasitized and thirty healthy sheep were selected based on severe cystic form observation in liver and lack of blood parasite along with no cystic conformation in carcass respectively. The results revealed a significant decrease (p<0.01) in albumin (Alb) and total plasma protein (TPP) and a significant increase (p<0.01) in HSP-27, MDA, total bilirubin and unconjugated bilirubin in the infected group compared with healthy ones.The results indicate low levels of TPP and Alb reveal liver damage in suffered sheep and MDA elevation demonstrates oxidative stress in infected group. In addition, HSP-27 enhancement may attribute to disease-induced stress conditions.Keywords: malondialdehyde, heat shock protein-27, Echinococcosis, blood parasites
Procedia PDF Downloads 60811468 Multi-Classification Deep Learning Model for Diagnosing Different Chest Diseases
Authors: Bandhan Dey, Muhsina Bintoon Yiasha, Gulam Sulaman Choudhury
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Chest disease is one of the most problematic ailments in our regular life. There are many known chest diseases out there. Diagnosing them correctly plays a vital role in the process of treatment. There are many methods available explicitly developed for different chest diseases. But the most common approach for diagnosing these diseases is through X-ray. In this paper, we proposed a multi-classification deep learning model for diagnosing COVID-19, lung cancer, pneumonia, tuberculosis, and atelectasis from chest X-rays. In the present work, we used the transfer learning method for better accuracy and fast training phase. The performance of three architectures is considered: InceptionV3, VGG-16, and VGG-19. We evaluated these deep learning architectures using public digital chest x-ray datasets with six classes (i.e., COVID-19, lung cancer, pneumonia, tuberculosis, atelectasis, and normal). The experiments are conducted on six-classification, and we found that VGG16 outperforms other proposed models with an accuracy of 95%.Keywords: deep learning, image classification, X-ray images, Tensorflow, Keras, chest diseases, convolutional neural networks, multi-classification
Procedia PDF Downloads 9211467 In vitro and invivo Antioxidant Studies of Grewia crenata Leaves Extract in Albino Rats
Authors: A. N.Ukwuani, A. K. Abdulfatah
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G. crenata is used locally for the treatment of fractured bones, wound healing and inflammatory conditions. In vitro and in vivo antioxidant activity of hydromethanolic extracts of the leaves of G. crenata were assessed. The phytochemical analysis shows the presence of phenols, flavonoids, saponins, cardiac glycosides and tannins. An in vitro quantitative analysis of phenols, flavonoids and tannins respectively were (164±1.20, 199±0.88 and 88.67±0.88 mg/100g FW). In vivo studies of hydromethanolic extract demonstrated a dose dependent increase in hepatic superoxide dismutase (1.14±0.14, 2.13±0.11, 2.55±0.11 U/mg Protein) with improvement in hepatic glutathione (6.98±0.42, 8.91±0.37, 11.07±0.46 µM/mg Protein) and Catalase (4.47±0.05, 6.24±0.02, 7.17±0.04 U/mg Protein) and Total protein (6.18±0.08, 6.69±0.18, 7.27±0.16 mg/ml) respectively at 100-300mg/kg body weight Grewia crenata leaves when compared to the control and standard drug. It can be concluded from the present findings of that G. crenata leaves possess antioxidant potential.Keywords: Grewia crenata, antioxidant, hydromethanolic extract, in vivo, in vitro
Procedia PDF Downloads 55311466 Functional Cell Surface Display Using Ice Nucleation Protein from Erwina ananas on Escherischia coli
Authors: Mei Yuin Joanne Wee, Rosli Md. Illias
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Cell surface display is the expression of a protein with an anchoring motif on the surface of the cell. This approach offers advantages when used in bioconversion in terms of easier purification steps and more efficient enzymatic reaction. A surface display system using ice nucleation protein (InaA) from Erwina ananas as an anchoring motif has been constructed to display xylanase (xyl) on the surface of Escherischia coli. The InaA was truncated so that it is made up of the N- and C-terminal domain (INPANC-xyl) and it has successfully directed xylanase to the surface of the cell. A study was also done on xylanase fused to two other ice nucleation proteins, InaK (INPKNC-xyl) and InaZ (INPZNC-xyl) from Pseudomonas syringae KCTC 1832 and Pseudomonas syringae S203 respectively. Surface localization of the fusion protein was verified using SDS-PAGE and Western blot on the cell fractions and all anchoring motifs were successfully displayed on the outer membrane of E. coli. Upon comparison, whole-cell activity of INPANC-xyl was more than six and five times higher than INPKNC-xyl and INPZNC-xyl respectively. Furthermore, the expression of INPANC-xyl on the surface of E. coli did not inhibit the growth of the cell. This is the first report of surface display system using ice nucleation protein, InaA from E. ananas. From this study, this anchoring motif offers an attractive alternative to the current surface display systems.Keywords: cell surface display, Escherischia coli, ice nucleation protein, xylanase
Procedia PDF Downloads 39011465 Fatigue Life Estimation of Tubular Joints - A Comparative Study
Authors: Jeron Maheswaran, Sudath C. Siriwardane
