Search results for: optical strain measurement
Commenced in January 2007
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Edition: International
Paper Count: 5622

Search results for: optical strain measurement

4032 Measuring Fundamental Growth Needs in a Youth Boatbuilding Context

Authors: Shane Theunissen, Rob Grandy

Abstract:

Historically and we would fairly conventionally within our formal schooling systems, we have convergent testing where all the students are expected to converge on the same answer, and that answer has been determined by an external authority that is reproducing knowledge of the hegemon. Many youths may not embody the cultural capital that's rewarded in formal schooling contexts as they aren't able to converge on the required answer that's being determined by the classroom teacher or the administrators. In this paper, we explore divergent processes that promote creative problem-solving. We embody this divergent process in our measurement of fundamental growth needs. To this end, we utilize the Mosaic Approach as a method for implementing the Outcomes That Matter framework. Outcomes That Matter is the name of the measurement tool built around the Circle of Courage framework, which is a way of identifying fundamental growth needs for young people. The Circle of Courage was developed by Martin-Broken-Leg and colleagues as a way to connect indigenous child-rearing philosophies with contemporary resilience and positive psychology research. The Outcomes that Matter framework puts forward four categories of growth needs for young people. These are: Belonging, which on a macro scale is acceptance into the greater community of practice, Mastery which includes a constellation of concepts including confidence, motivation, self-actualization, and self-determination, Independence refers to a sense of personal power into autonomy within a context where creativity and problem solving, and a personal voice can begin to emerge, and finally Generosity which includes interpersonal things like conflict resolution and teamwork. Outcomes of Matter puts these four domains into a measurement tool that facilitates collaborative assessment between the youth, teachers, and recreation therapists that allows for youth-led narratives pertaining to their fundamental growth outcomes. This application of the Outcomes That Matter framework is unique as it may be the first application of this framework in an educational boatbuilding context.

Keywords: collaboration, empowerment, outcomes that matter, mosaic approach, boat building

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4031 Self-Efficacy Perceptions and the Attitudes of Prospective Teachers towards Assessment and Evaluation

Authors: Münevver Başman, Ezel Tavşancıl

Abstract:

Making the right decisions about students depends on teachers’ use of the assessment and evaluation techniques effectively. In order to do that, teachers should have positive attitudes and adequate self-efficacy perception towards assessment and evaluation. The purpose of this study is to investigate relationship between self-efficacy perception and the attitudes of prospective teachers towards assessment and evaluation and what kind of differences these issues have in terms of a variety of demographic variables. The study group consisted of 277 prospective teachers who have been studying in different departments of Marmara University, Faculty of Education. In this study, ‘Personal Information Form’, ‘A Perceptual Scale for Measurement and Evaluation of Prospective Teachers Self-Efficacy in Education’ and ‘Attitudes toward Educational Measurement Inventory’ are applied. As a result, positive correlation was found between self-efficacy perceptions and the attitudes of prospective teachers towards assessment and evaluation. Considering different departments, there is a significant difference between the mean score of attitudes of prospective teachers and between the mean score of self-efficacy perceptions of them. However, considering variables of attending statistics class and the class types at the graduated high school, there is no significant difference between the mean score of attitudes of prospective teachers and between the mean score of self-efficacy perceptions of them.

Keywords: attitude, perception, prospective teacher, self-efficacy

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4030 Feasibility Study of Mine Tailing’s Treatment by Acidithiobacillus thiooxidans DSM 26636

Authors: M. Gómez-Ramírez, A. Rivas-Castillo, I. Rodríguez-Pozos, R. A. Avalos-Zuñiga, N. G. Rojas-Avelizapa

Abstract:

Among the diverse types of pollutants produced by anthropogenic activities, metals represent a serious threat, due to their accumulation in ecosystems and their elevated toxicity. The mine tailings of abandoned mines contain high levels of metals such as arsenic (As), zinc (Zn), copper (Cu), and lead (Pb), which do not suffer any degradation process, they are accumulated in environment. Abandoned mine tailings potentially could contaminate rivers and aquifers representing a risk for human health due to their high metal content. In an attempt to remove the metals and thereby mitigate the environmental pollution, an environmentally friendly and economical method of bioremediation has been introduced. Bioleaching has been actively studied over the last several years, and it is one of the bioremediation solutions used to treat heavy metals contained in sewage sludge, sediment and contaminated soil. Acidithiobacillus thiooxidans, an extremely acidophilic, chemolithoautotrophic, gram-negative, rod shaped microorganism, which is typically related to Cu mining operations (bioleaching), has been well studied for industrial applications. The sulfuric acid produced plays a major role in bioleaching. Specifically, Acidithiobacillus thiooxidans strain DSM 26636 has been able to leach Al, Ni, V, Fe, Mg, Si, and Ni contained in slags from coal combustion wastes. The present study reports the ability of A. thiooxidans DSM 26636 for the bioleaching of metals contained in two different mine tailing samples (MT1 and MT2). It was observed that Al, Fe, and Mn were removed in 36.3±1.7, 191.2±1.6, and 4.5±0.2 mg/kg for MT1, and in 74.5±0.3, 208.3±0.5, and 20.9±0.1 for MT2. Besides, < 1.5 mg/kg of Au and Ru were also bioleached from MT1; in MT2, bioleaching of Zn was observed at 55.7±1.3 mg/kg, besides removal of < 1.5 mg/kg was observed for As, Ir, Li, and 0.6 for Os in this residue. These results show the potential of strain DSM 26636 for the bioleaching of metals that came from different mine tailings.

Keywords: A. thiooxidans, bioleaching, metals, mine tailings

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4029 Design of an Instrumentation Setup and Data Acquisition System for a GAS Turbine Engine Using Suitable DAQ Software

Authors: Syed Nauman Bin Asghar Bukhari, Mohtashim Mansoor, Mohammad Nouman

Abstract:

Engine test-Bed system is a fundamental tool to measure dynamic parameters, economic performance, and reliability of an aircraft Engine, and its automation and accuracy directly influences the precision of acquired and analysed data. In this paper, we present the design of digital Data Acquisition (DAQ) system for a vintage aircraft engine test bed that lacks the capability of displaying all the analyzed parameters at one convenient location (one panel-one screen). Recording such measurements in the vintage test bed is not only time consuming but also prone to human errors. Digitizing such measurement system requires a Data Acquisition (DAQ) system capable of recording these parameters and displaying them on one screen-one panel monitor. The challenge in designing upgrade to the vintage systems arises with a need to build and integrate digital measurement system from scratch with a minimal budget and modifications to the existing vintage system. The proposed design not only displays all the key performance / maintenance parameters of the gas turbine engines for operator as well as quality inspector on separate screens but also records the data for further processing / archiving.

