Search results for: Loop-Mediated Isothermal Amplification (LAMP)
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 537

Search results for: Loop-Mediated Isothermal Amplification (LAMP)

537 Utility of the Loop-Mediated Isothermal Amplification Assay for the Diagnosis of Visceral Leishmaniasis from Blood Samples in Ethiopia

Authors: Dawit Gebreegzabher Hagos, Yazezew Kebede Kiro, Mahmud Abdulkader, Henk H. D. F. Schallig, Dawit Wolday

Abstract:

Rapid and accurate visceral leishmaniasis (VL) diagnosis is needed to initiate prompt treatment to reduce morbidity and mortality. Here, we evaluated the performance of loop-mediated isothermal amplification (LAMP) assay for the diagnosis of VL from blood in an endemic area in Ethiopia. LAMP was positive in 117/122 confirmed VL cases and negative in 149/152 controls, resulting in a sensitivity of 95.9% (95% CI: 90.69–98.66) and a specificity of 98.0% (95% CI: 94.34–99.59), respectively. The sensitivity of the LAMP assay was 95.0% (95% CI: 88.61–98.34) in HIV-negatives and 100% (95% CI: 85.18–100.0) in HIV-positives. Compared with microscopy, LAMP detected 82/87 (94.3%, 95% CI: 87.10–98.11) of the microscopy1 cases and was negative in 11/27 (40.7%, 95% CI: 22.39–61.20) of the microscopy2 cases. Compared with the rK39 serology, LAMP detected 113/120 (94.2%, 95% CI: 88.35–97.62) of the rK391 cases and was negative in 149/154 (96.8%, 95% CI: 92.59–98.94) of the rK392 cases. However, when compared with microscopy only, rK39 detected 83/87 (95.4%, 95% CI: 88.64–98.73) of the microscopy1 cases and negative in only 12/27 (44.4%, 95% CI: 25.48–64.67) of the microscopy– cases. There was an excellent agreement between rK39 and LAMP (Kappa 5 0.91, 95% CI: 0.86–0.96). Furthermore, an algorithm using rK39 followed by LAMP would yield a sensitivity of 99.2% (95%CI: 95.52–99.89) and a specificity of 98.0% (95% CI: 94.34–99.59). The findings demonstrate that the LAMP assay is an accurate and rapid molecular assay for VL diagnosis, including in HIV-1 co-infected patients, in an endemic setting.

Keywords: visceral leishmaniasis, HIV, diagnosis, LAMP, Ethiopia

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536 Development of Loop-Mediated Isothermal Amplification for Detection of Garlic in Food

Authors: Ting-Ying Su, Meng-Shiou Lee, Shyang-Chwen Sheu

Abstract:

Garlic is used commonly as a seasoning around the world. But some people suffer from allergy to garlic. Garlic may also cause burning of mouth, stomach, and throat. In some Buddhist traditions, consuming garlic is not allowed. The objective of this study is to develop a LAMP based method for detection of garlic in food. We designed specific primers targeted on ITS1-5.8S rRNA-ITS2 sequence of garlic DNA. The LAMP assay was performed using a set of four different primers F3, B3, FIP and BIP at 60˚C in less than 60 mins. Results showed that the primer was not cross-reactive to other commonly used spice including Chinese leek, Chinese onion, green onion, onion, pepper, basil, parsley, pepper and ginger. As low as 2% of garlic DNA could be detected. Garlic still could be detected by developed LAMP after boiled at 100˚C for 80 minutes and autoclaved at 121˚C for 60 minutes. Commercial products labeled with garlic ingredient could be identified by the developed method.

Keywords: garlic, loop-mediated isothermal amplification, processing, DNA

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535 Real-Time Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Human Papillomavirus 16 in Oral Squamous Cell Carcinoma

Authors: Suharni Mohamad Suharni Mohamad, Nurul Izzati Hamzan Nurul Izzati Hamzan, Norhayu Abdul Rahman Norhayu Abdul Rahman, Siti Suraiya Md Noor Siti Suraiya Md Noor

Abstract:

Human papillomavirus (HPV) is an important risk factor for development of oral cancer. HPV16 is the most common type found in HPV-positive squamous cell carcinoma. In the present study, we established a real-time loop-mediated isothermal amplification (real-time LAMP) for detection of HPV16. A set of six primers was specially designed to recognize eight distinct sequences of HPV16-E6. Detection and quantification was achieved by real-time monitoring using a real-time turbidimeter based on threshold time required for turbidity in the LAMP reaction. LAMP reagents (MgSO4, dNTPs, Bst polymerase concentrations) and various incubation times and temperatures were optimized. The sensitivity was determined using 10-fold serial dilutions of HPV16 standard strain. The specificity of was evaluated using other HPV genotypes. The optimized method was established with specifically designed primers by real-time detection in approximately 30 min at 65°C. The limit of detection of HPV16 using the LAMP assay was 10 pg/ml that could be detected in 30 min. The LAMP assay was 10 times more sensitive than the conventional PCR in detecting HPV16. No cross-reactivity with other HPV genotypes was observed. This quantitative real-time LAMP assay may improve diagnostic potential for the detection and quantification of HPV16 in clinical samples and epidemiological studies due to its rapidity, simplicity, high sensitivity and specificity. This assay will be further evaluated with HPV DNAs of saliva from patients with oral squamous cell carcinoma. Acknowledgement: This study was financially supported by the ScienceFund Grant, Ministry of Science, Technology and Innovation (305/PPSG/6113219).

