Search results for: mutation detection

Authors: Yi Li, Rui Lu, Lianjun Wang

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With the rapid development of chemical industry, the consumption of volatile organic compounds (VOCs) has increased extensively. In the process of VOCs production and application, plenty of them have been transferred to environment. As a result, it has led to pollution problems not only in soil and ground water but also to human beings. Thus, it is important to develop a sensitive and cost-effective analytical method for trace VOCs detection in environment. Surface-enhanced Raman Spectroscopy (SERS), as one of the most sensitive optical analytical technique with rapid response, pinpoint accuracy and noninvasive detection, has been widely used for ultratrace analysis. Based on the plasmon resonance on the nanoscale metallic surface, SERS technology can even detect single molecule due to abundant nanogaps (i.e. 'hot spots') on the nanosubstrate. In this work, a self-supported flexible silver nitrate (AgNO3)/ethylene-propylene copolymer (EPM) hybrid nanofibers was fabricated by electrospinning. After an in-situ chemical reduction using ice-cold sodium borohydride as reduction agent, numerous silver nanoparticles were formed on the nanofiber surface. By adjusting the reduction time and AgNO3 content, the morphology and dimension of silver nanoparticles could be controlled. According to the principles of solid-phase extraction, the hydrophobic substance is more likely to partition into the hydrophobic EPM membrane in an aqueous environment while water and other polar components are excluded from the analytes. By the enrichment of EPM fibers, the number of hydrophobic molecules located on the 'hot spots' generated from criss-crossed nanofibers is greatly increased, which further enhances SERS signal intensity. The as-prepared Ag/EPM hybrid nanofibers were first employed to detect common SERS probe molecule (p-aminothiophenol) with the detection limit down to 10-12 M, which demonstrated an excellent SERS performance. To further study the application of the fabricated substrate for monitoring hydrophobic substance in water, several typical VOCs, such as benzene, toluene and p-xylene, were selected as model compounds. The results showed that the characteristic peaks of these target analytes in the mixed aqueous solution could be distinguished even at a concentration of 10-6 M after multi-peaks gaussian fitting process, including C-H bending (850 cm-1), C-C ring stretching (1581 cm-1, 1600 cm-1) of benzene, C-H bending (844 cm-1 ,1151 cm-1), C-C ring stretching (1001 cm-1), CH3 bending vibration (1377 cm-1) of toluene, C-H bending (829 cm-1), C-C stretching (1614 cm-1) of p-xylene. The SERS substrate has remarkable advantages which combine the enrichment capacity from EPM and the Raman enhancement of Ag nanoparticles. Meanwhile, the huge specific surface area resulted from electrospinning is benificial to increase the number of adsoption sites and promotes 'hot spots' formation. In summary, this work provides powerful potential in rapid, on-site and accurate detection of trace VOCs using a portable Raman.

Keywords: electrospinning, ethylene-propylene copolymer, silver nanoparticles, SERS, VOCs

Procedia PDF Downloads 161

Authors: Mubashir Hussain, Mu Lv, Xiaohan Dong, Zhiyang Li, Bin Liu, Nongyue He

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Detection and classification of microbes have a vast range of applications in biomedical engineering especially in detection, characterization, and quantification of bacterial contaminants. For identification of pathogens, different techniques are emerging in the field of biomedical engineering. Latest technology uses light scattering, capable of identifying different pathogens without any need for biochemical processing. Bacterial Pathogens Identification System (BPIS) which uses a laser beam, passes through the sample and light scatters off. An assembly of photodetectors surrounded by the sample at different angles to detect the scattering of light. The algorithm of the system consists of two parts: (a) Library files, and (b) Comparator. Library files contain data of known species of bacterial microbes in the form of binned plots, while comparator compares data of unknown sample with library files. Using collected data of unknown bacterial species, highest voltage values stored in the form of peaks and arranged in 3D histograms to find the frequency of occurrence. Resulting data compared with library files of known bacterial species. If sample data matching with any library file of known bacterial species, sample identified as a matched microbe. An experiment performed to identify three different bacteria particles: Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli. By applying algorithm using library files of given samples, results were compromising. This system is potentially applicable to several biomedical areas, especially those related to cell morphology.

Keywords: microbial identification, laser scattering, peak identification, binned plots classification

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Authors: Beata Zima, Octavio A. Márquez Reyes, Masoud Mohammadgholiha, Jochen Moll, Luca de Marchi

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This document presents the approach of using compressed sensing in signal encoding and information transferring within a guided wave sensor network, comprised of specially designed frequency steerable acoustic transducers (FSATs). Wave propagation in a damaged plate was simulated using commercial FEM-based software COMSOL. Guided waves were excited by means of FSATs, characterized by the special shape of its electrodes, and modeled using PIC255 piezoelectric material. The special shape of the FSAT, allows for focusing wave energy in a certain direction, accordingly to the frequency components of its actuation signal, which makes available a larger monitored area. The process begins when a FSAT detects and records reflection from damage in the structure, this signal is then encoded and prepared for transmission, using a combined approach, based on Compressed Sensing Matching Pursuit and Quadrature Amplitude Modulation (QAM). After codification of the signal is in binary chars the information is transmitted between the nodes in the network. The message reaches the last node, where it is finally decoded and processed, to be used for damage detection and localization purposes. The main aim of the investigation is to determine the location of detected damage using reconstructed signals. The study demonstrates that the special steerable capabilities of FSATs, not only facilitate the detection of damage but also permit transmitting the damage information to a chosen area in a specific direction of the investigated structure.

Keywords: data compression, ultrasonic communication, guided waves, FEM analysis

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Authors: Won Young Choi, Kwan Kyu Park

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Recently, the road depression happening in the urban area is different from the cause of the sink hole and the generation mechanism occurring in the limestone area. The main cause of sinkholes occurring in the city center is the loss of soil due to the damage of old underground buried materials and groundwater discharge due to large underground excavation works. The method of detecting the sinkhole in the urban area is mostly using the Ground Penetration Radar (GPR). However, it is challenging to implement compact system and detecting watery state since it is based on electromagnetic waves. Although many ultrasonic underground detection studies have been conducted, near-ground detection (several tens of cm to several meters) has been developed for bulk systems using geophones as a receiver. The goal of this work is to fabricate a miniaturized sinkhole detecting system based on low-cost ultrasonic transducers of 40 kHz resonant frequency with high transmission pressure and receiving sensitivity. Motived by biomedical ultrasonic imaging methods, we detect air layers below the ground such as asphalt through the pulse-echo method. To improve image quality using multi-channel, linear array system is implemented, and image is acquired by classical synthetic aperture imaging method. We present the successful feasibility test of multi-channel sinkhole detector based on ultrasonic transducer. In this work, we presented and analyzed image results which are imaged by single channel pulse-echo imaging, synthetic aperture imaging.

