Search results for: thermally-activated delayed fluorescence
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 885

Search results for: thermally-activated delayed fluorescence

825 Fully Automated Methods for the Detection and Segmentation of Mitochondria in Microscopy Images

Authors: Blessing Ojeme, Frederick Quinn, Russell Karls, Shannon Quinn

Abstract:

The detection and segmentation of mitochondria from fluorescence microscopy are crucial for understanding the complex structure of the nervous system. However, the constant fission and fusion of mitochondria and image distortion in the background make the task of detection and segmentation challenging. In the literature, a number of open-source software tools and artificial intelligence (AI) methods have been described for analyzing mitochondrial images, achieving remarkable classification and quantitation results. However, the availability of combined expertise in the medical field and AI required to utilize these tools poses a challenge to its full adoption and use in clinical settings. Motivated by the advantages of automated methods in terms of good performance, minimum detection time, ease of implementation, and cross-platform compatibility, this study proposes a fully automated framework for the detection and segmentation of mitochondria using both image shape information and descriptive statistics. Using the low-cost, open-source python and openCV library, the algorithms are implemented in three stages: pre-processing, image binarization, and coarse-to-fine segmentation. The proposed model is validated using the mitochondrial fluorescence dataset. Ground truth labels generated using a Lab kit were also used to evaluate the performance of our detection and segmentation model. The study produces good detection and segmentation results and reports the challenges encountered during the image analysis of mitochondrial morphology from the fluorescence mitochondrial dataset. A discussion on the methods and future perspectives of fully automated frameworks conclude the paper.

Keywords: 2D, binarization, CLAHE, detection, fluorescence microscopy, mitochondria, segmentation

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824 Control of Chaotic Behaviour in Parallel-Connected DC-DC Buck-Boost Converters

Authors: Ammar Nimer Natsheh

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Chaos control is used to design a controller that is able to eliminate the chaotic behaviour of nonlinear dynamic systems that experience such phenomena. The paper describes the control of the bifurcation behaviour of a parallel-connected DC-DC buck-boost converter used to provide an interface between energy storage batteries and photovoltaic (PV) arrays as renewable energy sources. The paper presents a delayed feedback control scheme in a module converter comprises two identical buck-boost circuits and operates in the continuous-current conduction mode (CCM). MATLAB/SIMULINK simulation results show the effectiveness and robustness of the scheme.

Keywords: chaos, bifurcation, DC-DC Buck-Boost Converter, Delayed Feedback Control

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823 Peptide-Gold Nanocluster as an Optical Biosensor for Glycoconjugate Secreted from Leishmania

Authors: Y. A. Prada, Fanny Guzman, Rafael Cabanzo, John J. Castillo, Enrique Mejia-Ospino

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In this work, we show the important results about of synthesis of photoluminiscents gold nanoclusters using a small peptide as template for biosensing applications. Interestingly, we design one peptide (NBC2854) homologue to conservative domain from 215 250 residue of a galactolectin protein which can recognize the proteophosphoglycans (PPG) from Leishmania. Peptide was synthetized by multiple solid phase synthesis using FMoc group methodology in acid medium. Finally, the peptide was purified by High-Performance Liquid Chromatography using a Vydac C-18 preparative column and the detection was at 215 nm using a Photo Diode Array detector. Molecular mass of this peptide was confirmed by MALDI-TOF and to verify the α-helix structure we use Circular Dichroism. By means of the methodology used we obtained a novel fluorescents gold nanoclusters (AuNC) using NBC2854 as a template. In this work, we described an easy and fast microsonic method for the synthesis of AuNC with ≈ 3.0 nm of hydrodynamic size and photoemission at 630 nm. The presence of cysteine residue in the C-terminal of the peptide allows the formation of Au-S bond which confers stability to Peptide-based gold nanoclusters. Interactions between the peptide and gold nanoclusters were confirmed by X-ray Photoemission and Raman Spectroscopy. Notably, from the ultrafine spectra shown in the MALDI-TOF analysis which containing only 3-7 KDa species was assigned to Au₈-₁₈[NBC2854]₂ clusters. Finally, we evaluated the Peptide-gold nanocluster as an optical biosensor based on fluorescence spectroscopy and the fluorescence signal of PPG (0.1 µg-mL⁻¹ to 1000 µg-mL⁻¹) was amplified at the same wavelength emission (≈ 630 nm). This can suggest that there is a strong interaction between PPG and Pep@AuNC, therefore, the increase of the fluorescence intensity can be related to the association mechanism that take place when the target molecule is sensing by the Pep@AuNC conjugate. Further spectroscopic studies are necessary to evaluate the fluorescence mechanism involve in the sensing of the PPG by the Pep@AuNC. To our best knowledge the fabrication of an optical biosensor based on Pep@AuNC for sensing biomolecules such as Proteophosphoglycans which are secreted in abundance by parasites Leishmania.

Keywords: biosensing, fluorescence, Leishmania, peptide-gold nanoclusters, proteophosphoglycans

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822 Single-Molecule Analysis of Structure and Dynamics in Polymer Materials by Super-Resolution Technique

Authors: Hiroyuki Aoki

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The physical properties of polymer materials are dependent on the conformation and molecular motion of a polymer chain. Therefore, the structure and dynamic behavior of the single polymer chain have been the most important concerns in the field of polymer physics. However, it has been impossible to directly observe the conformation of the single polymer chain in a bulk medium. In the current work, the novel techniques to study the conformation and dynamics of a single polymer chain are proposed. Since a fluorescence method is extremely sensitive, the fluorescence microscopy enables the direct detection of a single molecule. However, the structure of the polymer chain as large as 100 nm cannot be resolved by conventional fluorescence methods because of the diffraction limit of light. In order to observe the single chains, we developed the labeling method of polymer materials with a photo-switchable dye and the super-resolution microscopy. The real-space conformational analysis of single polymer chains with the spatial resolution of 15-20 nm was achieved. The super-resolution microscopy enables us to obtain the three-dimensional coordinates; therefore, we succeeded the conformational analysis in three dimensions. The direct observation by the nanometric optical microscopy would reveal the detailed information on the molecular processes in the various polymer systems.