Keywords: Oral Squamous Cell Carcinoma (OSCC), Human Papillomavirus 16 (HPV16), Loop-Mediated Isothermal Amplification (LAMP), rapid detection

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534 Detection of Aflatoxin B1 Producing Aspergillus flavus Genes from Maize Feed Using Loop-Mediated Isothermal Amplification (LAMP) Technique

Authors: Sontana Mimapan, Phattarawadee Wattanasuntorn, Phanom Saijit

Abstract:

Aflatoxin contamination in maize, one of several agriculture crops grown for livestock feeding, is still a problem throughout the world mainly under hot and humid weather conditions like Thailand. In this study Aspergillus flavus (A. Flavus), the key fungus for aflatoxin production especially aflatoxin B1 (AFB1), isolated from naturally infected maize were identified and characterized according to colony morphology and PCR using ITS, Beta-tubulin and calmodulin genes. The strains were analysed for the presence of four aflatoxigenic biosynthesis genes in relation to their capability to produce AFB1, Ver1, Omt1, Nor1, and aflR. Aflatoxin production was then confirmed using immunoaffinity column technique. A loop-mediated isothermal amplification (LAMP) was applied as an innovative technique for rapid detection of target nucleic acid. The reaction condition was optimized at 65C for 60 min. and calcein flurescent reagent was added before amplification. The LAMP results showed clear differences between positive and negative reactions in end point analysis under daylight and UV light by the naked eye. In daylight, the samples with AFB1 producing A. Flavus genes developed a yellow to green color, but those without the genes retained the orange color. When excited with UV light, the positive samples become visible by bright green fluorescence. LAMP reactions were positive after addition of purified target DNA until dilutions of 10⁻⁶. The reaction products were then confirmed and visualized with 1% agarose gel electrophoresis. In this regards, 50 maize samples were collected from dairy farms and tested for the presence of four aflatoxigenic biosynthesis genes using LAMP technique. The results were positive in 18 samples (36%) but negative in 32 samples (64%). All of the samples were rechecked by PCR and the results were the same as LAMP, indicating 100% specificity. Additionally, when compared with the immunoaffinity column-based aflatoxin analysis, there was a significant correlation between LAMP results and aflatoxin analysis (r= 0.83, P < 0.05) which suggested that positive maize samples were likely to be a high- risk feed. In conclusion, the LAMP developed in this study can provide a simple and rapid approach for detecting AFB1 producing A. Flavus genes from maize and appeared to be a promising tool for the prediction of potential aflatoxigenic risk in livestock feedings.

Keywords: Aflatoxin B1, Aspergillus flavus genes, maize, loop-mediated isothermal amplification

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533 Duplex Real-Time Loop-Mediated Isothermal Amplification Assay for Simultaneous Detection of Beef and Pork

Authors: Mi-Ju Kim, Hae-Yeong Kim

Abstract:

Product mislabeling and adulteration have been increasing the concerns in processed meat products. Relatively inexpensive pork meat compared to meat such as beef was adulterated for economic benefit. These food fraud incidents related to pork were concerned due to economic, religious and health reasons. In this study, a rapid on-site detection method using loop-mediated isothermal amplification (LAMP) was developed for the simultaneous identification of beef and pork. Each specific LAMP primer for beef and pork was designed targeting on mitochondrial D-loop region. The LAMP assay reaction was performed at 65 ℃ for 40 min. The specificity of each primer for beef and pork was evaluated using DNAs extracted from 13 animal species including beef and pork. The sensitivity of duplex LAMP assay was examined by serial dilution of beef and pork DNAs, and reference binary mixtures. This assay was applied to processed meat products including beef and pork meat for monitoring. Each set of primers amplified only the targeted species with no cross-reactivity with animal species. The limit of detection of duplex real-time LAMP was 1 pg for each DNA of beef and pork and 1% pork in a beef-meat mixture. Commercial meat products that declared the presence of beef and/or pork meat on the label showed positive results for those species. This method was successfully applied to detect simultaneous beef and pork meats in processed meat products. The optimized duplex LAMP assay can identify simultaneously beef and pork meat within less than 40 min. A portable real-time fluorescence device used in this study is applicable for on-site detection of beef and pork in processed meat products. Thus, this developed assay was considered to be an efficient tool for monitoring meat products.

Keywords: beef, duplex real-time LAMP, meat identification, pork

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532 Isothermal Solid-Phase Amplification System for Detection of Yersinia pestis

Authors: Olena Mayboroda, Angel Gonzalez Benito, Jonathan Sabate Del Rio, Marketa Svobodova, Sandra Julich, Herbert Tomaso, Ciara K. O'Sullivan, Ioanis Katakis

Abstract:

DNA amplification is required for most molecular diagnostic applications but conventional PCR has disadvantages for field testing. Isothermal amplification techniques are being developed to respond to this problem. One of them is the Recombinase Polymerase Amplification (RPA) that operates at isothermal conditions without sacrificing specificity and sensitivity in easy-to-use formats. In this work RPA was used for the optical detection of solid-phase amplification of the potential biowarfare agent Yersinia pestis. Thiolated forward primers were immobilized on the surface of maleimide-activated microtitre plates for the quantitative detection of synthetic and genomic DNA, with elongation occurring only in the presence of the specific template DNA and solution phase reverse primers. Quantitative detection was achieved via the use of biotinylated reverse primers and post-amplification addition of streptavidin-HRP conjugate. The overall time of amplification and detection was less than 1 hour at a constant temperature of 37oC. Single-stranded and double-stranded DNA sequences were detected achieving detection limits of 4.04*10-13 M and 3.14*10-16 M, respectively. The system demonstrated high specificity with negligible responses to non-specific targets.