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In fatigue analysis, the structural detail of tubular joint has taken great attention among engineers. The DNV-RP-C203 is covering this topic quite well for simple and clear joint cases. For complex joint and geometry, where joint classification isn’t available and limitation on validity range of non-dimensional geometric parameters, the challenges become a fact among engineers. The classification of joint is important to carry out through the fatigue analysis. These joint configurations are identified by the connectivity and the load distribution of tubular joints. To overcome these problems to some extent, this paper compare the fatigue life of tubular joints in offshore jacket according to the stress concentration factors (SCF) in DNV-RP-C203 and finite element method employed Abaqus/CAE. The paper presents the geometric details, material properties and considered load history of the jacket structure. Describe the global structural analysis and identification of critical tubular joints for fatigue life estimation. Hence fatigue life is determined based on the guidelines provided by design codes. Fatigue analysis of tubular joints is conducted using finite element employed Abaqus/CAE [4] as next major step. Finally, obtained SCFs and fatigue lives are compared and their significances are discussed.Keywords: fatigue life, stress-concentration factor, finite element analysis, offshore jacket structure
Procedia PDF Downloads 45311464 Extraction of Rice Bran Protein Using Enzymes and Polysaccharide Precipitation
Authors: Sudarat Jiamyangyuen, Tipawan Thongsook, Riantong Singanusong, Chanida Saengtubtim
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Rice is a staple food as well as exported commodity of Thailand. Rice bran, a 10.5% constituent of rice grain, is a by-product of rice milling process. Rice bran is normally used as a raw material for rice bran oil production or sold as feed with a low price. Therefore, this study aimed to increase value of defatted rice bran as obtained after extracting of rice bran oil. Conventionally, the protein in defatted rice bran was extracted using alkaline extraction and acid precipitation, which results in reduction of nutritious components in rice bran. Rice bran protein concentrate is suitable for those who are allergenic of protein from other sources eg. milk, wheat. In addition to its hypoallergenic property, rice bran protein also contains good quantity of lysine. Thus it may act as a suitable ingredient for infant food formulations while adding variety to the restricted diets of children with food allergies. The objectives of this study were to compare properties of rice bran protein concentrate (RBPC) extracted from defatted rice bran using enzymes together with precipitation step using polysaccharides (alginate and carrageenan) to those of a control sample extracted using a conventional method. The results showed that extraction of protein from rice bran using enzymes exhibited the higher protein recovery compared to that extraction with alkaline. The extraction conditions using alcalase 2% (v/w) at 50 C, pH 9.5 gave the highest protein (2.44%) and yield (32.09%) in extracted solution compared to other enzymes. Rice bran protein concentrate powder prepared by a precipitation step using alginate (protein in solution: alginate 1:0.006) exhibited the highest protein (27.55%) and yield (6.62%). Precipitation using alginate was better than that of acid. RBPC extracted with alkaline (ALK) or enzyme alcalase (ALC), then precipitated with alginate (AL) (samples RBP-ALK-AL and RBP-ALC-AL) yielded the precipitation rate of 75% and 91.30%, respectively. Therefore, protein precipitation using alginate was then selected. Amino acid profile of control sample, and sample precipitated with alginate, as compared to casein and soy protein isolated, showed that control sample showed the highest content among all sample. Functional property study of RBP showed that the highest nitrogen solubility occurred in pH 8-10. There was no statically significant between emulsion capacity and emulsion stability of control and sample precipitated by alginate. However, control sample showed a higher of foaming and lower foam stability compared to those of sample precipitated with alginate. The finding was successful in terms of minimizing chemicals used in extraction and precipitation steps in preparation of rice bran protein concentrate. This research involves in a production of value-added product in which the double amount of protein (28%) compared to original amount (14%) contained in rice bran could be beneficial in terms of adding to food products eg. healthy drink with high protein and fiber. In addition, the basic knowledge of functional property of rice bran protein concentrate was obtained, which can be used to appropriately select the application of this value-added product from rice bran.Keywords: alginate, carrageenan, rice bran, rice bran protein
Procedia PDF Downloads 29511463 Experimental Study of Hyperparameter Tuning a Deep Learning Convolutional Recurrent Network for Text Classification
Authors: Bharatendra Rai
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The sequence of words in text data has long-term dependencies and is known to suffer from vanishing gradient problems when developing deep learning models. Although recurrent networks such as long short-term memory networks help to overcome this problem, achieving high text classification performance is a challenging problem. Convolutional recurrent networks that combine the advantages of long short-term memory networks and convolutional neural networks can be useful for text classification performance improvements. However, arriving at suitable hyperparameter values for convolutional recurrent networks is still a challenging task where fitting a model requires significant computing resources. This paper illustrates the advantages of using convolutional recurrent networks for text classification with the help of statistically planned computer experiments for hyperparameter tuning.Keywords: long short-term memory networks, convolutional recurrent networks, text classification, hyperparameter tuning, Tukey honest significant differences