Keywords: road depression, sinkhole, synthetic aperture imaging, ultrasonic transducer

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Authors: Faiza Babur Khan, Sohail Asghar

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With an emergence of distributed environment, cloud computing is proving to be the most stimulating computing paradigm shift in computer technology, resulting in spectacular expansion in IT industry. Many companies have augmented their technical infrastructure by adopting cloud resource sharing architecture. Cloud computing has opened doors to unlimited opportunities from application to platform availability, expandable storage and provision of computing environment. However, from a security viewpoint, an added risk level is introduced from clouds, weakening the protection mechanisms, and hardening the availability of privacy, data security and on demand service. Issues of trust, confidentiality, and integrity are elevated due to multitenant resource sharing architecture of cloud. Trust or reliability of cloud refers to its capability of providing the needed services precisely and unfailingly. Confidentiality is the ability of the architecture to ensure authorization of the relevant party to access its private data. It also guarantees integrity to protect the data from being fabricated by an unauthorized user. So in order to assure provision of secured cloud, a roadmap or model is obligatory to analyze a security problem, design mitigation strategies, and evaluate solutions. The aim of the paper is twofold; first to enlighten the factors which make cloud security critical along with alleviation strategies and secondly to propose an intrusion detection model that identifies the attackers in a preventive way using machine learning Random Forest classifier with an accuracy of 99.8%. This model uses less number of features. A comparison with other classifiers is also presented.

Keywords: cloud security, threats, machine learning, random forest, classification

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Authors: Habtamu Abdisa

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Spring water is particularly valuable due to its high mineral content, which is beneficial for human health. However, anthropogenic activities usually imbalance the natural levels of its composition, which can cause adverse health effects. Regular monitoring of a naturally given environmental resource is of great concern in the world today. The spectrophotometric application is one of the best methods for qualifying and quantifying the mineral contents of environmental water samples. This research was conducted to evaluate the quality of spring water concerning its heavy metal composition. A grab sampling technique was employed to collect representative samples, including duplicates. The samples were then treated with concentrated HNO3 to a pH level below 2 and stored at 4oC. The samples were digested and analyzed for cadmium (Cd), chromium (Cr), manganese (Mn), copper (Cu), iron (Fe), and zinc (Zn) following method validation. Atomic Absorption Spectrometry (AAS) was utilized for the sample analysis. Quality control measures, including blanks, duplicates, and certified reference materials (CRMs), were implemented to ensure the accuracy and precision of the analytical results. Of the metals analyzed in the water samples, Cd and Cr were found to be below the detection limit. However, the concentrations of Mn, Cu, Fe, and Zn ranged from mean values of 0.119-0.227 mg/L, 0.142-0.166 mg/L, 0.183-0.267 mg/L, and 0.074-0.181 mg/L, respectively. Sediment analysis revealed mean concentration ranges of 348.31-429.21 mg/kg, 0.23-0.28 mg/kg, 18.73-22.84 mg/kg, 2.76-3.15 mg/kg, 941.84-1128.56 mg/kg, and 42.39-66.53 mg/kg for Mn, Cd, Cu, Cr, Fe, and Zn, respectively. The study results established that the evaluated spring water and its associated sediment met the regulatory standards and guidelines for heavy metal concentrations. Furthermore, this research can enhance the quality assurance and control processes for environmental sample analysis, ensuring the generation of reliable data.

Keywords: method validation, heavy metal, spring water, sediment, method detection limit

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Authors: Duanghathai Saipinta, Tanittian Panyamongkol, Witaya Suriyasathaporn

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Animal management aims to provide a suitable environment for animals allowing maximal productivity in those animals. Prevention of disease is an important part of animal management. Foot-and-mouth disease (FMD) is a highly contagious viral disease in cattle and is an economically important animal disease worldwide. Monitoring the FMD virus in farms is useful management for the prevention of the FMD outbreak. A recent publication indicated collection samples from nasal swabs can be used for monitoring FMD in symptomatic cows. Therefore, the objectives of this study were to determine the FMD virus in asymptomatic dairy cattle using nasal swab samples during the absence of an FMD outbreak. The study was conducted from December 2020 to June 2021 using 185 asymptomatic signs of FMD dairy cattle in Chiang Mai Province, Thailand. By random cow selection, nasal mucosal swabs were used to collect samples from the selected cows and then were to evaluate the presence of FMD viruses using the real-time rt-PCR assay. In total, 4.9% of dairy cattle detected FMD virus, including 2 dairy farms in Mae-on (8 samples; 9.6%) and 1 farm in the Chai-Prakan district (1 sample; 1.2%). Interestingly, both farms in Mae-on were the outbreak of the FMD after this detection for 6 months. This indicated that the FMD virus presented in asymptomatic cattle might relate to the subsequent outbreak of FMD. The outbreak demonstrates the presence of the virus in the environment. In conclusion, monitoring of FMD can be performed by nasal swab collection. Further investigation is needed to show whether the FMD virus presented in asymptomatic FMD cattle could be the cause of the subsequent FMD outbreak or not.

Keywords: cattle, foot-and-mouth disease, nasal swab, real-time rt-PCR assay

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Authors: Catherine Thompson, Maryam Jalali, Peter Hills

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The allocation of visual attention is critical when driving and anything that limits attention will have a detrimental impact on safety. Engaging in a secondary task reduces the amount of attention directed to the road because drivers allocate resources towards this task, leaving fewer resources to process driving-relevant information. Yet the dangers associated with a secondary task do not end when the driver returns their attention to the road. Instead, the attentional settings adopted to complete a secondary task may persist to the road, affecting attention, and therefore affecting driver performance. This 'attentional inertia' effect was investigated in the current work. Forty drivers searched for hazards in driving video clips while their eye-movements were recorded. At varying intervals they were instructed to attend to a secondary task displayed on a tablet situated to their left-hand side. The secondary task consisted of three separate computer games that induced horizontal, vertical, and random eye movements. Visual search and hazard detection in the driving clips were compared across the three conditions of the secondary task. Results showed that the layout of information in the secondary task, and therefore the allocation of attention in this task, had an impact on subsequent search in the driving clips. Vertically presented information reduced the wide horizontal spread of search usually associated with accurate driving and had a negative influence on the detection of hazards. The findings show the additional dangers of engaging in a secondary task while driving. The attentional inertia effect has significant implications for semi-autonomous and autonomous vehicles in which drivers have greater opportunity to direct their attention away from the driving task.

