Search results for: multiplex ligation-dependent probe amplification

Authors: Zhengming Wu

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NanoFrazor lithography systems were developed as a first true alternative or extension to standard mask-less nanolithography methods like electron beam lithography (EBL). In contrast to EBL they are based on thermal scanning probe lithography (t-SPL). Here a heatable ultra-sharp probe tip with an apex of a few nm is used for patterning and simultaneously inspecting complex nanostructures. The heat impact from the probe on a thermal responsive resist generates those high-resolution nanostructures. The patterning depth of each individual pixel can be controlled with better than 1 nm precision using an integrated in-situ metrology method. Furthermore, the inherent imaging capability of the Nanofrazor technology allows for markerless overlay, which has been achieved with sub-5 nm accuracy as well as it supports stitching layout sections together with < 10 nm error. Pattern transfer from such resist features below 10 nm resolution were demonstrated. The technology has proven its value as an enabler of new kinds of ultra-high resolution nanodevices as well as for improving the performance of existing device concepts. The application range for this new nanolithography technique is very broad spanning from ultra-high resolution 2D and 3D patterning to chemical and physical modification of matter at the nanoscale. Nanometer-precise markerless overlay and non-invasiveness to sensitive materials are among the key strengths of the technology. However, while patterning at below 10 nm resolution is achieved, significantly increasing the patterning speed at the expense of resolution is not feasible by using the heated tip alone. Towards this end, an integrated laser write head for direct laser sublimation (DLS) of the thermal resist has been introduced for significantly faster patterning of micrometer to millimeter-scale features. Remarkably, the areas patterned by the tip and the laser are seamlessly stitched together and both processes work on the very same resist material enabling a true mix-and-match process with no developing or any other processing steps in between. The presentation will include examples for (i) high-quality metal contacting of 2D materials, (ii) tuning photonic molecules, (iii) generating nanofluidic devices and (iv) generating spintronic circuits. Some of these applications have been enabled only due to the various unique capabilities of NanoFrazor lithography like the absence of damage from a charged particle beam.

Keywords: nanofabrication, grayscale lithography, 2D materials device, nano-optics, photonics, spintronic circuits

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Authors: Matej Komanec, Tomas Nemecek, Dmytro Suslov, Petr Chvojka, Stanislav Zvanovec

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Nonlinear phenomena in the near- and mid-infrared region are attracting scientific attention mainly due to the supercontinuum generation possibilities and subsequent utilizations for ultra-wideband applications like e.g. absorption spectroscopy or optical coherence tomography. Thulium-based fiber lasers provide access to high-power ultrashort pump pulses in the vicinity of 2000 nm, which can be easily exploited for various nonlinear applications. The paper presents a simulation and experimental study of a pulsed thulium laser based for near-infrared (NIR) and mid-infrared (MIR) nonlinear applications in specialty optical fibers. In the first part of the paper the thulium laser is discussed. The thulium laser is based on a gain-switched seed-laser and a series of amplification stages for obtaining output peak powers in the order of kilowatts for pulses shorter than 200 ps in full-width at half-maximum. The pulsed thulium laser is first studied in a simulation software, focusing on seed-laser properties. Afterward, a pre-amplification thulium-based stage is discussed, with the focus of low-noise signal amplification, high signal gain and eliminating pulse distortions during pulse propagation in the gain medium. Following the pre-amplification stage a second gain stage is evaluated with incorporating a thulium-fiber of shorter length with increased rare-earth dopant ratio. Last a power-booster stage is analyzed, where the peak power of kilowatts should be achieved. Examples of analytical study are further validated by the experimental campaign. The simulation model is further corrected based on real components – parameters such as real insertion-losses, cross-talks, polarization dependencies, etc. are included. The second part of the paper evaluates the utilization of nonlinear phenomena, their specific features at the vicinity of 2000 nm, compared to e.g. 1550 nm, and presents supercontinuum modelling, based on the thulium laser pulsed output. Supercontinuum generation simulation is performed and provides reasonably accurate results, once fiber dispersion profile is precisely defined and fiber nonlinearity is known, furthermore input pulse shape and peak power must be known, which is assured thanks to the experimental measurement of the studied thulium pulsed laser. The supercontinuum simulation model is put in relation to designed and characterized specialty optical fibers, which are discussed in the third part of the paper. The focus is placed on silica and mainly on non-silica fibers (fluoride, chalcogenide, lead-silicate) in their conventional, microstructured or tapered variants. Parameters such as dispersion profile and nonlinearity of exploited fibers were characterized either with an accurate model, developed in COMSOL software or by direct experimental measurement to achieve even higher precision. The paper then combines all three studied topics and presents a possible application of such a thulium pulsed laser system working with specialty optical fibers.

Keywords: nonlinear phenomena, specialty optical fibers, supercontinuum generation, thulium laser

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Authors: Dali Gaganidze, Ekaterine Abashidze