Procedia PDF Downloads 12911462 Effects of Plasma Technology in Biodegradable Films for Food Packaging
Authors: Viviane P. Romani, Bradley D. Olsen, Vilásia G. Martins
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Biodegradable films for food packaging have gained growing attention due to environmental pollution caused by synthetic films and the interest in the better use of resources from nature. Important research advances were made in the development of materials from proteins, polysaccharides, and lipids. However, the commercial use of these new generation of sustainable materials for food packaging is still limited due to their low mechanical and barrier properties that could compromise the food quality and safety. Thus, strategies to improve the performance of these materials have been tested, such as chemical modifications, incorporation of reinforcing structures and others. Cold plasma is a versatile, fast and environmentally friendly technology. It consists of a partially ionized gas containing free electrons, ions, and radicals and neutral particles able to react with polymers and start different reactions, leading to the polymer degradation, functionalization, etching and/or cross-linking. In the present study, biodegradable films from fish protein prepared through the casting technique were plasma treated using an AC glow discharge equipment. The reactor was preliminary evacuated to ~7 Pa and the films were exposed to air plasma for 2, 5 and 8 min. The films were evaluated by their mechanical and water vapor permeability (WVP) properties and changes in the protein structure were observed using Scanning Electron Microscopy (SEM) and X-ray diffraction (XRD). Potential cross-links and elimination of surface defects by etching might be the reason for the increase in tensile strength and decrease in the elongation at break observed. Among the times of plasma application tested, no differences were observed when higher times of exposure were used. The X-ray pattern showed a broad peak at 2θ = 19.51º that corresponds to the distance of 4.6Å by applying the Bragg’s law. This distance corresponds to the average backbone distance within the α-helix. Thus, the changes observed in the films might indicate that the helical configuration of fish protein was disturbed by plasma treatment. SEM images showed surface damage in the films with 5 and 8 min of plasma treatment, indicating that 2 min was the most adequate time of treatment. It was verified that plasma removes water from the films once weight loss of 4.45% was registered for films treated during 2 min. However, after 24 h in 50% of relative humidity, the water lost was recovered. WVP increased from 0.53 to 0.65 g.mm/h.m².kPa after plasma treatment during 2 min, that is desired for some foods applications which require water passage through the packaging. In general, the plasma technology affects the properties and structure of fish protein films. Since this technology changes the surface of polymers, these films might be used to develop multilayer materials, as well as to incorporate active substances in the surface to obtain active packaging.Keywords: fish protein films, food packaging, improvement of properties, plasma treatment
Procedia PDF Downloads 16311461 Encapsulation and Protection of Bioactive Nutrients Based on Ligand-Binding Property of Milk Proteins
Authors: Hao Cheng, Yingzhou Ni, Amr M. Bakry, Li Liang
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Functional foods containing bioactive nutrients offer benefits beyond basic nutrition and hence the possibility of delaying and preventing chronic diseases. However, many bioactive nutrients degrade rapidly under food processing and storage conditions. Encapsulation can be used to overcome these limitations. Food proteins have been widely used as carrier materials for the preparation of nano/micro-particles because of their ability to form gels and emulsions and to interact with polysaccharides. The mechanisms of interaction between bioactive nutrients and proteins must be understood in order to develop protein-based lipid-free delivery systems. Beta-lactoglobulin, a small globular protein in milk whey, exhibits an affinity to a wide range of compounds. Alfa-tocopherol, resveratrol and folic acid were respectively bound to the central cavity, the outer surface near Trp19–Arg124 and the hydrophobic pocket in the groove between the alfa-helix and the beta-barrel of the protein. Beta-lactoglobulin could thus bind the three bioactive nutrients simultaneously to form protein-multi-ligand complexes. Beta-casein, an intrinsically unstructured but major milk protein, could also interact with resveratrol and folic acid to form complexes. These results suggest the potential to develop milk-protein-based complex carrier systems for encapsulation of multiple bioactive nutrients for functional food application and also pharmaceutical and medical uses.Keywords: milk protein, bioactive nutrient, interaction, protection
Procedia PDF Downloads 41211460 Quantitative Structure–Activity Relationship Analysis of Some Benzimidazole Derivatives by Linear Multivariate Method
Authors: Strahinja Z. Kovačević, Lidija R. Jevrić, Sanja O. Podunavac Kuzmanović