Keywords: polymer materials, single molecule, super-resolution techniques, conformation

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821 Conformational Switch of hRAGE upon Self-Association

Authors: Ikhlas Ahmed, Jamillah Zamoon

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The human receptor for advanced glycation end product is a plasma membrane receptor with an intrinsically disordered region. The protein consists of three extracellular domains, a single membrane spanning transmembrane domain, and a cytosolic domain which is intrinsically disordered and responsible for signaling. The disordered nature of the cytosolic domain allows it to be dynamic in solution. This receptor self-associates to higher forms. The association is triggered by ligand, metal or by the extracellular domain. Fluorescence spectroscopy technique is used to test the self-association of the different concentrations of the cytosolic domain. This work has concluded that the cytosolic domain of this receptor also self-associates. Moreover, the self-association does not require ligand or metal.

Keywords: fluorescence spectroscopy, hRAGE, IDP, Self-association

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820 Cadmium Telluride Quantum Dots (CdTe QDs)-Thymine Conjugate Based Fluorescence Biosensor for Sensitive Determination of Nucleobases/Nucleosides

Authors: Lucja Rodzik, Joanna Lewandowska-Lancucka, Michal Szuwarzynski, Krzysztof Szczubialka, Maria Nowakowska

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The analysis of nucleobases is of great importance for bioscience since their abnormal concentration in body fluids suggests the deficiency and mutation of the immune system, and it is considered to be an important parameter for diagnosis of various diseases. The presented conjugate meets the need for development of the effective, selective and highly sensitive sensor for nucleobase/nucleoside detection. The novel, highly fluorescent cadmium telluride quantum dots (CdTe QDs) functionalized with thymine and stabilized with thioglycolic acid (TGA) conjugates has been developed and thoroughly characterized. Successful formation of the material was confirmed by elemental analysis, and UV–Vis fluorescence and FTIR spectroscopies. The crystalline structure of the obtained product was characterized with X-ray diffraction (XRD) method. The composition of CdTe QDs and their thymine conjugate was also examined using X-ray photoelectron spectroscopy (XPS). The size of the CdTe-thymine was 3-6 nm as demonstrated using atomic force microscopy (AFM) and high resolution transmission electron microscopy (HRTEM) imaging. The plasmon resonance fluorescence band at 540 nm on excitation at 351 nm was observed for these nanoparticles. The intensity of this band increased with the increase in the amount of conjugated thymine with no shift in its position. Based on the fluorescence measurements, it was found that the CdTe-thymine conjugate interacted efficiently and selectively not only with adenine, a nucleobase complementary to thymine, but also with nucleosides and adenine-containing modified nucleosides, i.e., 5′-deoxy-5′-(methylthio)adenosine (MTA) and 2’-O-methyladenosine, the urinary tumor markers which allow monitoring of the disease progression. The applicability of the CdTe-thymine sensor for the real sample analysis was also investigated in simulated urine conditions. High sensitivity and selectivity of CdTe-thymine fluorescence towards adenine, adenosine and modified adenosine suggest that obtained conjugate can be potentially useful for development of the biosensor for complementary nucleobase/nucleoside detection.

Keywords: CdTe quantum dots, conjugate, sensor, thymine

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819 Using of Bimolecular Fluorescence Complementation (BiFC) Assays to Study Homo and/ or Heterodimerization of Laminin Receptor 37 LRP/ 67 LR with Galectin-3

Authors: Fulwah Alqahtani, Jafar Mahdavi, Lee Weldon, Nick Holliday, Dlawer Ala'Aldeen

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There are two isoforms of laminin receptor; monomeric 37 kDa laminin receptor precursor (37 LRP) and mature 67 kDa laminin receptor (67 LR). The relationship between the 67 LR and its precursor 37 LRP is not completely understood, but previous observations have suggested that 37 LRP can undergo homo- and/or hetero- dimerization with Galectin-3 (Gal-3) to form mature 67 LR. Gal-3 is the only member of the chimera-type group of galectins, and has one C-terminal carbohydrate recognition domain (CRD) that is responsible for binding the ß-galactoside moieties of mono- or oligosaccharides on several host and microbial molecules. The aim of this work was to investigate homo- and hetero-dimerization among the 37 LRP and Gal-3 to form mature 67 LR in mammalian cells using bimolecular fluorescence complementation (BiFC).

Keywords: 37 LRP, 67 LR, Gal-3, BiFC

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818 Fluorescing Aptamer-Gold Nanoparticle Complex for the Sensitive Detection of Bisphenol A

Authors: Eunsong Lee, Gae Baik Kim, Young Pil Kim

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Bisphenol A (BPA) is one of the endocrine disruptors (EDCs), which have been suspected to be associated with reproductive dysfunction and physiological abnormality in human. Since the BPA has been widely used to make plastics and epoxy resins, the leach of BPA from the lining of plastic products has been of major concern, due to its environmental or human exposure issues. The simple detection of BPA based on the self-assembly of aptamer-mediated gold nanoparticles (AuNPs) has been reported elsewhere, yet the detection sensitivity still remains challenging. Here we demonstrate an improved AuNP-based sensor of BPA by using fluorescence-combined AuNP colorimetry in order to overcome the drawback of traditional AuNP sensors. While the anti-BPA aptamer (full length or truncated ssDNA) triggered the self-assembly of unmodified AuNP (citrate-stabilized AuNP) in the presence of BPA at high salt concentrations, no fluorescence signal was observed by the subsequent addition of SYBR Green, due to a small amount of free anti-BPA aptamer. In contrast, the absence of BPA did not cause the self-assembly of AuNPs (no color change by salt-bridged surface stabilization) and high fluorescence signal by SYBP Green, which was due to a large amount of free anti-BPA aptamer. As a result, the quantitative analysis of BPA was achieved using the combination of absorption of AuNP with fluorescence intensity of SYBR green as a function of BPA concentration, which represented more improved detection sensitivity (as low as 1 ppb) than did in the AuNP colorimetric analysis. This method also enabled to detect high BPA in water-soluble extracts from thermal papers with high specificity against BPS and BPF. We suggest that this approach will be alternative for traditional AuNP colorimetric assays in the field of aptamer-based molecular diagnosis.

