Search results for: TaqMan probe
410 Analysis of OPG Gene Polymorphism T245G (rs3134069) in Slovak Postmenopausal Women
Authors: I. Boroňová, J. Bernasovská, J. Kľoc, Z. Tomková, E. Petrejčíková, S. Mačeková, J. Poráčová, M. M. Blaščáková
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Osteoporosis is a common multifactorial disease with a strong genetic component characterized by reduced bone mass and increased risk of fractures. Genetic factors play an important role in the pathogenesis of osteoporosis. The aim of our study was to identify the genotype and allele distribution of T245G polymorphism in OPG gene in Slovak postmenopausal women. A total of 200 unrelated Slovak postmenopausal women with diagnosed osteoporosis and 200 normal controls were genotyped for T245G (rs3134069) polymorphism of OPG gene. Genotyping was performed using the Custom Taqman®SNP Genotyping assays. Genotypes and alleles frequencies showed no significant differences (p=0.5551; p=0.6022). The results of the present study confirm the importance of T245G polymorphism in OPG gene in the pathogenesis of osteoporosis.Keywords: OPG gene, T245G polymorphism, osteoporosis, T245G polymorphism, real-time PCR
Procedia PDF Downloads 411409 Hohmann Transfer and Bi-Elliptic Hohmann Transfer in TRAPPIST-1 System
Authors: Jorge L. Nisperuza, Wilson Sandoval, Edward. A. Gil, Johan A. Jimenez
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In orbital mechanics, an active research topic is the calculation of interplanetary trajectories efficient in terms of energy and time. In this sense, this work concerns the calculation of the orbital elements for sending interplanetary probes in the extrasolar system TRAPPIST-1. Specifically, using the mathematical expressions of the circular and elliptical trajectory parameters, expressions for the flight time and the orbital transfer rate increase between orbits, the orbital parameters and the graphs of the trajectories of Hohmann and Hohmann bi-elliptic for sending a probe from the innermost planet to all the other planets of the studied system, are obtained. The relationship between the orbital transfer rate increments and the relationship between the flight times for the two transfer types is found. The results show that, for all cases under consideration, the Hohmann transfer results to be the least energy and temporary cost, a result according to the theory associated with Hohmann and Hohmann bi-elliptic transfers. Saving in the increase of the speed reaches up to 87% was found, and it happens for the transference between the two innermost planets, whereas the time of flight increases by a factor of up to 6.6 if one makes use of the bi-elliptic transfer, this for the case of sending a probe from the innermost planet to the outermost.Keywords: bi-elliptic Hohmann transfer, exoplanet, extrasolar system, Hohmann transfer, TRAPPIST-1
Procedia PDF Downloads 195408 Malate Dehydrogenase Enabled ZnO Nanowires as an Optical Tool for Malic Acid Detection in Horticultural Products
Authors: Rana Tabassum, Ravi Kant, Banshi D. Gupta
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Malic acid is an extensively distributed organic acid in numerous horticultural products in minute amounts which significantly contributes towards taste determination by balancing sugar and acid fractions. An enhanced concentration of malic acid is utilized as an indicator of fruit maturity. In addition, malic acid is also a crucial constituent of several cosmetics and pharmaceutical products. An efficient detection and quantification protocol for malic acid is thus highly demanded. In this study, we report a novel detection scheme for malic acid by synergistically collaborating fiber optic surface plasmon resonance (FOSPR) and distinctive features of nanomaterials favorable for sensing applications. The design blueprint involves the deposition of an assembly of malate dehydrogenase enzyme entrapped in ZnO nanowires forming the sensing route over silver coated central unclad core region of an optical fiber. The formation and subsequent decomposition of the enzyme-analyte complex on exposure of the sensing layer to malic acid solutions of diverse concentration results in modification of the dielectric function of the sensing layer which is manifested in terms of shift in resonance wavelength. Optimization of experimental variables such as enzyme concentration entrapped in ZnO nanowires, dip time of probe for deposition of sensing layer and working pH range of the sensing probe have been accomplished through SPR measurements. The optimized sensing probe displays high sensitivity, broad working range and a minimum limit of detection value and has been successfully tested for malic acid determination in real samples of fruit juices. The current work presents a novel perspective towards malic acid determination as the unique and cooperative combination of FOSPR and nanomaterials provides myriad advantages such as enhanced sensitivity, specificity, compactness together with the possibility of online monitoring and remote sensing.Keywords: surface plasmon resonance, optical fiber, sensor, malic acid