Keywords: Gas turbine engine, engine test cell, data acquisition, instrumentation

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4028 Antimicrobial Activity of Some Plant Extracts against Clinical Pathogen and Candida Species

Authors: Marwan Khalil Qader, Arshad Mohammad Abdullah

Abstract:

Antimicrobial resistance is a major cause of significant morbidity and mortality globally. Seven plant extracts (Plantago mediastepposa, Quercusc infectoria, Punic granatum, Thymus lcotschyana, Ginger officeinals, Rhus angustifolia and Cinnamon) were collected from different regions of Kurdistan region of Iraq. These plants’ extracts were dissolved in absolute ethanol and distillate water, after which they were assayed in vitro as an antimicrobial activity against Candida tropicalis, Candida albicanus, Candida dublinensis, Candida krusei and Candida glabrata also against 2 Gram-positive (Bacillus subtilis and Staphylococcus aureus) and 3 Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Klebsilla pneumonia). The antimicrobial activity was determined in ethanol extracts and distilled water extracts of these plants. The ethanolic extracts of Q. infectoria showed the maximum activity against all species of Candida fungus. The minimum inhibition zone of the Punic granatum ethanol extracts was 0.2 mg/ml for all microorganisms tested. Klebsilla pneumonia was the most sensitive bacterial strain to Quercusc infectoria and Rhus angustifolia ethanol extracts. Among both Gram-positive and Gram-negative bacteria tested with MIC of 0.2 mg/ml, the minimum inhibition zone of Ginger officeinals D. W. extracts was 0.2 mg/mL against Pseudomonas aeruginosa and Klebsilla pneumonia. The most sensitive bacterial strain to Thymus lcotschyana and Plantago mediastepposa D.W. extracts was S. aureus and E. coli.

Keywords: antimicrobial activity, pathogenic bacteria, plant extracts, chemical systems engineering

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4027 Mapping Method to Solve a Nonlinear Schrodinger Type Equation

Authors: Edamana Vasudevan Krishnan

Abstract:

This paper studies solitons in optical materials with the help of Mapping Method. Two types of nonlinear media have been investigated, namely, the cubic nonlinearity and the quintic nonlinearity. The soliton solutions, shock wave solutions and singular solutions have been derives with certain constraint conditions.

Keywords: solitons, integrability, metamaterials, mapping method

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4026 Implementation of Dozer Push Measurement under Payment Mechanism in Mining Operation

Authors: Anshar Ajatasatru

Abstract:

The decline of coal prices over past years have been significantly increasing the awareness of effective mining operation. A viable step must be undertaken in becoming more cost competitive while striving for best mining practice especially at Melak Coal Mine in East Kalimantan, Indonesia. This paper aims to show how effective dozer push measurement method can be implemented as it is controlled by contract rate on the unit basis of USD ($) per bcm. The method emerges from an idea of daily dozer push activity that continually shifts the overburden until final target design by mine planning. Volume calculation is then performed by calculating volume of each time overburden is removed within determined distance using cut and fill method from a high precision GNSS system which is applied into dozer as a guidance to ensure the optimum result of overburden removal. Accumulation of daily to weekly dozer push volume is found 95 bcm which is multiplied by average sell rate of $ 0,95, thus the amount monthly revenue is $ 90,25. Furthermore, the payment mechanism is then based on push distance and push grade. The push distance interval will determine the rates that vary from $ 0,9 - $ 2,69 per bcm and are influenced by certain push slope grade from -25% until +25%. The amount payable rates for dozer push operation shall be specifically following currency adjustment and is to be added to the monthly overburden volume claim, therefore, the sell rate of overburden volume per bcm may fluctuate depends on the real time exchange rate of Jakarta Interbank Spot Dollar Rate (JISDOR). The result indicates that dozer push measurement can be one of the surface mining alternative since it has enabled to refine method of work, operating cost and productivity improvement apart from exposing risk of low rented equipment performance. In addition, payment mechanism of contract rate by dozer push operation scheduling will ultimately deliver clients by almost 45% cost reduction in the form of low and consistent cost.

Keywords: contract rate, cut-fill method, dozer push, overburden volume

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4025 Optic Nerve Sheath Measurement in Children with Head Trauma

Authors: Sabiha Sahin, Kursad Bora Carman, Coskun Yarar

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Introduction: Measuring the diameter of the optic nerve sheath is a noninvasive and easy to use imaging technique to predict intracranial pressure in children and adults. The aim was to measure the diameter of the optic nerve sheath in pediatric head trauma. Methods: The study group consisted of 40 children with healthy and 40 patients with head trauma. Transorbital sonographic measurement of the optic nerve sheath diameter was performed. Conclusion: The mean diameters of the optic nerve sheath of right and left eyes were 0.408 ± 0.064 mm and 0.417 ± 0.065 mm, respectively, in the trauma group. These results were higher in patients than in control group. There was a negative correlation between optic nerve sheath diameters and Glasgow Coma Scales in patients with head trauma (p < 0.05). There was a positive correlation between optic nerve sheath diameters and positive CT findings, systolic blood pressure in patients with head trauma. The clinical status of the patients at admission, blood pH and lactate level were related to the optic nerve sheath diameter. Conclusion: Measuring the diameter of the optic nerve sheath is not an invasive technique and can be easily used to predict increased intracranial pressure and to prevent secondary brain injury.

Keywords: head trauma, intracranial pressure, optic nerve, sonography

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4024 Application of Interferometric Techniques for Quality Control Oils Used in the Food Industry

Authors: Andres Piña, Amy Meléndez, Pablo Cano, Tomas Cahuich

Abstract:

The purpose of this project is to propose a quick and environmentally friendly alternative to measure the quality of oils used in food industry. There is evidence that repeated and indiscriminate use of oils in food processing cause physicochemical changes with formation of potentially toxic compounds that can affect the health of consumers and cause organoleptic changes. In order to assess the quality of oils, non-destructive optical techniques such as Interferometry offer a rapid alternative to the use of reagents, using only the interaction of light on the oil. Through this project, we used interferograms of samples of oil placed under different heating conditions to establish the changes in their quality. These interferograms were obtained by means of a Mach-Zehnder Interferometer using a beam of light from a HeNe laser of 10mW at 632.8nm. Each interferogram was captured, analyzed and measured full width at half-maximum (FWHM) using the software from Amcap and ImageJ. The total of FWHMs was organized in three groups. It was observed that the average obtained from each of the FWHMs of group A shows a behavior that is almost linear, therefore it is probable that the exposure time is not relevant when the oil is kept under constant temperature. Group B exhibits a slight exponential model when temperature raises between 373 K and 393 K. Results of the t-Student show a probability of 95% (0.05) of the existence of variation in the molecular composition of both samples. Furthermore, we found a correlation between the Iodine Indexes (Physicochemical Analysis) and the Interferograms (Optical Analysis) of group C. Based on these results, this project highlights the importance of the quality of the oils used in food industry and shows how Interferometry can be a useful tool for this purpose.