Keywords: bisphenol A, colorimetric, fluoroscence, gold-aptamer nanobiosensor

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817 Application of a Modified Crank-Nicolson Method in Metallurgy

Authors: Kobamelo Mashaba

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The molten slag has a high substantial temperatures range between 1723-1923, carrying a huge amount of useful energy for reducing energy consumption and CO₂ emissions under the heat recovery process. Therefore in this study, we investigated the performance of the modified crank Nicolson method for a delayed partial differential equation on the heat recovery of molten slag in the metallurgical mining environment. It was proved that the proposed method converges quickly compared to the classic method with the existence of a unique solution. It was inferred from numerical result that the proposed methodology is more viable and profitable for the mining industry.

Keywords: delayed partial differential equation, modified Crank-Nicolson Method, molten slag, heat recovery, parabolic equation

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816 Comparison Between Vegans and Omnivores on the Recovery of Delayed Onset Muscle Soreness in Young Females

Authors: Njeim Pressila, Hajj-Boutros Guy, Antony D. Karelis

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Background: Acute resistance exercise is associated with an elevated inflammation response, which could lead to delayed onset muscle soreness (DOMS). There is evidence that suggests that ingesting foods that have anti-inflammation properties may help reduce DOMS. A vegan diet has also been shown to be an anti-inflammatory diet which could, in turn, decrease DOMS. Objective and hypothesis: The purpose of the present study will be to compare markers of DOMS between vegans and omnivores after acute resistance exercise in young females. We hypothesize that vegans will have a better recovery of DOMS markers after a resistance exercise session compared to omnivores. Methods: Population: We will recruit30 vegans and 30 omnivores to participate in this study. Allvolunteers will follow either a vegan or an omnivore diet for at least 2 years. Anthropometric measurements, body composition, musclestrength (leg and chest press), markers of DOMS (swelling, pain, and stiffness), and dietary factors, as well as a wellness and anxiety questionnaire will be measured. All participants will also perform an acute resistance exercise session in order to induce DOMS. Pertinence: This project will give us a better understanding on the recovery process of vegans after a resistance training session and, as such, provide useful information to health professionals and athletes/coaches (kinesiologists and nutritionists)

Keywords: vgeans, omnivores, delayed onset muscle soreness, pain, stifness

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815 Aggregation of Butanediyl-1,4-Bis(Tetradecyldimethylammonium Bromide) (14–4–14) Gemini Surfactants in Presence of Ethylene Glycol and Propylene Glycol

Authors: P. Ajmal Koya, Tariq Ahmad Wagay, K. Ismail

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One of the fundamental property of surfactant molecules are their ability to aggregate in water or binary mixtures of water and organic solvents as an effort to minimize their unfavourable interaction with the medium. In this work, influence two co-solvents (ethylene glycol (EG) and propylene glycol (PG)) on the aggregation properties of a cationic gemini surfactant, butanediyl-1,4-bis(tetradecyldimethylammonium bromide) (14–4–14), has been studied by conductance and steady state fluorescence at 298 K. The weight percentage of two co-solvents varied in between 0 and 50 % at an interval of 5 % up to 20 % and then 10 % up to 50 %. It was found that micellization process is delayed by the inclusion of both the co-solvents; consequently, a progressive increase was observed in critical micelle concentration (cmc) and Gibbs free energy of micellization (∆G0m), whereas a rough increase was observed in the values of degree of counter ion dissociation (α) and a decrease was obtained in values of average aggregation number (Nagg) and Stern-Volmer constant (KSV). At low weight percentage (up to 15 %) of co-solvents, 14–4–14 geminis were found to be almost equally prone to micellization both in EG–water (EG–WR) and in PG–water (PG–WR) mixed media while at high weight percentages they are more prone to micellization in EG–WR than in PG–WR mixed media.

Keywords: aggregation number, gemini surfactant, micellization, non aqueous solvent

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814 Rapid Detection System of Airborne Pathogens

Authors: Shigenori Togashi, Kei Takenaka

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We developed new processes which can collect and detect rapidly airborne pathogens such as the avian flu virus for the pandemic prevention. The fluorescence antibody technique is known as one of high-sensitive detection methods for viruses, but this needs up to a few hours to bind sufficient fluorescence dyes to viruses for detection. In this paper, we developed a mist-labeling can detect substitution viruses in a short time to improve the binding rate of fluorescent dyes and substitution viruses by the micro reaction process. Moreover, we developed the rapid detection system with the above 'mist labeling'. The detection system set with a sampling bag collecting patient’s breath and a cartridge can detect automatically pathogens within 10 minutes.

Keywords: viruses, sampler, mist, detection, fluorescent dyes, microreaction

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813 Time Delayed Susceptible-Vaccinated-Infected-Recovered-Susceptible Epidemic Model along with Nonlinear Incidence and Nonlinear Treatment

Authors: Kanica Goel, Nilam

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Infectious diseases are a leading cause of death worldwide and hence a great challenge for every nation. Thus, it becomes utmost essential to prevent and reduce the spread of infectious disease among humans. Mathematical models help to better understand the transmission dynamics and spread of infections. For this purpose, in the present article, we have proposed a nonlinear time-delayed SVIRS (Susceptible-Vaccinated-Infected-Recovered-Susceptible) mathematical model with nonlinear type incidence rate and nonlinear type treatment rate. Analytical study of the model shows that model exhibits two types of equilibrium points, namely, disease-free equilibrium and endemic equilibrium. Further, for the long-term behavior of the model, stability of the model is discussed with the help of basic reproduction number R₀ and we showed that disease-free equilibrium is locally asymptotically stable if the basic reproduction number R₀ is less than one and unstable if the basic reproduction number R₀ is greater than one for the time lag τ≥0. Furthermore, when basic reproduction number R₀ is one, using center manifold theory and Casillo-Chavez and Song theorem, we showed that the model undergoes transcritical bifurcation. Moreover, numerical simulations are being carried out using MATLAB 2012b to illustrate the theoretical results.

