Search results for: genotypic identification
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3002

Search results for: genotypic identification

2312 Comparing the Apparent Error Rate of Gender Specifying from Human Skeletal Remains by Using Classification and Cluster Methods

Authors: Jularat Chumnaul

Abstract:

In forensic science, corpses from various homicides are different; there are both complete and incomplete, depending on causes of death or forms of homicide. For example, some corpses are cut into pieces, some are camouflaged by dumping into the river, some are buried, some are burned to destroy the evidence, and others. If the corpses are incomplete, it can lead to the difficulty of personally identifying because some tissues and bones are destroyed. To specify gender of the corpses from skeletal remains, the most precise method is DNA identification. However, this method is costly and takes longer so that other identification techniques are used instead. The first technique that is widely used is considering the features of bones. In general, an evidence from the corpses such as some pieces of bones, especially the skull and pelvis can be used to identify their gender. To use this technique, forensic scientists are required observation skills in order to classify the difference between male and female bones. Although this technique is uncomplicated, saving time and cost, and the forensic scientists can fairly accurately determine gender by using this technique (apparently an accuracy rate of 90% or more), the crucial disadvantage is there are only some positions of skeleton that can be used to specify gender such as supraorbital ridge, nuchal crest, temporal lobe, mandible, and chin. Therefore, the skeletal remains that will be used have to be complete. The other technique that is widely used for gender specifying in forensic science and archeology is skeletal measurements. The advantage of this method is it can be used in several positions in one piece of bones, and it can be used even if the bones are not complete. In this study, the classification and cluster analysis are applied to this technique, including the Kth Nearest Neighbor Classification, Classification Tree, Ward Linkage Cluster, K-mean Cluster, and Two Step Cluster. The data contains 507 particular individuals and 9 skeletal measurements (diameter measurements), and the performance of five methods are investigated by considering the apparent error rate (APER). The results from this study indicate that the Two Step Cluster and Kth Nearest Neighbor method seem to be suitable to specify gender from human skeletal remains because both yield small apparent error rate of 0.20% and 4.14%, respectively. On the other hand, the Classification Tree, Ward Linkage Cluster, and K-mean Cluster method are not appropriate since they yield large apparent error rate of 10.65%, 10.65%, and 16.37%, respectively. However, there are other ways to evaluate the performance of classification such as an estimate of the error rate using the holdout procedure or misclassification costs, and the difference methods can make the different conclusions.

Keywords: skeletal measurements, classification, cluster, apparent error rate

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2311 Detect Circles in Image: Using Statistical Image Analysis

Authors: Fathi M. O. Hamed, Salma F. Elkofhaifee

Abstract:

The aim of this work is to detect geometrical shape objects in an image. In this paper, the object is considered to be as a circle shape. The identification requires find three characteristics, which are number, size, and location of the object. To achieve the goal of this work, this paper presents an algorithm that combines from some of statistical approaches and image analysis techniques. This algorithm has been implemented to arrive at the major objectives in this paper. The algorithm has been evaluated by using simulated data, and yields good results, and then it has been applied to real data.

Keywords: image processing, median filter, projection, scale-space, segmentation, threshold

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2310 Identification and Antibiotic Resistance Rates of Acinetobacter baumannii Strains Isolated from the Respiratory Tract Samples, Obtained from the Different Intensive Care Units

Authors: Recep Kesli, Gulşah Asik, Cengiz Demir, Onur Turkyilmaz

Abstract:

Objective: Acinetobacter baumannii (A. baumannii) can cause health-care associated infections, such as bacteremia, urinary tract and wound infections, endocarditis, meningitis, and pneumonia, particularly in intensive care unit patients. In this study, we aimed to evaluate A. baumannii production in sputum and bronchoalveolar lavage and susceptibilities for antibiotics in a 24 months period. Methods: Between October 2013 and September 2015, Acinetobacter baumannii isolated from respiratory tract speciments were evaluated retrospectively. The strains were isolated from the different intensive care units patients. A. baumannii strains were identified by both the conventional methods and aoutomated identification system -VITEK 2 (bio-Merieux, Marcy l’etoile, France). Antibiotic resistance testing was performed by Kirby-Bauer disc diffusion method according to CLSI criteria. Results: All the ninety isolates included in the study were from respiratory tract specimens. While of all the isolated 90 Acinetobacter baumannii strains were found to be resistant (100%), against ceftriaxone, ceftazidime, ciprofloxacin and piperacillin/ tazobactam, resistance rates against other tested antibiotics found as follows; meropenem 77, 86%, imipenem 75, 83%, trimethoprim-sulfamethoxazole (TMP-STX) 69, 76,6%, gentamicin 51, 56,6% and amikacin 48, 53,3%. Colistin was found as the most effective antibiotic against Acinetobacter baumannii, and there were not found any resistant (0%) strain against colistin. Conclusion: This study demonstrated that the no resistance was found in Acinetobacter baumannii against to colistin. High rates of resistance to carbapenems (imipenem and meropenem) and other tested antibiotics (ceftiaxone, ceftazidime, ciprofloxacine, piperacilline-tazobactam, TMP-STX gentamicin and amikacin) also have remarkable resistance rates. There was a significant relationship between demographic features of patients such as age, undergoing mechanical ventilation, length of hospital stay with resistance rates. High resistance rates against antibiotics require implementation of the infection control program and rational use of antibiotics. In the present study, while there were not found colistin resistance, panresistance were found against to ceftriaxone, ceftazidime, ciprofloxacin and piperacillin/ tazobactam.

Keywords: acinetobacter baumannii, antibiotic resistance, multi drug resistance, intensive care unit

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2309 Nontuberculous Mycobacterium Infection – Still An Important Disease Among People With Late HIV Diagnosis

Authors: Jakub Młoźniak, Adam Szymański, Gabriela Stondzik, Dagny Krankowska, Tomasz Mikuła