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Potato is one of the main agricultural crops in Georgia. Georgia produces early and late potato varieties in almost all regions. In traditional potato growing regions (Svaneti, Samckhet javaheti and Tsalka), the yield is higher than 30-35 t/ha. Among the plant pests that limit potato production and quality, the potato cyst nematodes (PCN) are harmful around the world. Yield losses caused by PCN are estimated up to 30%. Rout surveys conducted in two geographically distinct regions of Georgia producing potatoes - Samtskhe - Javakheti and Svaneti revealed potato cyst nematode Globodera rostochiensi. The aim of the study was the Phylogenetic analyses of Globodera rostochiensi revealed in Georgia by the amplification and sequencing of 28S gen in the D3 region and intergenic ITS1-15.8S-ITS2 region. Identification of all the samples from the two Globodera populations (Samtskhe - Javakheti and Svaneti), i.e., G. rostochiensis (20 isolates) were confirmed by conventional multiplex PCR with ITS 5 universal and PITSp4, PITSr3 specific primers of the cyst nematodes’ (G. pallida, G. rostochiensis). The size of PCR fragment 434 bp confirms that PCN samples from two populations, Samtskhe- Javakheti and Svaneti, belong to G. rostochiensi . The ITS1–5.8S-ITS2 regions were amplified using prime pairs: rDNA1 ( 5’ -TTGATTACGTCCCTGCCCTTT-3’ and rDNA2( 5’ TTTCACTCGCCGTTACTAAGG-3’), D3 expansion regions were amplified using primer pairs: D3A (5’ GACCCCTCTTGAAACACGGA-3’) and D3B (5’-TCGGAAGGAACCAGCTACTA-3’. PCR products of each region were cleaned up and sequenced using an ABI 3500xL Genetic Analyzer. Obtained sequencing results were analyzed by computer program BLASTN (https://blast.ncbi.nlm.nih.gov/Blast.cg). Phylogenetic analyses to resolve the relationships between the isolates were conducted in MEGA7 using both distance- and character-based methods. Based on analysis of G.rostochiensis isolate`s D3 expansion regions are grouped in three major clades (A, B and C) on the phylogenetic tree. Clade A is divided into three subclades; clade C is divided into two subclades. Isolates from the Samtckhet-javakheti population are in subclade 1 of clade A and isolates in subclade 1 of clade C. Isolates) from Svaneti populations are in subclade 2 of clade A and in clad B. In Clade C, subclade two is presented by three isolates from Svaneti and by one isolate (GL17) from Samckhet-Javakheti. . Based on analysis of G.rostochiensis isolate`s ITS1–5.8S-ITS2 regions are grouped in two main clades, the first contained 20 Georgian isolates of Globodera rostochiensis from Svaneti . The second clade contained 15 isolates of Globodera rostochiensis from Samckhet javakheti. Our investigation showed of high genetic variation of D3 and ITS1–5.8S-ITS2 region of rDNA of the isolates of G. rostochiensis from different geographic origins (Svameti, Samckhet-Javakheti) of Georgia. Acknowledgement: The research has been supported by the Shota Rustaveli National Scientific Foundation of Georgia : Project # FR17_235

Keywords: globodera rostochiensi, PCR, phylogenetic tree, sequencing

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Authors: Chariya Laohavich

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At the current, the trends in the lung disease at a university hospital are the treat and diagnosis by bronchoscopy. Bronchoscopic cryotherapy is a long time procedure 1-4 hours. The cryo probe is sensitive and easy to be damaged and expensive. We have this study management for protection the cryo probe, user satisfaction and qualities work. This study conducted in 4 stages: stage 1 for a survey of problems and assessment of user’s needs; stage 2 for designing and developing the Rama-chest cryo arm for a bronchoscopy process; stage 3 for test-implementing the Rama-chest cryo arm in real situations, studying its problems and obstacles, and evaluating the user satisfaction; and stage 4 for an overall assessment and improvement. The sample used in this study consisted of a total of 15 Ramathipbodi Hospital’s Bronchoscopist and bronchoscopist’s nurse who had used the Rama-chest cryo arm for bronchoscopic cryotherapy from January to June 2016. Objective: To study efficacy and user satisfaction on the Rama-chest cryo arm innovation for bronchoscopic cryotherapy. Data were collected using a Rama-chest cryo arm satisfaction assessment form and analysed based on mean and standard deviation. Result is the Rama-chest cryo arm was an innovation that accommodated during bronchoscopic cryotherapy. The subjects rated this the cryo arm as being most satisfactory (M = 4.86 ± , SD 0.48. Therefore we have developed a cryo arm that uses local material, practical and economic. Our innovation is not only flexible and sustainable development but also lean and seamless. This produced device can be used as effectively as the imported one, and thus can be eventually substituted.

Keywords: efficacy, satisfaction, Rama-chest cryo arm, innovation, bronchoscopic cryotherapy

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Authors: Jean Paul M. Milambo

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Purpose: Aromatase inhibitors (AIs) are indicated in the treatment of hormone-receptive breast cancer in postmenopausal women in various settings. Studies have shown cardiovascular events in some developed countries. To date the data is sparce for evidence-based recommendations in African clinical settings due to lack of cancer registries, capacity building and surveillance systems. Therefore, this study was conducted to assess the feasibility of HyBeacon® probe genotyping adjunctive to standard care for timely prediction and diagnosis of Aromatase inhibitors (AIs) associated adverse events in breast cancer survivors in Africa. Methods: Cross sectional study was conducted to assess the knowledge of POCT among six African countries using online survey and telephonically contacted. Incremental cost effectiveness ratio (ICER) was calculated, using diagnostic accuracy study. This was based on mathematical modeling. Results: One hundred twenty-six participants were considered for analysis (mean age = 61 years; SD = 7.11 years; 95%CI: 60-62 years). Comparison of genotyping from HyBeacon® probe technology to Sanger sequencing showed that sensitivity was reported at 99% (95% CI: 94.55% to 99.97%), specificity at 89.44% (95% CI: 87.25 to 91.38%), PPV at 51% (95%: 43.77 to 58.26%), and NPV at 99.88% (95% CI: 99.31 to 100.00%). Based on the mathematical model, the assumptions revealed that ICER was R7 044.55. Conclusion: POCT using HyBeacon® probe genotyping for AI-associated adverse events maybe cost effective in many African clinical settings. Integration of preventive measures for early detection and prevention guided by different subtype of breast cancer diagnosis with specific clinical, biomedical and genetic screenings may improve cancer survivorship. Feasibility of POCT was demonstrated but the implementation could be achieved by improving the integration of POCT within primary health cares, referral cancer hospitals with capacity building activities at different level of health systems. This finding is pertinent for a future envisioned implementation and global scale-up of POCT-based initiative as part of risk communication strategies with clear management pathways.

