Search results for: substrate specificity

Authors: Iman Farasat, Howard M. Salis

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The CRISPR/Cas9 system has revolutionized genome engineering by enabling site-directed and high-throughput genome editing, genome insertion, and gene knockdowns in several species, including bacteria, yeast, flies, worms, and human cell lines. This technology has the potential to enable human gene therapy to treat genetic diseases and cancer at the molecular level; however, the current CRISPR/Cas9 system suffers from seemingly sporadic off-target genome mutagenesis that prevents its use in gene therapy. A comprehensive mechanistic model that explains how the CRISPR/Cas9 functions would enable the rational design of the guide-RNAs responsible for target site selection while minimizing unexpected genome mutagenesis. Here, we present the first quantitative model of the CRISPR/Cas9 genome mutagenesis system that predicts how guide-RNA sequences (crRNAs) control target site selection and cleavage activity. We used statistical thermodynamics and law of mass action to develop a five-step biophysical model of cas9 cleavage, and examined it in vivo and in vitro. To predict a crRNA's binding specificities and cleavage rates, we then compiled a nearest neighbor (NN) energy model that accounts for all possible base pairings and mismatches between the crRNA and the possible genomic DNA sites. These calculations correctly predicted crRNA specificity across 5518 sites. Our analysis reveals that cas9 activity and specificity are anti-correlated, and, the trade-off between them is the determining factor in performing an RNA-mediated cleavage with minimal off-targets. To find an optimal solution, we first created a scheme of safe-design criteria for Cas9 target selection by systematic analysis of available high throughput measurements. We then used our biophysical model to determine the optimal Cas9 expression levels and timing that maximizes on-target cleavage and minimizes off-target activity. We successfully applied this approach in bacterial and mammalian cell lines to reduce off-target activity to near background mutagenesis level while maintaining high on-target cleavage rate.

Keywords: biophysical model, CRISPR, Cas9, genome editing

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Authors: Dace Grauda, Gunta Cekstere, Inta Belogrudova, Andis Karlsons, Isaak Rashal

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Trifolium repens L. (white or Dutch clover) is a perennial herb, belongs to legume family (Leguminosae Juss.), spread extensively by stolons and seeds. The species is cultivated worldwide and was naturalized in many countries in meadows, yards, gardens, along roads and streets etc., especially in temperate regions. It is widespread also in grasslands throughout Riga, the capital of Latvia. The goal of this study was to investigate genetic structure of white clover population in Riga and to evaluate influence of different salt concentration on plants. For this purpose universal retrotranspozone based IRAP (Inter-Retrotransposon Amplified Polymorphism) method was used. The plant material was collected in different regions of Riga and in several urban areas of Latvia. Plant DNA was isolated from in silicogel dried leaves of using 1% CTAB (cetyltrimet-ammonium bromide) buffer DNA extraction procedure. Genetic structure of city population and wild populations were compared. Soil salinization is an important issue associated with low water resources and highly urbanized areas in aride and semi-aride climate conditions, as well as de-icing salt application to prevent ice formation on roads in winter. The T. repens variety ‘Daile’ (form giganteum), one of the often used component of urban greeneries, was studied in this investigation. Plants were grown from seeds and cultivated in the light conditions (18-25 C, 16h/8h of day/night, light intensity 3000 lx) in plastic pots (200 ml), filled with commercial neutralized (pH 5.9 ± 0.3) peat substrate with mineral nutrients. To analyse the impact of increased soil salinity treatments with gradually rising NaCl (0; 20; 40; 60; 80; 100 mM) levels were arranged. Plants were watered when necessary with deionised water to provide optimum substrate moisture 60-70%. The experiment was terminated six weeks after establishment. For analysis of mineral nutrients, dry plant material (above ground part and roots) was used. Decrease of Na content can be significant under elevated salinity till 20 mM NaCl. High NaCl concentrations in the substrate increase Na, Cl, Cu, Fe, and Mn accumulation, but reduce S, Mg, K content in the plant above ground parts. Abiotic stresses generally changes the levels of DNA metilation. Several candidate gene for salt tolerance will be analysed for DNA metilation level using Pyromark-Q24 advanced.

Keywords: DNA metilation, IRAP, soil salinization, white clover

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Authors: Ahmad I. Ayesh

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This work reports on the fabrication of tin nanoclusters by sputtering and inert-gas condensation inside an ultra-high vacuum compatible system. This technique allows to fine tune the size and yield of nanoclusters by controlling the nanocluster source parameters. The produced nanoclusters are deposited on SiO2/Si substrate with pre-formed electrical electrodes to produce a nanocluster device. Those devices can be potentially used for gas sensor applications.

Keywords: tin, nanoclusters, inert-gas condensation, nanotechnology

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Authors: Parth Gupta, Ujjawal Singh, Shashank Kumar, Mansi Chandra, Arnab Sarkar

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Microfluidic devices have emerged as indispensable tools across various scientific disciplines, offering precise control and manipulation of fluids at the microscale. Their efficacy in flow-based research, spanning engineering, chemistry, and biology, relies heavily on the geometric design of microfluidic channels. This work introduces a novel approach to optimise these channels through Response Surface Methodology (RSM), departing from the conventional practice of addressing one parameter at a time. Traditionally, optimising microfluidic channels involved isolated adjustments to individual parameters, limiting the comprehensive understanding of their combined effects. In contrast, our approach considers the simultaneous impact of multiple parameters, employing RSM to efficiently explore the complex design space. The outcome is an innovative microfluidic channel that consumes an optimal sample volume and minimises flow time, enhancing overall efficiency. The relevance of geometric parameter optimization in microfluidic channels extends significantly in biomedical engineering. The flow characteristics of porous materials within these channels depend on many factors, including fluid viscosity, environmental conditions (such as temperature and humidity), and specific design parameters like sample volume, channel width, channel length, and substrate porosity. This intricate interplay directly influences the performance and efficacy of microfluidic devices, which, if not optimized, can lead to increased costs and errors in disease testing and analysis. In the context of biomedical applications, the proposed approach addresses the critical need for precision in fluid flow. it mitigate manufacturing costs associated with trial-and-error methodologies by optimising multiple geometric parameters concurrently. The resulting microfluidic channels offer enhanced performance and contribute to a streamlined, cost-effective process for testing and analyzing diseases. A key highlight of our methodology is its consideration of the interconnected nature of geometric parameters. For instance, the volume of the sample, when optimized alongside channel width, length, and substrate porosity, creates a synergistic effect that minimizes errors and maximizes efficiency. This holistic optimization approach ensures that microfluidic devices operate at their peak performance, delivering reliable results in disease testing. A key highlight of our methodology is its consideration of the interconnected nature of geometric parameters. For instance, the volume of the sample, when optimized alongside channel width, length, and substrate porosity, creates a synergistic effect that minimizes errors and maximizes efficiency. This holistic optimization approach ensures that microfluidic devices operate at their peak performance, delivering reliable results in disease testing. A key highlight of our methodology is its consideration of the interconnected nature of geometric parameters. For instance, the volume of the sample, when optimized alongside channel width, length, and substrate porosity, creates a synergistic effect that minimizes errors and maximizes efficiency. This holistic optimization approach ensures that microfluidic devices operate at their peak performance, delivering reliable results in disease testing.