Procedia PDF Downloads 380407 Cut-Off of CMV Cobas® Taqman® (CAP/CTM Roche®) for Introduction of Ganciclovir Pre-Emptive Therapy in Allogeneic Hematopoietic Stem Cell Transplant Recipients
Authors: B. B. S. Pereira, M. O. Souza, L. P. Zanetti, L. C. S. Oliveira, J. R. P. Moreno, M. P. Souza, V. R. Colturato, C. M. Machado
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Background: The introduction of prophylactic or preemptive therapies has effectively decreased the CMV mortality rates after hematopoietic stem cell transplantation (HSCT). CMV antigenemia (pp65) or quantitative PCR are methods currently approved for CMV surveillance in pre-emptive strategies. Commercial assays are preferred as cut-off levels defined by in-house assays may vary among different protocols and in general show low reproducibility. Moreover, comparison of published data among different centers is only possible if international standards of quantification are included in the assays. Recently, the World Health Organization (WHO) established the first international standard for CMV detection. The real time PCR COBAS Ampliprep/ CobasTaqMan (CAP/CTM) (Roche®) was developed using the WHO standard for CMV quantification. However, the cut-off for the introduction of antiviral has not been determined yet. Methods: We conducted a retrospective study to determine: 1) the sensitivity and specificity of the new CMV CAP/CTM test in comparison with pp65 antigenemia to detect episodes of CMV infection/reactivation, and 2) the cut-off of viral load for introduction of ganciclovir (GCV). Pp65 antigenemia was performed and the corresponding plasma samples were stored at -20°C for further CMV detection by CAP/CTM. Comparison of tests was performed by kappa index. The appearance of positive antigenemia was considered the state variable to determine the cut-off of CMV viral load by ROC curve. Statistical analysis was performed using SPSS software version 19 (SPSS, Chicago, IL, USA.). Results: Thirty-eight patients were included and followed from August 2014 through May 2015. The antigenemia test detected 53 episodes of CMV infection in 34 patients (89.5%), while CAP/CTM detected 37 episodes in 33 patients (86.8%). AG and PCR results were compared in 431 samples and Kappa index was 30.9%. The median time for first AG detection was 42 (28-140) days, while CAP/CTM detected at a median of 7 days earlier (34 days, ranging from 7 to 110 days). The optimum cut-off value of CMV DNA was 34.25 IU/mL to detect positive antigenemia with 88.2% of sensibility, 100% of specificity and AUC of 0.91. This cut-off value is below the limit of detection and quantification of the equipment which is 56 IU/mL. According to CMV recurrence definition, 16 episodes of CMV recurrence were detected by antigenemia (47.1%) and 4 (12.1%) by CAP/CTM. The duration of viremia as detected by antigenemia was shorter (60.5% of the episodes lasted ≤ 7 days) in comparison to CAP/CTM (57.9% of the episodes lasting 15 days or more). This data suggests that the use of antigenemia to define the duration of GCV therapy might prompt early interruption of antiviral, which may favor CMV reactivation. The CAP/CTM PCR could possibly provide a safer information concerning the duration of GCV therapy. As prolonged treatment may increase the risk of toxicity, this hypothesis should be confirmed in prospective trials. Conclusions: Even though CAP/CTM by ROCHE showed great qualitative correlation with the antigenemia technique, the fully automated CAP/CTM did not demonstrate increased sensitivity. The cut-off value below the limit of detection and quantification may result in delayed introduction of pre-emptive therapy.Keywords: antigenemia, CMV COBAS/TAQMAN, cytomegalovirus, antiviral cut-off
Procedia PDF Downloads 192406 Blood Oxygen Saturation Measurement System Using Broad-Band Light Source with LabVIEW Program
Authors: Myoung Ah Kim, Dong Ho Sin, Chul Gyu Song
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Blood oxygen saturation system is a well-established, noninvasive photoplethysmographic method to monitor vital signs. Conventional blood oxygen saturation measurements for the two LED light source is the ambiguity of the oxygen saturation measurement principle and the measurement results greatly influenced and heat and motion artifact. A high accuracy in order to solve these problems blood oxygen saturation measuring method has been proposed using a broadband light source that can be easily understood by the algorithm. The measurement of blood oxygen saturation based on broad-band light source has advantage of simple testing facility and easy understanding. Broadband light source based on blood oxygen saturation measuring program proposed in this paper is a combination of LabVIEW and MATLAB. Using the wavelength range of 450 nm-750 nm using a floating light absorption of oxyhemoglobin and deoxyhemoglobin to measure the blood oxygen saturation. Hand movement is to fix the probe to the motor stage in order to prevent oxygen saturation measurement that affect the sample and probe kept constant interval. Experimental results show that the proposed method noticeably increases the accuracy and saves time compared with the conventional methods.Keywords: oxygen saturation, broad-band light source, CCD, light reflectance theory