Keywords: breast cancer, diagnosis, point of care, South Africa, aromatase inhibitors

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Authors: H. Abu-Ali, A. Nabok, T. Smith

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Aptamers are single-strand of DNA or RNA nucleotides sequence which is designed in vitro using selection process known as SELEX (systematic evolution of ligands by exponential enrichment) were developed for the selective detection of many toxic materials. In this work, we have developed an electrochemical biosensor for highly selective and sensitive detection of Hg2+ and Pb2+ using a specific aptamer probe (SAP) labelled with ferrocene (or methylene blue) in (5′) end and the thiol group at its (3′) termini, respectively. The SAP has a specific coil structure that matching with G-G for Pb2+ and T-T for Hg2+ interaction binding nucleotides ions, respectively. Aptamers were immobilized onto surface of screen-printed gold electrodes via SH groups; then the cyclic voltammograms were recorded in binding buffer with the addition of the above metal salts in different concentrations. The resulted values of anode current increase upon binding heavy metal ions to aptamers and analyte due to the presence of electrochemically active probe, i.e. ferrocene or methylene blue group. The correlation between the anodic current values and the concentrations of Hg2+ and Pb2+ ions has been established in this work. To the best of our knowledge, this is the first example of using a specific DNA aptamers for electrochemical detection of heavy metals. Each increase in concentration of 0.1 μM results in an increase in the anode current value by simple DC electrochemical test i.e (Cyclic Voltammetry), thus providing an easy way of determining Hg2+ and Pb2+concentration.

Keywords: aptamer, based, biosensor, DNA, electrochemical, highly, specific

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Authors: Maxime Merheb, Rachel Matar

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Retrieving ancient DNA sequences which in return permit the entire genome sequencing from fossils have extraordinarily improved in recent years, thanks to sequencing technology and other methodological advances. In any case, the quest to search for ancient DNA is still obstructed by the damage inflicted on DNA which accumulates after the death of a living organism. We can characterize this damage into three main categories: (i) Physical abnormalities such as strand breaks which lead to the presence of short DNA fragments. (ii) Modified bases (mainly cytosine deamination) which cause errors in the sequence due to an incorporation of a false nucleotide during DNA amplification. (iii) DNA modifications referred to as blocking lesions, will halt the PCR extension which in return will also affect the amplification and sequencing process. We can clearly see that the issues arising from breakage and coding errors were significantly decreased in recent years. Fast sequencing of short DNA fragments was empowered by platforms for high-throughput sequencing, most of the coding errors were uncovered to be the consequences of cytosine deamination which can be easily removed from the DNA using enzymatic treatment. The methodology to repair DNA sequences is still in development, it can be basically explained by the process of reintroducing cytosine rather than uracil. This technique is thus restricted to amplified DNA molecules. To eliminate any type of damage (particularly those that block PCR) is a process still pending the complete repair methodologies; DNA detection right after extraction is highly needed. Before using any resources into extensive, unreasonable and uncertain repair techniques, it is vital to distinguish between two possible hypotheses; (i) DNA is none existent to be amplified to begin with therefore completely un-repairable, (ii) the DNA is refractory to PCR and it is worth to be repaired and amplified. Hence, it is extremely important to develop a non-enzymatic technique to detect the most degraded DNA.

Keywords: ancient DNA, DNA barcodong, enzymatic repair, PCR

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Authors: Moon Moon Devi, Ran Budnik

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The current large area cosmic ray detector surface arrays typically measure only the net flux and arrival-time of the charged particles produced in an extensive air shower (EAS). Measurement of the individual charged particles at a surface array will provide additional distinguishing parameters to identify the primary and to map the very high energy interactions in the upper layers of the atmosphere. In turn, these may probe anomalies in QCD interactions at energies beyond the reach of current accelerators. The recent attempts of studying the individual muon tracks are limited in their expandability to larger arrays and can only probe primary particles with energy up to about 10^15.5 eV. New developments in detector technology allow for a realistic cost of large area detectors, however with limitations on energy resolutions, directional information, and dynamic range. In this study, we perform a simulation study using CORSIKA to combine the energy spectrum and lateral spread of the muons with the longitudinal depth (Xmax) of an EAS initiated by a primary at ultra high energies (10¹⁶ – 10¹⁹) eV. Using proton and iron as the shower primaries, we show that the muon observables and Xmax together can be used to distinguish the primary. This study can be used to design a future detector for the surface array, which will be able to enhance our knowledge of primaries and QCD interactions.

Keywords: ultra high energy extensive air shower, muon tracking, air shower primaries, QCD interactions

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Authors: M. Irfan Fareed, Muhammad Tahir, Alvina Gul Kazi

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Castor bean (Ricinus communis; family Euphorbiaceae) is cultivated for the production of oil and as an ornamental plant throughout tropical regions. Leaf samples from castor bean plants with leaf curl and vein thickening were collected from areas around Okara (Pakistan) in 2011. PCR amplification using diagnostic primers showed the presence of a begomovirus and subsequently the specific pair (BurNF 5’- CCATGGTTGTGGCAGTTGATTGACAGATAC-3’, BurNR 5’- CCATGGATTCACGCACAGGGGAACCC-3’) was used to amplify and clone the whole genome of the virus. The complete nucleotide sequence was determined to be 2,759 nt (accession No. HE985227). Alignments showed the highest levels of nucleotide sequence identity (98.8%) with Cotton leaf curl Burewala virus (CLCuBuV; accession No. JF416947) No. JF416947). The virus in castor beans lacks on intact C2 gene, as is typical of CLCuBuV in cotton. An amplification product of ca. 1.4 kb was obtained in PCR with primers for betasatellites and the complete nucleotide sequence of a clone was determined to be 1373 nt (HE985228). The sequence showed 96.3% nucleotide sequence identity to the recombinant Cotton leaf curl Multan betasatellite (CLCuMB; JF502389). This is the first report of CLCuBuV and its betasatellite infecting castor bean, showing this plant species as an alternate host of the virus. Already many alternate host have been reported from different alternate host like tobacco, tomato, hibiscus, okra, ageratum, Digera arvensis, habiscus, Papaya and now in Ricinus communis. So, it is suggested that these alternate hosts should be avoided to grow near cotton growing regions.