Keywords: recombinase polymerase amplification, Yersinia pestis, solid-phase detection, ELONA

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531 Development of Loop Mediated Isothermal Amplification (Lamp) Assay for the Diagnosis of Ovine Theileriosis

Authors: Muhammad Fiaz Qamar, Uzma Mehreen, Muhammad Arfan Zaman, Kazim Ali

Abstract:

Ovine Theileriosis is a world-wide concern, especially in tropical and subtropical areas, due to having tick abundance that has received less awareness in different developed and developing areas due to less worth of sheep, low to the middle level of infection in different small ruminants herd. Across Asia, the prevalence reports have been conducted to provide equivalent calculation of flock and animal level prevalence of Theileriosisin animals. It is a challenge for veterinarians to timely diagnosis & control of Theileriosis and famers because of the nature of the organism and inadequacy of restricted plans to control. All most work is based upon the development of such a technique which should be farmer-friendly, less expensive, and easy to perform into the field. By the timely diagnosis of this disease will decrease the irrational use of the drugs, and other plan was to determine the prevalence of Theileriosis in District Jhang by using the conventional method, PCR and qPCR, and LAMP. We quantify the molecular epidemiology of T.lestoquardiin sheep from Jhang districts, Punjab, Pakistan. In this study, we concluded that the overall prevalence of Theileriosis was (32/350*100= 9.1%) in sheep by using Giemsa staining technique, whereas (48/350*100= 13%) is observed by using PCR technique (56/350*100=16%) in qPCR and the LAMP technique have shown up to this much prevalence percentage (60/350*100= 17.1%). The specificity and sensitivity also calculated in comparison with the PCR and LAMP technique. Means more positive results have been shown when the diagnosis has been done with the help of LAMP. And there is little bit of difference between the positive results of PCR and qPCR, and the least positive animals was by using Giemsa staining technique/conventional method. If we talk about the specificity and sensitivity of the LAMP as compared to PCR, The cross tabulation shows that the results of sensitivity of LAMP counted was 94.4%, and specificity of LAMP counted was 78%. Advances in scientific field must be upon reality based ideas which can lessen the gaps and hurdles in the way of scientific research; the lamp is one of such techniques which have done wonders in adding value and helping human at large. It is such a great biological diagnostic tools and has helped a lot in the proper diagnosis and treatment of certain diseases. Other methods for diagnosis, such as culture techniques and serological techniques, have exposed humans with great danger. However, with the help of molecular diagnostic technique like LAMP, exposure to such pathogens is being avoided in the current era Most prompt and tentative diagnosis can be made using LAMP. Other techniques like PCR has many disadvantages when compared to LAMP as PCR is a relatively expensive, time consuming, and very complicated procedure while LAMP is relatively cheap, easy to perform, less time consuming, and more accurate. LAMP technique has removed hurdles in the way of scientific research and molecular diagnostics, making it approachable to poor and developing countries.

Keywords: distribution, thelaria, LAMP, primer sequences, PCR

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530 Reflector Arrangement Effect on Ultraviolet Lamp Performance by CFX Simulation

Authors: William Sidharta, Chin-Tu Lu

Abstract:

Fluorescent ultraviolet lamp generates ultraviolet light which is commonly used in industrial field with certain purposes especially for curing process. Due to the value of inefficiency, there are changes in energy from electrical energy to the heat energy and this would make a defect on the industrial product caused by high temperature of lamp tube during ultraviolet light emission. The condition of industrial scale is further worsening, since commonly using dozens of fluorescent ultraviolet lamps to support huge production process and then it will generates much more heat energy. The maximum temperature of fluorescent ultraviolet lamp will get affected by arranging the lamp tube reflector and this study presents CFX simulation results of the maximum lamp tube temperature with some different reflector arrangements on purely natural convection phenomena. There exists certain spaces value of the reflector and the lamp tube to obtaining lower maximum temperature of the fluorescent ultraviolet lamp.

Keywords: CFX simulation, fluorescent UV lamp, lamp tube reflector, UV light

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529 A Microfluidic Biosensor for Detection of EGFR 19 Deletion Mutation Targeting Non-Small Cell Lung Cancer on Rolling Circle Amplification

Authors: Ji Su Kim, Bo Ram Choi, Ju Yeon Cho, Hyukjin Lee

Abstract:

Epidermal growth factor receptor (EGFR) 19 deletion mutation gene is over-expressed in carcinoma patient. EGFR 19 deletion mutation is known as typical biomarker of non-small cell lung cancer (NSCLC), which one section in the coding exon 19 of EGFR is deleted. Therefore, there have been many attempts over the years to detect EGFR 19 deletion mutation for replacing conventional diagnostic method such as PCR and tissue biopsy. We developed a simple and facile detection platform based on Rolling Circle Amplification (RCA), which provides highly amplified products in isothermal amplification of the ligated DNA template. Limit of detection (~50 nM) and a faster detection time (~30 min) could be achieved by introducing RCA.