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Nontuberculous mycobacteria (NTM) are bacterial species that cause diversely manifesting diseases mainly in immunocompromised patients. In people with HIV, NTM infection is an AIDS-defining disease and usually appears when the lymphocyte T CD4 count is below 50 cells/μl. The usage of antiretroviral therapy has decreased the prevalence of NTM among people with HIV, but the disease can still be observed especially among patients with late HIV diagnosis. Common presence in environment, human colonization, clinical similarity with tuberculosis and slow growth on culture makes NTM especially hard to diagnose. The study aimed to analyze the epidemiology and clinical course of NTM among patients with HIV. This study included patients with NTM and HIV admitted to our department between 2017 and 2023. Medical records of patients were analyzed and data on age, sex, median time from HIV diagnosis to identification of NTM infection, median CD4 count at NTM diagnosis, methods of determining NTM infection, type of species of mycobacteria identified, clinical symptoms and treatment course were gathered. Twenty-four patients (20 men, 4 women) with identified NTM were included in this study. Among them, 20 were HIV late presenters. The patients' median age was 40. The main symptoms which patients presented were fever, weight loss and cough. Pulmonary disease confirmed with positive cultures from sputum/bronchoalveolar lavage was present in 18 patients. M. avium was the most common species identified. M. marinum caused disseminated skin lesions in 1 patient. Out of all, 5 people were not treated for NTM caused by lack of symptoms and suspicion of colonization with mycobacterium. Concomitant tuberculosis was present in 6 patients. The median diagnostic time from HIV to NTM infections was 3.5 months. The median CD4 count at NTM identification was 69.5 cells/μl. Median NTM treatment time was 16 months but 7 patients haven’t finished their treatment yet. The most commonly used medications were ethambutol and clarithromycin. Among analyzed patients, 4 of them have died. NTM infections are still an important disease among patients who are HIV late presenters. This disease should be taken into consideration during the differential diagnosis of fever, weight loss and cough in people with HIV with lymphocyte T CD4 count <100 cells/μl. Presence of tuberculosis does not exclude nontuberculous mycobacterium coinfection.

Keywords: mycobacteriosis, HIV, late presenter, epidemiology

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2308 Identification and Classification of Fiber-Fortified Semolina by Near-Infrared Spectroscopy (NIR)

Authors: Amanda T. Badaró, Douglas F. Barbin, Sofia T. Garcia, Maria Teresa P. S. Clerici, Amanda R. Ferreira

Abstract:

Food fortification is the intentional addition of a nutrient in a food matrix and has been widely used to overcome the lack of nutrients in the diet or increasing the nutritional value of food. Fortified food must meet the demand of the population, taking into account their habits and risks that these foods may cause. Wheat and its by-products, such as semolina, has been strongly indicated to be used as a food vehicle since it is widely consumed and used in the production of other foods. These products have been strategically used to add some nutrients, such as fibers. Methods of analysis and quantification of these kinds of components are destructive and require lengthy sample preparation and analysis. Therefore, the industry has searched for faster and less invasive methods, such as Near-Infrared Spectroscopy (NIR). NIR is a rapid and cost-effective method, however, it is based on indirect measurements, yielding high amount of data. Therefore, NIR spectroscopy requires calibration with mathematical and statistical tools (Chemometrics) to extract analytical information from the corresponding spectra, as Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA). PCA is well suited for NIR, once it can handle many spectra at a time and be used for non-supervised classification. Advantages of the PCA, which is also a data reduction technique, is that it reduces the data spectra to a smaller number of latent variables for further interpretation. On the other hand, LDA is a supervised method that searches the Canonical Variables (CV) with the maximum separation among different categories. In LDA, the first CV is the direction of maximum ratio between inter and intra-class variances. The present work used a portable infrared spectrometer (NIR) for identification and classification of pure and fiber-fortified semolina samples. The fiber was added to semolina in two different concentrations, and after the spectra acquisition, the data was used for PCA and LDA to identify and discriminate the samples. The results showed that NIR spectroscopy associate to PCA was very effective in identifying pure and fiber-fortified semolina. Additionally, the classification range of the samples using LDA was between 78.3% and 95% for calibration and 75% and 95% for cross-validation. Thus, after the multivariate analysis such as PCA and LDA, it was possible to verify that NIR associated to chemometric methods is able to identify and classify the different samples in a fast and non-destructive way.

Keywords: Chemometrics, fiber, linear discriminant analysis, near-infrared spectroscopy, principal component analysis, semolina

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2307 Current Status of Inclusive Education for Students with Disabilities in Punjab, Pakistan

Authors: Muhammad Shahid Shah, Akram Maqbool, Samina Ashraf

Abstract:

Since start of this century, world has adopted inclusion as a trend in special education. To meet the challenges of inclusion response, the Punjab government has developed a progressive policy to implement inclusive education. The objectives of this research were to analyze the administration and implementation process by consideration on the management, student’s admission process, screening and assessment, adaptations in curriculum and instruction along with an evaluation, government and nonprofit organizations support. The sample consisted of 50 schools both public and private with a total of 3000 students, 9 percent of which (270) were students with disabilities. Among all the students with disabilities, 63 percent (170) were male and 37 percent (100) were female. The concluded remarks regarding management revealed that a large number of inclusive schools was lacking in terms of developing a certain model for inclusion, including the managerial breakup of staff, the involvement of stakeholders, and conducted frequent meetings. Many of schools are not able to restructure their school organizations due to lack of financial resources, consultations, and backup. As for as student’s admission/identification/assessment was concerned, only 12 percent schools applied a selection process regarding student admission, half of which used different procedures for disable candidates. Approximately 5 percent of inclusive schools had modified their curriculum, including a variety of standards. In terms of instruction, 25 percent of inclusive schools reported that they modified their instructional process. Only a few schools, however, provided special equipment for students with visual impairment, physical impairment, speech and hearing problems, students with mild intellectual disabilities, and autism. In a student evaluation, more than 45 percent reported that test items, administration, time allocations, and students’ reports were modified. For the primary board examination conducted by the Education Department of Government of Punjab, this number decreased dramatically. Finally, government and nonprofit organizations support in the forms of funding, coaching, and facilities were mostly provided by provincial governments and by Ghazali Education Trust.

Keywords: inclusion, identification, assessment, funding, facilities, evaluation

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2306 Inversely Designed Chipless Radio Frequency Identification (RFID) Tags Using Deep Learning

Authors: Madhawa Basnayaka, Jouni Paltakari

Abstract:

Fully passive backscattering chipless RFID tags are an emerging wireless technology with low cost, higher reading distance, and fast automatic identification without human interference, unlike already available technologies like optical barcodes. The design optimization of chipless RFID tags is crucial as it requires replacing integrated chips found in conventional RFID tags with printed geometric designs. These designs enable data encoding and decoding through backscattered electromagnetic (EM) signatures. The applications of chipless RFID tags have been limited due to the constraints of data encoding capacity and the ability to design accurate yet efficient configurations. The traditional approach to accomplishing design parameters for a desired EM response involves iterative adjustment of design parameters and simulating until the desired EM spectrum is achieved. However, traditional numerical simulation methods encounter limitations in optimizing design parameters efficiently due to the speed and resource consumption. In this work, a deep learning neural network (DNN) is utilized to establish a correlation between the EM spectrum and the dimensional parameters of nested centric rings, specifically square and octagonal. The proposed bi-directional DNN has two simultaneously running neural networks, namely spectrum prediction and design parameters prediction. First, spectrum prediction DNN was trained to minimize mean square error (MSE). After the training process was completed, the spectrum prediction DNN was able to accurately predict the EM spectrum according to the input design parameters within a few seconds. Then, the trained spectrum prediction DNN was connected to the design parameters prediction DNN and trained two networks simultaneously. For the first time in chipless tag design, design parameters were predicted accurately after training bi-directional DNN for a desired EM spectrum. The model was evaluated using a randomly generated spectrum and the tag was manufactured using the predicted geometrical parameters. The manufactured tags were successfully tested in the laboratory. The amount of iterative computer simulations has been significantly decreased by this approach. Therefore, highly efficient but ultrafast bi-directional DNN models allow rapid and complicated chipless RFID tag designs.