Keywords: microfluidic device, minitab, statistical optimization, response surface methodology

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Authors: Ankit Khurana

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The sufficient heat storage capacity or ability to dissipate heat is the most decisive parameter to have an effective and efficient functioning of Friction-based Brake Disc systems. The primary aim of the research was to analyse the effect of multiple coatings on lightweight disk rotors surface which not only alleviates the mass of vehicle & also, augments heat transfer. This research is projected to aid the automobile fraternity with an enunciated view over the thermal aspects in a braking system. The results of the project indicate that with the advent of modern coating technologies a brake system’s thermal curtailments can be removed and together with forced convection, heat transfer processes can see a drastic improvement leading to increased lifetime of the brake rotor. Other advantages of modifying the surface of a lightweight rotor substrate will be to reduce the overall weight of the vehicle, decrease the risk of thermal brake failure (brake fade and fluid vaporization), longer component life, as well as lower noise and vibration characteristics. A mathematical model was constructed in MATLAB which encompassing the various thermal characteristics of the proposed coatings and substrate materials required to approximate the heat flux values in a free and forced convection environment; resembling to a real-time braking phenomenon which could easily be modelled into a full cum scaled version of the alloy brake rotor part in ABAQUS. The finite element of a brake rotor was modelled in a constrained environment such that the nodal temperature between the contact surfaces of the coatings and substrate (Wrought Aluminum alloy) resemble an amalgamated solid brake rotor element. The initial results obtained were for a Plasma Electrolytic Oxidized (PEO) substrate wherein the Aluminum alloy gets a hard ceramic oxide layer grown on its transitional phase. The rotor was modelled and then evaluated in real-time for a constant ‘g’ braking event (based upon the mathematical heat flux input and convective surroundings), which reflected the necessity to deposit a conducting coat (sacrificial) above the PEO layer in order to inhibit thermal degradation of the barrier coating prematurely. Taguchi study was then used to bring out certain critical factors which may influence the maximum operating temperature of a multi-coated brake disc by simulating brake tests: a) an Alpine descent lasting 50 seconds; b) an Autobahn stop lasting 3.53 seconds; c) a Six–high speed repeated stop in accordance to FMVSS 135 lasting 46.25 seconds. Thermal Barrier coating thickness and Vane heat transfer coefficient were the two most influential factors and owing to their design and manufacturing constraints a final optimized model was obtained which survived the 6-high speed stop test as per the FMVSS -135 specifications. The simulation data highlighted the merits for preferring Wrought Aluminum alloy 7068 over Grey Cast Iron and Aluminum Metal Matrix Composite in coherence with the multiple coating depositions.

Keywords: lightweight brakes, surface modification, simulated braking, PEO, aluminum

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Authors: David Polcari, Samuel C. Perry, Loredano Pollegioni, Matthias Geissler, Janine Mauzeroll

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ᴅ-serine acts as an endogenous co-agonist for N-methyl-ᴅ-aspartate receptors in neuronal synapses. This makes it a key component in the development and function of a healthy brain, especially given its role in several neurodegenerative diseases such as Alzheimer’s disease and dementia. Despite such clear research motivations, the primary site and mechanism of ᴅ-serine release is still currently unclear. For this reason, we are developing a biosensor for the detection of ᴅ-serine utilizing a microelectrode in combination with a ᴅ-amino acid oxidase enzyme, which produces stoichiometric quantities of hydrogen peroxide in response to ᴅ-serine. For the fabrication of a biosensor with good selectivity, we use a permselective poly(meta-phenylenediamine) film to ensure only the target molecule is reacted, according to the size exclusion principle. In this work, we investigated the effect of the electrodeposition conditions used on the biosensor’s response time and selectivity. Careful optimization of the fabrication process allowed for enhanced biosensor response time. This allowed for the real time sensing of ᴅ-serine in a bulk solution, and also provided in means to map the efflux of ᴅ-serine in real time. This was done using scanning electrochemical microscopy (SECM) with the optimized biosensor to measure localized release of ᴅ-serine from an agar filled glass capillary sealed in an epoxy puck, which acted as a model system. The SECM area scan simultaneously provided information regarding the rate of ᴅ-serine flux from the model substrate, as well as the size of the substrate itself. This SECM methodology, which provides high spatial and temporal resolution, could be useful to investigate the primary site and mechanism of ᴅ-serine release in other biological samples.

Keywords: ᴅ-serine, enzymatic biosensor, microelectrode, scanning electrochemical microscopy

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Authors: Gizachew Belay Adugna

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Scalable and cost-effective device fabrication techniques are urgent to commercialize the perovskite solar cells (PSCs) for the next photovoltaic (PV) technology. Herein, large-area films of perovskite and hole-transporting materials (HTMs) were developed via a rapid and scalable thermal-assisting bar-coating process in the open air. High-quality and large crystalline grains of MAPbI₃ with homogenous morphology and thickness were obtained on a large-area (10 cm×10 cm) solution-sheared mp-TiO₂/c-TiO₂/FTO substrate. Encouraging photovoltaic performance of 19.02% was achieved for devices fabricated from the bar-coated perovskite film compared to that from the small-scale spin-coated film (17.27%) with 2,2′,7,7′-tetrakis-(N,N-di-p-methoxyphenylamine)-9,9′-spirobifluorene (spiro-OMeTAD) as an HTM whereas a higher power conversion efficiency of 19.89% with improved device stability was achieved by capping a fluorinated (HYC-2) HTM as an alternative to the traditional spiro-OMeTAD. The fluorinated exhibited better molecular packing in the HTM film and deeper HOMO level compared to the nonfluorinated counterpart; thus, improved hole mobility and overall charge extraction in the device were demonstrated. Furthermore, excellent film processability and an impressive PCE of 18.52% were achieved in the large area bar-coated HYC-2 prepared sequentially on the perovskite underlayer in the open atmosphere, compared to the bar-coated spiro-OMeTAD/perovskite (17.51%). This all-solution approach demonstrated the feasibility of high-quality films on a large-area substrate for PSCs, which is a vital step toward industrial-scale PV production.

Keywords: perovskite solar cells, hole transporting materials, up-scaling process, power conversion efficiency

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Authors: Muhammad Yasir Wadood, Fatemeh Babaeian

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By developing ultra-wideband (UWB) systems, there is a high demand for UWB filters with low insertion loss, wide bandwidth, and having a planar structure which is compatible with other components of the UWB system. A microstrip interdigital filter is a great option for designing UWB filters. However, the presence of via holes in this structure creates difficulties in the fabrication procedure of the filter. Especially in the higher frequency band, any misalignment of the drilled via hole with the Microstrip stubs causes large errors in the measurement results compared to the desired results. Moreover, in this case (high-frequency designs), the line width of the stubs are very narrow, so highly precise small via holes are required to be implemented, which increases the cost of fabrication significantly. Also, in this case, there is a risk of having fabrication errors. To combat this issue, in this paper, a via-less UWB microstrip filter is proposed which is designed based on a modification of a conventional inter-digital bandpass filter. The novel approaches in this filter design are 1) replacement of each via hole with a quarter-wavelength open circuit stub to avoid the complexity of manufacturing, 2) using a bend structure to reduce the unwanted coupling effects and 3) minimising the size. Using the proposed structure, a UWB filter operating in the frequency band of 3.9-6.6 GHz (1-dB bandwidth) is designed and fabricated. The promising results of the simulation and measurement are presented in this paper. The selected substrate for these designs was Rogers RO4003 with a thickness of 20 mils. This is a common substrate in most of the industrial projects. The compact size of the proposed filter is highly beneficial for applications which require a very miniature size of hardware.