Keywords: EGFR19, cancer, diagnosis, rolling circle amplification (RCA), hydrogel

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528 Design of DNA Origami Structures Using LAMP Products as a Combined System for the Detection of Extended Spectrum B-Lactamases

Authors: Kalaumari Mayoral-Peña, Ana I. Montejano-Montelongo, Josué Reyes-Muñoz, Gonzalo A. Ortiz-Mancilla, Mayrin Rodríguez-Cruz, Víctor Hernández-Villalobos, Jesús A. Guzmán-López, Santiago García-Jacobo, Iván Licona-Vázquez, Grisel Fierros-Romero, Rosario Flores-Vallejo

Abstract:

The group B-lactamic antibiotics include some of the most frequently used small drug molecules against bacterial infections. Nevertheless, an alarming decrease in their efficacy has been reported due to the emergence of antibiotic-resistant bacteria. Infections caused by bacteria expressing extended Spectrum B-lactamases (ESBLs) are difficult to treat and account for higher morbidity and mortality rates, delayed recovery, and high economic burden. According to the Global Report on Antimicrobial Resistance Surveillance, it is estimated that mortality due to resistant bacteria will ascend to 10 million cases per year worldwide. These facts highlight the importance of developing low-cost and readily accessible detection methods of drug-resistant ESBLs bacteria to prevent their spread and promote accurate and fast diagnosis. Bacterial detection is commonly done using molecular diagnostic techniques, where PCR stands out for its high performance. However, this technique requires specialized equipment not available everywhere, is time-consuming, and has a high cost. Loop-Mediated Isothermal Amplification (LAMP) is an alternative technique that works at a constant temperature, significantly decreasing the equipment cost. It yields double-stranded DNA of several lengths with repetitions of the target DNA sequence as a product. Although positive and negative results from LAMP can be discriminated by colorimetry, fluorescence, and turbidity, there is still a large room for improvement in the point-of-care implementation. DNA origami is a technique that allows the formation of 3D nanometric structures by folding a large single-stranded DNA (scaffold) into a determined shape with the help of short DNA sequences (staples), which hybridize with the scaffold. This research aimed to generate DNA origami structures using LAMP products as scaffolds to improve the sensitivity to detect ESBLs in point-of-care diagnosis. For this study, the coding sequence of the CTM-X-15 ESBL of E. coli was used to generate the LAMP products. The set of LAMP primers were designed using PrimerExplorerV5. As a result, a target sequence of 200 nucleotides from CTM-X-15 ESBL was obtained. Afterward, eight different DNA origami structures were designed using the target sequence in the SDCadnano and analyzed with CanDo to evaluate the stability of the 3D structures. The designs were constructed minimizing the total number of staples to reduce costs and complexity for point-of-care applications. After analyzing the DNA origami designs, two structures were selected. The first one was a zig-zag flat structure, while the second one was a wall-like shape. Given the sequence repetitions in the scaffold sequence, both were able to be assembled with only 6 different staples each one, ranging between 18 to 80 nucleotides. Simulations of both structures were performed using scaffolds of different sizes yielding stable structures in all the cases. The generation of the LAMP products were tested by colorimetry and electrophoresis. The formation of the DNA structures was analyzed using electrophoresis and colorimetry. The modeling of novel detection methods through bioinformatics tools allows reliable control and prediction of results. To our knowledge, this is the first study that uses LAMP products and DNA-origami in combination to delect ESBL-producing bacterial strains, which represent a promising methodology for diagnosis in the point-of-care.

Keywords: beta-lactamases, antibiotic resistance, DNA origami, isothermal amplification, LAMP technique, molecular diagnosis

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527 Product Form Bionic Design Based on Eye Tracking Data: A Case Study of Desk Lamp

Authors: Huan Lin, Liwen Pang

Abstract:

In order to reduce the ambiguity and uncertainty of product form bionic design, a product form bionic design method based on eye tracking is proposed. The eye-tracking experiment is designed to calculate the average time ranking of the specific parts of the bionic shape that the subjects are looking at. Key bionic shape is explored through the experiment and then applied to a desk lamp bionic design. During the design case, FAHP (Fuzzy Analytic Hierachy Process) and SD (Semantic Differential) method are firstly used to identify consumer emotional perception model toward desk lamp before product design. Through investigating different desk lamp design elements and consumer views, the form design factors on the desk lamp product are reflected and all design schemes are sequenced after caculation. Desk lamp form bionic design method is combined the key bionic shape extracted from eye-tracking experiment and priority of desk lamp design schemes. This study provides an objective and rational method to product form bionic design.

Keywords: Bionic design; Form; Eye tracking; FAHP; Desk lamp

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526 Comparative Study of Isothermal and Cyclic Oxidation on Titanium Alloys

Authors: Poonam Yadav, Dong Bok Lee

Abstract:

Isothermal oxidation at 800°C for 50h and Cyclic oxidation at 600°C and 800°C for 40h of Pure Ti and Ti64 were performed in a muffle furnace. In Cyclic oxidation, massive scale spallation occurred, and the oxide scale cracks and peels off were observed at high temperature, it represents oxide scale that formed during cyclic oxidation was spalled out owing to stresses due to thermal shock generated during repetitive oxidation and subsequent cooling. The thickness of scale is larger in cyclic oxidation than the isothermal case. This is due to inward diffusion of oxygen through oxide scales and/or pores and cracks in cyclic oxidation.