Keywords: artificial intelligence, chipless RFID, deep learning, machine learning

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2305 Automatic Detection of Sugarcane Diseases: A Computer Vision-Based Approach

Authors: Himanshu Sharma, Karthik Kumar, Harish Kumar

Abstract:

The major problem in crop cultivation is the occurrence of multiple crop diseases. During the growth stage, timely identification of crop diseases is paramount to ensure the high yield of crops, lower production costs, and minimize pesticide usage. In most cases, crop diseases produce observable characteristics and symptoms. The Surveyors usually diagnose crop diseases when they walk through the fields. However, surveyor inspections tend to be biased and error-prone due to the nature of the monotonous task and the subjectivity of individuals. In addition, visual inspection of each leaf or plant is costly, time-consuming, and labour-intensive. Furthermore, the plant pathologists and experts who can often identify the disease within the plant according to their symptoms in early stages are not readily available in remote regions. Therefore, this study specifically addressed early detection of leaf scald, red rot, and eyespot types of diseases within sugarcane plants. The study proposes a computer vision-based approach using a convolutional neural network (CNN) for automatic identification of crop diseases. To facilitate this, firstly, images of sugarcane diseases were taken from google without modifying the scene, background, or controlling the illumination to build the training dataset. Then, the testing dataset was developed based on the real-time collected images from the sugarcane field from India. Then, the image dataset is pre-processed for feature extraction and selection. Finally, the CNN-based Visual Geometry Group (VGG) model was deployed on the training and testing dataset to classify the images into diseased and healthy sugarcane plants and measure the model's performance using various parameters, i.e., accuracy, sensitivity, specificity, and F1-score. The promising result of the proposed model lays the groundwork for the automatic early detection of sugarcane disease. The proposed research directly sustains an increase in crop yield.

Keywords: automatic classification, computer vision, convolutional neural network, image processing, sugarcane disease, visual geometry group

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2304 Characterizing Nasal Microbiota in COVID-19 Patients: Insights from Nanopore Technology and Comparative Analysis

Authors: David Pinzauti, Simon De Jaegher, Maria D'Aguano, Manuele Biazzo

Abstract:

The COVID-19 pandemic has left an indelible mark on global health, leading to a pressing need for understanding the intricate interactions between the virus and the human microbiome. This study focuses on characterizing the nasal microbiota of patients affected by COVID-19, with a specific emphasis on the comparison with unaffected individuals, to shed light on the crucial role of the microbiome in the development of this viral disease. To achieve this objective, Nanopore technology was employed to analyze the bacterial 16s rRNA full-length gene present in nasal swabs collected in Malta between January 2021 and August 2022. A comprehensive dataset consisting of 268 samples (126 SARS-negative samples and 142 SARS-positive samples) was subjected to a comparative analysis using an in-house, custom pipeline. The findings from this study revealed that individuals affected by COVID-19 possess a nasal microbiota that is significantly less diverse, as evidenced by lower α diversity, and is characterized by distinct microbial communities compared to unaffected individuals. The beta diversity analyses were carried out at different taxonomic resolutions. At the phylum level, Bacteroidota was found to be more prevalent in SARS-negative samples, suggesting a potential decrease during the course of viral infection. At the species level, the identification of several specific biomarkers further underscores the critical role of the nasal microbiota in COVID-19 pathogenesis. Notably, species such as Finegoldia magna, Moraxella catarrhalis, and others exhibited relative abundance in SARS-positive samples, potentially serving as significant indicators of the disease. This study presents valuable insights into the relationship between COVID-19 and the nasal microbiota. The identification of distinct microbial communities and potential biomarkers associated with the disease offers promising avenues for further research and therapeutic interventions aimed at enhancing public health outcomes in the context of COVID-19.

Keywords: COVID-19, nasal microbiota, nanopore technology, 16s rRNA gene, biomarkers

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2303 Evaluation of the Incidence of Mycobacterium Tuberculosis Complex Associated with Soil, Hayfeed and Water in Three Agricultural Facilities in Amathole District Municipality in the Eastern Cape Province

Authors: Athini Ntloko

Abstract:

Mycobacterium bovis and other species of Mycobacterium tuberculosis complex (MTBC) can result to a zoonotic infection known as Bovine tuberculosis (bTB). MTBC has members that may contaminate an extensive range of hosts, including wildlife. Diverse wild species are known to cause disease in domestic livestock and are acknowledged as TB reservoirs. It has been a main study worldwide to deliberate on bTB risk factors as a result and some studies focused on particular parts of risk factors such as wildlife and herd management. The significance of the study was to determine the incidence of Mycobacterium tuberculosis complex that is associated with soil, hayfeed and water. Questionnaires were administered to thirty (30) smallholding farm owners in the two villages (kwaMasele and Qungqwala) and three (3) three commercial farms (Fort Hare dairy farm, Middledrift dairy farm and Seven star dairy farm). Detection of M. tuberculosis complex was achieved by Polymerase Chain Reaction using primers for IS6110; whereas a genotypic drug resistance mutation was detected using Genotype MTBDRplus assays. Nine percent (9%) of respondents had more than 40 cows in their herd, while 60% reported between 10 and 20 cows in their herd. Relationship between farm size and vaccination for TB differed from forty one percent (41%) being the highest to the least five percent (5%). The highest number of respondents who knew about relationship between TB cases and cattle location was ninety one percent (91%). Approximately fifty one percent (51%) of respondents had knowledge about wild life access to the farms. Relationship between import of cattle and farm size ranged from nine percent (9%) to thirty five percent (35%). Cattle sickness in relation to farm size differed from forty three (43%) being the highest to the least three percent (3%); while thirty three percent (33%) of respondents had knowledge about health management. Respondents with knowledge about the occurrence of TB infections in farms were forty-eight percent (48%). The frequency of DNA isolation from samples ranged from the highest forty-five percent (45%) from water to the least twenty two percent (22%) from soil. Fort Hare dairy farm had the highest number of positive samples, forty four percent (44%) from water samples; whereas Middledrift dairy farm had the lowest positive from water, seventeen percent (17%). Twelve (22%) out of 55 isolates showed resistance to INH and RIF that is, multi-drug resistance (MDR) and nine percent (9%) were sensitive to either INH or RIF. The mutations at rpoB gene differed from 58% being the highest to the least (23%). Fifty seven percent (57%) of samples showed a S315T1 mutation while only 14% possessed a S531L in the katG gene. The highest inhA mutations were detected in T8A (80 %) and the least was observed in A16G (17%). The results of this study reveal that risk factors for bTB in cattle and dairy farm workers are a serious issue abound in the Eastern Cape of South Africa; with the possibility of widespread dissemination of multidrug resistant determinants in MTBC from the environment.