Keywords: band-pass filters, inter-digital filter, microstrip, via-less

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Authors: Mohammed Panthakkal Abdul Muthalif, Shanmugasundaram Kanagaraj, Jumi Park, Hangyu Park, Youngson Choe

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The present study reports the incorporation of calcium ions into the CuS counter electrodes (CEs) in order to modify the photovoltaic performance of quantum dot-sensitized solar cells (QDSSCs). Metal ion-doped CuS thin film was prepared by the chemical bath deposition (CBD) method on FTO substrate and used directly as counter electrodes for TiO₂/CdS/CdSe/ZnS photoanodes based QDSSCs. For the Ca-doped CuS thin films, copper nitrate and thioacetamide were used as anionic and cationic precursors. Calcium nitrate tetrahydrate was used as doping material. The surface morphology of Ca-doped CuS CEs indicates that the fragments are uniformly distributed, and the structure is densely packed with high crystallinity. The changes observed in the diffraction patterns suggest that Ca dopant can introduce increased disorder into CuS material structure. EDX analysis was employed to determine the elemental identification, and the results confirmed the presence of Cu, S, and Ca on the FTO glass substrate. The photovoltaic current density – voltage characteristics of Ca-doped CuS CEs shows the specific improvements in open circuit voltage decay (Voc) and short-circuit current density (Jsc). Electrochemical impedance spectroscopy results display that Ca-doped CuS CEs have greater electrocatalytic activity and charge transport capacity than bare CuS. All the experimental results indicate that 20% Ca-doped CuS CE based QDSSCs exhibit high power conversion efficiency (η) of 4.92%, short circuit current density of 15.47 mA cm⁻², open circuit photovoltage of 0.611 V, and fill factor (FF) of 0.521 under illumination of one sun.

Keywords: Ca-doped CuS counter electrodes, surface morphology, chemical bath deposition method, electrocatalytic activity

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Authors: Katie Kilgour, Brendan Turner, Carly Catella, Michael Daniele, Stefano Menegatti

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In vivo, the extracellular matrix (ECM) provides biochemical and mechanical properties that are instructional to resident cells to form complex tissues with characteristics to develop and support vascular networks. In vitro, the development of vascular networks can be guided by biochemical patterning of substrates via spatial distribution and display of peptides and growth factors to prompt cell adhesion, differentiation, and proliferation. We have developed a technique utilizing peptide ligands that specifically bind vascular endothelial growth factor (VEGF), erythropoietin (EPO), or angiopoietin-1 (ANG1) to spatiotemporally distribute growth factors to cells. This allows for the controlled release of each growth factor, ultimately enhancing the formation of a vascular network. Our engineered tissue constructs (ETCs) are fabricated out of gelatin methacryloyl (GelMA), which is an ideal substrate for tailored stiffness and bio-functionality, and covalently patterned with growth factor specific peptides. These peptides mimic growth factor receptors, facilitating the non-covalent binding of the growth factors to the ETC, allowing for facile uptake by the cells. We have demonstrated in the absence of cells the binding affinity of VEGF, EPO, and ANG1 to their respective peptides and the ability for each to be patterned onto a GelMA substrate. The ability to organize growth factors on an ETC provides different functionality to develop organized vascular networks. Our results demonstrated a method to incorporate biochemical cues into ETCs that enable spatial and temporal control of growth factors. Future efforts will investigate the cellular response by evaluating gene expression, quantifying angiogenic activity, and measuring the speed of growth factor consumption.

Keywords: growth factor, hydrogel, peptide, angiogenesis, vascular, patterning

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Authors: Jana Andzane, Gunta Kunakova, Margarita Baitimirova, Mikelis Marnauza, Floriana Lombardi, Donats Erts

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Bismuth selenide (Bi₂Se₃) is known as a narrow band gap semiconductor with pronounced thermoelectric (TE) and topological insulator (TI) properties. Unique TI properties offer exciting possibilities for fundamental research as observing the exciton condensate and Majorana fermions, as well as practical application in spintronic and quantum information. In turn, TE properties of this material can be applied for wide range of thermoelectric applications, as well as for broadband photodetectors and near-infrared sensors. Nanostructuring of this material results in improvement of TI properties due to suppression of the bulk conductivity, and enhancement of TE properties because of increased phonon scattering at the nanoscale grains and interfaces. Regarding TE properties, crystallographic growth direction, as well as orientation of the nanostructures relative to the growth substrate, play significant role in improvement of TE performance of nanostructured material. For instance, Bi₂Se₃ layers consisting of randomly oriented nanostructures and/or of combination of them with planar nanostructures show significantly enhanced in comparison with bulk and only planar Bi₂Se₃ nanostructures TE properties. In this work, a catalyst-free vapour-solid deposition technique was applied for controlled obtaining of different types of Bi₂Se₃ nanostructures and continuous nanostructured layers for targeted applications. For example, separated Bi₂Se₃ nanoplates, nanobelts and nanowires can be used for investigations of TI properties; consisting from merged planar and/or randomly oriented nanostructures Bi₂Se₃ layers are useful for applications in heat-to-power conversion devices and infrared detectors. The vapour-solid deposition was carried out using quartz tube furnace (MTI Corp), equipped with an inert gas supply and pressure/temperature control system. Bi₂Se₃ nanostructures/nanostructured layers of desired type were obtained by adjustment of synthesis parameters (process temperature, deposition time, pressure, carrier gas flow) and selection of deposition substrate (glass, quartz, mica, indium-tin-oxide, graphene and carbon nanotubes). Morphology, structure and composition of obtained Bi₂Se₃ nanostructures and nanostructured layers were inspected using SEM, AFM, EDX and HRTEM techniques, as well as home-build experimental setup for thermoelectric measurements. It was found that introducing of temporary carrier gas flow into the process tube during the synthesis and deposition substrate choice significantly influence nanostructures formation mechanism. Electrical, thermoelectric, and topological insulator properties of different types of deposited Bi₂Se₃ nanostructures and nanostructured coatings are characterized as a function of thickness and discussed.