Keywords: cyclic, diffusion, isothermal, cyclic

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525 A Low-Power Comparator Structure with Arbitrary Pre-Amplification Delay

Authors: Ata Khorami, Mohammad Sharifkhani

Abstract:

In the dynamic comparators, the pre-amplifier amplifies the input differential voltage and when the output Vcm of the pre-amplifier becomes larger than Vth of the latch input transistors, the latch is activated and finalizes the comparison. As a result, the pre-amplification delay is fixed to a value and cannot be set at the minimum required delay, thus, significant power and delay are imposed. In this paper, a novel structure is proposed through which the pre-amplification delay can be set at any low value saving power and time. Simulations show that using the proposed structure, by setting the pre-amplification delay at the minimum required value the power and comparison delay can be reduced by 55% and 100ps respectively.

Keywords: dynamic comparator, low power comparator, analog to digital converter, pre-amplification delay

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524 Exact Formulas of the End-To-End Green’s Functions in Non-hermitian Systems

Authors: Haoshu Li, Shaolong Wan

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The recent focus has been on directional signal amplification of a signal input at one end of a one-dimensional chain and measured at the other end. The amplification rate is given by the end-to-end Green’s functions of the system. In this work, we derive the exact formulas for the end-to-end Green's functions of non-Hermitian single-band systems. While in the bulk region, it is found that the Green's functions are displaced from the prior established integral formula by O(e⁻ᵇᴸ). The results confirm the correspondence between the signal amplification and the non-Hermitian skin effect.

Keywords: non-Hermitian, Green's function, non-Hermitian skin effect, signal amplification

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523 Effect of Plastic Deformation on the Carbide-Free Bainite Transformation in Medium C-Si Steel

Authors: Mufath Zorgani, Carlos Garcia-Mateo, Mohammad Jahazi

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In this study, the influence of pre-strained austenite on the extent of isothermal bainite transformation in medium-carbon, high-silicon steel was investigated. Different amounts of deformations were applied at 600°C on the austenite right before quenching to the region, where isothermal bainitic transformation is activated. Four different temperatures of 325, 350, 375, and 400°C considering similar holding time 1800s at each temperature, were selected to investigate the extent of isothermal bainitic transformation. The results showed that the deformation-free austenite transforms to the higher volume fraction of CFB bainite when the isothermal transformation temperature reduced from 400 to 325°C, the introduction of plastic deformation in austenite prior to the formation of bainite invariably involves a delay of the same or identical isothermal treatment. On the other side, when the isothermal transformation temperature and deformation increases, the volume fraction and the plate thickness of bainite decreases and the amount of retained austenite increases. The shape of retained austenite is mostly representing blocky-shape one due to the less amount of transformed bainite. Moreover, the plate-like shape bainite cannot be resolved when the deformation amount reached 30%, and the isothermal transformation temperatures are of 375 and 400°C. The amount of retained austenite and the percentage of its transformation to martensite during the final cooling stage play a significant role in the variation of hardness level for different thermomechanical regimes.

Keywords: ausforming, carbide free bainite, dilatometry, microstructure

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522 Studies on Non-Isothermal Crystallization Kinetics of PP/SEBS-g-MA Blends

Authors: Rishi Sharma, S. N. Maiti

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The non-isothermal crystallization kinetics of PP/SEBS-g-MA blends up to 0-50% concentration of copolymer was studied by differential scanning calorimetry at four different cooling rates. Crystallization parameters were analyzed by Avrami and Jeziorny models. Primary and secondary crystallization processes were described by Avrami equation. Avrami model showed that all types of shapes grow from small dimensions during primary crystallization. However, three-dimensional crystal growth was observed during the secondary crystallization process. The crystallization peak and onset temperature decrease, however

Keywords: crystallization kinetics, non-isothermal, polypropylene, SEBS-g-MA

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521 Isothermal and Cyclic Oxidation of the Ti-6Al-4V Alloy

Authors: Poonam Yadav, Dong Bok Lee

Abstract:

In this study, the Ti-6Al-4V alloy was isothermally and cyclically oxidized at 800oC for 40 hours in air, and its oxidation behavior was characterized in terms of its oxidation rate, scaling rate, and scale spallation tendency. The isothermal oxidation tests indicated that Ti-6Al-4V oxidized fast and almost linearly, forming thick oxide scales. However, the scales that formed during isothermal oxidation were adherent. The cyclic oxidation tests indicated that the scales that formed on Ti-6Al-4V were highly susceptible to spallation owing to the large growth stress arisen and the thermal stress imposed during thermal cyclings. The formed scales frequently delaminated into several pieces owing to the excessive stress aroused by the repetitive thermal shock. Particularly, excessive oxidation and heavy spallation occurred at the edge of Ti-6Al-4V during cyclic oxidation.

Keywords: cyclic, isothermal, oxidation, spallation

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520 Design of Reinforced Concrete (RC) Walls Considering Shear Amplification by Nonlinear Dynamic Behavior

Authors: Sunghyun Kim, Hong-Gun Park

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In the performance-based design (PBD), by using the nonlinear dynamic analysis (NDA), the actual performance of the structure is evaluated. Unlike frame structures, in the wall structures, base shear force which is resulted from the NDA, is greatly amplified than that from the elastic analysis. This shear amplifying effect causes repeated designs which make designer difficult to apply the PBD. Therefore, in this paper, factors which affect shear amplification were studied. For the 20-story wall model, the NDA was performed. From the analysis results, the base shear amplification factor was proposed.