Keywords: food industry, interferometric, oils, quality control

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4023 Microscopic Visualization of the Ice Slurry Ice Particles

Authors: Juan José Milón Guzmán, Herbert Jesús Del Carpio Beltrán, Sergio Leal Braga

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Visualizations of ice particles of ice slurry are performed. The form and size of ice particles is investigated by optical microscopy. It permits to evaluate statistically the geometrical shapes of the ice crystals. The observed particle size corresponds with the different solutes (sugar, salt, propylene glycol).

Keywords: ice slurry, visualization, ice particles, solutes

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4022 Microscopic and Mesoscopic Deformation Behaviors of Mg-2Gd Alloy with or without Li Addition

Authors: Jing Li, Li Jin, Fulin Wang, Jie Dong, Wenjiang Ding

Abstract:

Mg-Li dual-phase alloy exhibits better combination of yield strength and elongation than the Mg single-phase alloy. To exploit its deformation behavior, the deformation mechanisms of Mg-2Gd alloy with or without Li addition, i.e., Mg-6Li-2Gd and Mg-2Gd alloy, have been studied at both microscale and mesoscale. EBSD-assisted slip trace, twin trace, and texture evolution analysis show that the α-Mg phase of Mg-6Li-2Gd alloy exhibits different microscopic deformation mechanisms with the Mg-2Gd alloy, i.e., mainly prismatic slip in the former one, while basal slip, prismatic slip and extension twin in the latter one. Further Schmid factor analysis results attribute this different intra-phase deformation mechanisms to the higher critical resolved shear stress (CRSS) value of extension twin and lower ratio of CRSSprismatic /CRSSbasal in the α-Mg phase of Mg-6Li-2Gd alloy. Additionally, Li addition can induce dual-phase microstructure in the Mg-6Li-2Gd alloy, leading to the formation of hetero-deformation induced (HDI) stress at the mesoscale. This can be evidenced by the hysteresis loops appearing during the loading-unloading-reloading (LUR) tensile tests and the activation of multiple slip activity in the α-Mg phase neighboring β-Li phase. The Mg-6Li-2Gd alloy shows higher yield strength is due to the harder α-Mg phase arising from solid solution hardening of Li addition, as well asthe strengthening of soft β-Li phase by the HDI stress during yield stage. Since the strain hardening rate of Mg-6Li-2Gd alloy is lower than that of Mg-2Gd alloy after ~2% strain, which is partly due to the weak contribution of HDI stress, Mg-6Li-2Gd alloy shows no obvious increase of uniform elongation than the Mg-2Gd alloy.But since the β-Li phase is effective in blunting the crack tips, the Mg-6Li-2Gd alloy shows ununiform elongation, which, thus, leads to the higher total elongation than the Mg-2Gd alloy.

Keywords: Mg-Li-Gd dual-phase alloy, phase boundary, HDI stress, dislocation slip activity, mechanical properties

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4021 Possible Involvement of DNA-methyltransferase and Histone Deacetylase in the Regulation of Virulence Potential of Acanthamoeba castellanii

Authors: Yi H. Wong, Li L. Chan, Chee O. Leong, Stephen Ambu, Joon W. Mak, Priyadashi S. Sahu

Abstract:

Background: Acanthamoeba is a free-living opportunistic protist which is ubiquitously distributed in the environment. Virulent Acanthamoeba can cause fatal encephalitis in immunocompromised patients and potential blinding keratitis in immunocompetent contact lens wearers. Approximately 24 species have been identified but only the A. castellanii, A. polyphaga and A. culbertsoni are commonly associated with human infections. Until to date, the precise molecular basis for Acanthamoeba pathogenesis remains unclear. Previous studies reported that Acanthamoeba virulence can be diminished through prolonged axenic culture but revived through serial mouse passages. As no clear explanation on this reversible pathogenesis is established, hereby, we postulate that the epigenetic regulators, DNA-methyltransferases (DNMT) and histone-deacetylases (HDAC), could possibly be involved in granting the virulence plasticity of Acanthamoeba spp. Methods: Four rounds of mouse passages were conducted to revive the virulence potential of the virulence-attenuated Acanthamoeba castellanii strain (ATCC 50492). Briefly, each mouse (n=6/group) was inoculated intraperitoneally with Acanthamoebae cells (2x 105 trophozoites/mouse) and incubated for 2 months. Acanthamoebae cells were isolated from infected mouse organs by culture method and subjected to subsequent mouse passage. In vitro cytopathic, encystment and gelatinolytic assays were conducted to evaluate the virulence characteristics of Acanthamoebae isolates for each passage. PCR primers which targeted on the 2 members (DNMT1 and DNMT2) and 5 members (HDAC1 to 5) of the DNMT and HDAC gene families respectively were custom designed. Quantitative real-time PCR (qPCR) was performed to detect and quantify the relative expression of the two gene families in each Acanthamoeba isolates. Beta-tubulin of A. castellanii (Genbank accession no: XP_004353728) was included as housekeeping gene for data normalisation. PCR mixtures were also analyzed by electrophoresis for amplicons detection. All statistical analyses were performed using the paired one-tailed Student’s t test. Results: Our pathogenicity tests showed that the virulence-reactivated Acanthamoeba had a higher degree of cytopathic effect on vero cells, a better resistance to encystment challenge and a higher gelatinolytic activity which was catalysed by serine protease. qPCR assay showed that DNMT1 expression was significantly higher in the virulence-reactivated compared to the virulence-attenuated Acanthamoeba strain (p ≤ 0.01). The specificity of primers which targeted on DNMT1 was confirmed by sequence analysis of PCR amplicons, which showed a 97% similarity to the published DNA-methyltransferase gene of A. castellanii (GenBank accession no: XM_004332804.1). Out of the five primer pairs which targeted on the HDAC family genes, only HDAC4 expression was significantly difference between the two variant strains. In contrast to DNMT1, HDAC4 expression was much higher in the virulence-attenuated Acanthamoeba strain. Conclusion: Our mouse passages had successfully restored the virulence of the attenuated strain. Our findings suggested that DNA-methyltransferase (DNMT1) and histone deacetylase (HDAC4) expressions are associated with virulence potential of Acanthamoeba spp.