Keywords: Ricinus communis, begomovirus, betasatellite, agriculture

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Authors: Mohamed ElMasry, Ahmad Ragheb, Tareq AbdelAziz, Mohamed Ghazy

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Choosing the suitable infrastructure system is becoming more challenging with the increase in demand for heavier structures contemporarily. This is the case where piled raft foundations have been widely used around the world to support heavy structures without extensive settlement. In the latter system, piles are rigidly connected to the raft, and most of the load goes to the soil layer on which the piles are bearing. In spite of that, when soil profiles contain thicker soft clay layers near the surface, or at relatively shallow depths, it is unfavorable to use the rigid piled raft foundation system. Consequently, the disconnected piled raft system was introduced as an alternative approach for the rigidly connected system. In this system, piles are disconnected from the raft using a cushion of soil, mostly of a granular interlayer. The cushion is used to redistribute the stresses among the piles and the subsoil. Piles are also used to stiffen the subsoil, and by this way reduce the settlement without being rigidly connected to the raft. However, the seismic loading effect on such disconnected foundation systems remains a problem, since the soil profiles may include thick clay layers which raise risks of amplification of the dynamic earthquake loads. In this paper, the effects of seismic behavior on the connected and disconnected piled raft systems are studied through a numerical model using Midas GTS NX Software. The study concerns the soil-structure interaction and the expected behavior of the systems. Advantages and disadvantages of each foundation approach are studied, and a comparison between the results are presented to show the effects of using disconnected piled raft systems in highly seismic zones. This was done by showing the excitation amplification in each of the foundation systems.

Keywords: soil-structure interaction, disconnected piled-raft, risks, seismic zones

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Authors: Hamzeh Alipour, Masoumeh Bagheri, Tahereh Karamzadeh, Abbasali Raz, Kourosh Azizi

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Netrin-A, a protein identified for conducting commissural axons, has a similar role in angiogenesis. In addition, studies have shown that one of the netrin-A receptors is expressed in the growing cells of small capillaries. It will be interesting to study this new group of molecules because their role in wound healing will become clearer in the future due to angiogenesis. The greenbottle blowfly Luciliasericata (L. sericata) larvae are increasingly used in maggot therapy of chronic wounds. This aim of this was the identification of moleculareatures of Netrin-A in L. sericata larvae. Larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericataNetrin-A (LSN-A) was then identified using Rapid Amplification of cDNA Ends (RACE) and Rapid Amplification of Genomic Ends (RAGE). Parts of the Netrin-A gene, including the middle, 3′-, and 5′-ends were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its molecular weight is estimated to be 78.6 kDa. The 3-D structure ofNetrin-A drawn by SWISS-MODEL revealed its similarity to the Netrin-1 of humans with 66.8% identity. The LSN-A protein conduces to repair the myelin membrane in neuronal cells. Ultimately, it can be an effective candidate in neural regeneration and wound healing. Furthermore, our next attempt is to deplore recombinant proteins for use in medical sciences.

Keywords: maggot therapy, netrin-A, RACE, RAGE, lucilia sericata

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Authors: Nidhi Chadha, A. K. Tiwari, Marilyn D. Milton, Anil K. Mishra

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Translocator protein (18 kDa) TSPO is highly expressed during microglia activation in neuroinflammation. Although a number of PET ligands have been developed for the visualization of activated microglia, one of the advantageous approaches is to develop potential optical imaging (OI) probe. Our study involves computational screening, synthesis and evaluation of TSPO ligand through various imaging modalities namely PET/SPECT/Optical. The initial computational screening involves pharmacophore modeling from the library designing having oxo-benzooxazol-3-yl-N-phenyl-acetamide groups and synthesis for visualization of efficacy of these compounds as multimodal imaging probes. Structure modeling of monomer, Ala147Thr mutated, parallel and anti-parallel TSPO dimers was performed and docking analysis was performed for distinct binding sites. Computational analysis showed pattern of variable binding profile of known diagnostic ligands and NBMP via interactions with conserved residues along with TSPO’s natural polymorphism of Ala147→Thr, which showed alteration in the binding affinity due to considerable changes in tertiary structure. Preliminary in vitro binding studies shows binding affinity in the range of 1-5 nm and selectivity was also certified by blocking studies. In summary, this skeleton was found to be potential probe for TSPO imaging due to ease in synthesis, appropriate lipophilicity and reach to specific region of brain.

Keywords: TSPO, molecular modeling, imaging, docking

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Authors: Mi-Ju Kim, Hae-Yeong Kim

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

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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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Authors: Zubeida Rossenkhan, Pervaiz K. Ahmed, Wee Chan Au

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In view of cross cultural career experiences, to the author’s best knowledge, the crucial role of culture and religious traditions in Asia remains understudied. Drawing on the notion of social capital as an invaluable resource needed for manager’s to progress, the purpose of this study is to probe the contextual experiences of Muslim women to elucidate unique challenges associated with social capital and career progress. Twenty-three in-depth interviews with top level Malay managers were conducted to probe experiences of upward career mobility and inequities in the workplace. Interpretive phenomenology was used to surface unique challenges and processes of creating and leveraging social capital. The study uncovers the unique challenges of Muslim women in Malaysia. Narratives of participants highlight not only generic forms of gender discrimination, but also culturally specific stereotypes and social expectations limiting their advancement. Interestingly, the findings identify a gender-religion handicap in the form of perceived inequality and restrictions rooted from the women manager’s gender and religion. The analysis also reveals how these Muslim women managers’ negotiate their challenges, especially how they access social capital and progress their careers. The research offers a unique perspective on the career experiences of Malay women managers’ in top management. The research provides insight into the unique processes of developing social capital utilized by this group of women for career success.