Procedia PDF Downloads 461405 Analysis of the Annual Proficiency Testing Procedure for Intermediate Reference Laboratories Conducted by the National Reference Laboratory from 2013 to 2017
Authors: Reena K., Mamatha H. G., Somshekarayya, P. Kumar
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Objectives: The annual proficiency testing of intermediate reference laboratories is conducted by the National Reference Laboratory (NRL) to assess the efficiency of the laboratories to correctly identify Mycobacterium tuberculosis and to determine its drug susceptibility pattern. The proficiency testing results from 2013 to 2017 were analyzed to determine laboratories that were consistent in reporting quality results and those that had difficulty in doing so. Methods: A panel of twenty cultures were sent out to each of these laboratories. The laboratories were expected to grow the cultures in their own laboratories, set up drug susceptibly testing by all the methods they were certified for and report the results within the stipulated time period. The turnaround time for reporting results, specificity, sensitivity positive and negative predictive values and efficiency of the laboratory in identifying the cultures were analyzed. Results: Most of the laboratories had reported their results within the stipulated time period. However, there was enormous delay in reporting results from few of the laboratories. This was mainly due to improper functioning of the biosafety level III laboratory. Only 40% of the laboratories had 100% efficiency in solid culture using Lowenstein Jensen medium. This was expected as a solid culture, and drug susceptibility testing is not used for diagnosing drug resistance. Rapid molecular methods such as Line probe assay and Genexpert are used to determine drug resistance. Automated liquid culture system such as the Mycobacterial growth indicator tube is used to determine prognosis of the patient while on treatment. It was observed that 90% of the laboratories had achieved 100% in the liquid culture method. Almost all laboratories had achieved 100% efficiency in the line probe assay method which is the method of choice for determining drug-resistant tuberculosis. Conclusion: Since the liquid culture and line probe assay technologies are routinely used for the detection of drug-resistant tuberculosis the laboratories exhibited higher level of efficiency as compared to solid culture and drug susceptibility testing which are rarely used. The infrastructure of the laboratory should be maintained properly so that samples can be processed safely and results could be declared on time.Keywords: annual proficiency testing, drug susceptibility testing, intermediate reference laboratory, national reference laboratory
Procedia PDF Downloads 182404 Study on the Non-Contact Sheet Resistance Measuring of Silver Nanowire Coated Film Using Terahertz Wave
Authors: Dong-Hyun Kim, Wan-Ho Chung, Hak-Sung Kim
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In this work, non-destructive evaluation was conducted to measure the sheet resistance of silver nanowire coated film and find a damage of that film using terahertz (THz) wave. Pulse type THz instrument was used, and the measurement was performed under transmission and pitch-catch reflection modes with 30 degree of incidence angle. In the transmission mode, the intensity of the THz wave was gradually increased as the conductivity decreased. Meanwhile, the intensity of THz wave was decreased as the conductivity decreased in the pitch-catch reflection mode. To confirm the conductivity of the film, sheet resistance was measured by 4-point probe station. Interaction formula was drawn from a relation between the intensity and the sheet resistance. Through substituting sheet resistance to the formula and comparing the resultant value with measured maximum THz wave intensity, measurement of sheet resistance using THz wave was more suitable than that using 4-point probe station. In addition, the damage on the silver nanowire coated film was detected by applying the THz image system. Therefore, the reliability of the entire film can be also be ensured. In conclusion, real-time monitoring using the THz wave can be applied in the transparent electrodes with detecting the damaged area as well as measuring the sheet resistance.Keywords: terahertz wave, sheet resistance, non-destructive evaluation, silver nanowire