Keywords: acanthamoeba, DNA-methyltransferase, histone deacetylase, virulence-associated proteins

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4020 Investigation of Ignition Delay for Low Molecular Hydrocarbon Fuel and Oxygen Mixture behind the Reflected Shock

Authors: K. R. Guna, Aldin Justin Sundararaj, B. C. Pillai, A. N. Subash

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A systematic study has been made for ignition delay times measurement behind a reflected shock wave for the low molecular weight hydrocarbon fuel in argon simulated gas mixtures. The low molecular hydrocarbon fuel–oxygen was diluted with argon for desired concentration is taken for the study. The suitability of the shock tube for measuring the ignition delay time is demonstrated by measuring the ignition delay for the liquefied petroleum gas for equivalence ratios (ф=0.5 & 1) in the temperature range 1150-1650 K. The pressure range was fixed from 5-15 bar. The ignition delay was measured by recording the ignition-induced pressure jump and emission from CH radical simultaneously. From conducting experiments, it was found that the ignition delay time for liquefied petroleum gas reduces with increase in temperature. The shock tube was calibrated for ethane-oxygen gas mixture and the results obtained from this study is compared with the earlier reported values and found to be comparably well suited for the measurement of ignition delay times. The above work was carried out using the shock tube facility at propulsion and high enthalpy laboratory, Karunya University.

Keywords: ignition delay, LPG, reflected shock, shock wave

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4019 Effect of Anisotropy on Steady Creep in a Whisker Reinforced Functionally Graded Composite Disc

Authors: V. K. Gupta, Tejeet Singh

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In many whisker reinforced composites, anisotropy may result due to material flow during processing operations such as forging, extrusion etc. The consequence of anisotropy, introduced during processing of disc material, has been investigated on the steady state creep deformations of the rotating disc. The disc material is assumed to undergo plastic deformations according to Hill’s anisotropic criterion. Steady state creep has been analyzed in a constant thickness rotating disc made of functionally graded 6061Al-SiCw (where the subscript ‘w’ stands for whisker) using Hill’s The content of reinforcement (SiCw) in the disc is assumed to decrease linearly from the inner to outer radius. The stresses and strain rates in the disc are estimated by solving the force equilibrium equation along with the constitutive equations describing multi-axial creep. The results obtained for anisotropic FGM disc have been compared with those estimated for isotropic FGM disc having the same average whisker content. The anisotropic constants, appearing in Hill’s yield criterion, have been obtained from the available experimental results. The results show that the presence of anisotropy reduces the tangential stress in the middle of the disc but near the inner and outer radii the tangential stress is higher when compared to isotropic disc. On the other hand, the steady state creep rates in the anisotropic disc are reduced significantly over the entire disc radius, with the maximum reduction observed at the inner radius. Further, in the presence of anisotropy the distribution of strain rate becomes relatively uniform over the entire disc, which may be responsible for reducing the extent of distortion in the disc.

Keywords: anisotropy, creep, functionally graded composite, rotating disc

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4018 Influence of Driving Speed on Bearing Capacity Measurement of Intra-Urban Roads with the Traffic Speed Deflectometer(Tsd)

Authors: Pahirangan Sivapatham, Barbara Esser, Andreas Grimmel

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In times of limited public funds and, in particular, an increased social, environmental awareness, as well as the limited availability of construction materials, sustainable and resource-saving pavement management system, is becoming more and more important. Therefore, the knowledge about the condition of the structural substances, particularly bearing capacity and its consideration while planning the maintenance measures of the subordinate network, i.e., state and municipal roads unavoidable. According to the experience, the recommended ride speed of the Traffic Speed Deflectometer (TSD) shall be higher than 40 km/h. Holding of this speed on the intra-urban roads is nearly not possible because of intersections and traffic lights as well as the speed limit. A sufficient background of experience for the evaluation of bearing capacity measurements with TSD in the range of lower speeds is not available yet. The aim of this study is to determine the possible lowest ride speed of the TSD while the bearing capacity measurement on the intra-urban roads. The manufacturer of the TSD used in this study states that the measurements can be conducted at a ride speed of higher than 5 km/h. It is well known that with decreasing ride speed, the viscous fractions in the response of the asphalt pavement increase. This must be taken into account when evaluating the bearing capacity data. In the scope of this study, several measurements were carried out at different speeds between 10 km/h and 60 km/h on the selected intra-urban roads with Pavement-Scanner of the University of Wuppertal, which is equipped with TSD. Pavement-Scanner is able to continuously determine the deflections of asphalt roads in flowing traffic at speeds of up to 80 km/h. The raw data is then aggregated to 10 m mean values so that, as a rule, a bearing capacity characteristic value can be determined for each 10 m road section. By means of analysing of obtained test results, the quality and validity of the determined data rate subject to the riding speed of TSD have been determined. Moreover, the data and pictures of the additional measuring systems of Pavement-Scanners such as High-Speed Road Monitor, Ground Penetration Radar and front cameras can be used to determine and eliminate irregularities in the pavement, which could influence the bearing capacity.

Keywords: bearing capacity measurement, traffic speed deflectometer, inter-urban roads, Pavement-Scanner, structural substance

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4017 3D Label-Free Bioimaging of Native Tissue with Selective Plane Illumination Optical Microscopy

Authors: Jing Zhang, Yvonne Reinwald, Nick Poulson, Alicia El Haj, Chung See, Mike Somekh, Melissa Mather

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Biomedical imaging of native tissue using light offers the potential to obtain excellent structural and functional information in a non-invasive manner with good temporal resolution. Image contrast can be derived from intrinsic absorption, fluorescence, or scatter, or through the use of extrinsic contrast. A major challenge in applying optical microscopy to in vivo tissue imaging is the effects of light attenuation which limits light penetration depth and achievable imaging resolution. Recently Selective Plane Illumination Microscopy (SPIM) has been used to map the 3D distribution of fluorophores dispersed in biological structures. In this approach, a focused sheet of light is used to illuminate the sample from the side to excite fluorophores within the sample of interest. Images are formed based on detection of fluorescence emission orthogonal to the illumination axis. By scanning the sample along the detection axis and acquiring a stack of images, 3D volumes can be obtained. The combination of rapid image acquisition speeds with the low photon dose to samples optical sectioning provides SPIM is an attractive approach for imaging biological samples in 3D. To date all implementations of SPIM rely on the use of fluorescence reporters be that endogenous or exogenous. This approach has the disadvantage that in the case of exogenous probes the specimens are altered from their native stage rendering them unsuitable for in vivo studies and in general fluorescence emission is weak and transient. Here we present for the first time to our knowledge a label-free implementation of SPIM that has downstream applications in the clinical setting. The experimental set up used in this work incorporates both label-free and fluorescent illumination arms in addition to a high specification camera that can be partitioned for simultaneous imaging of both fluorescent emission and scattered light from intrinsic sources of optical contrast in the sample being studied. This work first involved calibration of the imaging system and validation of the label-free method with well characterised fluorescent microbeads embedded in agarose gel. 3D constructs of mammalian cells cultured in agarose gel with varying cell concentrations were then imaged. A time course study to track cell proliferation in the 3D construct was also carried out and finally a native tissue sample was imaged. For each sample multiple images were obtained by scanning the sample along the axis of detection and 3D maps reconstructed. The results obtained validated label-free SPIM as a viable approach for imaging cells in a 3D gel construct and native tissue. This technique has the potential use in a near-patient environment that can provide results quickly and be implemented in an easy to use manner to provide more information with improved spatial resolution and depth penetration than current approaches.