Keywords: nonlinear incidence rate, nonlinear treatment rate, stability, time delayed SVIRS epidemic model

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812 Fluorescence in situ Hybridization (FISH) Detection of Bacteria and Archaea in Fecal Samples

Authors: Maria Nejjari, Michel Cloutier, Guylaine Talbot, Martin Lanthier

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The fluorescence in situ hybridization (FISH) is a staining technique that allows the identification, detection and quantification of microorganisms without prior cultivation by means of epifluorescence and confocal laser scanning microscopy (CLSM). Oligonucleotide probes have been used to detect bacteria and archaea that colonize the cattle and swine digestive systems. These bacterial strains have been obtained from fecal samples issued from cattle manure and swine slurry. The collection of these samples has been done at 3 different pit’s levels A, B and C with same height. Two collection depth levels have been taken in consideration, one collection level just under the pit’s surface and the second one at the bottom of the pit. Cells were fixed and FISH was performed using oligonucleotides of 15 to 25 nucleotides of length associated with a fluorescent molecule Cy3 or Cy5. The double hybridization using Cy3 probe targeting bacteria (Cy3-EUB338-I) along with a Cy5 probe targeting Archaea (Gy5-ARCH915) gave a better signal. The CLSM images show that there are more bacteria than archaea in swine slurry. However, the choice of fluorescent probes is critical for getting the double hybridization and a unique signature for each microorganism. FISH technique is an easy way to detect pathogens like E. coli O157, Listeria, Salmonella that easily contaminate water streams, agricultural soils and, consequently, food products and endanger human health.

Keywords: archaea, bacteria, detection, FISH, fluorescence

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811 Studying the Evolution of Soot and Precursors in Turbulent Flames Using Laser Diagnostics

Authors: Muhammad A. Ashraf, Scott Steinmetz, Matthew J. Dunn, Assaad R. Masri

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This study focuses on the evolution of soot and soot precursors in three different piloted diffusion turbulent flames. The fuel composition is as follow flame A (ethylene/nitrogen, 2:3 by volume), flame B (ethylene/air, 2:3 by volume), and flame C (pure methane). These flames are stabilized using a 4mm diameter jet surrounded by a pilot annulus with an outer diameter of 15 mm. The pilot issues combustion products from stoichiometric premixed flames of hydrogen, acetylene, and air. In all cases, the jet Reynolds number is 10,000, and air flows in the coflow stream at a velocity of 5 m/s. Time-resolved laser-induced fluorescence (LIF) is collected at two wavelength bands in the visible (445 nm) and UV regions (266 nm) along with laser-induced incandescence (LII). The combined results are employed to study concentration, size, and growth of soot and precursors. A set of four fast photo-multiplier tubes are used to record emission data in temporal domain. A 266nm laser pulse preferentially excites smaller nanoparticles which emit a fluorescence spectrum which is analysed to track the presence, evolution, and destruction of nanoparticles. A 1064nm laser pulse excites sufficiently large soot particles, and the resulting incandescence is collected at 1064nm. At downstream and outer radial locations, intermittency becomes a relevant factor. Therefore, data collected in turbulent flames is conditioned to account for intermittency so that the resulting mean profiles for scattering, fluorescence, and incandescence are shown for the events that contain traces of soot. It is found that in the upstream regions of the ethylene-air and ethylene-nitrogen flames, the presence of soot precursors is rather similar. However, further downstream, soot concentration grows larger in the ethylene-air flames.

Keywords: laser induced incandescence, laser induced fluorescence, soot, nanoparticles

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810 Luminescent Functionalized Graphene Oxide Based Sensitive Detection of Deadly Explosive TNP

Authors: Diptiman Dinda, Shyamal Kumar Saha

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In the 21st century, sensitive and selective detection of trace amounts of explosives has become a serious problem. Generally, nitro compound and its derivatives are being used worldwide to prepare different explosives. Recently, TNP (2, 4, 6 trinitrophenol) is the most commonly used constituent to prepare powerful explosives all over the world. It is even powerful than TNT or RDX. As explosives are electron deficient in nature, it is very difficult to detect one separately from a mixture. Again, due to its tremendous water solubility, detection of TNP in presence of other explosives from water is very challenging. Simple instrumentation, cost-effective, fast and high sensitivity make fluorescence based optical sensing a grand success compared to other techniques. Graphene oxide (GO), with large no of epoxy grps, incorporate localized nonradiative electron-hole centres on its surface to give very weak fluorescence. In this work, GO is functionalized with 2, 6-diamino pyridine to remove those epoxy grps. through SN2 reaction. This makes GO into a bright blue luminescent fluorophore (DAP/rGO) which shows an intense PL spectrum at ∼384 nm when excited at 309 nm wavelength. We have also characterized the material by FTIR, XPS, UV, XRD and Raman measurements. Using this as fluorophore, a large fluorescence quenching (96%) is observed after addition of only 200 µL of 1 mM TNP in water solution. Other nitro explosives give very moderate PL quenching compared to TNP. Such high selectivity is related to the operation of FRET mechanism from fluorophore to TNP during this PL quenching experiment. TCSPC measurement also reveals that the lifetime of DAP/rGO drastically decreases from 3.7 to 1.9 ns after addition of TNP. Our material is also quite sensitive to 125 ppb level of TNP. Finally, we believe that this graphene based luminescent material will emerge a new class of sensing materials to detect trace amounts of explosives from aqueous solution.