Keywords: attention, eye-movements, hazard perception, visual search

Procedia PDF Downloads 165

Authors: Jeremy Jenrette, Z. Y. C. Liu, Pranav Chimote, Edward Fox, Trevor Hastie, Francesco Ferretti

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Suitable shark conservation depends on well-informed population assessments. Direct methods such as scientific surveys and fisheries monitoring are adequate for defining population statuses, but species-specific indices of abundance and distribution coming from these sources are rare for most shark species. We can rapidly fill these information gaps by boosting media-based remote monitoring efforts with machine learning and automation. We created a database of shark images by sourcing 24,546 images covering 219 species of sharks from the web application spark pulse and the social network Instagram. We used object detection to extract shark features and inflate this database to 53,345 images. We packaged object-detection and image classification models into a Shark Detector bundle. We developed the Shark Detector to recognize and classify sharks from videos and images using transfer learning and convolutional neural networks (CNNs). We applied these models to common data-generation approaches of sharks: boosting training datasets, processing baited remote camera footage and online videos, and data-mining Instagram. We examined the accuracy of each model and tested genus and species prediction correctness as a result of training data quantity. The Shark Detector located sharks in baited remote footage and YouTube videos with an average accuracy of 89\%, and classified located subjects to the species level with 69\% accuracy (n =\ eight species). The Shark Detector sorted heterogeneous datasets of images sourced from Instagram with 91\% accuracy and classified species with 70\% accuracy (n =\ 17 species). Data-mining Instagram can inflate training datasets and increase the Shark Detector’s accuracy as well as facilitate archiving of historical and novel shark observations. Base accuracy of genus prediction was 68\% across 25 genera. The average base accuracy of species prediction within each genus class was 85\%. The Shark Detector can classify 45 species. All data-generation methods were processed without manual interaction. As media-based remote monitoring strives to dominate methods for observing sharks in nature, we developed an open-source Shark Detector to facilitate common identification applications. Prediction accuracy of the software pipeline increases as more images are added to the training dataset. We provide public access to the software on our GitHub page.

Keywords: classification, data mining, Instagram, remote monitoring, sharks

Procedia PDF Downloads 122

Authors: Motoki Seguchi, Fumiko Harada, Hiromitsu Shimakawa

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There have been many local companies in countryside that carefully produce and sell products, which include crafts and foods produced with traditional methods. These companies are likely to use commercials to advertise their products. However, it is difficult for companies to judge whether the commercials they create are having an impact on consumers. Therefore, to create effective commercials, this study researches what kind of gimmicks in commercials affect what kind of consumers. This study proposes a method for extracting psychological change points from the physiological characteristics of consumers while they are watching commercials and estimating the gimmicks in the commercial that affect consumer engagement. In this method, change point detection is applied to pupil size for estimating gimmicks that affect consumers’ emotional engagement, and to EDA for estimating gimmicks that affect cognitive engagement. A questionnaire is also used to estimate the commercials that influence behavioral engagement. As a result of estimating the gimmicks that influence consumer engagement using this method, it was found that there are some common features among the gimmicks. To influence cognitive engagement, it was found that it was useful to include flashback scenes, messages to be appealed to, the company’s name, and the company’s logos as gimmicks. It was also found that flashback scenes and story climaxes were useful in influencing emotional engagement. Furthermore, it was found that the use of storytelling commercials may or may not be useful, depending on which consumers are desired to take which behaviors. It also estimated the gimmicks that influence consumers for each target and found that the useful gimmicks are slightly different for students and working adults. By using this method, it can understand which gimmicks in the commercial affect which engagement of the consumers. Therefore, the results of this study can be used as a reference for the gimmicks that should be included in commercials when companies create their commercials in the future.

Keywords: change point detection, estimating engagement, physiological characteristics, psychological changes, watching commercials

Procedia PDF Downloads 187

Authors: Thidarat Duangyod, Chanida Palanuvej, Nijsiri Ruangrungsi

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Pentace burmanica Kurz., belonging to the Malvaceae family, is commonly used for anti-diarrhea in Thai traditional medicine. A method for quantification of (+)-catechin and (-)-epicatechin in P. burmanica stem bark from 12 different Thailand markets by reverse-phase high performance liquid chromatography (HPLC) was investigated and validated. The analysis was performed by a Shimadzu DGU-20A3 HPLC equipped with a Shimadzu SPD-M20A photo diode array detector. The separation was accomplished with an Inersil ODS-3 column (5 µm x 4.6 x 250 mm) using 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B) as mobile phase at the flow rate of 1 ml/min. The isocratic was set at 20% B for 15 min and the column temperature was maintained at 40 ºC. The detection was at the wavelength of 280 nm. Both (+)-catechin and (-)-epicatechin existed in the ethanolic extract of P. burmanica stem bark. The content of (-)-epicatechin was found as 59.74 ± 1.69 µg/mg of crude extract. In contrast, the quantitation of (+)-catechin content was omitted because of its small amount. The method was linear over a range of 5-200 µg/ml with good coefficients (r2 > 0.99) for (+)-catechin and (-)-epicatechin. Limit of detection values were found to be 4.80 µg/ml for (+)-catechin and 5.14 µg/ml for (-)-epicatechin. Limit of quantitation of (+)-catechin and (-)-epicatechin were of 14.54 µg/ml and 15.57 µg/ml respectively. Good repeatability and intermediate precision (%RSD < 3) were found in this study. The average recoveries of both (+)-catechin and (-)-epicatechin were obtained with good recovery in the range of 91.11 – 97.02% and 88.53 – 93.78%, respectively, with the %RSD less than 2. The peak purity indices of catechins were more than 0.99. The results suggested that HPLC method proved to be precise and accurate and the method can be conveniently used for (+)-catechin and (-)-epicatechin determination in ethanolic extract of P. burmanica stem bark. Moreover, the stem bark of P. burmanica was found to be a rich source of (-)-epicatechin.

Keywords: pentace burmanica, (+)-catechin, (-)-epicatechin, high performance liquid chromatography

Procedia PDF Downloads 455

Authors: Ruttayaporn Ngasaman, Saowakon Indouang, Usa Chethanond

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Leptospirosis is a public health concern zoonosis in Thailand. Human and animals are often infected by contact with contaminated water. The infected animals play an important role in leptospira infection for both human and other hosts via urine. In humans, it can cause a wide range of symptoms, some of which may present mild flu-like symptoms including fever, vomiting, and jaundice. Without treatment, Leptospirosis can lead to kidney damage, meningitis, liver failure, respiratory distress, and even death. The prevalence of leptospirosis in stray animals in Thailand is unknown. The aim of this study was to investigate leptospira infection in stray animals including dogs and cats in Songkhla province, Thailand. Total of 434 blood samples were collected from 370 stray dogs and 64 stray cats during the population control program from 2014 to 2018. Screening test using latex agglutination for the detection of antibodies against Leptospira interrogans in serum samples shows 29.26% (127/434) positive. There were 120 positive samples of stray dogs and 7 positive samples of stray cats. Detection by polymerase chain reaction specific to LipL32 gene of Leptospira interrogans showed 1.61% (7/434) positive. Stray cats (5/64) show higher prevalence than stray dogs (2/370). Although active infection was low detected, but seroprevalence was high. This result indicated that stray animals were not active infection during sample collection but they use to get infected or in a latent period of infection. They may act as a reservoir for domestic animals and human in which stay in the same environment. In order to prevent and reduce the risk of leptospira infection in a human, stray animals should be done health checking, vaccination, and disease treatment.