Keywords: bismuth seleinde, nanostructures, topological insulator, vapour-solid deposition

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Authors: Prasad Urvashi, Wynn Yezarni, Williams Shehan, Ravindran Arun

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Introduction: Primary health services are often the first point of contact that patients with mental illness have with the healthcare system. A number of tools have been developed to increase detection of depression in the context of primary care. However, one challenge amongst many includes utilizing these tools within the limited primary care consultation timeframe. Therefore, short questionnaires that screen for depression that are just as effective as more comprehensive diagnostic tools may be beneficial in improving detection rates of patients visiting a primary care setting. Objective: To develop and determine the sensitivity and specificity of a 2-Question Questionnaire (2-QQ) to screen for depression in in a suburban primary care clinic in Ragama, Sri Lanka. The purpose is to develop a short screening tool for depression that is culturally adapted in order to increase the detection of depression in the Sri Lankan patient population. Methods: This was a cross-sectional study involving two steps. Step one: verbal administration of 2-QQ to patients by their primary care physician. Step two: completion of the Peradeniya Depression Scale, a validated diagnostic tool for depression, the patient after their consultation with the primary care physician. The results from the PDS were then correlated to the results from the 2-QQ for each patient to determine sensitivity and specificity of the 2-QQ. Results: A score of 1/+ on the 2-QQ was most sensitive but least specific. Thus, setting the threshold at this level is effective for correctly identifying depressed patients, but also inaccurately captures patients who are not depressed. A score of 6 on the 2-QQ was most specific but least sensitive. Setting the threshold at this level is effective for correctly identifying patients without depression, but not very effective at capturing patients with depression. Discussion: In the context of primary care, it may be worthwhile setting the 2-QQ screen at a lower threshold for positivity (such as a score of 1 or above). This would generate a high test sensitivity and thus capture the majority of patients that have depression. On the other hand, by setting a low threshold for positivity, patients who do not have depression but score higher than 1 on the 2-QQ will also be falsely identified as testing positive for depression. However, the benefits of identifying patients who present with depression may outweigh the harms of falsely identifying a non-depressed patient. It is our hope that the 2-QQ will serve as a quick primary screen for depression in the primary care setting and serve as a catalyst to identify and treat individuals with depression.

Keywords: depression, primary care, screening tool, Sri Lanka

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Authors: John Justo Ambuchi, Zhaohan Zhang, Yujie Feng

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Quick development and usage of nanotechnology have resulted to massive use of various nanoparticles, such as iron oxide nanoparticles (IONPs) and multi-wall carbon nanotubes (MWCNTs). Thus, this study investigated the role of IONPs and MWCNTs in enhancing bioenergy recovery. Results show that IONPs at a concentration of 750 mg/L and MWCNTs at a concentration of 1500 mg/L induced faster substrate utilization and biogas production rates than the control. IONPs exhibited higher carbon oxygen demand (COD) removal efficiency than MWCNTs while on the contrary, MWCNT performance on biogas generation was remarkable than IONPs. Furthermore, scanning electron microscopy (SEM) investigation revealed extracellular polymeric substances (EPS) excretion from AGS had an interaction with nanoparticles. This interaction created a protective barrier to microbial consortia hence reducing their cytotoxicity. Microbial community analyses revealed genus predominance of bacteria of Anaerolineaceae and Longilinea. Their role in biodegradation of the substrate could have highly been boosted by nanoparticles. The archaea predominance of the genus level of Methanosaeta and Methanobacterium enhanced methanation process. The presence of bacteria of genus Geobacter was also reported. Their presence might have significantly contributed to direct interspecies electron transfer in the system. Exposure of AGS to nanoparticles promoted direct interspecies electron transfer among the anaerobic fermenting bacteria and their counterpart methanogens during the anaerobic digestion process. This results provide useful insightful information in understanding the response of microorganisms to IONPs and MWCNTs in the complex natural environment.

Keywords: anaerobic granular sludge, extracellular polymeric substances, iron oxide nanoparticles, multi-wall carbon nanotubes

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Authors: Renée M. Ripken, Johannes G. E. Gardeniers, Séverine Le Gac

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Controlling the multiphase behavior of aqueous biomass mixtures is essential when working in the biomass conversion industry. Here, the vapor/liquid equilibria (VLE) of ethylene glycol, glycerol, and xylitol were studied for temperatures between 25 and 200 °C and pressures of 1 to 10 bar. These experiments were performed in a microfluidic platform, which exhibits excellent heat transfer properties so that equilibrium is reached fast. Firstly, the saturated vapor pressure as a function of the temperature and the substrate mole fraction of the substrate was calculated using AspenPlus with a Redlich-Kwong-Soave Boston-Mathias (RKS-BM) model. Secondly, we developed a high-pressure and high-temperature microfluidic set-up for experimental validation. Furthermore, we have studied the multiphase flow pattern that occurs after the saturation temperature was achieved. A glass-silicon microfluidic device containing a 0.4 or 0.2 m long meandering channel with a depth of 250 μm and a width of 250 or 500 μm was fabricated using standard microfabrication techniques. This device was placed in a dedicated chip-holder, which includes a ceramic heater on the silicon side. The temperature was controlled and monitored by three K-type thermocouples: two were located between the heater and the silicon substrate, one to set the temperature and one to measure it, and the third one was placed in a 300 μm wide and 450 μm deep groove on the glass side to determine the heat loss over the silicon. An adjustable back pressure regulator and a pressure meter were added to control and evaluate the pressure during the experiment. Aqueous biomass solutions (10 wt%) were pumped at a flow rate of 10 μL/min using a syringe pump, and the temperature was slowly increased until the theoretical saturation temperature for the pre-set pressure was reached. First and surprisingly, a significant difference was observed between our theoretical saturation temperature and the experimental results. The experimental values were 10’s of degrees higher than the calculated ones and, in some cases, saturation could not be achieved. This discrepancy can be explained in different ways. Firstly, the pressure in the microchannel is locally higher due to both the thermal expansion of the liquid and the Laplace pressure that has to be overcome before a gas bubble can be formed. Secondly, superheating effects are likely to be present. Next, once saturation was reached, the flow pattern of the gas/liquid multiphase system was recorded. In our device, the point of nucleation can be controlled by taking advantage of the pressure drop across the channel and the accurate control of the temperature. Specifically, a higher temperature resulted in nucleation further upstream in the channel. As the void fraction increases downstream, the flow regime changes along the channel from bubbly flow to Taylor flow and later to annular flow. All three flow regimes were observed simultaneously. The findings of this study are key for the development and optimization of a microreactor for hydrogen production from biomass.

Keywords: biomass conversion, high pressure and high temperature microfluidics, multiphase, phase diagrams, superheating

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Authors: Tiffany Baetens, Sophie Halliez, Luc Buée, Emiliano Pallecchi, Vincent Thomy, Steve Arscott

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We demonstrate here a viable stretchable bio-microelectromechanical systems (BioMEMS) technology for use with biological studies concerned with the effect of high mechanical strains on living cells. An example of this is traumatic brain injury (TBI) where neurons are damaged with physical force to the brain during, e.g., accidents and sports. Robust, miniaturized integrated systems are needed by biologists to be able to study the effect of TBI on neuron cells in vitro. The major challenges in this area are (i) to develop micro, and nanofabrication processes which are based on stretchable substrates and to (ii) create systems which are robust and performant at very high mechanical strain values—sometimes as high as 100%. At the time of writing, such processes and systems were rapidly evolving subject of research and development. The BioMEMS which we present here is composed of an elastomer substrate (low Young’s modulus ~1 MPa) onto which is patterned robust electrodes and insulators. The patterning of the thin films is achieved using standard photolithography techniques directly on the elastomer substrate—thus making the process generic and applicable to many materials’ in based systems. The chosen elastomer used is commercial ‘Sylgard 184’ polydimethylsiloxane (PDMS). It is spin-coated onto a silicon wafer. Multistep ultra-violet based photolithography involving commercial photoresists are then used to pattern robust thin film metallic electrodes (chromium/gold) and insulating layers (parylene) on the top of the PDMS substrate. The thin film metals are deposited using thermal evaporation and shaped using lift-off techniques The BioMEMS has been characterized mechanically using an in-house strain-applicator tool. The system is composed of 12 electrodes with one reference electrode transversally-orientated to the uniaxial longitudinal straining of the system. The electrical resistance of the electrodes is observed to remain very stable with applied strain—with a resistivity approaching that of evaporated gold—up to an interline strain of ~50%. The mechanical characterization revealed some interesting original properties of such stretchable BioMEMS. For example, a Poisson effect induced electrical ‘self-healing’ of cracking was identified. Biocompatibility of the commercial photoresist has been studied and is conclusive. We will present the results of the BioMEMS, which has also characterized living cells with a commercial Multi Electrode Array (MEA) characterization tool (Multi Channel Systems, USA). The BioMEMS enables the cells to be strained up to 50% and then characterized electrically and optically.