Keywords: graphene, functionalization, fluorescence quenching, FRET, nitroexplosive detection

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809 Effect of Whey Proteins and Caffeic Acid Interactions on Antioxidant Activity and Protein Structure

Authors: Tassia Batista Pessato, Francielli Pires Ribeiro Morais, Fernanda Guimaraes Drummond Silva, Flavia Maria Netto

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Proteins and phenolic compounds can interact mainly by hydrophobic interactions. Those interactions may lead to structural changes in both molecules, which in turn could affect positively or negatively their functional and nutritional properties. Here, the structural changes of whey proteins (WPI) due to interaction with caffeic acid (CA) were investigated by intrinsic and extrinsic fluorescence. The effects of protein-phenolic compounds interactions on the total phenolic content and antioxidant activity were also assessed. The WPI-CA complexes were obtained by mixture of WPI and CA stock solutions in deionized water. The complexation was carried out at room temperature during 60 min, using 0.1 M NaOH to adjust pH at 7.0. The WPI concentration was fixed at 5 mg/mL, whereas the CA concentration varied in order to obtain four different WPI:CA molar relations (1:1; 2:1; 5:1; 10:1). WPI and phenolic solutions were used as controls. Intrinsic fluorescence spectra of the complexes (mainly due to Trp fluorescence emission) were obtained at λex = 280 nm and the emission intensities were measured from 290 to 500 nm. Extrinsic fluorescence was obtained as the measure of protein surface hydrophobicity (S0) using ANS as a fluorescence probe. Total phenolic content was determined by Folin-Ciocalteau and the antioxidant activity by FRAP and ORAC methods. Increasing concentrations of CA resulted in decreasing of WPI intrinsic fluorescence. The emission band of WPI red shifted from 332 to 354 nm as the phenolic concentration increased, which is related to the exposure of Trp residue to the more hydrophilic environment and unfolding of protein structure. In general, the complexes presented lower S0 values than WPI, suggesting that CA hindered ANS binding to hydrophobic sites of WPI. The total phenolic content in the complexes was lower than the sum of two compounds isolated. WPI showed negligible AA measured by FRAP. However, as the relative concentration of CA increased in the complexes, the FRAP values enhanced, indicating that AA measure by this technique comes mainly from CA. In contrast, the WPI ORAC value (82.3 ± 1.5 µM TE/g) suggest that its AA is related to the capacity of H+ transfer. The complexes exhibited no important improvement of AA measured by ORAC in relation to the isolated components, suggesting complexation partially suppressed AA of the compounds. The results hereby presented indicate that interaction of WPI and CA occurred, and this interaction caused a structural change in the proteins. The complexation can either hide or expose antioxidant sites of both components. In conclusion, although the CA can undergo an AA suppression due to the interaction with proteins, the AA of WPI could be enhanced due to protein unfolding and exposure of antioxidant sites.

Keywords: bioactive properties, milk proteins, phenolic acids, protein-phenolic compounds complexation

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808 Early Versus Delayed Antiretroviral Therapy in HIV‐positive People with Tuberculosis

Authors: Mohhamed El Habib Labdouni

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Introduction: Co-infection with VIH and tuberculosis poses one of the major ongoing challenges for global TB and AIDS prevention and control. The objective of this study is to raise the issue of the resurgence of TB, in People living with VIH supported in a referent center in western Algeria. Its epidemiological, clinical, biological and radiological new trends, and to compare the mortality rate between early and delayed ART. Methods: It was a prospective study, during 36 months from the 01st/01/2012 to 31st/12/2014, by identifying and analyzing cases of TB-VIH co-infection. Our population was devised in two groups/ early ART and delayed ART. The primary and secondary endpoints were analyzed with Kaplan-Meier curves and log-rank test the period of follow up, which was fixed at 300 weeks. Results: Sixty cases of co-infection TB -VIH were enrolled in our study: 78.3% had pulmonary tuberculosis associated with extra-pulmonary, 13.3% had only pulmonary tuberculosis and 08.3% presented strictly extra-pulmonary TB. The clinical particularity of this co-infection is the frequency of serious localization such us: pleural 23.3%, peritoneal 31.7%, and meningeal suffusion 13.3%.y-.biologicaly we notice the predominance both of pancytopenia and leucoanemia, hyponatremia in 38,6% and hypokalemia in 19,3%. By analyzing Kaplan-Meier survival curves, we notice that early ART initiation is associated with a significant reduction of all-cause mortality (p = 0,000), and we have identified several prognostic factors such as hypokalemia hyponatremia, leukocytosis thrombopenemia leucothrombopenia (p = 0,005). Conclusion: Our study confirms most of the results reported in the literature. Early ART initiation reduces the rate of all-cause mortality, despite the probability of the occurrence of TB-IRIS.

Keywords: TB-HIV co-infection, early ART, hyponatremia, extrapulmonary tuberculosis

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807 Release of Calcein from Liposomes Using Low and High Frequency Ultrasound

Authors: Ghaleb A. Husseini, Salma E. Ahmed, Hesham G. Moussa, Ana M. Martins, Mohammad Al-Sayah, Nasser Qaddoumi

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This abstract aims to investigate the use of targeted liposomes as anticancer drug carriers in vitro in combination with ultrasound applied as drug trigger; in order to reduce the side effects caused by traditional chemotherapy. Pegylated liposomes were used to encapsulate calcein and then release this model drug when 20-kHz, 40-kHz, 1-MHz and 3-MHz ultrasound were applied at different acoustic power densities. Fluorescence techniques were then used to measure the percent drug release of calcein from these targeted liposomes. Results showed that as the power density increases, at the four frequencies studied, the release of calcein also increased. Based on these results, we believe that ultrasound can be used to increase the rate and amount of chemotherapeutics release from liposomes.