Procedia PDF Downloads 491403 Fluorescence in situ Hybridization (FISH) Detection of Bacteria and Archaea in Fecal Samples
Authors: Maria Nejjari, Michel Cloutier, Guylaine Talbot, Martin Lanthier
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The fluorescence in situ hybridization (FISH) is a staining technique that allows the identification, detection and quantification of microorganisms without prior cultivation by means of epifluorescence and confocal laser scanning microscopy (CLSM). Oligonucleotide probes have been used to detect bacteria and archaea that colonize the cattle and swine digestive systems. These bacterial strains have been obtained from fecal samples issued from cattle manure and swine slurry. The collection of these samples has been done at 3 different pit’s levels A, B and C with same height. Two collection depth levels have been taken in consideration, one collection level just under the pit’s surface and the second one at the bottom of the pit. Cells were fixed and FISH was performed using oligonucleotides of 15 to 25 nucleotides of length associated with a fluorescent molecule Cy3 or Cy5. The double hybridization using Cy3 probe targeting bacteria (Cy3-EUB338-I) along with a Cy5 probe targeting Archaea (Gy5-ARCH915) gave a better signal. The CLSM images show that there are more bacteria than archaea in swine slurry. However, the choice of fluorescent probes is critical for getting the double hybridization and a unique signature for each microorganism. FISH technique is an easy way to detect pathogens like E. coli O157, Listeria, Salmonella that easily contaminate water streams, agricultural soils and, consequently, food products and endanger human health.Keywords: archaea, bacteria, detection, FISH, fluorescence
Procedia PDF Downloads 388402 Equation for Predicting Inferior Vena Cava Diameter as a Potential Pointer for Heart Failure Diagnosis among Adult in Azare, Bauchi State, Nigeria
Authors: M. K. Yusuf, W. O. Hamman, U. E. Umana, S. B. Oladele
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Background: Dilatation of the inferior vena cava (IVC) is used as the ultrasonic diagnostic feature in patients suspected of congestive heart failure. The IVC diameter has been reported to vary among the various body mass indexes (BMI) and body shape indexes (ABSI). Knowledge of these variations is useful in precision diagnoses of CHF by imaging scientists. Aim: The study aimed to establish an equation for predicting the ultrasonic mean diameter of the IVC among the various BMI/ABSI of inhabitants of Azare, Bauchi State-Nigeria. Methodology: Two hundred physically healthy adult subjects of both sexes were classified into under, normal, over, and obese weights using their BMIs after selection using a structured questionnaire following their informed consent for an abdominal ultrasound scan. The probe was placed on the midline of the body, halfway between the xiphoid process and the umbilicus, with the marker on the probe directed towards the patient's head to obtain a longitudinal view of the IVC. The maximum IVC diameter was measured from the subcostal view using the electronic caliper of the scan machine. The mean value of each group was obtained, and the results were analysed. Results: A novel equation {(IVC Diameter = 1.04 +0.01(X) where X= BMI} has been generated for determining the IVC diameter among the populace. Conclusion: An equation for predicting the IVC diameter from individual BMI values in apparently healthy subjects has been established.Keywords: equation, ultrasonic, IVC diameter, body adiposities
Procedia PDF Downloads 72401 NanoFrazor Lithography for advanced 2D and 3D Nanodevices
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
Procedia PDF Downloads 72400 Efficacy and User Satisfaction on the Rama-Chest Cryo Arm Innovation for Bronchoscopic Cryotherapy
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
Procedia PDF Downloads 244399 Prediction of Cardiovascular Markers Associated With Aromatase Inhibitors Side Effects Among Breast Cancer Women in Africa
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
Procedia PDF Downloads 79398 Influence of the Local External Pressure on Measured Parameters of Cutaneous Microcirculation
Authors: Irina Mizeva, Elena Potapova, Viktor Dremin, Mikhail Mezentsev, Valeri Shupletsov