Keywords: bioimaging, optics, selective plane illumination microscopy, tissue imaging

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4016 Experimental Uniaxial Tensile Characterization of One-Dimensional Nickel Nanowires

Authors: Ram Mohan, Mahendran Samykano, Shyam Aravamudhan

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Metallic nanowires with sub-micron and hundreds of nanometer diameter have a diversity of applications in nano/micro-electromechanical systems (NEMS/MEMS). Characterizing the mechanical properties of such sub-micron and nano-scale metallic nanowires are tedious; require sophisticated and careful experimentation to be performed within high-powered microscopy systems (scanning electron microscope (SEM), atomic force microscope (AFM)). Also, needed are nanoscale devices for placing the nanowires; loading them with the intended conditions; obtaining the data for load–deflection during the deformation within the high-powered microscopy environment poses significant challenges. Even picking the grown nanowires and placing them correctly within a nanoscale loading device is not an easy task. Mechanical characterizations through experimental methods for such nanowires are still very limited. Various techniques at different levels of fidelity, resolution, and induced errors have been attempted by material science and nanomaterial researchers. The methods for determining the load, deflection within the nanoscale devices also pose a significant problem. The state of the art is thus still at its infancy. All these factors result and is seen in the wide differences in the characterization curves and the reported properties in the current literature. In this paper, we discuss and present our experimental method, results, and discussions of uniaxial tensile loading and the development of subsequent stress–strain characteristics curves for Nickel nanowires. Nickel nanowires in the diameter range of 220–270 nm were obtained in our laboratory via an electrodeposition method, which is a solution based, template method followed in our present work for growing 1-D Nickel nanowires. Process variables such as the presence of magnetic field, its intensity; and varying electrical current density during the electrodeposition process were found to influence the morphological and physical characteristics including crystal orientation, size of the grown nanowires1. To further understand the correlation and influence of electrodeposition process variables, associated formed structural features of our grown Nickel nanowires to their mechanical properties, careful experiments within scanning electron microscope (SEM) were conducted. Details of the uniaxial tensile characterization, testing methodology, nanoscale testing device, load–deflection characteristics, microscopy images of failure progression, and the subsequent stress–strain curves are discussed and presented.

Keywords: uniaxial tensile characterization, nanowires, electrodeposition, stress-strain, nickel

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4015 Unveiling the Impact of Ultra High Vacuum Annealing Levels on Physico-Chemical Properties of Bulk ZnSe Semiconductor

Authors: Kheira Hamaida, Mohamed Salah Halati

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In this current paper, our aim work is to link as possible the obtained simulation results and the other experimental ones, just focusing on the electronic and optical properties of ZnSe. The predictive spectra of the total and partial densities of states using the Full Potential Linearized/Augmented Plane Wave method with the newly Tran-Blaha (TB) modified Becke-Johnson (mBJ) exchange-correlation potential (EXC). So the upper valence energy (UVE) levels contain the relative contribution of Se-(4p and 3d) states with considerable contribution from the electrons of Zn-2s orbital. The dielectric function of w-ZnSe, with its two parts, appears with a noticeable anisotropy character. The microscopic origins of the electronic states that are responsible for the observed peaks in the spectrum are determined through the decomposition of the spectrum to the individual contributions of the electronic transitions between the pairs of bands, where Vi is an occupied state in the valence band, and Ci is an unoccupied state in the conduction band. X-PES (X Ray-Photo Electron Spectroscopy) is an important technique used to probe the homogeneity, stoichiometry, and purity state of the title compound. In order to check the electron transitions derived from simulations and the others from Reflected Electron Energy Loss Spectroscopy (REELS) technique which was of great sensitivity, is used to determine the interband electronic transitions. In the optical window (Eg), all the electron energy states created were also determined through the specific gaussian deconvolution of the photoluminescence spectrum (PLS) that probed under a room temperature (RT).

Keywords: spectroscopy, WIEN2K, IIB-VIA semiconductors, dielectric function

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4014 Risk Measurement and Management Strategies in Poultry Farm Enterprises in Imo State, Nigeria

Authors: Donatus Otuiheoma Ohajianya, Augusta Onyekachi Unamba

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This study analyzed risk among poultry farm enterprises in Imo State of Nigeria. Specifically, it examined sources of risks, the major risks associated with poultry farm enterprise, and the risk-reducing strategies among the poultry farm enterprises in the study area. Primary data collected in 2015 with validated questionnaire from 120 proportionately and randomly selected poultry farm enterprises were used for the study. The data were analyzed with descriptive statistics and W-Statistic that was validated with Pearson Criterion (X2). The results showed that major risk sources affecting poultry farm enterprises were production, marketing, financial and political in that order. The results found a W-Statistic value of 0.789, which was verified by Pearson Criterion to obtain X2-Calculated value of 4.65 which is lower that X2-Critical value of 11.07 at 5% significant level. The risk-reducing strategies were found to be diversification, savings, co-operative marketing, borrowing, and insurance. It was recommended that government and donor agencies should make policies aimed at encouraging poultry farm enterprises adopt the highlighted risk-reducing strategies in risk management to improve their productivity and farm income.

Keywords: risk, measurement, management, poultry farm, Imo State

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4013 CFD Simulation and Investigation of Critical Two-Phase Flow Rate in Wellhead Choke

Authors: Alireza Rafie Boldaji, Ahmad Saboonchi

Abstract:

Chokes are commonly used in oil and gas production systems. A choke is a restriction basically designed to control flow rates of oil and gas wells, to prevent the downstream disturbances from propagating upstream (critical flow), and to protect the surface equipment facilities against slugging at high flowing pressures. There are different methods to calculate the multiphase flow rate, one of the multiphase flow measurement methods is the separation and measurement by on¬e-phaseFlow meter, another common method is the use of movable separator, their operations are very labor-intensive and costly. The current method used is based on the flow differential pressure on both sides of choke. Three groups of correlations describing two-phase flow through wellhead chokes were examined. The first group involved simple empirical equations similar to those of Gilbert, the second group comprised derived equations of two-phase flow incorporating PVT properties, and third group is computational method. In the article we calculate the flow of oil and gas through choke with simulation of this two phase flow bye computational fluid dynamic method, we use Ansys- fluent for this simulation and finally compared results of computational simulation whit empirical equations, the results show good agreement between experimental and numerical results.