Keywords: performance based design, shear amplification factor, nonlinear dynamic analysis, RC shear wall

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519 Experimental Demonstration of Broadband Erbium-Doped Fiber Amplifier

Authors: Belloui Bouzid

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In this paper, broadband design of erbium doped fiber amplifier (EDFA) is demonstrated and proved experimentally. High and broad gain is covered in C and L bands. The used technique combines, in one configuration, two double passes with split band structure for the amplification of two traveled signals one for the C band and the other for L band. This new topology is to investigate the trends of high gain and wide amplification at different status of pumping power, input wavelength, and input signal power. The presented paper is to explore the performance of EDFA gain using what it can be called double pass double branch wide band amplification configuration. The obtained results show high gain and wide broadening range of 44.24 dB and 80 nm amplification respectively.

Keywords: erbium doped fiber amplifier, erbium doped fiber laser, optical amplification, fiber laser

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518 Improvement of Mechanical Properties and Corrosion Resistance of AA7056 Aluminum Alloys by the Non-isothermal Aging Process

Authors: Tse-An Pan, Sheng-Long Lee

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The effect of non-isothermal aging on the mechanical properties and corrosion resistance of Al-9Zn-2.3Mg-1.9Cu (AA7056) alloys was investigated. The results revealed that thick materials were limited to retrogression and re-aging treatment (RRA). It could not reach the retrogression temperature in the RRA treatment. Compared with the RRA treatment, the non-isothermal aging (NIA) treatment produced discontinuous precipitates at grain boundaries, while the intragranular precipitates were fine and dense. The strength was similar to that of the RRA treatment; the corrosion resistance of the alloy was significantly improved by NIA aging. NIA treatment was less affected by the thickness of the alloy. The difference between the actual temperature and the setting temperature of the alloy is minimal during the aging process. The combination of properties could overcome the fact that RRA treatment cannot handle thick materials.

Keywords: Al-Zn-Mg-Cu alloy, corrosion, retrogression, re-aging, non-isothermal aging

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517 Effect of Scarp Topography on Seismic Ground Motion

Authors: Haiping Ding, Rongchu Zhu, Zhenxia Song

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Local irregular topography has a great impact on earthquake ground motion. For scarp topography, using numerical simulation method, the influence extent and scope of the scarp terrain on scarp's upside and downside ground motion are discussed in case of different vertical incident SV waves. The results show that: (1) The amplification factor of scarp's upside region is greater than that of the free surface, while the amplification factor of scarp's downside part is less than that of the free surface; (2) When the slope angle increases, for x component, amplification factors of the scarp upside also increase, while the downside part decrease with it. For z component, both of the upside and downside amplification factors will increase; (3) When the slope angle changes, the influence scope of scarp's downside part is almost unchanged, but for the upside part, it slightly becomes greater with the increase of slope angle; (4) Due to the existence of the scarp, the z component ground motion appears at the surface. Its amplification factor increases for larger slope angle, and the peaks of the surface responses are related with incident waves. However, the input wave has little effects on the x component amplification factors.

Keywords: scarp topography, ground motion, amplification factor, vertical incident wave

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516 Dynamic Amplification Factors of Some City Bridges

Authors: I. Paeglite, A. Paeglitis

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The paper presents a study of dynamic effects obtained from the dynamic load testing of the city highway bridges in Latvia carried out from 2005 to 2012. 9 pre-stressed concrete bridges and 4 composite bridges were considered. 11 of 13 bridges were designed according to the Eurocodes but two according to the previous structural codes used in Latvia (SNIP 2.05.03-84). The dynamic properties of the bridges were obtained by heavy vehicles passing the bridge roadway with different driving speeds and with or without even pavement. The obtained values of the Dynamic amplification factor (DAF) and bridge natural frequency were analyzed and compared to the values of built-in traffic load models provided in Eurocode 1. The actual DAF values for even bridge deck in the most cases are smaller than the value adopted in Eurocode 1. Vehicle speed for uneven pavements significantly influence Dynamic amplification factor values.

Keywords: bridge, dynamic effects, load testing, dynamic amplification factor

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515 A Low-Cost Disposable PDMS Microfluidic Cartridge with Reagent Storage Silicone Blisters for Isothermal DNA Amplification

Authors: L. Ereku, R. E. Mackay, A. Naveenathayalan, K. Ajayi, W. Balachandran

Abstract:

Over the past decade the increase of sexually transmitted infections (STIs) especially in the developing world due to high cost and lack of sufficient medical testing have given rise to the need for a rapid, low cost point of care medical diagnostic that is disposable and most significantly reproduces equivocal results achieved within centralised laboratories. This paper present the development of a disposable PDMS microfluidic cartridge incorporating blisters filled with reagents required for isothermal DNA amplification in clinical diagnostics and point-of-care testing. In view of circumventing the necessity for external complex microfluidic pumps, designing on-chip pressurised fluid reservoirs is embraced using finger actuation and blister storage. The fabrication of the blisters takes into consideration three proponents that include: material characteristics, fluid volume and structural design. Silicone rubber is the chosen material due to its good chemical stability, considerable tear resistance and moderate tension/compression strength. The case of fluid capacity and structural form go hand in hand as the reagent need for the experimental analysis determines the volume size of the blisters, whereas the structural form has to be designed to provide low compression stress when deformed for fluid expulsion. Furthermore, the top and bottom section of the blisters are embedded with miniature polar opposite magnets at a defined parallel distance. These magnets are needed to lock or restrain the blisters when fully compressed so as to prevent unneeded backflow as a result of elasticity. The integrated chip is bonded onto a large microscope glass slide (50mm x 75mm). Each part is manufactured using a 3D printed mould designed using Solidworks software. Die-casting is employed, using 3D printed moulds, to form the deformable blisters by forcing a proprietary liquid silicone rubber through the positive mould cavity. The set silicone rubber is removed from the cast and prefilled with liquid reagent and then sealed with a thin (0.3mm) burstable layer of recast silicone rubber. The main microfluidic cartridge is fabricated using classical soft lithographic techniques. The cartridge incorporates microchannel circuitry, mixing chamber, inlet port, outlet port, reaction chamber and waste chamber. Polydimethylsiloxane (PDMS, QSil 216) is mixed and degassed using a centrifuge (ratio 10:1) is then poured after the prefilled blisters are correctly positioned on the negative mould. Heat treatment of about 50C to 60C in the oven for about 3hours is needed to achieve curing. The latter chip production stage involves bonding the cured PDMS to the glass slide. A plasma coroner treater device BD20-AC (Electro-Technic Products Inc., US) is used to activate the PDMS and glass slide before they are both joined and adequately compressed together, then left in the oven over the night to ensure bonding. There are two blisters in total needed for experimentation; the first will be used as a wash buffer to remove any remaining cell debris and unbound DNA while the second will contain 100uL amplification reagents. This paper will present results of chemical cell lysis, extraction using a biopolymer paper membrane and isothermal amplification on a low-cost platform using the finger actuated blisters for reagent storage. The platform has been shown to detect 1x105 copies of Chlamydia trachomatis using Recombinase Polymerase Amplification (RPA).

Keywords: finger actuation, point of care, reagent storage, silicone blisters

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514 Non-Isothermal Stationary Laminar Oil Flow Numerical Simulation

Authors: Daniyar Bossinov

Abstract:

This paper considers a non-isothermal stationary waxy crude oil flow in a two-dimensional axisymmetric pipe with the transition of a Newtonian fluid to a non-Newtonian fluid. The viscosity and yield stress of waxy crude oil are highly dependent on temperature changes. During the hot pumping of waxy crude oil through a buried pipeline, a non-isothermal flow occurs due to heat transfer to the surrounding soil. This leads to a decrease in flow temperature, an increase in viscosity, the appearance of yield stress, the crystallization of wax, and the deposition of solid particles on the pipeline's inner wall. The deposition of oil solid particles reduces a pipeline flow area and leads to the appearance of a stagnant zone with thermal insulation in the near-wall area. Waxy crude oil properties change. A Newtonian fluid at low temperatures transits to a non-Newtonian fluid. The one-dimensional modeling of a non-isothermal waxy crude oil flow in a two-dimensional axisymmetric pipeline by traditional averaging of temperature and velocity over the pipeline cross-section does not allow for explaining a physics phenomenon. Therefore, in this work, a two-dimensional flow model and the heat transfer of waxy oil are constructed. The calculated data show the transition of a Newtonian fluid to a non-Newtonian fluid due to the heat exchange of waxy oil with the environment.

Keywords: non-isothermal laminar flow, waxy crude oil, stagnant zone, yield stress

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513 Anterior Chamber Depth Measured with Orbscan and Pentacam Compared with Smith Method in 102 Phakic Eyes

Authors: Mohammad Ghandehari Motlagh

Abstract:

Purpose: Comparing anterior chamber depth (ACD) measured with Orbscan II and Pentacam HR compared with the Smith method results. Methods: Smith method (1979) is a reliable method of measuring ACD only with help of slit lamp. In this study 102 phakic eyes as PRK candidates were imaged with both OrbScan and Pentacam and finally ACD was measured thru Smith method with slit lamp. ACD measured with Smith method was presumed as the gold standard and was compared with ACD of the 2 imaging devices. Contraindication cases for PRK and pseudophakic eyes have been excluded from the study. Results: Mean age of the patients was 35.2 ±14.8 yrs/old including 56 M(54.9%)and 46 F(45.09%).Acceptable correlation of ACD measured thru Smith method with Orbscan and Pentacam are R=0.958 and R=0.942 respectively and so Orbscan results can be used in procedures relying on ACD. Conclusion: ACDs measured with OrbScan is more precise than Pentacam and so can be more useful in some surgery procedures relying ACD results such as phakic IOLs and in cycloplegia contraindications.