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The relationship between antibacterial activity of eighteen different substituted benzimidazole derivatives and their molecular characteristics was studied using chemometric QSAR (Quantitative Structure–Activity Relationships) approach. QSAR analysis has been carried out on inhibitory activity towards Staphylococcus aureus, by using molecular descriptors, as well as minimal inhibitory activity (MIC). Molecular descriptors were calculated from the optimized structures. Principal component analysis (PCA) followed by hierarchical cluster analysis (HCA) and multiple linear regression (MLR) was performed in order to select molecular descriptors that best describe the antibacterial behavior of the compounds investigated, and to determine the similarities between molecules. The HCA grouped the molecules in separated clusters which have the similar inhibitory activity. PCA showed very similar classification of molecules as the HCA, and displayed which descriptors contribute to that classification. MLR equations, that represent MIC as a function of the in silico molecular descriptors were established. The statistical significance of the estimated models was confirmed by standard statistical measures and cross-validation parameters (SD = 0.0816, F = 46.27, R = 0.9791, R2CV = 0.8266, R2adj = 0.9379, PRESS = 0.1116). These parameters indicate the possibility of application of the established chemometric models in prediction of the antibacterial behaviour of studied derivatives and structurally very similar compounds.Keywords: antibacterial, benzimidazole, molecular descriptors, QSAR
Procedia PDF Downloads 36411459 An Automatic Generating Unified Modelling Language Use Case Diagram and Test Cases Based on Classification Tree Method
Authors: Wassana Naiyapo, Atichat Sangtong
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The processes in software development by Object Oriented methodology have many stages those take time and high cost. The inconceivable error in system analysis process will affect to the design and the implementation process. The unexpected output causes the reason why we need to revise the previous process. The more rollback of each process takes more expense and delayed time. Therefore, the good test process from the early phase, the implemented software is efficient, reliable and also meet the user’s requirement. Unified Modelling Language (UML) is the tool which uses symbols to describe the work process in Object Oriented Analysis (OOA). This paper presents the approach for automatically generated UML use case diagram and test cases. UML use case diagram is generated from the event table and test cases are generated from use case specifications and Graphic User Interfaces (GUI). Test cases are derived from the Classification Tree Method (CTM) that classify data to a node present in the hierarchy structure. Moreover, this paper refers to the program that generates use case diagram and test cases. As the result, it can reduce work time and increase efficiency work.Keywords: classification tree method, test case, UML use case diagram, use case specification
Procedia PDF Downloads 16211458 Performance Evaluation of Contemporary Classifiers for Automatic Detection of Epileptic EEG
Authors: K. E. Ch. Vidyasagar, M. Moghavvemi, T. S. S. T. Prabhat
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Epilepsy is a global problem, and with seizures eluding even the smartest of diagnoses a requirement for automatic detection of the same using electroencephalogram (EEG) would have a huge impact in diagnosis of the disorder. Among a multitude of methods for automatic epilepsy detection, one should find the best method out, based on accuracy, for classification. This paper reasons out, and rationalizes, the best methods for classification. Accuracy is based on the classifier, and thus this paper discusses classifiers like quadratic discriminant analysis (QDA), classification and regression tree (CART), support vector machine (SVM), naive Bayes classifier (NBC), linear discriminant analysis (LDA), K-nearest neighbor (KNN) and artificial neural networks (ANN). Results show that ANN is the most accurate of all the above stated classifiers with 97.7% accuracy, 97.25% specificity and 98.28% sensitivity in its merit. This is followed closely by SVM with 1% variation in result. These results would certainly help researchers choose the best classifier for detection of epilepsy.Keywords: classification, seizure, KNN, SVM, LDA, ANN, epilepsy
Procedia PDF Downloads 52011457 Application of Computational Chemistry for Searching Anticancer Derivatives of 2-Phenazinamines as Bcr-Abl Tyrosine Kinase Inhibitors
Authors: Gajanan M. Sonwane
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The computational studies on 2-phenazinamines with their protein targets have been carried out to design compounds with potential anticancer activity. This strategy of designing compounds possessing selectivity over specific tyrosine kinase has been achieved through G-QSAR and molecular docking studies. The objective of this research has been to design newer 2-phenazinamine derivatives as Bcr-Abl tyrosine kinase inhibitors by G-QSAR, molecular docking studies followed by wet-lab studies along with evaluation of their anticancer potential. Computational chemistry was done by using VLife MDS 4.3 and Autodock 4.2 followed by wet-lab experiments for synthesizing 2-phenazinamine derivatives. The chemical structures of ligands in 2D were drawn by employing Chemdraw 2D Ultra 8.0 and were converted into 3D. These were optimized by using a semi-empirical method called MOPAC. The protein structure was retrieved from RCSC protein data bank as a PDB file. The binding interactions of protein and ligands were done by using PYMOL. The molecular properties of the designed compounds were predicted in silico by using Osiris property explorer. The parent compound 2-phenazinamine was synthesized by reduction of 2, 4-dinitro-N-phenyl-benzenamine in the presence of tin chloride followed by cyclization in the presence of nitrobenzene and magnesium sulfate. The derivatization at the amino function of 2-phenazinamine was performed by treating parent compound with various aldehydes in the presence of dicyclohexylcarbodiimide (DCC) and urea to afford 2-(2-chlorophenyl)-3-(phenazine-2-yl) thiazolidine-4-one. Synthesized 39 novel derivatives of 2-phenazinamine and performed antioxidant activity, anti antiproliferative on the bulb of onion and anticancer activity on cell line showing significant competition with marked blockbuster drug imatinib.Keywords: computer-aided drug design, tyrosin kinases, anticancer, docking