Keywords: career success, gender discrimination, malaysia, Muslim women, social capital

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Authors: Hongquan Zhang

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Inspired by endogenous protein motors, researchers have constructed various synthetic DNA motors based on the specificity and predictability of Watson-Crick base pairing. However, the application of DNA motors to signal amplification and biosensing is limited because of low mobility and difficulty in real-time monitoring of the walking process. The objective of our work was to construct a new type of DNA motor termed target-triggered DNA motors that can walk for hundreds of steps in response to a single target binding event. To improve the mobility and processivity of DNA motors, we used gold nanoparticles (AuNPs) as scaffolds to build high-density, three-dimensional tracks. Hundreds of track strands are conjugated to a single AuNP. To enable DNA motors to respond to specific protein and nucleic acid targets, we adapted the binding-induced DNA assembly into the design of the target-triggered DNA motors. In response to the binding of specific target molecules, DNA motors are activated to autonomously walk along AuNP, which is powered by a nicking endonuclease or DNAzyme-catalyzed cleavage of track strands. Each moving step restores the fluorescence of a dye molecule, enabling monitoring of the operation of DNA motors in real time. The motors can translate a single binding event into the generation of hundreds of oligonucleotides from a single nanoparticle. The motors have been applied to amplify the detection of proteins and nucleic acids in test tubes and live cells. The motors were able to detect low pM concentrations of specific protein and nucleic acid targets in homogeneous solutions without the need for separation. Target-triggered DNA motors are significant for broadening applications of DNA motors to molecular sensing, cell imagining, molecular interaction monitoring, and controlled delivery and release of therapeutics.

Keywords: biosensing, DNA motors, gold nanoparticles, signal amplification

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Authors: Aminah Salim, Idrees Ahmed Nasir, Abdul Qayyum Rao, Muhammad Ali, Muhammad Sohail Anjum, Ayesha Hameed, Bushra Tabassum, Anwar Khan, Arfan Ali, Mariyam Zameer, Tayyab Husnain

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Risk assessment of transgenic herbicide tolerant sugarcane having CEMB codon optimized cp4EPSPS gene was done in present study. Fifteen days old chicks taken from K&Ns Company were randomly assorted into four groups with eight chicks in each group namely control chicken group fed with commercial diet, non-transgenic group fed with non-experimental sugarcane and transgenic group fed with transgenic sugarcane with minimum and maximum level. Body weights, biochemical analysis for Urea, alkaline phosphatase, alanine transferase, aspartate transferase, creatinine and bilirubin determination and histological examination of chicks fed with four types of feed was taken at fifteen days interval and no significant difference was observed in body weight biochemical and histological studies of all four groups. Protein isolated from the serum sample was analyzed through dipstick and SDS-PAGE, showing the absence of transgene protein in the serum sample of control and experimental groups. Moreover the amplification of cp4EPSPS gene with gene specific primers of DNA isolated from chicks blood and also from commercial diet was done to determine the presence and mobility of any nucleotide fragment of the transgene in/from feed and no amplification was obtained in feed as well as in blood extracted DNA of any group. Also no mRNA expression of cp4EPSPS gene was obtained in any tissue of four groups of chicks. From the results it is clear that there is no deleterious or harmful effect of the CEMB codon optimized transgenic cp4EPSPS sugarcane on the chicks health.

Keywords: chicks, cp4EPSPS, glyphosate, sugarcane

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Authors: Frantisek Odvarka

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Based on the data from the Moravian Karst, the influence of the calcite speleothem growth by selected meteorological factors was evaluated. External temperature was determined as one of the main factors influencing speleothem growth in Moravian Karst. This factor significantly influences the CO₂ concentration in soil/epikarst, and cave atmosphere in the Moravian Karst and significantly contributes to the changes in the CO₂ partial pressure differences between soil/epikarst and cave atmosphere in Moravian Karst, which determines the drip water supersaturation with respect to the calcite and quantity of precipitated calcite in the Moravian Karst cave environment. External air temperatures and cave air temperatures were measured using a COMET S3120 data logger, which can measure temperatures in the range from -30 to +80 °C with an accuracy of ± 0.4 °C. CO₂ concentrations in the cave and soils were measured with a FT A600 CO₂H Ahlborn probe (value range 0 ppmv to 10,000 ppmv, accuracy 1 ppmv), which was connected to the data logger ALMEMO 2290-4, V5 Ahlborn. The soil temperature was measured with a FHA646E1 Ahlborn probe (temperature range -20 to 70 °C, accuracy ± 0.4 °C) connected to an ALMEMO 2290-4 V5 Ahlborn data logger. The airflow velocities into and out of the cave were monitored by a FVA395 TH4 Thermo anemometer (speed range from 0.05 to 2 m s⁻¹, accuracy ± 0.04 m s⁻¹), which was connected to the ALMEMO 2590-4 V5 Ahlborn data logger for recording. The flow was measured in the lower and upper entrance of the Imperial Cave. The data were analyzed in MS Office Excel 2019 and PHREEQC.

Keywords: speleothem growth, carbon dioxide partial pressure, Moravian Karst, external temperature

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Authors: Norul Husna A. Kasim, Siva K. Balasundram

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The gap between ‘actual yield’ and ‘potential yield’ has remained a problem in the Malaysian oil palm industry. Ineffective maturity assessment and untimely harvesting have compounded this problem. Typically, the traditional method of palm oil quality and yield assessment is destructive, costly and laborious. Fluorescence-sensing offers a new means of assessing palm oil quality and yield non-destructively. This work describes the estimation of palm oil quality and yield using a multi-parametric fluorescence sensor (Multiplex®) to quantify the concentration of secondary metabolites, such as anthocyanin and flavonoid, in fresh fruit bunches across three different palm ages (6, 9, and 12 years-old). Results show that fluorescence sensing is an effective means of assessing FFB maturity, in terms of palm oil quality and yield quantifications.