Keywords: orbscan, pentacam, anterior chamber depth, slit lamp

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512 Tribocorrosion Behavior of Austempered Ductile Iron Microalloyed with Boron

Authors: S. Gvazava, N. Khidasheli, G. Gordeziani, A. DL. Batako

Abstract:

The work presented in this paper studied the tribological characteristics (wear resistance, friction coefficient) of austempered ductile iron (ADI) with different combinations of structural composition (upper bainite, lower bainite, retained austenite) in dry sliding friction. A range of structural states of the metal matrix was obtained by changing the regimes of isothermal quenching of high-strength cast iron. The tribological tests were carried out using two sets of isothermal quenched cast irons. After austenitization at 900°С for 60 minutes, the specimens from the first group were isothermally quenched at the 300°С temperature and the specimens from the second set – at 400°С. The investigations showed that the isothermal quenching increases the friction coefficient of high-strength cast irons. The friction coefficient was found to be in the range from 0.4 to 0.55 for cast irons, depending on the structures of the metal matrix. The quenched cast irons having lower bainite demonstrate higher wear resistance in dry friction conditions. The dependence of wear resistance on the amount of retained austenite in isothermal quenched cast irons has a nonlinear characteristic and reaches its maximum value when the content of retained austenite is about 15-22%. The boron micro-additives allowed to reduce the friction coefficient of ADI and increase their wear resistance by 1.5-1.7 times.

Keywords: wear resistance, dry sliding, austempering, ADI, friction coefficient, retained austenite, isothermal quenching

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511 Effect of Site Amplification on Seismic Safety Evaluation of Flyover Pier

Authors: Mohammad Raihan Mukhlis, M. Abdur Rahman Bhuiyan

Abstract:

Bangladesh is a developing country in which a lot of multi-span simply/continuous supported flyovers are being constructed in its major cities. Being situated in a seismically active region, seismic safety evaluation of flyovers is essential for seismic risk reduction. Effects of site amplification on seismic safety evaluation of flyover piers are the main concern of this study. In this regard, failure mode, lateral strength and displacement ductility of piers of a typical multi-span simply supported flyover have been evaluated by Japan Road Association (JRA) recommended guidelines, with and without considering site amplification. Ultimate flexural strengths of piers have been computed using the pushover analysis results. Shear capacity of piers has been calculated using the guidelines of JRA. Lateral strengths have been determined depending on the failure modes of the piers. Displacement ductility of piers has been computed using yield and ultimate displacements of the piers obtained from the pushover analysis results. Selected earthquake time history is used in seismic safety evaluation of the flyover piers. Finally, the ductility design method is used to conduct the seismic safety evaluation of the piers with and without considering site amplification. From the numerical results, it has been revealed that the effects of site amplification on seismic safety evaluation of bridge structures should be carefully taken into account.

Keywords: displacement ductility, flyover pier, lateral strength, safety evaluation, site amplification

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510 Analysis of Building Response from Vertical Ground Motions

Authors: George C. Yao, Chao-Yu Tu, Wei-Chung Chen, Fung-Wen Kuo, Yu-Shan Chang

Abstract:

Building structures are subjected to both horizontal and vertical ground motions during earthquakes, but only the horizontal ground motion has been extensively studied and considered in design. Most of the prevailing seismic codes assume the vertical component to be 1/2 to 2/3 of the horizontal one. In order to understand the building responses from vertical ground motions, many earthquakes records are studied in this paper. System identification methods (ARX Model) are used to analyze the strong motions and to find out the characteristics of the vertical amplification factors and the natural frequencies of buildings. Analysis results show that the vertical amplification factors for high-rise buildings and low-rise building are 1.78 and 2.52 respectively, and the average vertical amplification factor of all buildings is about 2. The relationship between the vertical natural frequency and building height was regressed to a suggested formula in this study. The result points out an important message; the taller the building is, the greater chance of resonance of vertical vibration on the building will be.

Keywords: vertical ground motion, vertical amplification factor, natural frequency, component

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509 Visual Detection of Escherichia coli (E. coli) through Formation of Beads Aggregation in Capillary Tube by Rolling Circle Amplification

Authors: Bo Ram Choi, Ji Su Kim, Juyeon Cho, Hyukjin Lee

Abstract:

Food contaminated by bacteria (E.coli), causes food poisoning, which occurs to many patients worldwide annually. We have introduced an application of rolling circle amplification (RCA) as a versatile biosensor and developed a diagnostic platform composed of capillary tube and microbeads for rapid and easy detection of Escherichia coli (E. coli). When specific mRNA of E.coli is extracted from cell lysis, rolling circle amplification (RCA) of DNA template can be achieved and can be visualized by beads aggregation in capillary tube. In contrast, if there is no bacterial pathogen in sample, no beads aggregation can be seen. This assay is possible to detect visually target gene without specific equipment. It is likely to the development of a genetic kit for point of care testing (POCT) that can detect target gene using microbeads.

Keywords: rolling circle amplification (RCA), Escherichia coli (E. coli), point of care testing (POCT), beads aggregation, capillary tube

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508 The Spectral Power Amplification on the Regular Lattices

Authors: Kotbi Lakhdar, Hachi Mostefa

Abstract:

We show that a simple transformation between the regular lattices (the square, the triangular, and the honeycomb) belonging to the same dimensionality can explain in a natural way the universality of the critical exponents found in phase transitions and critical phenomena. It suffices that the Hamiltonian and the lattice present similar writing forms. In addition, it appears that if a property can be calculated for a given lattice then it can be extrapolated simply to any other lattice belonging to the same dimensionality. In this study, we have restricted ourselves on the spectral power amplification (SPA), we note that the SPA does not have an effect on the critical exponents but does have an effect by the criticality temperature of the lattice; the generalisation to other lattice could be shown according to the containment principle.

Keywords: ising model, phase transitions, critical temperature, critical exponent, spectral power amplification

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