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The local tissue perfusion is regulated by the microvascular tone which is under the control of a number of physiological mechanisms. Laser Doppler flowmetry (LDF) together with wavelet analyses is the most commonly used technique to study the regulatory mechanisms of cutaneous microcirculation. External factors such as temperature, local pressure of the probe on the skin, etc. influence on the blood flow characteristics and are used as physiological tests to evaluate microvascular regulatory mechanisms. Local probe pressure influences on the microcirculation parameters measured by optical methods: diffuse reflectance spectroscopy, fluorescence spectroscopy, and LDF. Therefore, further study of probe pressure effects can be useful to improve the reliability of optical measurement. During pressure tests variation of the mean perfusion measured by means of LDF usually is estimated. An additional information concerning the physiological mechanisms of the vascular tone regulation system in response to local pressure can be obtained using spectral analyses of LDF samples. The aim of the present work was to develop protocol and algorithm of data processing appropriate for study physiological response to the local pressure test. Involving 6 subjects (20±2 years) and providing 5 measurements for every subject we estimated intersubject and-inter group variability of response of both averaged and oscillating parts of the LDF sample on external surface pressure. The final purpose of the work was to find special features which further can be used in wider clinic studies. The cutaneous perfusion measurements were carried out by LAKK-02 (SPE LAZMA Ltd., Russia), the skin loading was provided by the originally designed device which allows one to distribute the pressure around the LDF probe. The probe was installed on the dorsal part of the distal finger of the index figure. We collected measurements continuously for one hour and varied loading from 0 to 180mmHg stepwise with a step duration of 10 minutes. Further, we post-processed the samples using the wavelet transform and traced the energy of oscillations in five frequency bands over time. Weak loading leads to pressure-induced vasodilation, so one should take into account that the perfusion measured under pressure conditions will be overestimated. On the other hand, we revealed a decrease in endothelial associated fluctuations. Further loading (88 mmHg) induces amplification of pulsations in all frequency bands. We assume that such loading leads to a higher number of closed capillaries, higher input of arterioles in the LDF signal and as a consequence more vivid oscillations which mainly are formed in arterioles. External pressure higher than 144 mmHg leads to the decrease of oscillating components, after removing the loading very rapid restore of the tissue perfusion takes place. In this work, we have demonstrated that local skin loading influence on the microcirculation parameters measured by optic technique; this should be taken into account while developing portable electronic devices. The proposed protocol of local loading allows one to evaluate PIV as far as to trace dynamic of blood flow oscillations. This study was supported by the Russian Science Foundation under project N 18-15-00201.Keywords: blood microcirculation, laser Doppler flowmetry, pressure-induced vasodilation, wavelet analyses blood
Procedia PDF Downloads 151397 Highly Specific DNA-Aptamer-Based Electrochemical Biosensor for Mercury (II) and Lead (II) Ions Detection in Water Samples
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
Procedia PDF Downloads 162396 Gene Expression Profiling of Iron-Related Genes of Pasteurella multocida Serotype A Strain PMTB2.1
Authors: Shagufta Jabeen, Faez Jesse Firdaus Abdullah, Zunita Zakaria, Nurulfiza Mat Isa, Yung Chie Tan, Wai Yan Yee, Abdul Rahman Omar
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Pasteurella multocida is associated with acute, as well as, chronic infections in avian and bovine such as pasteurellosis and hemorrhagic septicemia (HS) in cattle and buffaloes. Iron is one of the most important nutrients for pathogenic bacteria including Pasteurella and acts as a cofactor or prosthetic group in several essential enzymes and is needed for amino acid, pyrimidine, and DNA biosynthesis. In our recent study, we showed that 2% of Pasteurella multocida serotype A strain PMTB2.1 encode for iron regulating genes (Accession number CP007205.1). Genome sequencing of other Pasteurella multocida serotypes namely PM70 and HB01 also indicated up to 2.5% of the respective genome encode for iron regulating genes, suggesting that Pasteurella multocida genome comprises of multiple systems for iron uptake. Since P. multocida PMTB2.1 has more than 40 CDs out of 2097 CDs (approximately 2%), encode for iron-regulated. The gene expression profiling of four iron-regulating genes namely fbpb, yfea, fece and fur were characterized under iron-restricted environment. The P. multocida strain PMTB2.1 was grown in broth with and without iron chelating agent and samples were collected at different time points. Relative mRNA expression profile of these genes was determined using Taqman probe based real-time PCR assay. The data analysis, normalization with two house-keeping genes and the quantification of fold changes were carried out using Bio-Rad CFX manager software version 3.1. Results of this study reflect that iron reduced environment has significant effect on expression profile of iron regulating genes (p < 0.05) when compared to control (normal broth) and all evaluated genes act