Keywords: BioMEMS, elastomer, electrical impedance measurements of living cells, high mechanical strain, microfabrication, stretchable systems, thin films, traumatic brain injury

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Authors: Pradakshina Sharma, Jagriti Narang

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Due to the critical significance in the early identification of infectious diseases, electrochemical sensors have garnered considerable interest. Here, we develop a detection platform for the chikungunya virus by rationally implementing the extremely high charge-transfer efficiency of a ternary nanocomposite of graphene oxide, silver, and gold (G/Ag/Au) (CHIKV). Because paper is an inexpensive substrate and can be produced in large quantities, the use of electrochemical paper analytical device (EPAD) origami further enhances the sensor's appealing qualities. A cost-effective platform for point-of-care diagnostics is provided by paper-based testing. These types of sensors are referred to as eco-designed analytical tools due to their efficient production, usage of the eco-friendly substrate, and potential to reduce waste management after measuring by incinerating the sensor. In this research, the paper's foldability property has been used to develop and create 3D multifaceted biosensors that can specifically detect the CHIKVX-ray diffraction, scanning electron microscopy, UV-vis spectroscopy, and transmission electron microscopy (TEM) were used to characterize the produced nanoparticles. In this work, aptamers are used since they are thought to be a unique and sensitive tool for use in rapid diagnostic methods. Cyclic voltammetry (CV) and linear sweep voltammetry (LSV), which were both validated with a potentiostat, were used to measure the analytical response of the biosensor. The target CHIKV antigen was hybridized with using the aptamer-modified electrode as a signal modulation platform, and its presence was determined by a decline in the current produced by its interaction with an anionic mediator, Methylene Blue (MB). Additionally, a detection limit of 1ng/ml and a broad linear range of 1ng/ml-10µg/ml for the CHIKV antigen were reported.

Keywords: biosensors, ePAD, arboviral infections, point of care

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Authors: Rwei-Ching Chang, Bo-Yu Su

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Adhesion of copper thin films deposited on polyethylene terephthAdhesion of copper thin films deposited on polyethylene terephthalate substrate by direct current sputtering with different sputtering parameters is discussed in this work. The effects of plasma treatment with 0, 5, and 10 minutes on the thin film properties are investigated first. Various argon flow rates at 40, 50, 60 standard cubic centimeters per minute (sccm), deposition power at 30, 40, 50 W, and film thickness at 100, 200, 300 nm are also discussed. The 3-dimensional surface profilometer, micro scratch machine, and optical microscope are used to characterize the thin film properties. The results show that the increase of the plasma treatment time on the polyethylene terephthalate surface affects the roughness and critical load of the films. The critical load increases as the plasma treatment time increases. When the plasma treatment time was adjusted from 5 minutes to 10 minutes, the adhesion increased from 8.20 mN to 13.67 mN. When the argon flow rate is decreased from 60 sccm to 40 sccm, the adhesion increases from 8.27 mN to 13.67 mN. The adhesion is also increased by the condition of higher power, where the adhesion increased from 13.67 mN to 25.07 mN as the power increases from 30 W to 50 W. The adhesion of the film increases from 13.67 mN to 21.41mN as the film thickness increases from 100 nm to 300 nm. Comparing all the deposition parameters, it indicates the change of the power and thickness has much improvement on the film adhesion.alate substrate by direct current sputtering with different sputtering parameters is discussed in this work. The effects of plasma treatment with 0, 5, and 10 minutes on the thin film properties are investigated first. Various argon flow rates at 40, 50, 60 standard cubic centimeters per minute (sccm), deposition power at 30, 40, 50 W, and film thickness at 100, 200, 300 nm are also discussed. The 3-dimensional surface profilometer, micro scratch machine, and optical microscope are used to characterize the thin film properties. The results show that the increase of the plasma treatment time on the polyethylene terephthalate surface affects the roughness and critical load of the films. The critical load increases as the plasma treatment time increases. When the plasma treatment time was adjusted from 5 minutes to 10 minutes, the adhesion increased from 8.20 mN to 13.67 mN. When the argon flow rate is decreased from 60 sccm to 40 sccm, the adhesion increases from 8.27 mN to 13.67 mN. The adhesion is also increased by the condition of higher power, where the adhesion increased from 13.67 mN to 25.07 mN as the power increases from 30 W to 50 W. The adhesion of the film increases from 13.67 mN to 21.41mN as the film thickness increases from 100 nm to 300 nm. Comparing all the deposition parameters, it indicates the change of the power and thickness has much improvement on the film adhesion.

Keywords: flexible substrate, sputtering, adhesion, copper thin film

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Authors: Himanshu Singh, Rishi Kant, Shantanu Bhattacharya

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This paper adopts a top-down approach for mathematical modeling to predict the size reduction from micro to nano-scale through persistent etching. The process is simulated using a finite difference approach. Previously, various researchers have simulated the etching process for 1-D and 2-D substrates. It consists of two processes: 1) Convection-Diffusion in the etchant domain; 2) Chemical reaction at the surface of the particle. Since the process requires analysis along moving boundary, partial differential equations involved cannot be solved using conventional methods. In 1-D, this problem is very similar to Stefan's problem of moving ice-water boundary. A fixed grid method using finite volume method is very popular for modelling of etching on a one and two dimensional substrate. Other popular approaches include moving grid method and level set method. In this method, finite difference method was used to discretize the spherical diffusion equation. Due to symmetrical distribution of etchant, the angular terms in the equation can be neglected. Concentration is assumed to be constant at the outer boundary. At the particle boundary, the concentration of the etchant is assumed to be zero since the rate of reaction is much faster than rate of diffusion. The rate of reaction is proportional to the velocity of the moving boundary of the particle. Modelling of the above reaction was carried out using Matlab. The initial particle size was taken to be 50 microns. The density, molecular weight and diffusion coefficient of the substrate were taken as 2.1 gm/cm3, 60 and 10-5 cm2/s respectively. The etch-rate was found to decline initially and it gradually became constant at 0.02µ/s (1.2µ/min). The concentration profile was plotted along with space at different time intervals. Initially, a sudden drop is observed at the particle boundary due to high-etch rate. This change becomes more gradual with time due to declination of etch rate.

Keywords: particle size reduction, micromixer, FDM modelling, wet etching

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Authors: S. T. Kumbhar, M. S. Bhatia, R. C. Khairate

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An effective nanodrug delivery system was developed by using chitosan for increased encapsulation efficiency and retarded release of curcumin. Potential ionotropic gelation method was used for the development of chitosan nanoparticles with TPP as cross-linker. The characterization was done for analysis of size, structure, surface morphology, and thermal behavior of synthesized chitosan nanoparticles. The encapsulation efficiency was more than 80%, with improved drug loading capacity. The in-vitro drug release study showed that curcumin release rate was decreased significantly. These chitosan nanoparticles could be a suitable platform for co-delivery of curcumin and anticancer agent for enhanced cytotoxic effect on tumor cells.