Keywords: leptospirosis, stray animals, risk reduction, Thailand

Procedia PDF Downloads 135

Authors: Arpit K. Shrivastava, Nirmal K. Mohakud, Subrat Kumar, Priyadarshi S. Sahu

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Acute diarrhea is one of the major causes of morbidity and mortality among children less than five years of age. Multiple etiologies have been implicated for infectious gastroenteritis causing acute diarrhea. In our study fecal samples (n=165) were collected from children (<5 years) presenting with symptoms of acute diarrhea. Samples were screened for viral, bacterial, and parasitic etiologies such as Rotavirus, Adenovirus, Diarrhoeagenic Escherichia coli (EPEC, EHEC, STEC, O157, O111), Shigella spp., Salmonella spp., Vibrio cholera, Cryptosporidium spp., and Giardia spp. The overall results from our study showed that 57% of children below 5 years of age with acute diarrhea were positive for at least one infectious etiology. Diarrhoeagenic Escherichia coli was detected to be the major etiological agent (29.09%) followed by Rotavirus (24.24%), Shigella (21.21%), Adenovirus (5.45%), Cryptosporidium (2.42%), and Giardia (0.60%). Among the different DEC strains, EPEC was detected significantly higher in <2 years children in comparison to >2 years age group (p =0.001). Concurrent infections with two or more pathogens were observed in 47 of 160 (28.48%) cases with a predominant incidence particularly in <2-year-old children (66.66%) compared to children of 2 to 5 years age group. Co-infection of Rotavirus with Shigella was the most frequent combination, which was detected in 17.94% cases, followed by Rotavirus with EPEC (15.38%) and Shigella with STEC (12.82%). Detection of multiple infectious etiologies and diagnosis of the right causative agent(s) can immensely help in better management of acute childhood diarrhea. In future more studies focusing on the detection of cases with concurrent infections must be carried out, as we believe that the etiological agents might be complementing each other’s strategies of pathogenesis resulting in severe diarrhea.

Keywords: children, co-infection, infectious diarrhea, Odisha

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Authors: Adel M. Al-Shutairi, Ahmed H. Al-Zahrani

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Tea is a popular beverage drunk by millions of people throughout the globe. Tea has considerable health advantages, in-cluding antioxidant, antibacterial, antiviral, chemopreventive, and anticarcinogenic properties. As a result of environmental pollution (atmospheric deposition) and the production process, tealeaves may also include a variety of dangerous substances, such as polycyclic aromatic hydrocarbons (PAHs). In this study, graphene oxide reinforced chitosan/poly-nitroaniline polymer was prepared to develop a sensitive and reliable solid phase extraction method (SPE) for extraction of PAH7 in tea samples, followed by high-performance liquid chromatography- fluorescence detection. The prepared adsorbent was validated in terms of linearity, the limit of detection, the limit of quantification, recovery (%), accuracy (%), and precision (%) for the determination of the PAH7 (benzo[a]pyrene, benzo[a]anthracene, benzo[b]fluoranthene, chrysene, benzo[b]fluoranthene, Dibenzo[a,h]anthracene and Benzo[g,h,i]perylene) in tea samples. The concentration was determined in two types of tea commercially available in Saudi Arabia, including black tea and green tea. The maximum mean of Σ7PAHs in black tea samples was 68.23 ± 0.02 ug kg-1 and 26.68 ± 0.01 ug kg-1 in green tea samples. The minimum mean of Σ7PAHs in black tea samples was 37.93 ± 0.01 ug kg-1 and 15.26 ± 0.01 ug kg-1 in green tea samples. The mean value of benzo[a]pyrene in black tea samples ranged from 6.85 to 12.17 ug kg-1, where two samples exceeded the standard level (10 ug kg-1) established by the European Union (UE), while in green tea ranged from 1.78 to 2.81 ug kg-1. Low levels of Σ7PAHs in green tea samples were detected in comparison with black tea samples.

Keywords: polycyclic aromatic hydrocarbons, CS, PNA and GO, black/green tea, solid phase extraction, Saudi Arabia

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Authors: Gajanan M. Sonwane

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Nowadays, the entire pharmaceutical industry is facing the challenge of increasing efficiency and innovation. The major hurdles are the growing cost of research and development and a concurrent stagnating number of new chemical entities (NCEs). Hence, the challenge is to select the most druggable targets and to search the equivalent drug-like compounds, which also possess specific pharmacokinetic and toxicological properties that allow them to be developed as drugs. The present research work includes the studies of developing new anticancer heterocycles by using molecular modeling techniques. The heterocycles synthesized through such methodology are much effective as various physicochemical parameters have been already studied and the structure has been optimized for its best fit in the receptor. Hence, on the basis of the literature survey and considering the need to develop newer anticancer agents, new phenazinamine derivatives were designed by subjecting the nucleus to molecular modeling, viz., GQSAR analysis and docking studies. Simultaneously, these designed derivatives were subjected to in silico prediction of biological activity through PASS studies and then in silico toxicity risk assessment studies. In PASS studies, it was found that all the derivatives exhibited a good spectrum of biological activities confirming its anticancer potential. The toxicity risk assessment studies revealed that all the derivatives obey Lipinski’s rule. Amongst these series, compounds 4c, 5b and 6c were found to possess logP and drug-likeness values comparable with the standard Imatinib (used for anticancer activity studies) and also with the standard drug methotrexate (used for antimitotic activity studies). One of the most notable mutations is the threonine to isoleucine mutation at codon 315 (T315I), which is known to be resistant to all currently available TKI. Enzyme assay planned for confirmation of target selective activity.

Keywords: drug design, tyrosine kinases, anticancer, Phenazinamine

Procedia PDF Downloads 117

Authors: Hsiao-Chuan Huang, King-Lung Kuo, Mei-Hsin Lo, Hsiao-Yun Chou, Yusen Lin

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The most robust and economical method for laboratory diagnosis of TB is to identify mycobacterial bacilli (AFB) under acid-fast staining despite its disadvantages of low sensitivity and labor-intensive. Though digital pathology becomes popular in medicine, an automated microscopic system for microbiology is still not available. A new AI-assisted automated microscopic system, consisting of a microscopic scanner and recognition program powered by big data and deep learning, may significantly increase the sensitivity of TB smear microscopy. Thus, the objective is to evaluate such an automatic system for the identification of AFB. A total of 5,930 smears was enrolled for this study. An intelligent microscope system (TB-Scan, Wellgen Medical, Taiwan) was used for microscopic image scanning and AFB detection. 272 AFB smears were used for transfer learning to increase the accuracy. Referee medical technicians were used as Gold Standard for result discrepancy. Results showed that, under a total of 1726 AFB smears, the automated system's accuracy, sensitivity and specificity were 95.6% (1,650/1,726), 87.7% (57/65), and 95.9% (1,593/1,661), respectively. Compared to culture, the sensitivity for human technicians was only 33.8% (38/142); however, the automated system can achieve 74.6% (106/142), which is significantly higher than human technicians, and this is the first of such an automated microscope system for TB smear testing in a controlled trial. This automated system could achieve higher TB smear sensitivity and laboratory efficiency and may complement molecular methods (eg. GeneXpert) to reduce the total cost for TB control. Furthermore, such an automated system is capable of remote access by the internet and can be deployed in the area with limited medical resources.