Keywords: anthocyanin, flavonoid fluorescence sensor, palm oil yield and quality

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Authors: Rama Bhargava, Surabhi Nishad

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The infusion of nanofluids has dramatically enhanced the heat-carrying capacity of the fluids, applicable to many engineering and medical process where the temperature below freezing is required. Cryosurgery is an efficient therapy for the treatment of cancer, but sometimes the excessive cooling may harm the nearby healthy cells. Efforts are therefore done to develop a model which can cause to generate the low temperature as required. In the present study, a mathematical model is developed based on the bioheat transfer equation to simulate the heat transfer from the probe on a tumor (with irregular domain) using the hybrid technique consisting of element free Galerkin method with αα-family of approximation. The probe is loaded will nano-particles. The effects of different nanoparticles, namely Al₂O₃, Fe₃O₄, Au on the heat-producing rate, is obtained. It is observed that the temperature can be brought to (60°C)-(-30°C) at a faster freezing rate on the infusion of different nanoparticles. Besides increasing the freezing rate, the volume of the nanoparticle can also control the size and growth of ice crystals formed during the freezing process. The study is also made to find the time required to achieve the desired temperature. The problem is further extended for multi tumors of different shapes and sizes. The irregular shape of the frozen domain and the direction of ice growth are very sensitive issues, posing a challenge for simulation. The Meshfree method has been one of the accurate methods in such problems as a domain is naturally irregular. The discretization is done using the nodes only. MLS approximation is taken in order to generate the shape functions. Sufficiently accurate results are obtained.

Keywords: cryosurgery, EFGM, hybrid, nanoparticles

Procedia PDF Downloads 100

Authors: Aminu Bashir Mohammad, Agwu Ezera, Abdulrazaq G. Habib, Garba Iliyasu

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Background: Drug resistance tuberculosis (DR-TB) continues to be a challenge in developing countries with poor resources. Routine screening for primary DR-TB before commencing treatment is not done in public hospitals in Nigeria, even with the large body of evidence that shows a high prevalence of primary DR-TB. Data on drug resistance and its genetic determinant among follow up TB patients is lacking in Nigeria. Hence the aim of this study was to determine the prevalence and genetic determinant of drug resistance among follow up TB patients in a tertiary hospital in Nigeria. Methods: This was a cross-sectional laboratory-based study conducted on 384 sputum samples collected from consented follow-up tuberculosis patients. Standard microbiology methods (Zeil-Nielsen staining and microscopy) and PCR (Line Probe Assay)] were used to analyze the samples collected. Person’s Chi-square was used to analyze the data generated. Results: Out of three hundred and eighty-four (384) sputum samples analyzed for mycobacterium tuberculosis (MTB) and DR-TB twenty-five 25 (6.5%) were found to be AFB positive. These samples were subjected to PCR (Line Probe Assay) out of which 18(72%) tested positive for DR-TB. Mutations conferring resistance to rifampicin (rpo B) and isoniazid (katG, and or inhA) were detected in 12/18(66.7%) and 6/18(33.3%), respectively. Transmission dynamic of DR-TB was not significantly (p>0.05) dependent on demographic characteristics. Conclusion: There is a need to strengthened the laboratory capacity for diagnosis of TB and drug resistance testing and make these services available, affordable, and accessible to the patients who need them.

Keywords: drug resistance tuberculosis, genetic determinant, intensive phase, Nigeria

Procedia PDF Downloads 258

Authors: Amar F. Siddique

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The Guwahati city is one of the fastest growing cities of the north-eastern region of India, situated on the South Bank of the Brahmaputra River falls in the highest seismic zone level V. The city has witnessed many high magnitude earthquakes in the past decades. The Assam earthquake occurred on August 15, 1950, of moment magnitude 8.7 epicentered near Rima, Tibet was one of the major earthquakes which caused a serious structural damage and widespread soil liquefaction in and around the region. Hence the study of ground motion characteristics of Guwahati city is very essential. In this present work 1D equivalent linear ground response analysis (GRA) has been adopted using Deep soil software. The analysis has been done for two typical sites namely, Panbazar and Azara comprising total four boreholes location in Guwahati city of India. GRA of the sites is carried out by using an input motion recorded at Nongpoh station (recorded PGA 0.048g) and Nongstoin station (recorded PGA 0.047g) of 1997 Indo-Burma earthquake. In comparison to motion recorded at Nongpoh, different amplifications of bedrock peak ground acceleration (PGA) are obtained for all the boreholes by the motion recorded at Nongstoin station; although, the Fourier amplitude ratios (FAR) and fundamental frequencies remain almost same. The difference in recorded duration and frequency content of the two motions mainly influence the amplification of motions thus getting different surface PGA and amplification factor keeping a constant bedrock PGA. From the results of response spectra, it is found that at the period of less than 0.2 sec the ground motion recorded at Nongpoh station will give a high spectral acceleration (SA) on the structures than at Nongstoin station. Again for a period greater than 0.2 sec the ground motion recorded at Nongstoin station will give a high SA on the structures than at Nongpoh station.

Keywords: fourier amplitude ratio, ground response analysis, peak ground acceleration, spectral acceleration

Procedia PDF Downloads 152

Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

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Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

Procedia PDF Downloads 224

Authors: Shringika Soni, Utkarsh Jain, Nidhi Chauhan

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Aptamers are recently discovered ~80-100 bp long artificial oligonucleotides that not only demonstrated their applications in therapeutics; it is tremendously used in diagnostic and sensing application to detect different biomarkers and drugs. Synthesizing aptamers for proteins or genomic template is comparatively feasible in laboratory, but drugs or other chemical target based aptamers require major specification and proper optimization and validation. One has to optimize all selection, amplification, and characterization steps of the end product, which is extremely time-consuming. Therefore, we performed asymmetric PCR (polymerase chain reaction) for random oligonucleotides pool synthesis, and further use them in Systematic evolution of ligands by exponential enrichment (SELEX) for anti-psychotic drugs based aptamers synthesis. Anti-psychotic drugs are major tranquilizers to control psychosis for proper cognitive functions. Though their low medical use, their misuse may lead to severe medical condition as addiction and can promote crime in social and economical impact. In this work, we have approached the in-vitro SELEX method for ssDNA synthesis for anti-psychotic drugs (in this case ‘target’) based aptamer synthesis. The study was performed in three stages, where first stage included synthesis of random oligonucleotides pool via asymmetric PCR where end product was analyzed with electrophoresis and purified for further stages. The purified oligonucleotide pool was incubated in SELEX buffer, and further partition was performed in the next stage to obtain target specific aptamers. The isolated oligonucleotides are characterized and quantified after each round of partition, and significant results were obtained. After the repetitive partition and amplification steps of target-specific oligonucleotides, final stage included sequencing of end product. We can confirm the specific sequence for anti-psychoactive drugs, which will be further used in diagnostic application in clinical and forensic set-up.