Keywords: Curcumin, chitosan, nanoparticles, anticancer activity

Procedia PDF Downloads 151

Authors: E. Bartkiene, V. Krungleviciute, J. Kucinskiene, R. Antanaitis, A. Kucinskas

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In calves less than 10-day-old, infection commonly cause severe diarrhoea and high mortality. To prevention of calves diseases a common practice is to treat calves with prophylactic antibiotics, in this case the use of lactic acid bacteria (LAB) is promising. Often LAB strains are incubated in comercial de Man-Rogosa-Sharpe (MRS) medium, the culture are centrifuged, the cells are washing with sterile water, and this suspension is used as a starter culture for animal health care. Juice of potatoe tubers is industrial wastes, wich may constitute a source of digestible nutrients for microorganisms. In our study the ability of LAB to utilize potatoe tubers juice in cell synthesis without external nutrient supplement was investigated, and the influence of multiplied LAB on calves blood parameters was evaluated. Calves were selected based on the analogy principle (treatment group (n=6), control group (n=8)). For the treatment group 14 days was given a 50 ml of fermented potatoe tubers juice containing 9.6 log10 cfu/ml of LAB. Blood parameters (gas and biochemical) were assessed by use of an auto-analyzers (Hitachi 705 and EPOC). Before the experiment, blood pH of treatment group calves was 7.33, control – 7.36, whereas, after 14 days, 7.28 and 7.36, respectively. Calves blood pH in the treatment group remained stable over the all experiment period. Concentration of PCO2 in control calves group blood increased from 63.95 to 70.93, whereas, in the treatment group decreased from 63.08 to 60.71. Concentration of lactate in the treatment group decreased from 3.20 mmol/l to 2.64 mmol/l, whereas, in control - increased from 3.95 mmol/l to 4.29 mmol/l. Concentration of AST in the control calves group increased from 50.18 IU/L to 58.9 IU/L, whereas, in treatment group decreased from 49.82 IU/L to 33.1 IU/L. We conclude that the 50 ml of fermented potatoe tubers juice containing 9.6 log10 cfu/ml of LAB per day, by using 14 days, reduced risk of developing acidosis (stabilizes blood pH (p < 0.05)), reduces lactates and PCO2 concentration (p < 0.05) and risk of liver lesions (reduces AST concentration (p < 0.005)) in blood of calves.

Keywords: alternative substrate, blood parameters, calves, lactic acid bacteria

Procedia PDF Downloads 286

Authors: Meng-Jie Hsiao, Cam Nguyen

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Wide-band devices in Ku band (12-18 GHz) and K band (18-27 GHz) have received significant attention for high-data-rate communications and high-resolution sensing. Especially, devices operating around 24 GHz is attractive due to the 24-GHz unlicensed applications. One of the most important components in RF systems is power amplifier (PA). Various PAs have been developed in the Ku and K bands on GaAs, InP, and silicon (Si) processes. Although the PAs using GaAs or InP process could have better power handling and efficiency than those realized on Si, it is very hard to integrate the entire system on the same substrate for GaAs or InP. Si, on the other hand, facilitates single-chip systems. Hence, good PAs on Si substrate are desirable. Especially, Si-based PA having good linearity is necessary for next generation communication protocols implemented on Si. We report a 16.5 to 25.5 GHz Si-based PA having flat saturated power of 19.5 ± 1.5 dBm, output 1-dB power compression (OP1dB) of 16.5 ± 1.5 dBm, and 15-23 % power added efficiency (PAE). The PA consists of a drive amplifier, two main amplifiers, and lump-element Wilkinson power divider and combiner designed and fabricated in TowerJazz 0.18µm SiGe BiCMOS process having unity power gain frequency (fMAX) of more than 250 GHz. The PA is realized as a cascode amplifier implementing both heterojunction bipolar transistor (HBT) and n-channel metal–oxide–semiconductor field-effect transistor (NMOS) devices for gain, frequency response, and linearity consideration. Particularly, a body-floating technique is utilized for the NMOS devices to improve the voltage swing and eliminate parasitic capacitances. The developed PA has measured flat gain of 20 ± 1.5 dB across 16.5-25.5 GHz. At 24 GHz, the saturated power, OP1dB, and maximum PAE are 20.8 dBm, 18.1 dBm, and 23%, respectively. Its high performance makes it attractive for use in Ku/K-band, especially 24 GHz, communication and radar systems. This paper was made possible by NPRP grant # 6-241-2-102 from the Qatar National Research Fund (a member of Qatar Foundation). The statements made herein are solely the responsibility of the authors.

Keywords: power amplifiers, amplifiers, communication systems, radar systems

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Authors: Aulanni’am Aulanni’am

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Incidence of Diabetes Mellitus (DM) progressively increasing it became a serious problem in Indonesia and it is a disease that government is priority to be addressed. The longer a person is suffering from diabetes the more likely to develop complications particularly diabetic patients who are not well maintained. Therefore, Incidence of Diabetes Mellitus needs to be done in the early diagnosis of pre-phase of the disease. In this pre-phase disease, already happening destruction of pancreatic beta cells and declining in beta cell function and the sign autoimmunity reactions associated with beta cell destruction. Type 1 DM is a multifactorial disease triggered by genetic and environmental factors, which leads to the destruction of pancreatic beta cells. Early marker of "beta cell autoreactivity" is the synthesis of autoantibodies against 65-kDa protein, which can be a molecule that can be detected early in the disease pathomechanism. The importance of early diagnosis of diabetic patients held in the phase of pre-disease is to determine the progression towards the onset of pancreatic beta cell destruction and take precautions. However, the price for this examination is very expensive ($ 150/ test), the anti-GAD65 abs examination cannot be carried out routinely in most or even in all laboratories in Indonesia. Therefore, production-based Rapid Test Recombinant Human Protein GAD65 with "Reverse Flow Immunchromatography Technique" in Indonesia is believed to reduce costs and improve the quality of care of patients with diabetes in Indonesia. Rapid Test Product innovation is very simple and suitable for screening and routine inspection of GAD65 autoantibodies. In the blood serum of patients with diabetes caused by autoimmunity, autoantibody-GAD65 is a major serologic marker to detect autoimmune reaction because their concentration level of stability.GAD65 autoantibodies can be found 10 years before clinical symptoms of diabetes. Early diagnosis is more focused to detect the presence autontibodi-GAD65 given specification and high sensitivity. Autoantibodies- GAD65 that circulates in the blood is a major indicator of the destruction of the islet cells of the pancreas. Results of research in collaboration with Biofarma has produced GAD65 autoantibodies based Rapid Test had conducted the soft launch of products and has been tested with the results of a sensitivity of 100 percent and a specificity between 90 and 96% compared with the gold standard (import product) which worked based on ELISA method.