Keywords: liposomes, calcein release, high frequency ultrasound, low frequency ultrasound, fluorescence techniques

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806 Comparative Analysis of Short and Long Term Salt Stress on the Photosynthetic Apparatus and Chloroplast Ultrastructure of Thellungiella salsuginea

Authors: Rahma Goussi, Walid Derbali, Arafet Manaa, Simone Cantamessa, Graziella Berta, Chedly Abdelly, Roberto Barbato

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Salinity is one of the most important abiotic affecting plant growth and productivity worldwide. Photosynthesis, together with cell growth, is among the primary processes to be affected by salinity. Here, we report the effects of salinity stress on the primary processes of photosynthesis in a model halophyte Thellungiella Salsuginea. Plants were cultivated in hydroponic system with different NaCl concentrations (0, 100, 200 and 400 mM) during 2 weeks. The obtained results showed an obvious change in the photosynthetic efficiency of photosystem I (PSI) and phostosytem II (PSII), related to NaCl concentration supplemented to the medium and the stress duration considered. With moderate salinity (100 and 200 mM NaCl), no significant variation was observed in photosynthetic parameters of PSI and PSII and Chl fluorescence whatever the time of stress application. Also, the photosynthesis apparatus Fo, Fm and Fv fluorescence, as well as Fv/Fm were not affected by salt stress. While a significant decrease was observed on quantum yields Y(I), Y(II) and electron transport rate ETR(I), ETR(II) under high salt treatment (400 mM NaCl) with prolonged period (15 days). This reduction is quantitatively compensated by a corresponding increase of energy dissipation Y(NPQ) and a progressive decrease in Fv/Fm under salt treatment. The intensity of the OJIP fluorescence transient decreased with increase in NaCl concentration, with a major effect observed during prolonged period of salt stress. Ultrastructural analysis with Light Microscopy and Transmission Electron Microscopy of T. salsuginea chloroplasts showed some cellular changes, such as the shape of the mesophyll cells and number of chloroplast/cell only under higher NaCl concentration. Salt-stress caused the swelling of thylakoids in T. Salsuginea mesophyll with more accumulation of starch as compared to control plant.

Keywords: fluorescence, halophyte, photosynthesis, salt stress

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805 Temporary Ureteric Catheterization after Ureteropyeloscopy: Experience from Regional Australia

Authors: Jake Tempo, Jack Crozier, Huay Ann Chia, Philip Tan

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Purpose: A prospective study was performed to determine whether temporary ureteric catheterization should be eliminated as a prophylactic method for preventing ureteric obstruction after uncomplicated ureteropyeloscopic lithotripsy. Material and Methods: From 2010 to 2014, 227 patients underwent uncomplicated ureteroscopic and/or pyeloscopic lithotripsy. Three patient-groups based on postoperative drainage method were analysed: temporary uretericcatheter (TUC), -ureteric JJ stent, and no-stent groups. Exclusion criteria included urosepsis, ureteric injury, and non-surgical complications delaying hospital-discharge. Outcome measures included parenteral analgesic requirements, prolonged hospitalization ≥2 days due to postoperative-pain, and readmissions rate. Results: Delayed discharge was reported in 14.5%(9 of 62) patients in the TUC group compared to 3.4%(4 of 119) in theureteric JJ stent group and 8.7%(4 of 46) in the no-drainage-group (p=0.02). Odds ratio for delayed-discharge between catheter- versus-ureteric JJ stent is 4.9 (95% CI = 1.6-15.0; p < 0.01). Parenteral analgesic requirements in the TUC group (12.9%) was also significantly higher than theureteric JJ stent group (1.7%; p=0.003). Readmissions were negligible between groups. Conclusions: Patients with ureteric catheters after uncomplicated ureteroscopy have a prolonged hospital stay with increased pain and parenteral analgesic requirements. There is a 7.6-fold increased requirement for parenteral analgesia and a 4.2-fold increased risk of delayed-discharge compared to a patient with a ureteric JJ stent.

Keywords: ureteric catheter, ureteric stent, ureteroscopy, pyeloscopy

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804 Timing of Ileostomy Closure Following Rectal Cancer Surgery at an Australian Regional Hospital

Authors: Tedman Cheuk-Yiu Chau, Xavier Harvey, Hung Nguyen

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Defunctioning ileostomies are frequently used as an adjunct to low anterior resection in the surgical treatment of rectal cancer. Despite reducing the rate of clinically relevant anastomotic leak, the burden of defunctioning ileostomy is significant, with up to two-thirds of patients reporting stoma-related morbidity. International data have demonstrated an increased risk of bowel dysfunction and lower quality of life in patients with delayed closure (greater than six months post-surgery). While timely reversal is safe and cost-effective, the time to the reversal in Australian and New Zealand public hospitals is not described in the published literature. Thus, it is important to assess the current timeliness of ileostomy closure in the Australian regional context and examine the reasons for the delay. A retrospective analysis of ileostomy closure in Launceston General Hospital (LGH) patients treated with low/ultra low anterior resection for rectal cancer between 2012 and 2019 was undertaken. 94 cases of rectal adenocarcinoma undergoing ultralow anterior resection were examined over the years between 2012-2019. Amongst these, 21 cases (22.3%) were not reversed due to disease progress, death prior to reversal, or surgical complication. Demographics, disease status, surgical technique, and hospital inpatient events of these cases were examined. An average waiting time of 213.2 days was noted. Reasons for the delay include non-specified/prolonged hospital waiting time (54%), delayed or complicated chemotherapy course (13%), surgical complication (11%), advanced age, and frailty(5%). Complication of a delayed ileostomy reversal includes post-operation ileus and the development of an incisional hernia. We conclude that a delayed reversal of ileostomy can contribute to a higher incidence of stoma-related co-morbidities and contribute to a longer hospital stay and therefore use of public hospital resources.