Procedia PDF Downloads 14011456 3D Receiver Operator Characteristic Histogram
Authors: Xiaoli Zhang, Xiongfei Li, Yuncong Feng
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ROC curves, as a widely used evaluating tool in machine learning field, are the tradeoff of true positive rate and negative rate. However, they are blamed for ignoring some vital information in the evaluation process, such as the amount of information about the target that each instance carries, predicted score given by each classification model to each instance. Hence, in this paper, a new classification performance method is proposed by extending the Receiver Operator Characteristic (ROC) curves to 3D space, which is denoted as 3D ROC Histogram. In the histogram, theKeywords: classification, performance evaluation, receiver operating characteristic histogram, hardness prediction
Procedia PDF Downloads 31311455 Protein Quality of Game Meat Hunted in Latvia
Authors: Vita Strazdina, Aleksandrs Jemeljanovs, Vita Sterna
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Not all proteins have the same nutritional value, since protein quality strongly depends on its amino acid composition and digestibility. The meat of game animals could be a high protein source because of its well-balanced essential amino acids composition. Investigations about biochemical composition of game meat such as wild boar (Sus scrofa scrofa), roe deer (Capreolus capreolus) and beaver (Castor fiber) are not very much. Therefore, the aim of the investigation was evaluate protein composition of game meat hunted in Latvia. The biochemical analysis, evaluation of connective tissue and essential amino acids in meat samples were done, the amino acids score were calculate. Results of analysis showed that protein content 20.88-22.05% of all types of meat samples is not different statistically. The content of connective tissue from 1.3% in roe deer till 1.5% in beaver meat allowed classified game animal as high quality meat. The sum of essential amino acids in game meat samples were determined 7.05–8.26g100g-1. Roe deer meat has highest protein content and lowest content of connective tissues among game meat hunted in Latvia. Concluded that amino acid score for limiting amino acids phenylalanine and tyrosine is high and shows high biological value of game meat.Keywords: dietic product, game meat, amino acids, scores
Procedia PDF Downloads 32111454 Impact of Dietary L-Threonine Supplementation on Performance and Health of Broiler Chickens, a Review
Authors: Mandana Hoseini
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During last decades, intensive selection for higher growth rate in broiler chickens has accelerated daily body weight gain, which this has changed/increased the trends and amounts of nutrient requirements in the diet. As a result, considerable studies have been focused on the better determination of protein/amino acids requirements in modern broiler diets. One approach to minimize dietary crude protein inclusion levels is substitution of some of the dietary crude protein with synthetic amino acids. In addition, using synthetic forms of limiting essential amino acids in the diet could help better coincidence of dietary protein with ideal protein concept, which this in turn, minimizes nitrogen dissipation and environmental pollution. Threonine is usually considered as the third limiting amino acid in broiler diets. Recent studies have been demonstrated that dietary supplemental threonine would optimize growth performance, immune system, intestinal morphology, as well as oxidative defense in broiler chickens. In this review, threonine metabolism and its effects in relation with different aspects of broiler performance have been discussed.Keywords: immune system, intestine, performance, requirement, threonine
Procedia PDF Downloads 10811453 Combined Odd Pair Autoregressive Coefficients for Epileptic EEG Signals Classification by Radial Basis Function Neural Network
Authors: Boukari Nassim
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This paper describes the use of odd pair autoregressive coefficients (Yule _Walker and Burg) for the feature extraction of electroencephalogram (EEG) signals. In the classification: the radial basis function neural network neural network (RBFNN) is employed. The RBFNN is described by his architecture and his characteristics: as the RBF is defined by the spread which is modified for improving the results of the classification. Five types of EEG signals are defined for this work: Set A, Set B for normal signals, Set C, Set D for interictal signals, set E for ictal signal (we can found that in Bonn university). In outputs, two classes are given (AC, AD, AE, BC, BD, BE, CE, DE), the best accuracy is calculated at 99% for the combined odd pair autoregressive coefficients. Our method is very effective for the diagnosis of epileptic EEG signals.Keywords: epilepsy, EEG signals classification, combined odd pair autoregressive coefficients, radial basis function neural network
Procedia PDF Downloads 34511452 Use of Segmentation and Color Adjustment for Skin Tone Classification in Dermatological Images
Authors: Fernando Duarte
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The work aims to evaluate the use of classical image processing methodologies towards skin tone classification in dermatological images. The skin tone is an important attribute when considering several factor for skin cancer diagnosis. Currently, there is a lack of clear methodologies to classify the skin tone based only on the dermatological image. In this work, a recent released dataset with the label for skin tone was used as reference for the evaluation of classical methodologies for segmentation and adjustment of color space for classification of skin tone in dermatological images. It was noticed that even though the classical methodologies can work fine for segmentation and color adjustment, classifying the skin tone without proper control of the aquisition of the sample images ended being very unreliable.Keywords: segmentation, classification, color space, skin tone, Fitzpatrick