Keywords: hayfeed, isoniazid, multi-drug resistance, mycobacterium tuberculosis complex, polymerase chain reaction, rifampicin, soil, water

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2302 Identification of Bioactive Substances of Opuntia ficus-indica By-Products

Authors: N. Chougui, R. Larbat

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The first economic importance of Opuntia ficus-indica relies on the production of edible fruits. This food transformation generates a large amount of by-products (seeds and peels) in addition to cladodes produced by the plant. Several studies showed the richness of these products with bioactive substances like phenolics that have potential applications. Indeed, phenolics have been associated with protection against oxidation and several biological activities responsible of different pathologies. Consequently, there has been a growing interest in identifying natural antioxidants from plants. This study falls within the framework of the industrial exploitation of by-products of the plant. The study aims to investigate the metabolic profile of three by-products (cladodes, peel seeds) regarding total phenolic content by liquid chromatography coupled to mass spectrometry approach (LC-MSn). The byproducts were first washed, crushed and stored at negative temperature. The total phenolic compounds were then extracted by aqueous-ethanolic solvent in order to be quantified and characterized by LC-MS. According to the results obtained, the peel extract was the richest in phenolic compounds (1512.58 mg GAE/100 g DM) followed by the cladode extract (629.23 GAE/100 g DM) and finally by the seed extract (88.82 GAE/100 g DM) which is mainly used for its oil. The LC-MS analysis revealed diversity in phenolics in the three extracts and allowed the identification of hydroxybenzoic acids, hydroxycinnamic acids and flavonoids. The highest complexity was observed in the seed phenolic composition; more than twenty compounds were detected that belong to acids esters among which three feruloyl sucrose isomers. Sixteen compounds belonging to hydroxybenzoic acids, hydroxycinnamic acids and flavonoids were identified in the peel extract, whereas, only nine compounds were found in the cladode extract. It is interesting to highlight that the phenolic composition of the cladode extract was closer to that of the peel exact. However, from a quantitative viewpoint, the peel extract presented the highest amounts. Piscidic and eucomic acids were the two most concentrated molecules, corresponding to 271.3 and 121.6 mg GAE/ 100g DM respectively. The identified compounds were known to have high antioxidant and antiradical potential with the ability to inhibit lipid peroxidation and to exhibit a wide range of biological and therapeutic properties. The findings highlight the importance of using the Opuntia ficus-indica by-products.

Keywords: characterization, LC-MSn analysis, Opuntia ficus-indica, phenolics

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2301 Incorporation of Safety into Design by Safety Cube

Authors: Mohammad Rajabalinejad

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Safety is often seen as a requirement or a performance indicator through the design process, and this does not always result in optimally safe products or systems. This paper suggests integrating the best safety practices with the design process to enrich the exploration experience for designers and add extra values for customers. For this purpose, the commonly practiced safety standards and design methods have been reviewed and their common blocks have been merged forming Safety Cube. Safety Cube combines common blocks for design, hazard identification, risk assessment and risk reduction through an integral approach. An example application presents the use of Safety Cube for design of machinery.

Keywords: safety, safety cube, product, system, machinery, design

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2300 Proposition of an Ontology of Diseases and Their Signs from Medical Ontologies Integration

Authors: Adama Sow, Abdoulaye Guiss´e, Oumar Niang

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To assist medical diagnosis, we propose a federation of several existing and open medical ontologies and terminologies. The goal is to merge the strengths of all these resources to provide clinicians the access to a variety of shared knowledges that can facilitate identification and association of human diseases and all of their available characteristic signs such as symptoms and clinical signs. This work results to an integration model loaded from target known ontologies of the bioportal platform such as DOID, MESH, and SNOMED for diseases selection, SYMP, and CSSO for all existing signs.

Keywords: medical decision, medical ontologies, ontologies integration, linked data, knowledge engineering, e-health system

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2299 Preliminary Study of Gold Nanostars/Enhanced Filter for Keratitis Microorganism Raman Fingerprint Analysis

Authors: Chi-Chang Lin, Jian-Rong Wu, Jiun-Yan Chiu

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Myopia, ubiquitous symptom that is necessary to correct the eyesight by optical lens struggles many people for their daily life. Recent years, younger people raise interesting on using contact lens because of its convenience and aesthetics. In clinical, the risk of eye infections increases owing to the behavior of incorrectly using contact lens unsupervised cleaning which raising the infection risk of cornea, named ocular keratitis. In order to overcome the identification needs, new detection or analysis method with rapid and more accurate identification for clinical microorganism is importantly needed. In our study, we take advantage of Raman spectroscopy having unique fingerprint for different functional groups as the distinct and fast examination tool on microorganism. As we know, Raman scatting signals are normally too weak for the detection, especially in biological field. Here, we applied special SERS enhancement substrates to generate higher Raman signals. SERS filter we designed in this article that prepared by deposition of silver nanoparticles directly onto cellulose filter surface and suspension nanoparticles - gold nanostars (AuNSs) also be introduced together to achieve better enhancement for lower concentration analyte (i.e., various bacteria). Research targets also focusing on studying the shape effect of synthetic AuNSs, needle-like surface morphology may possible creates more hot-spot for getting higher SERS enhance ability. We utilized new designed SERS technology to distinguish the bacteria from ocular keratitis under strain level, and specific Raman and SERS fingerprint were grouped under pattern recognition process. We reported a new method combined different SERS substrates can be applied for clinical microorganism detection under strain level with simple, rapid preparation and low cost. Our presenting SERS technology not only shows the great potential for clinical bacteria detection but also can be used for environmental pollution and food safety analysis.