Keywords: TB smears, automated microscope, artificial intelligence, medical imaging

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Authors: Irene Alevizos, Jessica Lewis, Marina Harper, John Boyce

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Acinetobacter baumannii causes a range of severe nosocomial-acquired infections, and many strains are multi-drug resistant. A. baumannii possesses survival mechanisms allowing it to thrive in competitive polymicrobial environments, including a Type VI Secretion System (T6SS) that injects effector proteins into other bacteria to give a competitive advantage. The effects of T6SS firing are broad and depend entirely on the effector that is delivered. Effects can include toxicity against prokaryotic or eukaryotic cells and the acquisition of essential nutrients. The T6SS of some species can deliver ‘specialised effectors’ that are fused directly to T6SS components, such as PAAR proteins. PAAR proteins are predicted to form the piercing tip of the T6SS and are essential for T6SS function. Although no specialised effectors have been identified in A. baumannii, many strains encode multiple PAAR proteins. Analysis of PAAR proteins across the species identified 12 families of PAAR proteins with distinct C-terminal extensions. A. baumannii AB307-0294 encodes two PAAR proteins, one of which has a C-terminal extension. Mutation of one or both of the PAAR-encoding genes in this strain showed that expression of either PAAR protein was sufficient for T6SS function. We employed a heterologous expression approach and determined that PAAR proteins from different A. baumannii strains, as well as the closely related A. baylyi species, could complement the A. baumannii ∆paar mutant and restore T6SS function. Furthermore, we showed that PAAR fusions could be used to deliver artificially cloned protein fragments by generating Histidine- and Streptavidin- tagged PAAR specialised effectors, which restored T6SS activity. This provides evidence that the fusion of protein fragments onto PAAR proteins in A. baumannii is compatible with a functional T6SS. Successful delivery by this mechanism extends the scope of what the T6SS can deliver, including user designed proteins.

Keywords: A. baumannii, effectors, PAAR, T6SS

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Authors: I. Schäfer, B. Kohn, M. Volkmann, E. Müller

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Introduction: Blood-feeding arthropods transmit parasitic, bacterial, or viral pathogens to domestic animals and wildlife. Vector-borne infections are gaining significance due to the increase of travel, import of domestic animals from abroad, and the changing climate in Europe. Aims of the study: The main objective of this retrospective study was to assess the prevalence of vector-borne infections in cats in which a ‘Feline Travel Profile’ had been conducted. Material and Methods: This retrospective study included test results from cats for which a ‘Feline Travel Profile’ established by LABOKLIN had been requested by veterinarians between April 2012 and December 2019. This profile contains direct detection methods via polymerase chain reaction (PCR) for Hepatozoon spp. and Dirofilaria spp. as well as indirect detection methods via immunofluorescence antibody test (IFAT) for Ehrlichia spp. and Leishmania spp. This profile was expanded to include an IFAT for Rickettsia spp. from July 2015 onwards. The prevalence of the different vector-borne infectious agents was calculated. Results: A total of 602 cats were tested using the ‘Feline Travel Profile’. Positive test results were as follows: Rickettsia spp. IFAT 54/442 (12.2%), Ehrlichia spp. IFAT 68/602 (11.3%), Leishmania spp. IFAT 21/602 (3.5%), Hepatozoon spp. PCR 51/595 (8.6%), and Dirofilaria spp. PCR 1/595 cats (0.2%). Co-infections with more than one pathogen could be detected in 22/602 cats. Conclusions: 170/602 cats (28.2%) were tested positive for at least one vector-borne pathogen. Infections with multiple pathogens could be detected in 3.7% of the cats. The data emphasizes the importance of considering vector-borne infections as potential differential diagnoses in cats.

Keywords: arthopod-transmitted infections, feline vector-borne infections, Germany, laboratory diagnostics

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Authors: Mirco Tarozzi, Giacomo Pignagnoli, Andrea Benedetti

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Structural systems are often subjected to degradation processes due to different kind of phenomena like unexpected loadings, ageing of the materials and fatigue cycles. This is true especially for bridges, in which their safety evaluation is crucial for the purpose of a design of planning maintenance. This paper discusses the experimental evaluation of the stiffness reduction from frequency changes due to uniform damage scenario. For this purpose, a 1:4 scaled bridge has been built in the laboratory of the University of Bologna. It is made of concrete and its cross section is composed by a slab linked to four beams. This concrete deck is 6 m long and 3 m wide, and its natural frequencies have been identified dynamically by exciting it with an impact hammer, a dropping weight, or by walking on it randomly. After that, a set of loading cycles has been applied to this bridge in order to produce a uniformly distributed crack pattern. During the loading phase, either cracking moment and yielding moment has been reached. In order to define the relationship between frequency variation and loss in stiffness, the identification of the natural frequencies of the bridge has been performed, before and after the occurrence of the damage, corresponding to each load step. The behavior of breathing cracks and its effect on the natural frequencies has been taken into account in the analytical calculations. By using a sort of exponential function given from the study of lot of experimental tests in the literature, it has been possible to predict the stiffness reduction through the frequency variation measurements. During the load test also crack opening and middle span vertical displacement has been monitored.

Keywords: concrete bridge, damage detection, dynamic test, frequency shifts, operational modal analysis

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Authors: Keita Nishio, Takashi Kaburagi, Yosuke Kurihara

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Recent advances in medical technology have served to enhance average life expectancy. However, the total time for which the patients are prescribed complete bedrest has also increased. With patients being required to maintain a constant lying posture- also called bedsore- development of a system to detect patient roll-over becomes imperative. For this purpose, extant studies have proposed the use of cameras, and favorable results have been reported. Continuous on-camera monitoring, however, tends to violate patient privacy. We have proposed unconstrained bio-signal measurement system that could detect body-motion during sleep and does not violate patient’s privacy. Therefore, in this study, we propose a roll-over detection method by the date obtained from the bi-signal measurement system. Signals recorded by the sensor were assumed to comprise respiration, pulse, body motion, and noise components. Compared the body-motion and respiration, pulse component, the body-motion, during roll-over, generate large vibration. Thus, analysis of the body-motion component facilitates detection of the roll-over tendency. The large vibration associated with the roll-over motion has a great effect on the Root Mean Square (RMS) value of time series of the body motion component calculated during short 10 s segments. After calculation, the RMS value during each segment was compared to a threshold value set in advance. If RMS value in any segment exceeded the threshold, corresponding data were considered to indicate occurrence of a roll-over. In order to validate the proposed method, we conducted experiment. A bi-directional microphone was adopted as a high-sensitivity pressure sensor and was placed between the mattress and bedframe. Recorded signals passed through an analog Band-pass Filter (BPF) operating over the 0.16-16 Hz bandwidth. BPF allowed the respiration, pulse, and body-motion to pass whilst removing the noise component. Output from BPF was A/D converted with the sampling frequency 100Hz, and the measurement time was 480 seconds. The number of subjects and data corresponded to 5 and 10, respectively. Subjects laid on a mattress in the supine position. During data measurement, subjects—upon the investigator's instruction—were asked to roll over into four different positions—supine to left lateral, left lateral to prone, prone to right lateral, and right lateral to supine. Recorded data was divided into 48 segments with 10 s intervals, and the corresponding RMS value for each segment was calculated. The system was evaluated by the accuracy between the investigator’s instruction and the detected segment. As the result, an accuracy of 100% was achieved. While reviewing the time series of recorded data, segments indicating roll-over tendencies were observed to demonstrate a large amplitude. However, clear differences between decubitus and the roll-over motion could not be confirmed. Extant researches possessed a disadvantage in terms of patient privacy. The proposed study, however, demonstrates more precise detection of patient roll-over tendencies without violating their privacy. As a future prospect, decubitus estimation before and after roll-over could be attempted. Since in this paper, we could not confirm the clear differences between decubitus and the roll-over motion, future studies could be based on utilization of the respiration and pulse components.