Procedia PDF Downloads 3511451 Expression of Human Papillomavirus Type 18 L1 Virus-Like Particles in Methylotropic Yeast, Pichia Pastoris
Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko
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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology
Procedia PDF Downloads 40711450 Interaction between Kazal-Type Serine Proteinase Inhibitor SPIPm2 and Cyclophilin A from the Black Tiger Shrimp Penaeus monodon
Authors: Sirikwan Ponprateep, Anchalee Tassanakajon, Vichien Rimphanitchayakit
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A Kazal-type serine proteinase inhibitor, SPIPm2, was abundantly expressed in the hemocytes and secreted into shrimp plasma has anti-viral property against white spot syndrome virus (WSSV). To discover the molecular mechanism of antiviral activity, the binding assay showed that SPIPm2 bind to the components of viral particle and shrimp hemocyte. From our previous report, viral target protein of SPIPm2 was identified, namely WSV477 using yeast two-hybrid screening. WSV477 is an early gene product of WSSV and involved in viral propagation. In this study, the co-immunoprecipitation technique and Tandem Mass Spectrometry (LC-MS/MS) was used to identify the target protein of SPIPm2 from shrimp hemocyte. The target protein of SPIPm2 was cyclophilin A. In vertebrate, cyclophilin A or peptidylprolyl isomerase A was reported to be the immune suppressor interacted with cyclosporin A involved in immune defense response. The recombinant cyclophilin A from Penaeus monodon (rPmCypA) was produced in E.coli system and purified using Ni-NTA column to confirm the protein-protein interaction. In vitro pull-down assay showed the interaction between rSPIPm2 and rPmCypA. To study the biological function of these proteins, the expression analysis of immune gene in shrimp defense pathways will be investigated after rPmCypA administration.Keywords: cyclophilin A, protein-protein interaction, Kazal-type serine proteinase inhibitor, Penaeus monodon
Procedia PDF Downloads 23611449 The Effect of the Earthworm (Lumbricus rubellus) as the Source of Protein Feed and Pathogen Antibacterial for Broiler
Authors: Waode Nurmayani, Nikmatul Riswanda
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Broilers are chickens which are kept with the most efficient time and hoped get a good body weight. All things are done, for example with the improvement of feed and use antibiotics. Feed cost is the most cost to be spent. Nearly 80% of the cost is spent just for buy feed. Earthworm (Lumbricus rubellus) is a good choice to reduce the cost of feed protein source. The Earthworm has a high crude protein content of about 48.5%-61.9%, rich with proline amino acid about 15% of the 62 amino acids. Not only about protein, this earthworm also has a role in disease prevention. Prevention of disease in livestock usual with use feed supplement. Earthworm (Lumbricus rubellus) is one of the natural materials used as feed. In addition, several types of earthworms that have been known to contain active substances about antibacterial pathogens namely Lumbricus rubellus. The earthworm could be used as an antibiotic because it contain the antibody of Lumbricine active substance. So that, this animal feed from Lumbricus rubellus could improve the performance of broilers. Bioactive of anti-bacterial is called Lumbricine able to inhibit the growth of pathogenic bacteria in the intestinal wall so that the population of pathogenic bacteria is reduced. The method of write in this scientific writing is divided into 3 techniques, namely data completion, data analysis, and thinking pan from various literature about earthworm (Lumbricus rubellus) as broiler feed. It is expected that innovation of feed material of earthworm (Lumbricus rubellus) could reduce the cost of protein feed and the use of chemical antibiotics.Keywords: earthworm, broiler, protein, antibiotic
Procedia PDF Downloads 15711448 Proteomic Analysis of Excretory Secretory Antigen (ESA) from Entamoeba histolytica HM1: IMSS
Authors: N. Othman, J. Ujang, M. N. Ismail, R. Noordin, B. H. Lim
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Amoebiasis is caused by the Entamoeba histolytica and still endemic in many parts of the tropical region, worldwide. Currently, there is no available vaccine against amoebiasis. Hence, there is an urgent need to develop a vaccine. The excretory secretory antigen (ESA) of E. histolytica is a suitable biomarker for the vaccine candidate since it can modulate the host immune response. Hence, the objective of this study is to identify the proteome of the ESA towards finding suitable biomarker for the vaccine candidate. The non-gel based and gel-based proteomics analyses were performed to identify proteins. Two kinds of mass spectrometry with different ionization systems were utilized i.e. LC-MS/MS (ESI) and MALDI-TOF/TOF. Then, the functional proteins classification analysis was performed using PANTHER software. Combination of the LC -MS/MS for the non-gel based and MALDI-TOF/TOF for the gel-based approaches identified a total of 273 proteins from the ESA. Both systems identified 29 similar proteins whereby 239 and 5 more proteins were identified by LC-MS/MS and MALDI-TOF/TOF, respectively. Functional classification analysis showed the majority of proteins involved in the metabolic process (24%), primary metabolic process (19%) and protein metabolic process (10%). Thus, this study has revealed the proteome the E. histolytica ESA and the identified proteins merit further investigations as a vaccine candidate.Keywords: E. histolytica, ESA, proteomics, biomarker