Keywords: anterior resection, colorectal surgery, ileostomy reversal, rectal cancer

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803 Further Analysis of Global Robust Stability of Neural Networks with Multiple Time Delays

Authors: Sabri Arik

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In this paper, we study the global asymptotic robust stability of delayed neural networks with norm-bounded uncertainties. By employing the Lyapunov stability theory and Homeomorphic mapping theorem, we derive some new types of sufficient conditions ensuring the existence, uniqueness and global asymptotic stability of the equilibrium point for the class of neural networks with discrete time delays under parameter uncertainties and with respect to continuous and slopebounded activation functions. An important aspect of our results is their low computational complexity as the reported results can be verified by checking some properties symmetric matrices associated with the uncertainty sets of network parameters. The obtained results are shown to be generalization of some of the previously published corresponding results. Some comparative numerical examples are also constructed to compare our results with some closely related existing literature results.

Keywords: neural networks, delayed systems, lyapunov functionals, stability analysis

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802 Evaluation of Real Time PCR Methods for Food Safety

Authors: Ergun Sakalar, Kubra Bilgic

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In the last decades, real-time PCR has become a reliable tool preferred to use in many laboratories for pathogen detection. This technique allows for monitoring target amplification via fluorescent molecules besides admit of quantitative analysis by enabling of convert outcomes of thermal cycling to digital data. Sensitivity and traceability of real-time PCR are based on measuring of fluorescence that appears only when fluorescent reporter dye bound to specific target DNA.The fluorescent reporter systems developed for this purpose are divided into two groups. The first group consists of intercalator fluorescence dyes such as SYBR Green, EvaGreen which binds to double-stranded DNA. On the other hand, the second group includes fluorophore-labeled oligonucleotide probes that are separated into three subgroups due to differences in mechanism of action; initial primer-probes such as Cyclicons, Angler®, Amplifluor®, LUX™, Scorpions, and the second one hydrolysis probes like TaqMan, Snake assay, finally hybridization probes, for instance, Molecular Beacons, Hybprobe/FRET, HyBeacon™, MGB-Eclipse, ResonSense®, Yin-Yang, MGB-Pleiades. In addition nucleic acid analogues, an increase of probe affinity to target site is also employed with fluorescence-labeled probes. Consequently, abundant real-time PCR detection chemistries are chosen by researcher according to the field of application, mechanism of action, advantages, and proper structures of primer/probes.

Keywords: fluorescent dye, food safety, molecular probes, nucleic acid analogues

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801 Temporal Progression of Episodic Memory as Function of Encoding Condition and Age: Further Investigation of Action Memory in School-Aged Children

Authors: Farzaneh Badinlou, Reza Kormi-Nouri, Monika Knopf

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Studies of adults' episodic memory have found that enacted encoding not only improve recall performance but also retrieve faster during the recall period. The current study focused on exploring the temporal progression of different encoding conditions in younger and older school children. 204 students from two age group of 8 and 14 participated in this study. During the study phase, we studied action encoding in two forms; participants performed the phrases by themselves (SPT), and observed the performance of the experimenter (EPT), which were compared with verbal encoding; participants listened to verbal action phrases (VT). At test phase, we used immediate and delayed free recall tests. We observed significant differences in memory performance as function of age group, and encoding conditions in both immediate and delayed free recall tests. Moreover, temporal progression of recall was faster in older children when compared with younger ones. The interaction of age-group and encoding condition was only significant in delayed recall displaying that younger children performed better in EPT whereas older children outperformed in SPT. It was proposed that enactment effect in form of SPT enhances item-specific processing, whereas EPT improves relational information processing and this differential processes are responsible for the results achieved in younger and older children. The role of memory strategies and information processing methods in younger and older children were considered in this study. Moreover, the temporal progression of recall was faster in action encoding in the form of SPT and EPT compared with verbal encoding in both immediate and delayed free recall and size of enactment effect was constantly increased throughout the recall period. The results of the present study provide further evidence that the action memory is explained with an emphasis on the notion of information processing and strategic views. These results also reveal the temporal progression of recall as a new dimension of episodic memory in children.

Keywords: action memory, enactment effect, episodic memory, school-aged children, temporal progression

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800 Photoswitchable and Polar-Dependent Fluorescence of Diarylethenes

Authors: Sofia Lazareva, Artem Smolentsev

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Fluorescent photochromic materials collect strong interest due to their possible application in organic photonics such as optical logic systems, optical memory, visualizing sensors, as well as characterization of polymers and biological systems. In photochromic fluorescence switching systems the emission of fluorophore is modulated between ‘on’ and ‘off’ via the photoisomerization of photochromic moieties resulting in effective resonance energy transfer (FRET). In current work, we have studied both photochromic and fluorescent properties of several diarylethenes. It was found that coloured forms of these compounds are not fluorescent because of the efficient intramolecular energy transfer. Spectral and photochromic parameters of investigated substances have been measured in five solvents having different polarity. Quantum yields of photochromic transformation A↔B ΦA→B and ΦB→A as well as B isomer extinction coefficients were determined by kinetic method. It was found that the photocyclization reaction quantum yield of all compounds decreases with the increase of solvent polarity. In addition, the solvent polarity is revealed to affect fluorescence significantly. Increasing of the solvent dielectric constant was found to result in a strong shift of emission band position from 450 nm (nhexane) to 550 nm (DMSO and ethanol) for all three compounds. Moreover, the emission intensive in polar solvents becomes weak and hardly detectable in n-hexane. The only one exception in the described dependence is abnormally low fluorescence quantum yield in ethanol presumably caused by the loss of electron-donating properties of nitrogen atom due to the protonation. An effect of the protonation was also confirmed by the addition of concentrated HCl in solution resulting in a complete disappearance of the fluorescent band. Excited state dynamics were investigated by ultrafast optical spectroscopy methods. Kinetic curves of excited states absorption and fluorescence decays were measured. Lifetimes of transient states were calculated from the data measured. The mechanism of ring opening reaction was found to be polarity dependent. Comparative analysis of kinetics measured in acetonitrile and hexane reveals differences in relaxation dynamics after the laser pulse. The most important fact is the presence of two decay processes in acetonitrile, whereas only one is present in hexane. This fact supports an assumption made on the basis of steady-state preliminary experiments that in polar solvents occur stabilization of TICT state. Thus, results achieved prove the hypothesis of two channel mechanism of energy relaxation of compounds studied.