Keywords: diabetes mellitus, GAD65 autoantibodies, rapid test, sensitivity, specificity

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Authors: Joseph A. Bentil, Anders Thygesen, Lene Langea, Moses Mensah, Anne Meyer

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The study investigated the saccharification potential of in-house enzymes produced from a white-rot basidiomycete strain, Polyporus ciliatus CBS 366.74. The in-house enzymes were produced by growing the fungus on mono and composite substrates of cocoa pod husk (CPH) and green seaweed (GS) (Ulva lactuca sp.) with and without the addition of 25mM ammonium nitrate at 4%w/v substrate concentration in submerged condition for 144 hours. The crude enzyme extracts preparations (CEE 1-5 and CEE 1-5+AN) obtained from the fungal cultivation process were sterile-filtered and used as enzyme sources for enzymatic hydrolysis of hydrothermally pretreated corn stover using a commercial cocktail enzyme, Cellic Ctec3, as benchmark. The hydrolysis was conducted at 50ᵒC with 50mM sodium acetate buffer, pH 5 based on enzyme dosages of 5 and 10 CMCase Units/g biomass at 1%w/v dry weight substrate concentration at time points of 6, 24, and 72 hours. The enzyme activity profile of the in-house enzymes varied among the growth substrates with the composite substrates (50-75% GS and AN inclusion), yielding better enzyme activities, especially endoglucanases (0.4-0.5U/mL), β-glucosidases (0.1-0.2 U/mL), and xylanases (3-10 U/mL). However, nitrogen supplementation had no significant effect on enzyme activities of crude extracts from 100% GS substituted substrates. From the enzymatic hydrolysis, it was observed that the in-house enzymes were capable of hydrolysing the pretreated corn stover at varying degrees; however, the saccharification yield was less than 10%. Consequently, theoretical glucose yield was ten times lower than Cellic Ctec3 at both dosage levels. There was no linear correlation between glucose yield and enzyme dosage for the in-house enzymes, unlike the benchmark enzyme. It is therefore recommended that the in-house enzymes are used to complement the dosage of commercial enzymes to reduce the cost of biomass saccharification.

Keywords: enzyme production, hydrolysis yield, feedstock, enzyme blend, Polyporus ciliatus

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Authors: M. Umar Dahot, G. S. Mangrio

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In this study Agro-industrial waste was used as a carbon source, which is a low cost substrate. Along with this, various sugars and molasses of 2.5% and 5% were investigated as substrate/carbon source for the growth of A.niger and Pectinase production. Different nitrogen sources were also used. An overview of results obtained show that 5% sucrose, 5% molasses and 0.4% (NH4)2SO4 were found the best carbon and nitrogen sources for the production of pectinase by A. niger. The maximum production of pectinase (26.87units/ml) was observed at pH 6.0 after 72 hrs incubation. The optimum temperature for the maximum production of pectinase was achieved at 35ºC when maximum production of pectinase was obtained as 28.25Units/ml.Pectinase enzyme was purified with ammonium sulphate precipitation and dialyzed sample was finally applied on gel filtration chromatography (Sephadex G-100) and Ion Exchange DEAE A-50. The enzyme was purified 2.5 fold by gel chromatography on Sephadex G-100 and Four fractions were obtained, Fraction 1, 2, 4 showed single band while Fraction -3 showed multiple bands on SDS Page electrophoresis. Fraction -3 was pooled, dialyzed and separated on Sephdex A-50 and two fractions 3a and 3b showed single band. The molecular weights of the purified fractions were detected in the range of 33000 ± 2000 and 38000± 2000 Daltons. The purified enzyme was specifically most active with pure pectin, while pectin, Lemon pectin and orange peel given lower activity as compared to (control). The optimum pH and temperature for pectinase activity was found between pH 5.0 and 6.0 and 40°- 50°C, respectively. The enzyme was stable over the pH range 3.0-8.0. The thermostability of was determined and it was observed that the pectinase activity is heat stable and retains activity more than 40% when incubated at 90°C for 10 minutes. The pectinase activity of F3a and F3b was increased with different metal ions. The Pectinase activity was stimulated in the presence of CaCl2 up to 10-30%. ZnSO4, MnSO4 and Mg SO4 showed higher activity in fractions F3a and F3b, which indicates that the pectinase belongs to metalo-enzymes. It is concluded that A. niger is capable to produce pH stable and thermostable pectinase, which can be used for industrial purposes.

Keywords: pectinase, a. niger, production, purification, characterization

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Authors: A. Natsir, M. Nadir, S. Syahrir, A. Mujnisa

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In tropical countries such as in Indonesia, rice straw plays an important role in fulfilling the needs of feed for ruminant, especially during the dry season in which the availability of forage is very limited. However, the main problem of using rice straw as a feedstuff is low digestibility due to the existence of the links between lignin and cellulose or hemicellulose, and imbalance of its minerals content. One alternative to solve this problem is by application of biodecomposer (BS) derived from rumen bacterial of the ruminant. This study was designed to assess the effects of BS application on the changes of the chemical composition of rice straw. Four adults local buffalo raised under typical feeding conditions were used as a source of inoculum for BS development. The animal was fed for a month with a diet consisted of rice straw and elephant grass before taking rumen fluid samples. Samples of rumen fluid were inoculated in the carboxymethyl cellulose (CMC) media under anaerobic condition for 48 hours at 37°C. The mixture of CMC media and microbes are ready to be used as a biodecomposer following incubation of the mixture under anaerobic condition for 7 days at 45°C. The effectiveness of BS then assessed by applying the BS on the straw according to completely randomized design consisted of four treatments and three replication. One hundred g of ground coarse rice straw was used as the substrate. The BS was applied to the rice straw substrate with the following composition: Rice straw without BS (P0), rice straw + 5% BS (P1), rice straw +10% BS (P2), and rice straw + 15% BS. The mixture of rice straw and BS then fermented under anaerobic for four weeks. Following the fermentation, the chemical composition of rice straw was evaluated. The results indicated that the crude protein content of rice straw significantly increased (P < 0.05) as the level of BS increased. On the other hand, the concentration of crude fiber of the rice straw was significantly decreased (P < 0.05) as the level of BS increased. Other nutrients such as minerals did not change (P > 0.05) due to the treatments. In conclusion, application of BS developed from rumen bacterial of buffalo has a promising prospect to be used as a biological agent to improve the quality of rice straw as feeding for ruminant.

Keywords: biodecomposer, local buffalo, rumen microbial, chemical composition

Procedia PDF Downloads 173

Authors: Tesnim Charrad, Kaouther Nouira, Ahmed Ferchichi

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In order to reduce the number of deaths due to heart problems, we propose the use of Hierarchical Temporal Memory Algorithm (HTM) which is a real time anomaly detection algorithm. HTM is a cortical learning algorithm based on neocortex used for anomaly detection. In other words, it is based on a conceptual theory of how the human brain can work. It is powerful in predicting unusual patterns, anomaly detection and classification. In this paper, HTM have been implemented and tested on ECG datasets in order to detect cardiac anomalies. Experiments showed good performance in terms of specificity, sensitivity and execution time.