Procedia PDF Downloads 34311447 Plasma-Induced Modification of Biomolecules: A Tool for Analysis of Protein Structures
Authors: Yuting Wu, Faraz Choudhury, Daniel Benjamin, James Whalin, Joshua Blatz, Leon Shohet, Michael Sussman, Mark Richards
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Plasma-Induced Modification of Biomolecules (PLIMB) has been developed as a technology, which, together with mass spectrometry, measures three-dimensional structural characteristics of proteins. This technique uses hydroxyl radicals generated by atmospheric-pressure plasma discharge to react with the solvent-accessible side chains of protein in an aqueous solution. In this work, we investigate the three-dimensional structure of hemoglobin and myoglobin using PLIMB. Additional modifications to these proteins, such as oxidation, fragmentations, and conformational changes caused by PLIMB are also explored. These results show that PLIMB, coupled with mass spectrometry, is an effective way to determine solvent access to hemoproteins. Furthermore, we show that many factors, including pH and the electrical parameters used to generate the plasma, have a significant influence on solvent accessibility.Keywords: plasma, hemoglobin, myoglobin, solvent access
Procedia PDF Downloads 19311446 Apparent Ileal and Excreta Digestibility of Protein Poultry By-Product Meal in 21 to 28 Days of Age Broiler Chicken
Authors: N. Mahmoudnia, M. Khormali
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This experiment was conducted to determine the apparent protein digestibility of poultry byproduct meal (PBPM) from two industrial poultry slaughter-houses on Ross 308 male broiler chickens in independent comparisons. The experiment consisted of seven dietary treatments and three replicates per treatment with three broiler chickens per replicate in a completely randomized design. Dietary treatments consisted of a control corn- soybean diet, and levels 3, 6, and 9% PBPM produced by slaughter-house 1 and levels 3, 6, and 9% PBPM produced by slaughter house 2. Chromic oxide was added to the experimental diets as an indigestible marker. The apparent protein digestibility of each diet were determined with two methods of sample collection of ileum and excreta in 21-28 d of age. The results this experiment showed that use of PBPM had no significant effect on the performance of broiler chicks during period of experiments. The apparent protein digestibility of PBPM groups was significantly higher than control group by excreta sampling procedure (P<0.05). Using of PBPM 2 significantly (P<0.05) decreased the apparent protein digestibility values based on ileum sampling procedure vs control (85.21 vs. 90.14).Based results of this experiment,it is possible to use of PBPM 1 in broiler chicken.Keywords: poultry by-product meal, apparent protein digestibility, independed comparison, broiler chicken
Procedia PDF Downloads 49211445 Evaluation of Coagulation Efficiency of Protein Extracts from Lupinus Albus L., Moringa Stenopetala Cufod., Trigonella Foenum-Graecum L. And Vicia Faba L. For Water Purification
Authors: Neway Adele, Adey Feleke
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Access to clean drinking water is a basic human right. However, an estimated 1.2 billion people across the world consume unclean water daily. Interest has been growing in natural coagulants as the health and environmental concerns of conventional chemical coagulants are rising. Natural coagulants have the potential to serve as alternative water treatment agents. In this study, Lupinus albus, Moringa stenopetala, Trigonella foenum-graecum and Vicia faba protein extracts were evaluated as natural coagulants for water treatment. The protein extracts were purified from crude extracts using a protein purifier, and protein concentrations were determined by the spectrophotometric method. Small-volume coagulation efficiency tests were conducted on raw water taken from the Legedadi water treatment plant. These were done using a completely randomized design (CRD) experiment with settling times of 0 min (initial time), 90 min, 180 min and 270 min and protein extract doses of 5 mg/L, 10 mg/L, 15 mg/L and 20 mg/L. Raw water as negative control and polyelectrolyte as positive control were also included. The optical density (OD) values were measured for all the samples. At 270 min and 20 mg/L, the coagulation efficiency percentages for Lupinus albus, Moringa stenopetala, Trigonella foenum-graecum and Vicia faba protein extracts were 71%, 89%, 12% and 67% in the water sample collected in April 2019 respectively. Similarly, Lupinus albus, Moringa stenopetala and Vicia faba achieved 17%, 92% and 12% at 270 min settling times and 5 mg/L, 20 mg/L and 10 mg/L concentration in the water sample collected from August 2019, respectively. Negative control (raw water) and polyelectrolyte (positive control) were also 6 − 10% and 89 − 94% at 270 min settling time in April and August 2019, respectively. Among the four protein extracts, Moringa stenopetala showed the highest coagulation efficiency, similar to polyelectrolyte. This study concluded that Moringa stenopetala protein extract could be used as a natural coagulant for water purification in both sampling times.Keywords: coagulation efficiency, extraction, natural coagulant, protein extract
Procedia PDF Downloads 6811444 New Kinetic Approach to the Enzymatic Hydrolysis of Proteins: A Case of Thermolysin-Catalyzed Albumin
Authors: Anna Trusek-Holownia, Andrzej Noworyta