Keywords: bedsore, high-sensitivity pressure sensor, roll-over, unconstrained bio-signal measurement

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Authors: Moez Elsaadani, Noel Durand, Brice Sorli, Didier Montet

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Governments and international instances are trying to improve the food safety system to prevent, reduce or avoid the increase of food borne diseases. This food risk is one of the major concerns for the humanity. The contamination by mycotoxins is a threat to the health and life of humans and animals. One of the most common mycotoxin contaminating feed and foodstuffs is Ochratoxin A (OTA), which is a secondary metabolite, produced by Aspergillus and Penicillium strains. OTA has a chronic toxic effect and proved to be mutagenic, nephrotoxic, teratogenic, immunosuppressive, and carcinogenic. On the other side, because of their high stability, specificity, affinity, and their easy chemical synthesis, aptamer based methods are applied to OTA biosensing as alternative to traditional analytical technique. In this work, five aptamers have been tested to confirm qualitatively and quantitatively their binding with OTA. In the same time, three different analytical methods were tested and compared based on their ability to detect and quantify the OTA. The best protocol that was established to quantify free OTA from linked OTA involved an ultrafiltration method in green coffee solution with. OTA was quantified by HPLC-FLD to calculate the binding percentage of all five aptamers. One aptamer (The most effective with 87% binding with OTA) has been selected to be our biorecognition element to study its electrical response (variation of electrical properties) in the presence of OTA in order to be able to make a pairing with a radio frequency identification (RFID). This device, which is characterized by its low cost, speed, and a simple wireless information transmission, will implement the knowledge on the mycotoxins molecular sensors (aptamers), an electronic device that will link the information, the quantification and make it available to operators.

Keywords: aptamer, aptasensor, detection, Ochratoxin A

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Authors: Nasim Hashemi, Abbas Sheikhtaheri

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Telemedicine is the use of telecommunication and information technologies to provide health care services that would often not be consistently available in distant rural communities to people at these remote areas. Teleophthalmology is a branch of telemedicine that delivers eye care through digital medical equipment and telecommunications technology. Thus, teleophthalmology can overcome geographical barriers and improve quality, access, and affordability of eye health care services. Since teleophthalmology has been widespread applied in recent years, the aim of this study was to determine the different applications of teleophthalmology in the world. To this end, three bibliographic databases (Medline, ScienceDirect, Scopus) were comprehensively searched with these keywords: eye care, eye health care, primary eye care, diagnosis, detection, and screening of different eye diseases in conjunction with telemedicine, telehealth, teleophthalmology, e-services, and information technology. All types of papers were included in the study with no time restriction. We conducted the search strategies until 2015. Finally 70 articles were surveyed. We classified the results based on the’type of eye problems covered’ and ‘the type of telemedicine services’. Based on the review, from the ‘perspective of health care levels’, there are three level for eye health care as primary, secondary and tertiary eye care. From the ‘perspective of eye care services’, the main application of teleophthalmology in primary eye care was related to the diagnosis of different eye diseases such as diabetic retinopathy, macular edema, strabismus and aged related macular degeneration. The main application of teleophthalmology in secondary and tertiary eye care was related to the screening of eye problems i.e. diabetic retinopathy, astigmatism, glaucoma screening. Teleconsultation between health care providers and ophthalmologists and also education and training sessions for patients were other types of teleophthalmology in world. Real time, store–forward and hybrid methods were the main forms of the communication from the perspective of ‘teleophthalmology mode’ which is used based on IT infrastructure between sending and receiving centers. In aspect of specialists, early detection of serious aged-related ophthalmic disease in population, screening of eye disease processes, consultation in an emergency cases and comprehensive eye examination were the most important benefits of teleophthalmology. Cost-effectiveness of teleophthalmology projects resulted from reducing transportation and accommodation cost, access to affordable eye care services and receiving specialist opinions were also the main advantages of teleophthalmology for patients. Teleophthalmology brings valuable secondary and tertiary care to remote areas. So, applying teleophthalmology for detection, treatment and screening purposes and expanding its use in new applications such as eye surgery will be a key tool to promote public health and integrating eye care to primary health care.

Keywords: applications, telehealth, telemedicine, teleophthalmology

Procedia PDF Downloads 375

Authors: Yuta Nakano, Kozo Kajiwara, Atsushi Hori, Takeshi Fujita

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In association with the wave of electric vehicles (EV), low energy consumption systems have become more and more important. One of the key technologies to realize low energy consumption is a dynamic vision sensor (DVS), or we can call it an event sensor, neuromorphic vision sensor and so on. This sensor has several features, such as high temporal resolution, which can achieve 1 Mframe/s, and a high dynamic range (120 DB). However, the point that can contribute to low energy consumption the most is its sparsity; to be more specific, this sensor only captures the pixels that have intensity change. In other words, there is no signal in the area that does not have any intensity change. That is to say, this sensor is more energy efficient than conventional sensors such as RGB cameras because we can remove redundant data. On the other side of the advantages, it is difficult to handle the data because the data format is completely different from RGB image; for example, acquired signals are asynchronous and sparse, and each signal is composed of x-y coordinate, polarity (two values: +1 or -1) and time stamp, it does not include intensity such as RGB values. Therefore, as we cannot use existing algorithms straightforwardly, we have to design a new processing algorithm to cope with DVS data. In order to solve difficulties caused by data format differences, most of the prior arts make a frame data and feed it to deep learning such as Convolutional Neural Networks (CNN) for object detection and recognition purposes. However, even though we can feed the data, it is still difficult to achieve good performance due to a lack of intensity information. Although polarity is often used as intensity instead of RGB pixel value, it is apparent that polarity information is not rich enough. Considering this context, we proposed to use the timestamp information as a data representation that is fed to deep learning. Concretely, at first, we also make frame data divided by a certain time period, then give intensity value in response to the timestamp in each frame; for example, a high value is given on a recent signal. We expected that this data representation could capture the features, especially of moving objects, because timestamp represents the movement direction and speed. By using this proposal method, we made our own dataset by DVS fixed on a parked car to develop an application for a surveillance system that can detect persons around the car. We think DVS is one of the ideal sensors for surveillance purposes because this sensor can run for a long time with low energy consumption in a NOT dynamic situation. For comparison purposes, we reproduced state of the art method as a benchmark, which makes frames the same as us and feeds polarity information to CNN. Then, we measured the object detection performances of the benchmark and ours on the same dataset. As a result, our method achieved a maximum of 7 points greater than the benchmark in the F1 score.