differently with response to iron reduction in media. The highest relative fold change of fece gene was observed at early stage of treatment indicating that PMTB2.1 may utilize its periplasmic protein at early stage to acquire iron. Furthermore, down-regulation expression of fece with the elevated expression of other genes at later time points suggests that PMTB2.1 control their iron requirements in response to iron availability by down-regulating the expression of iron proteins. Moreover, significantly high relative fold change (p ≤ 0.05) of fbpb gene is probably associated with the ability of P. multocida to directly use host iron complex such as hem, hemoglobin. In addition, the significant increase (p ≤ 0.05) in fbpb and yfea expressions also reflects the utilization of multiple iron systems in P. multocida strain PMTB2.1. The findings of this study are very much important as relative scarcity of free iron within hosts creates a major barrier to microbial growth inside host and utilization of outer-membrane proteins system in iron acquisition probably occurred at early stage of infection with P. multocida. In conclusion, the presence and utilization of multiple iron system in P. multocida strain PMTB2.1 revealed the importance of iron in the survival of P. multocida.Keywords: iron-related genes, real-time PCR, gene expression profiling, fold changes
Procedia PDF Downloads 462395 Signal Amplification Using Graphene Oxide in Label Free Biosensor for Pathogen Detection
Authors: Agampodi Promoda Perera, Yong Shin, Mi Kyoung Park
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The successful detection of pathogenic bacteria in blood provides important information for early detection, diagnosis and the prevention and treatment of infectious diseases. Silicon microring resonators are refractive-index-based optical biosensors that provide highly sensitive, label-free, real-time multiplexed detection of biomolecules. We demonstrate the technique of using GO (graphene oxide) to enhance the signal output of the silicon microring optical sensor. The activated carboxylic groups in GO molecules bind directly to single stranded DNA with an amino modified 5’ end. This conjugation amplifies the shift in resonant wavelength in a real-time manner. We designed a capture probe for strain Staphylococcus aureus of 21 bp and a longer complementary target sequence of 70 bp. The mismatched target sequence we used was of Streptococcus agalactiae of 70 bp. GO is added after the complementary binding of the probe and target. GO conjugates to the unbound single stranded segment of the target and increase the wavelength shift on the silicon microring resonator. Furthermore, our results show that GO could successfully differentiate between the mismatched DNA sequences from the complementary DNA sequence. Therefore, the proposed concept could effectively enhance sensitivity of pathogen detection sensors.Keywords: label free biosensor, pathogenic bacteria, graphene oxide, diagnosis
Procedia PDF Downloads 469394 Probing Extensive Air Shower Primaries and Their Interactions by Combining Individual Muon Tracks and Shower Depth
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
Procedia PDF Downloads 229393 Design and Preliminary Evaluation of Benzoxazolone-Based Agents for Targeting Mitochondrial-Located Translocator Protein
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
Procedia PDF Downloads 462392 Challenges in Creating Social Capital: A Perspective of Muslim Female Managers in Malaysia
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
Procedia PDF Downloads 130391 Impact of External Temperature on the Speleothem Growth in the Moravian Karst
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
Procedia PDF Downloads 144390 Numerical Simulation of Phase Transfer during Cryosurgery for an Irregular Tumor Using Hybrid Approach
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 125389 Prevalence and Genetic Determinant of Drug Resistant Tuberculosis among Patients Completing Intensive Phase of Treatment in a Tertiary Referral Center in Nigeria
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 288388 Association of Selected Polymorphisms of BER Pathway with the Risk of Colorectal Cancer in the Polish Population
Authors: Jacek Kabzinski, Karolina Przybylowska, Lukasz Dziki, Adam Dziki, Ireneusz Majsterek