Keywords: bacteria, gold nanostars, Raman spectroscopy surface-enhanced Raman scattering filter

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2298 Isolation and Identification of Salmonella spp and Salmonella enteritidis, from Distributed Chicken Samples in the Tehran Province using Culture and PCR Techniques

Authors: Seyedeh Banafsheh Bagheri Marzouni, Sona Rostampour Yasouri

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Salmonella is one of the most important common pathogens between humans and animals worldwide. Globally, the prevalence of the disease in humans is due to the consumption of food contaminated with animal-derived Salmonella. These foods include eggs, red meat, chicken, and milk. Contamination of chicken and its products with Salmonella may occur at any stage of the chicken processing chain. Salmonella infection is usually not fatal. However, its occurrence is considered dangerous in some individuals, such as infants, children, the elderly, pregnant women, or individuals with weakened immune systems. If Salmonella infection enters the bloodstream, the possibility of contamination of tissues throughout the body will arise. Therefore, determining the potential risk of Salmonella at various stages is essential from the perspective of consumers and public health. The aim of this study is to isolate and identify Salmonella from chicken samples distributed in the Tehran market using the Gold standard culture method and PCR techniques based on specific genes, invA and ent. During the years 2022-2023, sampling was performed using swabs from the liver and intestinal contents of distributed chickens in the Tehran province, with a total of 120 samples taken under aseptic conditions. The samples were initially enriched in buffered peptone water (BPW) for pre-enrichment overnight. Then, the samples were incubated in selective enrichment media, including TT broth and RVS medium, at temperatures of 37°C and 42°C, respectively, for 18 to 24 hours. Organisms that grew in the liquid medium and produced turbidity were transferred to selective media (XLD and BGA) and incubated overnight at 37°C for isolation. Suspicious Salmonella colonies were selected for DNA extraction, and PCR technique was performed using specific primers that targeted the invA and ent genes in Salmonella. The results indicated that 94 samples were Salmonella using the PCR technique. Of these, 71 samples were positive based on the invA gene, and 23 samples were positive based on the ent gene. Although the culture technique is the Gold standard, PCR is a faster and more accurate method. Rapid detection through PCR can enable the identification of Salmonella contamination in food items and the implementation of necessary measures for disease control and prevention.

Keywords: culture, PCR, salmonella spp, salmonella enteritidis

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2297 Analyzing How Working From Home Can Lead to Higher Job Satisfaction for Employees Who Have Care Responsibilities Using Structural Equation Modeling

Authors: Christian Louis Kühner, Florian Pfeffel, Valentin Nickolai

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Taking care of children, dependents, or pets can be a difficult and time-consuming task. Especially for part- and full-time employees, it can feel exhausting and overwhelming to meet these obligations besides working a job. Thus, working mostly at home and not having to drive to the company can save valuable time and stress. This study aims to show the influence that the working model has on the job satisfaction of employees with care responsibilities in comparison to employees who do not have such obligations. Using structural equation modeling (SEM), the three work models, “work from home”, “working remotely”, and a hybrid model, have been analyzed based on 13 influencing constructs on job satisfaction. These 13 factors have been further summarized into three groups “classic influencing factors”, “influencing factors changed by remote working”, and “new remote working influencing factors”. Based on the influencing factors on job satisfaction, an online survey was conducted with n = 684 employees from the service sector. Here, Cronbach’s alpha of the individual constructs was shown to be suitable. Furthermore, the construct validity of the constructs was confirmed by face validity, content validity, convergent validity (AVE > 0.5: CR > 0.7), and discriminant validity. In addition, confirmatory factor analysis (CFA) confirmed the model fit for the investigated sample (CMIN/DF: 2.567; CFI: 0.927; RMSEA: 0.048). The SEM-analysis has shown that the most significant influencing factor on job satisfaction is “identification with the work” with β = 0.540, followed by “Appreciation” (β = 0.151), “Compensation” (β = 0.124), “Work-Life-Balance” (β = 0.116), and “Communication and Exchange of Information” (β = 0.105). While the significance of each factor can vary depending on the work model, the SEM-analysis shows that the identification with the work is the most significant factor in all three work models and, in the case of the traditional office work model, it is the only significant influencing factor. The study shows that among the employees with care responsibilities, the higher the proportion of working from home in comparison to working from the office, the more satisfied the employees are with their job. Since the work models that meet the requirements of comprehensive care led to higher job satisfaction amongst employees with such obligations, adapting as a company to such private obligations by employees can be crucial to sustained success. Conversely, the satisfaction level of the working model where employees work at the office is higher for workers without caregiving responsibilities.

Keywords: care responsibilities, home office, job satisfaction, structural equation modeling

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2296 A Risk-Based Approach to Construction Management

Authors: Chloe E. Edwards, Yasaman Shahtaheri

Abstract:

Risk management plays a fundamental role in project planning and delivery. The purpose of incorporating risk management into project management practices is to identify and address uncertainties related to key project-related activities. The uncertainties, known as risk events, can relate to project deliverables that are quantifiable and are often measured by impact to project schedule, cost, or environmental impact. Risk management should be incorporated as an iterative practice throughout the planning, execution, and commissioning phases of a project. This paper specifically examines how risk management contributes to effective project planning and delivery through a case study of a transportation project. This case study focused solely on impacts to project schedule regarding three milestones: readiness for delivery, readiness for testing and commissioning, and completion of the facility. The case study followed the ISO 31000: Risk Management – Guidelines. The key factors that are outlined by these guidelines include understanding the scope and context of the project, conducting a risk assessment including identification, analysis, and evaluation, and lastly, risk treatment through mitigation measures. This process requires continuous consultation with subject matter experts and monitoring to iteratively update the risks accordingly. The risk identification process led to a total of fourteen risks related to design, permitting, construction, and commissioning. The analysis involved running 1,000 Monte Carlo simulations through @RISK 8.0 Industrial software to determine potential milestone completion dates based on the project baseline schedule. These dates include the best case, most likely case, and worst case to provide an estimated delay for each milestone. Evaluation of these results provided insight into which risks were the highest contributors to the projected milestone completion dates. Based on the analysis results, the risk management team was able to provide recommendations for mitigation measures to reduce the likelihood of risks occurring. The risk management team also provided recommendations for managing the identified risks and project activities moving forward to meet the most likely or best-case milestone completion dates.