Keywords: event camera, dynamic vision sensor, deep learning, data representation, object recognition, low energy consumption

Procedia PDF Downloads 101

Authors: F. M. Pisano, M. Ciminello

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Aerospace, mechanical, and civil engineering infrastructures can take advantages from damage detection and identification strategies in terms of maintenance cost reduction and operational life improvements, as well for safety scopes. The challenge is to detect so called “barely visible impact damage” (BVID), due to low/medium energy impacts, that can progressively compromise the structure integrity. The occurrence of any local change in material properties, that can degrade the structure performance, is to be monitored using so called Structural Health Monitoring (SHM) systems, in charge of comparing the structure states before and after damage occurs. SHM seeks for any "anomalous" response collected by means of sensor networks and then analyzed using appropriate algorithms. Independently of the specific analysis approach adopted for structural damage detection and localization, textual reports, tables and graphs describing possible outlier coordinates and damage severity are usually provided as artifacts to be elaborated for information extraction about the current health conditions of the structure under investigation. Visual Analytics can support the processing of monitored measurements offering data navigation and exploration tools leveraging the native human capabilities of understanding images faster than texts and tables. Herein, a SHM system enrichment by integration of a Visual Analytics component is investigated. Analytical dashboards have been created by combining worksheets, so that a useful Visual Analytics tool is provided to structural analysts for exploring the structure health conditions examined by a Principal Component Analysis based algorithm.

Keywords: interactive dashboards, optical fibers, structural health monitoring, visual analytics

Procedia PDF Downloads 125

Authors: Masoud Soltanialvar, Ali Bagherpour

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Documented cases of human infection with H9N2 avian influenza viruses, first detected in 1999 in Hong Kong and China, indicate that these viruses can be directly transmitted from birds to humans. In this study, we characterized the mutation in the Hemagglutinin (HA) genes and proteins that correlates with a shift in affinity of the Hemagglutinin (HA) protein from the “avian” type sialic receptors to the “human” type in 10 Iranian isolates. We delineated the genomes and receptor binding profile of HA gene of some field isolates and established their phylogenetic relationship to the other Asian H9N2 sub lineages. A total of 1200 tissue samples collected from 40 farms located in various states of Iran during 2008 – 2010 as part of a program to monitor Avian Influenza Viruses (AIV) infection. To determine the genetic relationship of Iranian viruses, the Hemagglutinin (HA) genes from ten isolates were amplified and sequenced (by RT-PCR method). Nucleotide sequences (orf) of the (HA) genes were used for phylogenetic tree construction. Deduced amino acid sequences showed the presence of L226 (234 in H9 numbering) in all ten Iranian isolates which indicates a preference to binding of α (2–6) sialic acid receptors, so these Iranian H9N2 viruses have the potential to infect human beings. These isolates showed high degree of homology with 2 human H9N2 isolates A/HK/1073/99, A/HK/1074/99. Phylogenetic analysis of showed that all the HA genes of the Iranian H9N2 viruses fall into a single group within a G1-like sublineage which had contributed as donor of six internal genes to H5N1 highly pathogenic avian influenza. The results of this study indicated that all Iranian viruses have the potential to emerge as highly pathogenic influenza virus, and considering the homology of these isolates with human H9N2 strains, it seems that the potential of these avian influenza isolates to infect human should not be overlooked.

Keywords: influenza virus, hemagglutinin, neuraminidase, Iran

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Authors: Samah Jawad Jabari

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Water is the most important and valuable resource not only for human life but also for all living things on the planet. The water supply utilities should fulfill the water requirement quantitatively and qualitatively. Drinking water systems are exposed to both natural (hurricanes and flood) and manmade hazards (risks) that are common in Palestine. Non-Revenue Water (NRW) is a manmade risk which remains a major concern in Palestine, as the NRW levels are estimated to be at a high level. In this research, Hebron city water distribution network was taken as a case study to estimate and audit the NRW levels. The research also investigated the state of the existing water distribution system in the study area by investigating the water losses and obtained more information on NRW prevention and management practices. Data and information have been collected from the Palestinian Water Authority (PWA) and Hebron Municipality (HM) archive. In addition to that, a questionnaire has been designed and administered by the researcher in order to collect the necessary data for water auditing. The questionnaire also assessed the views of stakeholder in PWA and HM (staff) on the current status of the NRW in the Hebron water distribution system. The important result obtained by this research shows that NRW in Hebron city was high and in excess of 30%. The main factors that contribute to NRW were the inaccuracies in billing volumes, unauthorized consumption, and the method of estimating consumptions through faulty meters. Policy for NRW reduction is available in Palestine; however, it is clear that the number of qualified staff available to carry out the activities related to leak detection is low, and that there is a lack of appropriate technologies to reduce water losses and undertake sufficient system maintenance, which needs to be improved to enhance the performance of the network and decrease the level of NRW losses.

Keywords: non-revenue water, water auditing, leak detection, water meters

Procedia PDF Downloads 299

Authors: Sung-Hyeon Park, Jin-Wook Jang, Hak-Sung Kim

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In recent years, Terahertz (THz) time-domain spectroscopy (TDS) imaging method has been received considerable interest as a promising non-destructive technique for detection of internal defects. In comparison to other non-destructive techniques such as x-ray inspection method, scanning acoustic tomograph (SAT) and microwave inspection method, THz-TDS imaging method has many advantages: First, it can measure the exact thickness and location of defects. Second, it doesn’t require the liquid couplant while it is very crucial to deliver that power of ultrasonic wave in SAT method. Third, it didn’t damage to materials and be harmful to human bodies while x-ray inspection method does. Finally, it exhibits better spatial resolution than microwave inspection method. However, this technology couldn’t be applied to IC package because THz radiation can penetrate through a wide variety of materials including polymers and ceramics except of metals. Therefore, it is difficult to detect the defects in IC package which are composed of not only epoxy and semiconductor materials but also various metals such as copper, aluminum and gold. In this work, we proposed a special method for detecting the void in the IC package using THz-TDS imaging system. The IC package specimens for this study are prepared by Packaging Engineering Team in Samsung Electronics. Our THz-TDS imaging system has a special reflection mode called pitch-catch mode which can change the incidence angle in the reflection mode from 10 o to 70 o while the others have transmission and the normal reflection mode or the reflection mode fixed at certain angle. Therefore, to find the voids in the IC package, we investigated the appropriate angle as changing the incidence angle of THz wave emitter and detector. As the results, the voids in the IC packages were successfully detected using our THz-TDS imaging system.