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Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.Keywords: peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides
Procedia PDF Downloads 43711443 Automatic Classification Using Dynamic Fuzzy C Means Algorithm and Mathematical Morphology: Application in 3D MRI Image
Authors: Abdelkhalek Bakkari
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Image segmentation is a critical step in image processing and pattern recognition. In this paper, we proposed a new robust automatic image classification based on a dynamic fuzzy c-means algorithm and mathematical morphology. The proposed segmentation algorithm (DFCM_MM) has been applied to MR perfusion images. The obtained results show the validity and robustness of the proposed approach.Keywords: segmentation, classification, dynamic, fuzzy c-means, MR image
Procedia PDF Downloads 47811442 C-eXpress: A Web-Based Analysis Platform for Comparative Functional Genomics and Proteomics in Human Cancer Cell Line, NCI-60 as an Example
Authors: Chi-Ching Lee, Po-Jung Huang, Kuo-Yang Huang, Petrus Tang
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Background: Recent advances in high-throughput research technologies such as new-generation sequencing and multi-dimensional liquid chromatography makes it possible to dissect the complete transcriptome and proteome in a single run for the first time. However, it is almost impossible for many laboratories to handle and analysis these “BIG” data without the support from a bioinformatics team. We aimed to provide a web-based analysis platform for users with only limited knowledge on bio-computing to study the functional genomics and proteomics. Method: We use NCI-60 as an example dataset to demonstrate the power of the web-based analysis platform and data delivering system: C-eXpress takes a simple text file that contain the standard NCBI gene or protein ID and expression levels (rpkm or fold) as input file to generate a distribution map of gene/protein expression levels in a heatmap diagram organized by color gradients. The diagram is hyper-linked to a dynamic html table that allows the users to filter the datasets based on various gene features. A dynamic summary chart is generated automatically after each filtering process. Results: We implemented an integrated database that contain pre-defined annotations such as gene/protein properties (ID, name, length, MW, pI); pathways based on KEGG and GO biological process; subcellular localization based on GO cellular component; functional classification based on GO molecular function, kinase, peptidase and transporter. Multiple ways of sorting of column and rows is also provided for comparative analysis and visualization of multiple samples.Keywords: cancer, visualization, database, functional annotation
Procedia PDF Downloads 61811441 Classification of Construction Projects
Authors: M. Safa, A. Sabet, S. MacGillivray, M. Davidson, K. Kaczmarczyk, C. T. Haas, G. E. Gibson, D. Rayside
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To address construction project requirements and specifications, scholars and practitioners need to establish a taxonomy according to a scheme that best fits their need. While existing characterization methods are continuously being improved, new ones are devised to cover project properties which have not been previously addressed. One such method, the Project Definition Rating Index (PDRI), has received limited consideration strictly as a classification scheme. Developed by the Construction Industry Institute (CII) in 1996, the PDRI has been refined over the last two decades as a method for evaluating a project's scope definition completeness during front-end planning (FEP). The main contribution of this study is a review of practical project classification methods, and a discussion of how PDRI can be used to classify projects based on their readiness in the FEP phase. The proposed model has been applied to 59 construction projects in Ontario, and the results are discussed.Keywords: project classification, project definition rating index (PDRI), risk, project goals alignment
Procedia PDF Downloads 67711440 Determination of Biomolecular Interactions Using Microscale Thermophoresis
Authors: Lynn Lehmann, Dinorah Leyva, Ana Lazic, Stefan Duhr, Philipp Baaske
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Characterization of biomolecular interactions, such as protein-protein, protein-nucleic acid or protein-small molecule, provides critical insights into cellular processes and is essential for the development of drug diagnostics and therapeutics. Here we present a novel, label-free, and tether-free technology to analyze picomolar to millimolar affinities of biomolecular interactions by Microscale Thermophoresis (MST). The entropy of the hydration shell surrounding molecules determines thermophoretic movement. MST exploits this principle by measuring interactions using optically generated temperature gradients. MST detects changes in the size, charge and hydration shell of molecules and measures biomolecule interactions under close-to-native conditions: immobilization-free and in bioliquids of choice, including cell lysates and blood serum. Thus, MST measures interactions under close-to-native conditions, and without laborious sample purification. We demonstrate how MST determines the picomolar affinities of antibody::antigen interactions, and protein::protein interactions measured from directly from cell lysates. MST assays are highly adaptable to fit to the diverse requirements of different and complex biomolecules. NanoTemper´s unique technology is ideal for studies requiring flexibility and sensitivity at the experimental scale, making MST suitable for basic research investigations and pharmaceutical applications.Keywords: biochemistry, biophysics, molecular interactions, quantitative techniques
Procedia PDF Downloads 523