Keywords: construction management, monte carlo simulation, project delivery, risk assessment, transportation engineering

Procedia PDF Downloads 107
2295 Differentiated Instruction for All Learners: Strategies for Full Inclusion

Authors: Susan Dodd

Abstract:

This presentation details the methodology for teachers to identify and support a population of students who have historically been overlooked in regards to their educational needs. The twice exceptional (2e) student is a learner who is considered gifted and also has a learning disability, as defined by the Individuals with Disabilities Education Act (IDEA). Many of these students remain underserved throughout their educational careers because their exceptionalities may mask each other, resulting in a special population of students who are not achieving to their fullest potential. There are three common scenarios that may make the identification of a 2e student challenging. First, the student may have been identified as gifted, and her disability may go unnoticed. She could also be considered an under-achiever, or she may be able to compensate for her disability under the school works becomes more challenging. In the second scenario, the student may be identified as having a learning disability and is only receiving remedial services where his giftedness will not be highlighted. His overall IQ scores may be misleading because they were impacted by his learning disability. In the third scenario, the student is able to compensate for her ability well enough to maintain average scores, and she goes undetected as both gifted and learning disabled. Research in the area identifies the complexity involved in identifying 2e students, and how multiple forms of assessment are required. It is important for teachers to be aware of the common characteristics exhibited by many 2e students, so these learners can be identified and appropriately served. Once 2e students have been identified, teachers are then challenged to meet the varying needs of these exceptional learners. Strength-based teaching entails simultaneously providing gifted instruction as well as individualized accommodations for those students. Research in this field has yielded strategies that have proven helpful for teaching 2e students, as well as other students who may be struggling academically. Differentiated instruction, while necessary in all classrooms, is especially important for 2e students, as is encouragement for academic success. Teachers who take the time to really know their students will have a better understanding of each student’s strengths and areas for growth, and therefore tailor instruction to extend the intellectual capacities for optimal achievement. Teachers should also understand that some learning activities can prove very frustrating to students, and these activities can be modified based on individual student needs. Because 2e students can often become discouraged by their learning challenges, it is especially important for teachers to assist students in recognizing their own strengths and maintaining motivation for learning. Although research on the needs of 2e students has spanned across two decades, this population remains underserved in many educational institutions. Teacher awareness of the identification of and the support strategies for 2e students is critical for their success.

Keywords: gifted, learning disability, special needs, twice exceptional

Procedia PDF Downloads 178
2294 Parameter Estimation in Dynamical Systems Based on Latent Variables

Authors: Arcady Ponosov

Abstract:

A novel mathematical approach is suggested, which facilitates a compressed representation and efficient validation of parameter-rich ordinary differential equation models describing the dynamics of complex, especially biology-related, systems and which is based on identification of the system's latent variables. In particular, an efficient parameter estimation method for the compressed non-linear dynamical systems is developed. The method is applied to the so-called 'power-law systems' being non-linear differential equations typically used in Biochemical System Theory.

Keywords: generalized law of mass action, metamodels, principal components, synergetic systems

Procedia PDF Downloads 355
2293 Identification of COVID-SARS Variants Based on Lactate Test Results

Authors: Zoltan Horvath, Dora Nagy

Abstract:

In this research, it was examined whether individual COVID variants cause differences in the lactate curve of cyclists. After all, the virus variants attacked different organs in our body during the infections. During our tests, we used a traditional lactate step test, the results of which were compared with the values before the infection. In the tests, it has been proven that different virus variants show unique lactate curves. In this way, based on the lactate curve, it is possible to identify which variant caused the disease. Thanks to this, it has been shorten the return time, because we can apply the best return protocol after infection to the competitors.

Keywords: COVID-Sars19, lactate, virus mutation, lactate profile

Procedia PDF Downloads 66
2292 Internet as a Marketing Tool for Tourism Promotion

Authors: Emeka Okonkwo

Abstract:

The Information Technology (IT) has prevailed over all functions of strategic and operational management. The Internet (a product of information technology) has increasingly become a popular medium for marketing. This paper examines the potentials of Internet for tourism marketing. To achieve this, the paper x-rays the characteristics of tourism marketing and examines the application of the Internet in tourism marketing. It is argued that the use of Internet for tourism marketing will not only reach a broad audience and reduce the cost of transaction (by conventional methods used by travel agents in times past), but, will also alleviate the problems of identification, authentication and confirmation of travels/package tours by tourists as well as promotion of tourism industry.

Keywords: internet, marketing, tourism, tourism management

Procedia PDF Downloads 418
2291 Design of a Fuzzy Luenberger Observer for Fault Nonlinear System

Authors: Mounir Bekaik, Messaoud Ramdani

Abstract:

We present in this work a new technique of stabilization for fault nonlinear systems. The approach we adopt focus on a fuzzy Luenverger observer. The T-S approximation of the nonlinear observer is based on fuzzy C-Means clustering algorithm to find local linear subsystems. The MOESP identification approach was applied to design an empirical model describing the subsystems state variables. The gain of the observer is given by the minimization of the estimation error through Lyapunov-krasovskii functional and LMI approach. We consider a three tank hydraulic system for an illustrative example.

Keywords: nonlinear system, fuzzy, faults, TS, Lyapunov-Krasovskii, observer

Procedia PDF Downloads 331
2290 Modeling of Crack Growth in Railway Axles under Static Loading

Authors: Zellagui Redouane, Bellaouar Ahmed, Lachi Mohammed

Abstract:

The railway axles are the essential parts in the bogie of train, and its failure creates a big problem in the railway transport; during the work of this parts we noticed a premature deterioration. The aim has been presented a predictive model allowing the identification of the probable causes that are the cause of these premature deterioration. The results are employed for predicting fatigue crack growth in the railway axle, Also we want to present the variation value of stress intensity factor in different positions of elliptical crack tip. The modeling of axle in performed by the SOLID WORKS software and imported into ANSYS.

Keywords: crack growth, static load, railway axle, lifetime

Procedia PDF Downloads 364
2289 Isolation, Identification and Screening of Pectinase Producing Fungi Isolated from Apple (Malus Domestica)

Authors: Shameel Pervez, Saad Aziz Durrani, Ibatsam Khokhar

Abstract:

Pectinase is an enzyme that breaks down pectin, a compound responsible for structural integrity of the plant. Pectin is difficult to break down mechanically and the cost is very high, that is why many industries including food industries use pectinase enzyme produced by microbes for pectin breakdown. Apple (Malus domestica) is an important fruit in terms of market value. Every year, millions of apples are wasted due to post-harvest rot caused by fungi. Fungi are natural decomposers of our ecosystem and are infamous for post-harvest rot of apple fruit but at the same time they are prized for their high production of valuable extracellular enzymes such as pectinase. In this study, fungi belonging to different genus were isolated from rotten apples. Rotten samples of apple were picked from different markets of Lahore. After surface sterilization, the rotten parts were cut into small pieces and placed onto MEA media plates for three days. Afterwards, distinct colonies were picked and purified by sub-culturing. The isolates were identified to genus level through the study of basic colony morphology and microscopic features. The isolates were then subjected to screening for pectinase activity on MS media to compare pectinase production and were then subsequently tested for pathogenic activity through wound suspension method to evaluate the pathogenic activity of isolates in comparison with their pectinolytic activity. A total of twelve fungal strains were isolates from rotten apples. They were belonging to genus Penicillium, Alternaria, Paecilomyces and Rhizopus. Upon screening for pectinolytic activity, isolates Pen 1, Pen 4, and Rz showed high pectinolytic activity and were further subjected to DNA isolation and partial sequencing for species identification. The results of partial sequencing were combined with in-depth study of morphological features revealing Pen 1 as Penicillium janthinellum, Pen 4 as Penicillium griseofulvum, and Rz as Rhizopus microsporus. Pathogenic activity of all twelve isolates was evaluated. Penicillium spp. were highly pathogenic and destructive and same was the case with Paecilomyces sp. and Rhizopus sp. However, Alternaria spp. were found to be more consistent in their pathogenic activity, on all types of apples.

Keywords: apple, pectinase, fungal pathogens, penicillium, rhizopus

Procedia PDF Downloads 63
2288 Identification and Characterization of in Vivo, in Vitro and Reactive Metabolites of Zorifertinib Using Liquid Chromatography Lon Trap Mass Spectrometry

Authors: Adnan A. Kadi, Nasser S. Al-Shakliah, Haitham Al-Rabiah

Abstract:

Zorifertinib is a novel, potent, oral, a small molecule used to treat non-small cell lung cancer (NSCLC). zorifertinib is an Epidermal Growth Factor Receptor (EGFR) inhibitor and has good blood–brain barrier permeability for (NSCLC) patients with EGFR mutations. zorifertinibis currently at phase II/III clinical trials. The current research reports the characterization and identification of in vitro, in vivo and reactive intermediates of zorifertinib. Prediction of susceptible sites of metabolism and reactivity pathways (cyanide and GSH) of zorifertinib were performed by the Xenosite web predictor tool. In-vitro metabolites of zorifertinib were performed by incubation with rat liver microsomes (RLMs) and isolated perfused rat liver hepatocytes. Extraction of zorifertinib and it's in vitro metabolites from the incubation mixtures were done by protein precipitation. In vivo metabolism was done by giving a single oral dose of zorifertinib(10 mg/Kg) to Sprague Dawely rats in metabolic cages by using oral gavage. Urine was gathered and filtered at specific time intervals (0, 6, 12, 18, 24, 48, 72,96and 120 hr) from zorifertinib dosing. A similar volume of ACN was added to each collected urine sample. Both layers (organic and aqueous) were injected into liquid chromatography ion trap mass spectrometry(LC-IT-MS) to detect vivozorifertinib metabolites. N-methyl piperizine ring and quinazoline group of zorifertinib undergoe metabolism forming iminium and electro deficient conjugated system respectively, which are very reactive toward nucleophilic macromolecules. Incubation of zorifertinib with RLMs in the presence of 1.0 mM KCN and 1.0 Mm glutathione were made to check reactive metabolites as it is often responsible for toxicities associated with this drug. For in vitro metabolites there were nine in vitro phase I metabolites, four in vitro phase II metabolites, eleven reactive metabolites(three cyano adducts, five GSH conjugates metabolites, and three methoxy metabolites of zorifertinib were detected by LC-IT-MS. For in vivo metabolites, there were eight in vivo phase I, tenin vivo phase II metabolitesofzorifertinib were detected by LC-IT-MS. In vitro and in vivo phase I metabolic pathways wereN- demthylation, O-demethylation, hydroxylation, reduction, defluorination, and dechlorination. In vivo phase II metabolic reaction was direct conjugation of zorifertinib with glucuronic acid and sulphate.

Keywords: in vivo metabolites, in vitro metabolites, cyano adducts, GSH conjugate

Procedia PDF Downloads 198
2287 Realization of Autonomous Guidance Service by Integrating Information from NFC and MEMS

Authors: Dawei Cai

Abstract:

In this paper, we present an autonomous guidance service by combining the position information from NFC and the orientation information from a 6 axis acceleration and terrestrial magnetism sensor. We developed an algorithm to calculate the device orientation based on the data from acceleration and terrestrial magnetism sensor. If visitors want to know some explanation about an exhibit in front of him, what he has to do is just lift up his mobile device. The identification program will automatically identify the status based on the information from NFC and MEMS, and start playing explanation content for him. This service may be convenient for old people or disables or children.

Keywords: NFC, ubiquitous computing, guide sysem, MEMS

Procedia PDF Downloads 409
2286 Differential Expression Analysis of Busseola fusca Larval Transcriptome in Response to Cry1Ab Toxin Challenge

Authors: Bianca Peterson, Tomasz J. Sańko, Carlos C. Bezuidenhout, Johnnie Van Den Berg

Abstract:

Busseola fusca (Fuller) (Lepidoptera: Noctuidae), the maize stem borer, is a major pest in sub-Saharan Africa. It causes economic damage to maize and sorghum crops and has evolved non-recessive resistance to genetically modified (GM) maize expressing the Cry1Ab insecticidal toxin. Since B. fusca is a non-model organism, very little genomic information is publicly available, and is limited to some cytochrome c oxidase I, cytochrome b, and microsatellite data. The biology of B. fusca is well-described, but still poorly understood. This, in combination with its larval-specific behavior, may pose problems for limiting the spread of current resistant B. fusca populations or preventing resistance evolution in other susceptible populations. As part of on-going research into resistance evolution, B. fusca larvae were collected from Bt and non-Bt maize in South Africa, followed by RNA isolation (15 specimens) and sequencing on the Illumina HiSeq 2500 platform. Quality of reads was assessed with FastQC, after which Trimmomatic was used to trim adapters and remove low quality, short reads. Trinity was used for the de novo assembly, whereas TransRate was used for assembly quality assessment. Transcript identification employed BLAST (BLASTn, BLASTp, and tBLASTx comparisons), for which two libraries (nucleotide and protein) were created from 3.27 million lepidopteran sequences. Several transcripts that have previously been implicated in Cry toxin resistance was identified for B. fusca. These included aminopeptidase N, cadherin, alkaline phosphatase, ATP-binding cassette transporter proteins, and mitogen-activated protein kinase. MEGA7 was used to align these transcripts to reference sequences from Lepidoptera to detect mutations that might potentially be contributing to Cry toxin resistance in this pest. RSEM and Bioconductor were used to perform differential gene expression analysis on groups of B. fusca larvae challenged and unchallenged with the Cry1Ab toxin. Pairwise expression comparisons of transcripts that were at least 16-fold expressed at a false-discovery corrected statistical significance (p) ≤ 0.001 were extracted and visualized in a hierarchically clustered heatmap using R. A total of 329,194 transcripts with an N50 of 1,019 bp were generated from the over 167.5 million high-quality paired-end reads. Furthermore, 110 transcripts were over 10 kbp long, of which the largest one was 29,395 bp. BLAST comparisons resulted in identification of 157,099 (47.72%) transcripts, among which only 3,718 (2.37%) were identified as Cry toxin receptors from lepidopteran insects. According to transcript expression profiles, transcripts were grouped into three subclusters according to the similarity of their expression patterns. Several immune-related transcripts (pathogen recognition receptors, antimicrobial peptides, and inhibitors) were up-regulated in the larvae feeding on Bt maize, indicating an enhanced immune status in response to toxin exposure. Above all, extremely up-regulated arylphorin genes suggest that enhanced epithelial healing is one of the resistance mechanisms employed by B. fusca larvae against the Cry1Ab toxin. This study is the first to provide a resource base and some insights into a potential mechanism of Cry1Ab toxin resistance in B. fusca. Transcriptomic data generated in this study allows identification of genes that can be targeted by biotechnological improvements of GM crops.