Keywords: terahertz, non-destructive technique, void, IC package

Procedia PDF Downloads 474

Authors: K. Sathishkumar, V. Thiagarasu

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Due to recent advances in DNA microarray technology, it is now feasible to obtain gene expression profiles of tissue samples at relatively low costs. Many scientists around the world use the advantage of this gene profiling to characterize complex biological circumstances and diseases. Microarray techniques that are used in genome-wide gene expression and genome mutation analysis help scientists and physicians in understanding of the pathophysiological mechanisms, in diagnoses and prognoses, and choosing treatment plans. DNA microarray technology has now made it possible to simultaneously monitor the expression levels of thousands of genes during important biological processes and across collections of related samples. Elucidating the patterns hidden in gene expression data offers a tremendous opportunity for an enhanced understanding of functional genomics. However, the large number of genes and the complexity of biological networks greatly increase the challenges of comprehending and interpreting the resulting mass of data, which often consists of millions of measurements. A first step toward addressing this challenge is the use of clustering techniques, which is essential in the data mining process to reveal natural structures and identify interesting patterns in the underlying data. This work presents an analysis of several clustering algorithms proposed to deals with the gene expression data effectively. The existing clustering algorithms like Support Vector Machine (SVM), K-means algorithm and evolutionary algorithm etc. are analyzed thoroughly to identify the advantages and limitations. The performance evaluation of the existing algorithms is carried out to determine the best approach. In order to improve the classification performance of the best approach in terms of Accuracy, Convergence Behavior and processing time, a hybrid clustering based optimization approach has been proposed.

Keywords: microarray technology, gene expression data, clustering, gene Selection

Procedia PDF Downloads 325

Authors: Dharshika Rajalingam, Jeffery W. Peng

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Acinetobacter baumannii is a challenging threat to global health, recognized as a multidrug-resistant pathogen. -lactamase is one of the principal resistant mechanisms developed by A. baumannii to survive against -lactam antibiotics. OXA24/40 is one of the types of -lactamases, a well-documented carbapenem hydrolyzing class D -lactamases (CHDL). It was revealed that OXA24/40 showed resistivity against doripenem, one of the carbapenems, by two different mechanisms as hydrolysis and -lactonization. Furthermore, it undergoes genetic mutations to broaden the -lactamase activity to survive against antibiotic environments. One of the crucial characterizations of prokaryotes to develop adaptation is post-translational modification (PTM), mainly phosphorylation. However, the PTM of OXA24/40 is an unknown feature, and the impact of PTM on antibiotic resistivity is yet to be explored. We approached these hypotheses using NMR and MS techniques and found that the OXA24/40 could be phosphorylated in vitro. The Ser81 at the active STFK motif of OXA24/40 of catalytic pocket was identified as the site of phosphorylation using 1D 31P NMR experiment, whereas S81 is required to form an acyl-enzyme complex between enzyme and -lactam antibiotics. The activity of completely phosphorylated OXA24/40 wild type against doripenem revealed that the phosphorylation of active Ser inactivates the -lactamases activity of OXA24/40. The 1D 1H CPMG NMR-based activity assay of phosphorylated OXA24/40 against doripenem confirmed that both deactivating mechanisms are inhibited by phosphorylation. Carbamylated Lysine at the active STFK motif is one of the critical features of CHDL required for the acylation and deacylation reactions of the enzyme. The 1D 13C NMR experiment confirmed that the K84 of phosphorylated OXA24/40 is de-carbamylated. Phosphorylation of OXA24/40 affects both active S81 and carbamylated K84 of OXA24 that are required for the resistivity of -lactamase. So, phosphorylation could be one of the reasons for the genetic mutation of OXA24/40 for the development of antibiotic resistivity. Further research can lead to an understanding of the effect of phosphorylation on the clinical mutants of the OXA24-like -lactamase family on the broadening of -lactamase activity.

Keywords: OXA24/40, phosphorylation, clinical mutants, resistivity

Procedia PDF Downloads 79

Authors: Béla Kovács, Éva Bódi, Farzaneh Garousi, Szilvia Várallyay, Áron Soós, Xénia Vágó, Dávid Andrási

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In agriculture for analysis of elements in different food and food raw materials, moreover environmental samples generally flame atomic absorption spectrometers (FAAS), graphite furnace atomic absorption spectrometers (GF-AAS), inductively coupled plasma optical emission spectrometers (ICP-OES) and inductively coupled plasma mass spectrometers (ICP-MS) are routinely applied. An inductively coupled plasma mass spectrometer (ICP-MS) is capable for analysis of 70-80 elements in multielemental mode, from 1-5 cm3 volume of a sample, moreover the detection limits of elements are in µg/kg-ng/kg (ppb-ppt) concentration range. All the analytical instruments have different physical and chemical interfering effects analysing the above types of samples. The smaller the concentration of an analyte and the larger the concentration of the matrix the larger the interfering effects. Nowadays there is very important to analyse growingly smaller concentrations of elements. From the above analytical instruments generally the inductively coupled plasma mass spectrometer is capable of analysing the smallest concentration of elements. The applied ICP-MS instrument has Collision Cell Technology (CCT) also. Using CCT mode certain elements have better (smaller) detection limits with 1-3 magnitudes comparing to a normal ICP-MS analytical method. The CCT mode has better detection limits mainly for analysis of selenium, arsenic, germanium, vanadium and chromium. To elaborate an analytical method for trace elements with an inductively coupled plasma mass spectrometer the most important interfering effects (problems) were evaluated: 1) Physical interferences; 2) Spectral interferences (elemental and molecular isobaric); 3) Effect of easily ionisable elements; 4) Memory interferences. Analysing food and food raw materials, moreover environmental samples an other (new) interfering effect emerged in ICP-MS, namely the effect of various matrixes having different evaporation and nebulization effectiveness, moreover having different quantity of carbon content of food and food raw materials, moreover environmental samples. In our research work the effect of different water-soluble compounds furthermore the effect of various quantity of carbon content (as sample matrix) were examined on changes of intensity of the applied elements. So finally we could find “opportunities” to decrease or eliminate the error of the analyses of applied elements (Cr, Co, Ni, Cu, Zn, Ge, As, Se, Mo, Cd, Sn, Sb, Te, Hg, Pb, Bi). To analyse these elements in the above samples, the most appropriate inductively coupled plasma mass spectrometer is a quadrupole instrument applying a collision cell technique (CCT). The extent of interfering effect of carbon content depends on the type of compounds. The carbon content significantly affects the measured concentration (intensities) of the above elements, which can be corrected using different internal standards.

Keywords: elements, environmental and food samples, ICP-MS, interference effects

Procedia PDF Downloads 504