Keywords: cardiac anomalies, ECG, HTM, real time anomaly detection

Procedia PDF Downloads 191

Authors: Zeynep Yazgan, Gökhan Aygün, Reyhan Çalışkan

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Objective: Invasive fungal infections are a serious threat in terms of morbidity and mortality, especially in immunocompromised patients. The most frequently isolated agents are Aspergillus genus fungi, and sensitivity to azoles, which are the first choice in treatment, decreases. In our study, we aimed to investigate the use of the agar plate screening method as a fast, easy, and practical method in determining azole resistance in Aspergillus spp. species. Methods: Our study was conducted with 125 Aspergillus spp. isolates produced from various clinical samples. Aspergillus spp. isolates were identified by conventional methods and azole resistance was determined by gradient test and agar plate screening method. Broth microdilution method was applied to resistant isolates, and CypA-L98H and CypA-M220 mutations in the cyp51A gene were investigated. Results: In our study, 55 A. fumigatus complex (44%), 42 A. flavus (33.6%), 6 A. terreus (5%), 4 A. niger (3%) and 18 Aspergillus spp. (14%) were identified. With the gradient test method, resistance to VOR and POS was detected in 1 (1.8%) of A.fumigatus isolates, and resistance to ITR was detected in 3 (5.45%). With the agar plate method, 1 of the A.fumigatus isolates (1.8%) had VOR, ITR, POS, 1 of the A.terreus isolates (16.7%) had VOR, 1 of the A.niger isolates (25%) had ITR. Resistance to VOR and POS was detected in 2 Aspergillus spp. isolates (11%), and resistance to ITR was detected in 1 (5.6%). Sensitivity and specificity were determined as 100% for VOR and POS in A. fumigatus species, 33.3% and 100% for ITR, respectively, 100% for ITR in A. flavus species, and 100% for ITR and POS in A. terreus species. By broth microdilution method in 7 isolates in which resistance was detected by gradient test and/or agar plate screening method; 1 A.fumigatus resistant to ITR, VOR, POS, 2 A.fumigatus resistant to ITR, 2 Aspergillus spp. ITR, VOR, POS MICs were determined as 2µg/ml and 8µg/ml, 8µg/ml and >32µg/ml, 0.5µg/ml and 4µg/ml, respectively. CypA-L98H mutations were detected in 5 of these isolates, CypA-M220 mutations were detected in 6, and no mutation was detected in 1. CypA-L98H and CypA-M220 mutations were detected in 1 isolate for which resistance was not detected. Conclusion: The need for rapid antifungal susceptibility screening tests is increasing in the treatment of aspergillosis. Although the sensitivity of the agar plate method was determined to be 33.3% for A.fumigatus ITR in our study, its sensitivity and specificity were determined to be 100% for ITR, VOR, and POS in other species. The low sensitivity value detected for A.fumigatus showed that agar plate drug concentrations should be updated in accordance with the latest regulations of EUCAST guidelines. The CypA-L98H and CypA-M220 mutations detected in our study suggested that the distribution of azole resistance-related mutations in different regions in our country should be investigated. In conclusion, it is thought that the agar plate method, which can be easily applied to detect azole resistance, is a fast and practical method in routine use and can contribute to both the determination of effective treatment strategies and the generation of epidemiological data.

Keywords: Aspergillus, agar plate, azole resistance, cyp51A, cypA-L98H, cypA-M220

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Authors: P. Gayathri, G. P. Jeyanthi

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The bark of Polyalthia longifolia is used in ayurvedic system of medicine for the manangement of various ailments including diabetes mellitus. The bark of P. longifolia extracts was extracted using various polar and non-polar solvents and tested for inhibition of alpha-amylase and alpha-glucosidase among which the ethanolic extracts were found to be more potent. The ethanolic extracts of the bark were tested for the in vitro inhibition of alpha-amylase using starch as substrate and alpha-glucosidase using p-nitro phenyl alpha-D-gluco pyranoside as substrate to establish its in vitro antidiabetic effect. The mechanism of inhibition was determined by Dixon plot and Cornish-Bowden plot. The cytotoxic effect of the extract was tested on L6 and Vero cell-lines. The extract was partially purified by TLC. The individual effect of the ethanolic extract, TLC fractions and its combinatorial effect with insulin and glibenclamide on glucose uptake by rat hemi-diaphragm were studied.Results revealed that the ethanolic extracts of Polyalthia longifolia bark exhibited competitive inhibition of alpha-amylase and alpha-glucosidase. The extracts were also found not to be cytotoxic at the highest dose of 1 mg/mL. Glucose uptake study revealed that the extract alone and when combined with insulin, decreased the glucose uptake when compared to insulin control, however the purified TLC fractions exhibited significantly higher (p<0.05) glucose uptake by the rat hemi-diaphragm when compared to insulin. The study shows various possible mechanism of in vitro antidiabetic effect of the P. longifolia bark.

Keywords: alpha-amylase, alpha-glucosidase, dixon, cornish-bowden, L6 , Vero cell-lines, glucose uptake, polyalthia longifolia bark, ethanolic extract, TLC fractions

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Authors: Janneth González, George Barreto, Ludis Morales, Angélica Sabogal

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Sea anemone neurotoxins are peptides that interact with Na+ and K+ channels, resulting in specific alterations on their functions. Some of these neurotoxins (1ROO, 1BGK, 2K9E, 1BEI) are important for the treatment of nearly eighty autoimmune disorders due to their specificity for Kv1.3 channel. The aim of this study was to identify the common residues among these neurotoxins by computational methods, and establish whether there is a pattern useful for the future generation of a treatment for autoimmune diseases. Our results showed eight new key common residues between the studied neurotoxins interacting with a histidine ring and the selectivity filter of the receptor, thus showing a possible pattern of interaction. This knowledge may serve as an input for the design of more promising drugs for autoimmune treatments.

Keywords: neurotoxins, potassium channel, Kv1.3, computational methods, autoimmune diseases

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Authors: Sellidj Abdelaziz, Lebaili Soltane

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A cobalt-based alloy type Co-Cr-Ni-WC was deposited by plasma transferred arc projection (PTA) on a stainless steel valve. The alloy is characterized at the equilibrium by a solid solution Co (γ) mainly dendritic, and eutectic carbides M₇C₃ and ηM₆C. At the deposit/substrate interface, this microstructure is modified by the fast cooling mode of the alloy when applied in the liquid state on the relatively cold steel substrate. The structure formed in this case is heterogeneous and metastable phases can occur and evolve over temperature service. Coating properties and reliability are directly related to microstructures formed during deposition. We were interested more particularly in this microstructure formed during the solidification of the deposit in the region of the interface joining the soldered couple and its evolution during cyclic heat treatments at temperatures similar to those of the thermal environment of the valve. The characterization was carried out by SEM-EDS microprobe CAMECA, XRD, and micro hardness profiles. The deposit obtained has a linear and regular appearance that is free of cracks and with little porosity. The morphology of the microstructure represents solidification stages that are relatively fast with a temperature gradient high at the beginning of the interface by forming a plane front solid solution Co (γ). It gradually changes with the decreasing temperature gradient by getting farther from the junction towards the outer limit of the deposit. The matrix takes the forms: cellular, mixed (cells and dendrites) and dendritic. Dendritic growth is done according to primary ramifications in the direction of the heat removal which takes place in the direction perpendicular to the interface, towards the external surface of the deposit, following secondary and tertiary undeveloped arms. The eutectic carbides M₇C₃ and ηM₆C formed are very thin and are located in the intercellular and interdendritic spaces of the solid solution Co (γ).

Keywords: Co-Ni-Cr-W-C alloy, solid deposit, microstructure, carbides, cyclic heat treatment

Procedia PDF Downloads 96