Keywords: epithelial healing, Lepidoptera, resistance, transcriptome

Procedia PDF Downloads 201
2285 Valorizing Traditional Greek Wheat Varieties: Use of DNA Barcoding for Species Identification and Biochemical Analysis of Their Nutritional Value

Authors: Niki Mougiou, Spyros Didos, Ioanna Bouzouka, Athina Theodorakopoulou, Michael Kornaros, Anagnostis Argiriou

Abstract:

Grains from traditional old Greek cereal varieties were evaluated and compared to commercial cultivars, like Simeto and Mexicali 81, in an effort to valorize local products and assess the nutritional benefits of ancient grains. The samples studied in this research included common wheat, durum wheat, emmer (Triticum dicoccum) and einkorn (Triticum monococcum), as well as barley, oats and rye grains. The Internal Transcribed Spacer 2 (ITS2) nuclear region was amplified and sequenced as a barcode for species identification, allowing the verification of the label of each product. After that, the total content of bound and free polyphenols and flavonoids, as well as the antioxidant activity of bound and free compounds, was measured by classic colorimetric assays using Folin- Ciocalteu, AlCl₃ and DPPH‧ (2,2-diphenyl-1-picrylhydrazyl) reagents, respectively. Moreover, the level of variation of fatty acids was determined in all samples by gas chromatography. The results showed that local old landraces of emmer and einkorn had the highest polyphenol content, 2.4 and 3.3 times higher than the average value of 5 durum wheat samples, respectively. Regarding the total flavonoid content, einkorn had 2.6-fold and emmer 2-fold higher values than common wheat. The antioxidant activity of free or bound compounds was at the same level, at about 20-30% higher in both einkorn and emmer compared to common wheat. Five main fatty acids were detected in all samples, in order of decreasing amounts: linoleic (C18:2) > palmitic (C16:0) ≈ , oleic (C18:1) > eicosenoic (C20:1, cis-11) > stearic (C18:0). Emmer and einkorn showed a higher diversity of fatty acids and a higher content of mono-unsaturated fatty acids compared to common wheat. The results of this study demonstrate the high nutritional value of old local landraces that have been put aside by more productive, yet with lower qualitative characteristics, commercial cultivars, underlining the importance of maintaining sustainable agricultural practices to ensure their continued cultivation.

Keywords: biochemical analysis, nutritional value, plant barcoding, wheat

Procedia PDF Downloads 85
2284 Inverse Scattering of Two-Dimensional Objects Using an Enhancement Method

Authors: A.R. Eskandari, M.R. Eskandari

Abstract:

A 2D complete identification algorithm for dielectric and multiple objects immersed in air is presented. The employed technique consists of initially retrieving the shape and position of the scattering object using a linear sampling method and then determining the electric permittivity and conductivity of the scatterer using adjoint sensitivity analysis. This inversion algorithm results in high computational speed and efficiency, and it can be generalized for any scatterer structure. Also, this method is robust with respect to noise. The numerical results clearly show that this hybrid approach provides accurate reconstructions of various objects.

Keywords: inverse scattering, microwave imaging, two-dimensional objects, Linear Sampling Method (LSM)

Procedia PDF Downloads 387
2283 Potential Serological Biomarker for Early Detection of Pregnancy in Cows

Authors: Shveta Bathla, Preeti Rawat, Sudarshan Kumar, Rubina Baithalu, Jogender Singh Rana, Tushar Kumar Mohanty, Ashok Kumar Mohanty

Abstract:

Pregnancy is a complex process which includes series of events such as fertilization, formation of blastocyst, implantation of embryo, placental formation and development of fetus. The success of these events depends on various interactions which are synchronized by endocrine interaction between a receptive dam and competent embryo. These interactions lead to change in expression of hormones and proteins. But till date no protein biomarker is available which can be used to detect successful completion of these events. We employed quantitative proteomics approach to develop putative serological biomarker which has diagnostic applicability for early detection of pregnancy in cows. For this study, sera were collected from control (non-pregnant, n=6) and pregnant animals on successive days of pregnancy (7, 19, 45, n=6). The sera were subjected to depletion for removal of albumin using Norgen depletion kit. The tryptic peptides were labeled with iTRAQ. The peptides were pooled and fractionated using bRPLC over 80 min gradient. Then 12 fractions were injected to nLC for identification and quantitation in DDA mode using ESI. Identification using Mascot search revealed 2056 proteins out of which 352 proteins were differentially expressed. Twenty proteins were upregulated and twelve proteins were down-regulated with fold change > 1.5 and < 0.6 respectively (p < 0.05). The gene ontology studies of DEPs using Panther software revealed that majority of proteins are actively involved in catalytic activities, binding and enzyme regulatory activities. The DEP'S such as NF2, MAPK, GRIPI, UGT1A1, PARP, CD68 were further subjected to pathway analysis using KEGG and Cytoscape plugin Cluego that showed involvement of proteins in successful implantation, maintenance of pluripotency, regulation of luteal function, differentiation of endometrial macrophages, protection from oxidative stress and developmental pathways such as Hippo. Further efforts are continuing for targeted proteomics, western blot to validate potential biomarkers and development of diagnostic kit for early pregnancy diagnosis in cows.

Keywords: bRPLC, Cluego, ESI, iTRAQ, KEGG, Panther

Procedia PDF Downloads 460