Keywords: anti-psychotic drugs, aptamer, biosensor, ssDNA, SELEX

Procedia PDF Downloads 108

Authors: E. S. Manuylovich, V. A. Astapenko, P. A. Golovinsky

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The dispersion relation binding the constant of propagation and frequency is calculated for silver cone. The evolution of the electric field of ultrashort pulse during its propagation in conical structure is considered. Increasing of electric field during pulse propagation to the top of the cone is observed. Reduction of the pulse duration at a certain distance is observed. The dependence of minimum pulse duration on initial chirp and cone angle is investigated.

Keywords: ultrashort pulses, surface plasmon polariton, dispersion, silver cone

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Authors: Narek Margaryan, Zhozef Panosyan

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Due to their unique properties, carbon nanofilms have become the object of general attention and intensive research. In this case it plays a very important role to study surface properties of these films. It is also important to study processes of forming of this films, which is accompanied by a process of self-organization at the nano and micro levels. For more detailed investigation, we examined diamond-like carbon (DLC) layers deposited by chemical vapor deposition (CVD) method on Ge substrate and hydro-generated grapheme layers obtained on surface of colloidal solution using grouping method. In this report surface transformation of these CVD nanolayers is studied by atomic force microscopy (AFM) upon deposition time. Also, it can be successfully used to study surface properties of self-assembled grapheme layers. In turn, it is possible to sketch out their boundary line, which enables one to draw an idea of peculiarities of formation of these layers. Images obtained by AFM are investigated as a mathematical set of numbers and fractal and roughness analysis were done. Fractal dimension, Regne’s fractal coefficient, histogram, Fast Fourier transformation, etc. were obtained. The dependence of fractal parameters on the deposition duration for CVD films and on temperature of solution tribolayers was revealed. As an important surface parameter for our carbon films, surface energy was calculated as function of Regne’s fractal coefficient. Surface potential was also measured with Kelvin probe method using semi-contacting AFM. The dependence of surface potential on the deposition duration for CVD films and on temperature of solution for hydro-generated graphene was found as well. Results obtained by fractal analysis method was related with purly esperimental results for number of samples.

Keywords: nanostructured films, self-assembled grapheme, diamond-like carbon, surface potential, Kelvin probe method, fractal analysis

Procedia PDF Downloads 243

Authors: Shang-Yu Chiang, Yu-Shan Tsai, Shih-Hsien Sung, Chung-Ming Lo

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Mitral regurgitation (MR) is a heart disorder which the mitral valve does not close properly when the heart pumps out blood. MR is the most common form of valvular heart disease in the adult population. The diagnostic echocardiographic finding of MR is straightforward due to the well-known clinical evidence. In the determination of MR severity, quantification of sonographic findings would be useful for clinical decision making. Clinically, the vena contracta is a standard for MR evaluation. Vena contracta is the point in a blood stream where the diameter of the stream is the least, and the velocity is the maximum. The quantification of vena contracta, i.e. the vena contracta width (VCW) at mitral valve, can be a numeric measurement for severity assessment. However, manually delineating the VCW may not accurate enough. The result highly depends on the operator experience. Therefore, this study proposed an automatic method to quantify VCW to evaluate MR severity. Based on color Doppler ultrasound, VCW can be observed from the blood flows to the probe as the appearance of red or yellow area. The corresponding brightness represents the value of the flow rate. In the experiment, colors were firstly transformed into HSV (hue, saturation and value) to be closely align with the way human vision perceives red and yellow. Using ellipse to fit the high flow rate area in left atrium, the angle between the mitral valve and the ultrasound probe was calculated to get the vertical shortest diameter as the VCW. Taking the manual measurement as the standard, the method achieved only 0.02 (0.38 vs. 0.36) to 0.03 (0.42 vs. 0.45) cm differences. The result showed that the proposed automatic VCW extraction can be efficient and accurate for clinical use. The process also has the potential to reduce intra- or inter-observer variability at measuring subtle distances.

Keywords: mitral regurgitation, vena contracta, color doppler, image processing

Procedia PDF Downloads 347

Authors: Narges Razmjoo, Hamid Sefidari, Michael Strand

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Biomass combustion is a growing technique for heat and power production due to the increasing stringent regulations with CO2 emissions. Grate-fired systems have been regarded as a common and popular combustion technology for burning woody biomass. However, some grate furnaces are not well optimized and may emit significant amount of unwanted compounds such as dust, NOx, CO, and unburned gaseous components. The combustion characteristics inside the fuel bed are of practical interest, as they are directly related to the release of volatiles and affect the stability and the efficiency of the fuel bed combustion. Although numerous studies have been presented on the grate firing of biomass, to the author’s knowledge, none of them have conducted a detailed experimental study within the fuel bed. It is difficult to conduct measurements of temperature and gas species inside the burning bed of the fuel in full-scale boilers. Results from such inside bed measurements can also be applied by the numerical experts for modeling the fuel bed combustion. The current work presents an experimental investigation into the combustion behavior of wet woody biomass (53 %) in a 4 MW reciprocating grate boiler, by focusing on the gas species distribution along the height of the fuel bed. The local concentrations of gases (CO, CO2, CH4, NO, and O2) inside the fuel bed were measured through a glass port situated on the side wall of the furnace. The measurements were carried out at five different heights of the fuel bed, by means of a bent stainless steel probe containing a type-k thermocouple. The sample gas extracted from the fuel bed, through the probe, was filtered and dried and then was analyzed using two infrared spectrometers. Temperatures of about 200-1100 °C were measured close to the grate, indicating that char combustion is occurring at the bottom of the fuel bed and propagates upward. The CO and CO2 concentration varied in the range of 15-35 vol % and 3-16 vol %, respectively, and NO concentration varied between 10-140 ppm. The profile of the gas concentrations distribution along the bed height provided a good overview of the combustion sub-processes in the fuel bed.