Keywords: CFD, two-phase, choke, critical

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4012 A Customize Battery Management Approach for Satellite

Authors: Muhammad Affan, Muhammad Ilyas Raza, Muhammad Harris Hashmi

Abstract:

This work is attributed to the battery management unit design of student Satellites under Pakistan National Student Satellite Program (PNSSP). The aim has been to design a customized, low-cost, efficient, reliable and less-complex battery management scheme for the Satellite. Nowadays, Lithium Ion (Li-ion) batteries have become the de-facto standard for remote applications, especially for satellites. Li-ion cells are selected for secondary storage. The design also addresses Li-ion safety requirements by monitoring, balancing and protecting cells for safe and prolonged operation. Accurate voltage measurement of individual cells was the main challenge because all the actions triggered were based on the digital voltage measurement. For this purpose, a resistive-divider network is used to maintain simplicity and cost-effectiveness. To cater the problem of insufficient i/o pins on microcontroller, fast multiplexers and de-multiplexers were used. The discrepancy inherited in the given design is the dissipation of heat due to the dissipative resistors. However, it is still considered to be the optimum adoption, considering the simple and cost-effective nature of the passive balancing technique. Furthermore, it is a completely unique solution, customized to meet specific requirements. However, there is still an option for a more advanced and expensive design.

Keywords: satellite, battery module, passive balancing, dissipative

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4011 Machine Learning Classification of Fused Sentinel-1 and Sentinel-2 Image Data Towards Mapping Fruit Plantations in Highly Heterogenous Landscapes

Authors: Yingisani Chabalala, Elhadi Adam, Khalid Adem Ali

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Mapping smallholder fruit plantations using optical data is challenging due to morphological landscape heterogeneity and crop types having overlapped spectral signatures. Furthermore, cloud covers limit the use of optical sensing, especially in subtropical climates where they are persistent. This research assessed the effectiveness of Sentinel-1 (S1) and Sentinel-2 (S2) data for mapping fruit trees and co-existing land-use types by using support vector machine (SVM) and random forest (RF) classifiers independently. These classifiers were also applied to fused data from the two sensors. Feature ranks were extracted using the RF mean decrease accuracy (MDA) and forward variable selection (FVS) to identify optimal spectral windows to classify fruit trees. Based on RF MDA and FVS, the SVM classifier resulted in relatively high classification accuracy with overall accuracy (OA) = 0.91.6% and kappa coefficient = 0.91% when applied to the fused satellite data. Application of SVM to S1, S2, S2 selected variables and S1S2 fusion independently produced OA = 27.64, Kappa coefficient = 0.13%; OA= 87%, Kappa coefficient = 86.89%; OA = 69.33, Kappa coefficient = 69. %; OA = 87.01%, Kappa coefficient = 87%, respectively. Results also indicated that the optimal spectral bands for fruit tree mapping are green (B3) and SWIR_2 (B10) for S2, whereas for S1, the vertical-horizontal (VH) polarization band. Including the textural metrics from the VV channel improved crop discrimination and co-existing land use cover types. The fusion approach proved robust and well-suited for accurate smallholder fruit plantation mapping.

Keywords: smallholder agriculture, fruit trees, data fusion, precision agriculture

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4010 The Application of Whole-Cell Luminescent Biosensors for Assessing Bactericidal Properties of Medicinal Plants

Authors: Yuliya Y. Gavrichenko

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Background and Aims: The increasing bacterial resistance to almost all the available antibiotics has encouraged scientists to search for alternative sources of antibacterial agents. Nowadays, it is known that many plant secondary metabolites have diverse biological activity. These compounds can be potentially active against human bacterial and viral infections. Extended research has been carried out to explore the use of the luminescent bacterial test as a rapid, accurate and inexpensive method to assess the antibacterial properties and to predict the biological activity spectra for plant origin substances. Method: Botanical material of fifteen species was collected from their natural and cultural habitats on the Crimean peninsula. The aqueous extracts of following plants were tested: Robinia pseudoacacia L., Sideritis comosa, Cotinus coggygria Scop., Thymus serpyllum L., Juglans regia L., Securigera varia L., Achillea millefolium L., Phlomis taurica, Corylus avellana L., Sambucus nigra L., Helichrysum arenarium L., Glycyrrhiza glabra L., Elytrigia repens L., Echium vulgare L., Conium maculatum L. The test was carried out using luminous strains of marine bacteria Photobacterium leiognathi, which was isolated from the Sea of Azov as well as four Escherichia coli MG1655 recombinant strains harbouring Vibrio fischeri luxCDABE genes. Results: The bactericidal capacity of plant extracts showed significant differences in the study. Cotinus coggygria, Phlomis taurica, Juglans regia L. proved to be the most toxic to P. leiognathi. (EC50 = 0.33 g dried plant/l). Glycyrrhiza glabra L., Robinia pseudoacacia L., Sideritis comosa and Helichrysum arenarium L. had moderate inhibitory effects (EC50 = 3.3 g dried plant/l). The rest of the aqueous extracts have decreased the luminescence of no more than 50% at the lowest concentration (16.5 g dried plant/l). Antibacterial activity of herbal extracts against constitutively luminescent E. coli MG1655 (pXen7-lux) strain was observed at approximately the same level as for P. leiognathi. Cotinus coggygria and Conium maculatum L. extracts have increased light emission in the mutant E. coli MG1655 (pFabA-lux) strain which is associated with cell membranes damage. Sideritis comosa, Phlomis taurica, Juglans regia induced SOS response in E. coli (pColD-lux) strain. Glycyrrhiza glabra L. induced protein damage response in E. coli MG1655 (pIbpA-lux) strain. Conclusion: The received results have shown that the plants’ extracts had nonspecific antimicrobial effects against both E. coli (pXen7-lux) and P. leiognathi biosensors. Mutagenic, cytotoxic and protein damage effects have been observed. In general, the bioluminescent inhibition test result correlated with the traditional use of screened plants. It leads to the following conclusion that whole-cell luminescent biosensors could be the indicator of overall plants antibacterial capacity. The results of the investigation have shown a possibility of bioluminescent method in medicine and pharmacy as an approach to research the antibacterial properties of medicinal plants.