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The incidence of colorectal cancer (CRC) is increasing from year to year. Despite intensive research CRC etiology remains unknown. Studies suggest that at the basis of the process of carcinogenesis can lie reduced efficiency of DNA repair mechanisms, often caused by polymorphisms in DNA repair genes. The aim of the study was to determine the relationship between gene polymorphisms Pro242Arg of PolB gene and Arg780His of Lig3 gene and modulation of the risk of colorectal cancer in the Polish population. Determination of the molecular basis of carcinogenesis process and predicting increased risk will allow qualifying patients to increased risk group and including them in preventive program. We used blood collected from 110 patients diagnosed with colorectal cancer. The control group consisted of equal number of healthy people. Genotyping was performed by TaqMan method. The obtained results indicate that the genotype 780Arg/His of Lig3 gene is associated with an increased risk of colorectal cancer. On the basis of these results, we conclude that Lig3 gene polymorphism Arg780His may be associated with an increased risk of colorectal cancer.Keywords: BER, colorectal cancer, PolB, Lig3, polymorphisms
Procedia PDF Downloads 454387 Invistigation of Surface Properties of Nanostructured Carbon Films
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 268386 The Association Between COL4A3 Variant RS55703767 With the Susceptibility to Diabetic Kidney Disease in Patients with Type 2 Diabetes Mellitus: Results from the Cohort Study
Authors: Zi-Han Li, Zi-Jun Sun, Dong-Yuan Chang, Li Zhu, Min Chen, Ming-Hui Zhao
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Aims: A genome-wide association study (GWAS) reported that patients with the rs55703767 minor allele in collagen type IV α3 chain encoding gene COL4A3 showed protection against diabetic kidney disease (DKD) in type 1 diabetes mellitus (T1DM). However, the role of rs55703767 in type 2 DKD has not been elucidated. The aim of the current study was to investigate the association between COL4A3 variant rs55703767 and DKD risk in Chinese patients with type 2 diabetes mellitus (T2DM). Methods: This nested case-control study was performed on 1311 patients who had T2DM for at least 10 years, including 580 with DKD and 731 without DKD. We detected the genotypes of all patients by TaqMan SNP Genotyping Assay and analyzed the association between COL4A3 variant rs55703767 and DKD risk. Results: Genetic analysis revealed that there was no significant difference between T2DM patients with DKD and those without DKD regarding allele or genotype frequencies of rs55703767, and the effect of this variant was not hyperglycemia specific. Conclusion: Our findings suggested that there was no detectable association between the COL4A3 variant rs55703767 and the susceptibility to DKD in the Chinese T2DM population.Keywords: collagen type IV α3 chain, gene polymorphism, type 2 diabetes, diabetic kidney disease
Procedia PDF Downloads 112385 Quantitative Evaluation of Mitral Regurgitation by Using Color Doppler Ultrasound
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 371384 The Influence of Polymorphisms of NER System Genes on the Risk of Colorectal Cancer in the Polish Population
Authors: Ireneusz Majsterek, Karolina Przybylowska, Lukasz Dziki, Adam Dziki, Jacek Kabzinski
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Colorectal cancer (CRC) is one of the deadliest cancers. Every year we see an increase in the number of cases, and in spite of intensive research etiology of the disease remains unknown. For many years, researchers are seeking to associate genetic factors with an increased risk of CRC, so far it has proved to be a compelling link between the MMR system of DNA repair and hereditary nonpolyposis colorectal cancers (HNPCC). Currently, research is focused on finding the relationship between the remaining DNA repair systems and an increased risk of developing colorectal cancer. The aim of the study was to determine the relationship between gene polymorphisms Ser835Ser of XPF gene and Gly23Ala of XPA gene–elements of NER DNA repair system, and modulation of the risk of colorectal cancer in the Polish population. Determination of the molecular basis of carcinogenesis process and predicting increased risk will allow qualifying patients to increased risk group and including them in preventive program. We used blood collected from 110 patients diagnosed with colorectal cancer. The control group consisted of equal number of healthy people. Genotyping was performed by TaqMan method. The obtained results indicate that the genotype 23Gly/Ala of XPA gene is associated with an increased risk of colorectal cancer, while 23Ala/Ala as well as TCT allele of Ser835Ser of XPF gene may reduce the risk of CRC.Keywords: NER, colorectal cancer, XPA, XPF, polymorphisms
Procedia PDF Downloads 568383 Combustion Characteristics of Wet Woody Biomass in a Grate Furnace: Including Measurements within the Bed
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 327382 The Fast Diagnosis of Acanthamoeba Keratitis Using Real-Time PCR Assay
Authors: Fadime Eroglu
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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 121381 Remote BioMonitoring of Mothers and Newborns for Temperature Surveillance Using a Smart Wearable Sensor: Techno-Feasibility Study and Clinical Trial in Southern India
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
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