Keywords: experimental, fuel bed, grate firing, wood combustion

Procedia PDF Downloads 301

Authors: Fadime Eroglu

Abstract:

Acanthamoeba genus belongs to kingdom protozoa, and it is known as free-living amoebae. Acanthamoeba genus has been isolated from human bodies, swimming pools, bottled mineral water, contact lens solutions, dust, and soil. The members of the genus Acanthamoeba causes Acanthamoeba Keratitis which is a painful sight-threatening disease of the eyes. In recent years, the prevalence of Acanthamoeba keratitis has been high rate reported. The eight different Acanthamoeba species are known to be effective in Acanthamoeba keratitis. These species are Acanthamoeba castellanii, Acanthamoeba polyphaga, Acanthamoeba griffini, Acanthamoeba hatchetti, Acanthamoeba culbertsoni and Acanhtamoeba rhysodes. The conventional diagnosis of Acanthamoeba Keratitis has relied on cytological preparations and growth of Acanthamoeba in culture. However molecular methods such as real-time PCR has been found to be more sensitive. The real-time PCR has now emerged as an effective method for more rapid testing for the diagnosis of infectious disease in decade. Therefore, a real-time PCR assay for the detection of Acanthamoeba keratitis and Acanthamoeba species have been developed in this study. The 18S rRNA sequences from Acanthamoeba species were obtained from National Center for Biotechnology Information and sequences were aligned with MEGA 6 programme. Primers and probe were designed using Custom Primers-OligoPerfectTMDesigner (ThermoFisherScientific, Waltham, MA, USA). They were also assayed for hairpin formation and degree of primer-dimer formation with Multiple Primer Analyzer ( ThermoFisherScientific, Watham, MA, USA). The eight different ATCC Acanthamoeba species were obtained, and DNA was extracted using the Qiagen Mini DNA extraction kit (Qiagen, Hilden, Germany). The DNA of Acanthamoeba species were analyzed using newly designed primer and probe set in real-time PCR assay. The early definitive laboratory diagnosis of Acanthamoeba Keratitis and the rapid initiation of suitable therapy is necessary for clinical prognosis. The results of the study have been showed that new primer and probes could be used for detection and distinguish for Acanthamoeba species. These new developing methods are helpful for diagnosis of Acanthamoeba Keratitis.

Keywords: Acathamoeba Keratitis, Acanthamoeba species, fast diagnosis, Real-Time PCR

Procedia PDF Downloads 93

Authors: Prem K. Mony, Bharadwaj Amrutur, Prashanth Thankachan, Swarnarekha Bhat, Suman Rao, Maryann Washington, Annamma Thomas, N. Sheela, Hiteshwar Rao, Sumi Antony

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The disease burden among mothers and newborns is caused mostly by a handful of avoidable conditions occurring around the time of childbirth and within the first month following delivery. Real-time monitoring of vital parameters of mothers and neonates offers a potential opportunity to impact access as well as the quality of care in vulnerable populations. We describe the design, development and testing of an innovative wearable device for remote biomonitoring (RBM) of body temperatures in mothers and neonates in a hospital in southern India. The architecture consists of: [1] a low-cost, wearable sensor tag; [2] a gateway device for ‘real-time’ communication link; [3] piggy-backing on a commercial GSM communication network; and [4] an algorithm-based data analytics system. Requirements for the device were: long battery-life upto 28 days (with sampling frequency 5/hr); robustness; IP 68 hermetic sealing; and human-centric design. We undertook pre-clinical laboratory testing followed by clinical trial phases I & IIa for evaluation of safety and efficacy in the following sequence: seven healthy adult volunteers; 18 healthy mothers; and three sets of babies – 3 healthy babies; 10 stable babies in the Neonatal Intensive Care Unit (NICU) and 1 baby with hypoxic ischaemic encephalopathy (HIE). The 3-coin thickness, pebble-design sensor weighing about 8 gms was secured onto the abdomen for the baby and over the upper arm for adults. In the laboratory setting, the response-time of the sensor device to attain thermal equilibrium with the surroundings was 4 minutes vis-a-vis 3 minutes observed with a precision-grade digital thermometer used as a reference standard. The accuracy was ±0.1°C of the reference standard within the temperature range of 25-40°C. The adult volunteers, aged 20 to 45 years, contributed a total of 345 hours of readings over a 7-day period and the postnatal mothers provided a total of 403 paired readings. The mean skin temperatures measured in the adults by the sensor were about 2°C lower than the axillary temperature readings (sensor =34.1 vs digital = 36.1); this difference was statistically significant (t-test=13.8; p<0.001). The healthy neonates provided a total of 39 paired readings; the mean difference in temperature was 0.13°C (sensor =36.9 vs digital = 36.7; p=0.2). The neonates in the NICU provided a total of 130 paired readings. Their mean skin temperature measured by the sensor was 0.6°C lower than that measured by the radiant warmer probe (sensor =35.9 vs warmer probe = 36.5; p < 0.001). The neonate with HIE provided a total of 25 paired readings with the mean sensor reading being not different from the radian warmer probe reading (sensor =33.5 vs warmer probe = 33.5; p=0.8). No major adverse events were noted in both the adults and neonates; four adult volunteers reported mild sweating under the device/arm band and one volunteer developed mild skin allergy. This proof-of-concept study shows that real-time monitoring of temperatures is technically feasible and that this innovation appears to be promising in terms of both safety and accuracy (with appropriate calibration) for improved maternal and neonatal health.

Keywords: public health, remote biomonitoring, temperature surveillance, wearable sensors, mothers and newborns

Procedia PDF Downloads 178