Keywords: antibacterial property, bioluminescence, medicinal plants, whole-cell biosensors

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4009 Traditional Values and Their Adaptation in Social Housing Design: Towards a New Typology and Establishment of 'Airhouse' Standard in Malaysia

Authors: Mohd Firrdhaus Mohd Sahabuddin, Cristina Gonzalez-Longo

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Large migration from rural areas to urban areas like Kuala Lumpur has led to some implications for economic, social and cultural development. This high population has placed enormous demand on the existing housing stocks, especially for low-income groups. However, some issues arise, one of which is overheated indoor air temperature. This problem contributes to the high-energy usage that forces huge sums of money to be spent on cooling the house by using mechanical equipment. Therefore, this study focuses on thermal comfort in social housing, and incorporates traditional values into its design to achieve a certain measurement of natural ventilation in a house. From the study, the carbon emission and energy consumption for an air-conditioned house is 67%, 66% higher than a naturally ventilated house. Therefore, this research has come up with a new typology design, which has a large exposed wall area and full-length openings on the opposite walls to increase cross ventilation. At the end of this research, the measurement of thermal comfort for a naturally ventilated building called ‘AirHouse’ has been identified.

Keywords: tropical architecture, natural ventilation, passive design, AirHouse, social housing design

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4008 Crack Initiation Assessment during Fracture of Heat Treated Duplex Stainless Steels

Authors: Faraj Ahmed E. Alhegagi, Anagia M. Khamkam Mohamed, Bassam F. Alhajaji

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Duplex stainless steels (DSS) are widely employed in industry for apparatus working with sea water in petroleum, refineries and in chemical plants. Fracture of DSS takes place by cleavage of the ferrite phase and the austenite phase ductile tear off. Pop-in is an important feature takes place during fracture of DSS. The procedure of Pop-ins assessment plays an important role in fracture toughness studies. In present work, Zeron100 DSS specimens were heat treated at different temperatures, cooled and pulled to failure to assess the pop-ins criterion in crack initiation prediction. The outcome results were compared to the British Standard (BS 7448) and the ASTEM standard (E1290) for Crack-Tip Opening Displacement (CTOD) fracture toughness measurement. Pop-in took place during specimens loading specially for those specimens heat treated at higher temperatures. The standard BS7448 was followed to check specimen validity for fractured toughness assessment by direct determination of KIC. In most cases, specimens were invalid for KIC measurement. The two procedures were equivalent only when single pop-ins were assessed. A considerable contrast in fracture toughness value between was observed where multiple pop-ins were assessed.

Keywords: fracture toughness, stainless steels, pop ins, crack assessment

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4007 Estimating X-Ray Spectra for Digital Mammography by Using the Expectation Maximization Algorithm: A Monte Carlo Simulation Study

Authors: Chieh-Chun Chang, Cheng-Ting Shih, Yan-Lin Liu, Shu-Jun Chang, Jay Wu

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With the widespread use of digital mammography (DM), radiation dose evaluation of breasts has become important. X-ray spectra are one of the key factors that influence the absorbed dose of glandular tissue. In this study, we estimated the X-ray spectrum of DM using the expectation maximization (EM) algorithm with the transmission measurement data. The interpolating polynomial model proposed by Boone was applied to generate the initial guess of the DM spectrum with the target/filter combination of Mo/Mo and the tube voltage of 26 kVp. The Monte Carlo N-particle code (MCNP5) was used to tally the transmission data through aluminum sheets of 0.2 to 3 mm. The X-ray spectrum was reconstructed by using the EM algorithm iteratively. The influence of the initial guess for EM reconstruction was evaluated. The percentage error of the average energy between the reference spectrum inputted for Monte Carlo simulation and the spectrum estimated by the EM algorithm was -0.14%. The normalized root mean square error (NRMSE) and the normalized root max square error (NRMaSE) between both spectra were 0.6% and 2.3%, respectively. We conclude that the EM algorithm with transmission measurement data is a convenient and useful tool for estimating x-ray spectra for DM in clinical practice.

Keywords: digital mammography, expectation maximization algorithm, X-Ray spectrum, X-Ray

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4006 Austempered Compacted Graphite Irons: Influence of Austempering Temperature on Microstructure and Microscratch Behavior

Authors: Rohollah Ghasemi, Arvin Ghorbani

Abstract:

This study investigates the effect of austempering temperature on microstructure and scratch behavior of the austempered heat-treated compacted graphite irons. The as-cast was used as base material for heat treatment practices. The samples were extracted from as-cast ferritic CGI pieces and were heat treated under austenitising temperature of 900°C for 60 minutes which followed by quenching in salt-bath at different austempering temperatures of 275°C, 325°C and 375°C. For all heat treatments, an austempering holding time of 30 minutes was selected for this study. Light optical microscope (LOM) and scanning electron microscope (SEM) and electron back scattered diffraction (EBSD) analysis confirmed the ausferritic matrix formed in all heat-treated samples. Microscratches were performed under the load of 200, 600 and 1000 mN using a sphero-conical diamond indenter with a tip radius of 50 μm and induced cone angle 90° at a speed of 10 μm/s at room temperature ~25°C. An instrumented nanoindentation machine was used for performing nanoindentation hardness measurement and microscratch testing. Hardness measurements and scratch resistance showed a significant increase in Brinell, Vickers, and nanoindentation hardness values as well as microscratch resistance of the heat-treated samples compared to the as-cast ferritic sample. The increase in hardness and improvement in microscratch resistance are associated with the formation of the ausferrite matrix consisted of carbon-saturated retained austenite and acicular ferrite in austempered matrix. The maximum hardness was observed for samples austempered at 275°C which resulted in the formation of very fine acicular ferrite. In addition, nanohardness values showed a quite significant variation in the matrix due to the presence of acicular ferrite and carbon-saturated retained austenite. It was also observed that the increase of austempering temperature resulted in increase of volume of the carbon-saturated retained austenite and decrease of hardness values.

Keywords: austempered CGI, austempering, scratch testing, scratch plastic deformation, scratch hardness

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4005 Autophagy in the Midgut Epithelium of Spodoptera exigua Hübner (Lepidoptera: Noctuidae) Larvae Exposed to Various Cadmium Concentration - 6-Generational Exposure

Authors: Magdalena Maria Rost-Roszkowska, Alina Chachulska-Żymełka, Monika Tarnawska, Maria Augustyniak, Alina Kafel, Agnieszka Babczyńska

Abstract:

Autophagy is a form of cell remodeling in which an internalization of organelles into vacuoles that are called autophagosomes occur. Autophagosomes are the targets of lysosomes, thus causing digestion of cytoplasmic components. Eventually, it can lead to the death of the entire cell. However, in response to several stress factors, e.g., starvation, heavy metals (e.g., cadmium) autophagy can also act as a pro-survival factor, protecting the cell against its death. The main aim of our studies was to check if the process of autophagy, which could appear in the midgut epithelium after Cd treatment, can be fixed during the following generations of insects. As a model animal, we chose the beet armyworm Spodoptera exigua Hübner (Lepidoptera: Noctuidae), a well-known polyphagous pest of many vegetable crops. We analyzed specimens at final larval stage (5th larval stage), due to its hyperfagy, resulting in great amount of cadmium assimilate. The culture consisted of two strains: a control strain (K) fed a standard diet, and a cadmium strain (Cd), fed on standard diet supplemented with cadmium (44 mg Cd per kg of dry weight of food) for 146 generations, both strains. In addition, the control insects were transferred to the Cd supplemented diet (5 mg Cd per kg of dry weight of food, 10 mg Cd per kg of dry weight of food, 20 mg Cd per kg of dry weight of food, 44 mg Cd per kg of dry weight of food). Therefore, we obtained Cd1, Cd2, Cd3 and KCd experimental groups. Autophagy has been examined using transmission electron microscope. During this process, degenerated organelles were surrounded by a membranous phagophore and enclosed in an autophagosome. Eventually, after the autophagosome fused with a lysosome, an autolysosome was formed and the process of the digestion of organelles began. During the 1st year of the experiment, we analyzed specimens of 6 generations in all the lines. The intensity of autophagy depends significantly on the generation, tissue and cadmium concentration in the insect rearing medium. In the Ist, IInd, IIIrd, IVth, Vth and VIth generation the intensity of autophagy in the midguts from cadmium-exposed strains decreased gradually according to the following order of strains: Cd1, Cd2, Cd3 and KCd. The higher amount of cells with autophagy was observed in Cd1 and Cd2. However, it was still higher than the percentage of cells with autophagy in the same tissues of the insects from the control and multigenerational cadmium strain. This may indicate that during 6-generational exposure to various Cd concentration, a preserved tolerance to cadmium was not maintained. The study has been financed by the National Science Centre Poland, grant no 2016/21/B/NZ8/00831.

Keywords: autophagy, cell death, digestive system, ultrastructure

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4004 A Finite Element Study of Laminitis in Horses

Authors: Naeim Akbari Shahkhosravi, Reza Kakavand, Helen M. S. Davies, Amin Komeili

Abstract:

Equine locomotion and performance are significantly affected by hoof health. One of the most critical diseases of the hoof is laminitis, which can lead to horse lameness in a severe condition. This disease exhibits the mechanical properties degradation of the laminar junction tissue within the hoof. Therefore, it is essential to investigate the biomechanics of the hoof, focusing specifically on excessive and cumulatively accumulated stresses within the laminar junction tissue. For this aim, the current study generated a novel equine hoof Finite Element (FE) model under dynamic physiological loading conditions and employing a hyperelastic material model. Associated tissues of the equine hoof were segmented from computed tomography scans of an equine forelimb, including the navicular bone, third phalanx, sole, frog, laminar junction, digital cushion, and medial- dorsal- lateral wall areas. The inner tissues were connected based on the hoof anatomy, and the hoof was under a dynamic loading over cyclic strides at the trot. The strain distribution on the hoof wall of the model was compared with the published in vivo strain measurements to validate the model. Then the validated model was used to study the development of laminitis. The ultimate stress tolerated by the laminar junction before rupture was considered as a stress threshold. The tissue damage was simulated through iterative reduction of the tissue’s mechanical properties in the presence of excessive maximum principal stresses. The findings of this investigation revealed how damage initiates from the medial and lateral sides of the tissue and propagates through the hoof dorsal area.

Keywords: horse hoof, laminitis, finite element model, continuous damage

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4003 A Novel Concept of Optical Immunosensor Based on High-Affinity Recombinant Protein Binders for Tailored Target-Specific Detection

Authors: Alena Semeradtova, Marcel Stofik, Lucie Mareckova, Petr Maly, Ondrej Stanek, Jan Maly

Abstract:

Recently, novel strategies based on so-called molecular evolution were shown to be effective for the production of various peptide ligand libraries with high affinities to molecular targets of interest comparable or even better than monoclonal antibodies. The major advantage of these peptide scaffolds is mainly their prevailing low molecular weight and simple structure. This study describes a new high-affinity binding molecules based immunesensor using a simple optical system for human serum albumin (HSA) detection as a model molecule. We present a comparison of two variants of recombinant binders based on albumin binding domain of the protein G (ABD) performed on micropatterned glass chip. Binding domains may be tailored to any specific target of interest by molecular evolution. Micropatterened glass chips were prepared using UV-photolithography on chromium sputtered glasses. Glass surface was modified by (3-aminopropyl)trietoxysilane and biotin-PEG-acid using EDC/NHS chemistry. Two variants of high-affinity binding molecules were used to detect target molecule. Firstly, a variant is based on ABD domain fused with TolA chain. This molecule is in vivo biotinylated and each molecule contains one molecule of biotin and one ABD domain. Secondly, the variant is ABD domain based on streptavidin molecule and contains four gaps for biotin and four ABD domains. These high-affinity molecules were immobilized to the chip surface via biotin-streptavidin chemistry. To eliminate nonspecific binding 1% bovine serum albumin (BSA) or 6% fetal bovine serum (FBS) were used in every step. For both variants range of measured concentrations of fluorescently labelled HSA was 0 – 30 µg/ml. As a control, we performed a simultaneous assay without high-affinity binding molecules. Fluorescent signal was measured using inverse fluorescent microscope Olympus IX 70 with COOL LED pE 4000 as a light source, related filters, and camera Retiga 2000R as a detector. The fluorescent signal from non-modified areas was substracted from the signal of the fluorescent areas. Results were presented in graphs showing the dependence of measured grayscale value on the log-scale of HSA concentration. For the TolA variant the limit of detection (LOD) of the optical immunosensor proposed in this study is calculated to be 0,20 µg/ml for HSA detection in 1% BSA and 0,24 µg/ml in 6% FBS. In the case of streptavidin-based molecule, it was 0,04 µg/ml and 0,07 µg/ml respectively. The dynamical range of the immunosensor was possible to estimate just in the case of TolA variant and it was calculated to be 0,49 – 3,75 µg/ml and 0,73-1,88 µg/ml respectively. In the case of the streptavidin-based the variant we didn´t reach the surface saturation even with the 480 ug/ml concentration and the upper value of dynamical range was not estimated. Lower value was calculated to be 0,14 µg/ml and 0,17 µg/ml respectively. Based on the obtained results, it´s clear that both variants are useful for creating the bio-recognizing layer on immunosensors. For this particular system, it is obvious that the variant based on streptavidin molecule is more useful for biosensing on glass planar surfaces. Immunosensors based on this variant would exhibit better limit of detection and wide dynamical range.

Keywords: high affinity binding molecules, human serum albumin, optical immunosensor, protein G, UV-photolitography

Procedia PDF Downloads 357