Keywords: diarylethenes, fluorescence switching, FRET, photochromism, TICT state

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799 Segmental Dynamics of Poly(Alkyl Methacrylate) Chain in Ultra-Thin Spin-Cast Films

Authors: Hiroyuki Aoki

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Polymeric materials are often used in a form of thin film such as food wrap and surface coating. In such the applications, polymer films thinner than 100 nm have been often used. The thickness of such the ultra-thin film is less than the unperturbed size of a polymer chain; therefore, the polymer chain in an ultra-thin film is strongly constrained. However, the details on the constrained dynamics of polymer molecules in ultra-thin films are still unclear. In the current study, the segmental dynamics of single polymer chain was directly investigated by fluorescence microscopy. The individual chains of poly(alkyl methacrylate) labeled by a perylenediimide dye molecule were observed by a highly sensitive fluorescence microscope in a defocus condition. The translational and rotational diffusion of the center segment in a single polymer chain was directly analyzed. The segmental motion in a thin film with a thickness of 10 nm was found to be suppressed compared to that in a bulk state. The detailed analysis of the molecular motion revealed that the diffusion rate of the in-plane rotation was similar to the thin film and the bulk; on the other hand, the out-of-plane motion was restricted in a thin film. This result indicates that the spatial restriction in an ultra-thin film thinner than the unperturbed chain dimension alters the dynamics of individual molecules in a polymer system.

Keywords: polymer materials, single molecule, molecular motion, fluorescence microscopy, super-resolution techniques

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798 Photophysics and Photochemistry of Cross-Conjugated Y-Shaped Enediyne Fluorophores

Authors: Anuja Singh, Avik K. Pati, Ashok K. Mishra

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Organic fluorophores with π-conjugated scaffolds are important because of their interesting optoelectronic properties. In recent years, our lab has been engaged in understanding the photophysics of small diacetylene bridged fluorophores and found the diynes as a promising class of π-conjugated fluorophores. Building on this understanding, recently we have focused on the photophysics of a less explored class of cross-conjugated Y-shaped enediynes (one double and two triple bonds). Here we present the photophysical properties of such enediynes which show interesting photophysical properties that include dual emissions from locally excited (LE) and intramolecular charge transfer (ICT) states and ring size dependent aggregate fluorescence in non-aqueous media. The dyes also show prominent aggregate fluorescence in mixed-aqueous solvents and solid powder form. We further show that the solid state fluorescence can be reversibly switched multiple of cycles by external stimuli, highlighting their potential applications in solid states. The enediynes with push-pull electronic substituents/moieties exhibit high contrast fluorescence color switching upon continuous photon illumination. The intriguing photophysical outcomes of the enediynyl fluorophores are judiciously exploited to generate single-component white light emission in binary solvent mixtures and sense polar aprotic vapor in polymer film matrices. The photophysical behavior of the dyes is further successfully utilized to monitor the microenvironment changes of biologically relevant anisotropic media such as bile salts. In summary, the newly introduced cross-conjugated enediynes enrich the toolbox of organic fluorophores and vouch to display versatile applications.

Keywords: aggregation in solution and solid state, enediynes, physical photochemistry and photophysics, vapor sensing and white light emission

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797 Anti-Fibrillation Propensity of a Flavonoid Baicalein against the Fibrils of Hen Egg White Lysozyme: Potential Therapeutics for Lysozyme Amyloidosis

Authors: Naveed Ahmad Fazili

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More than 20 human diseases involve the fibrillation of a specific protein/peptide which forms pathological deposits at various sites. Hereditary lysozyme amyloidosis is a systemic disorder which mostly affects liver, spleen and kidney. This conformational disorder is featured by lysozyme fibril formation. In vivo lysozyme fibrillation was simulated under in vitro conditions using a strong denaturant GdHCl at 3M concentration. Sharp decline in the ANS fluorescence intensity compared to the partially unfolded states, almost 20 fold increase in ThT fluorescence intensity, increase in absorbance at 450 nm suggesting turbidity, negative ellipticity peak in the far-UVCD at 217 nm, red shift of 50 nm compared to the native state in congo red assay and appearance of a network of long rope like fibrils in TEM analysis suggested HEWL fibrillation. Anti-fibrillation potency of baicalein against the preformed fibrils of HEWL was investigated following ThT assay in which there was a dose dependent decrease in ThT fluorescence intensity compared to the fibrillar state of HEWL with the maximum effect observed at 150 μM baicalein concentration, loss of negative ellipticity peak in the far-UVCD region, dip in the Rayleigh scattering intensity and absorbance at 350 nm and 450 nm respectively together with a reduction in the density of fibrillar structure in TEM imaging. Thus, it could be suggested that baicalein could prove to be a positive therapeutics for hereditary human lysozyme amyloidosis.

Keywords: amyloid fibrils, baicalein, congo red, negative ellipticity, therapeutics

Procedia PDF Downloads 271
796 Inducing Flow Experience in Mobile Learning: An Experiment Using a Spanish Learning Mobile Application

Authors: S. Jonsson, D. Millard, C. Bokhove

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Smartphones are ubiquitous and frequently used as learning tools, which makes the design of educational apps an important area of research. A key issue is designing apps to encourage engagement while maintaining a focus on the educational aspects of the app. Flow experience is a promising method for addressing this issue, which refers to a mental state of cognitive absorption and positive emotion. Flow experience has been shown to be associated with positive emotion and increased learning performance. Studies have shown that immediate feedback is an antecedent to Flow. This experiment investigates the effect of immediate feedback on Flow experience. An app teaching Spanish phrases was developed, and 30 participants completed both a 10min session with immediate feedback and a 10min session with delayed feedback. The app contained a task where the user assembles Spanish phrases by pressing bricks with Spanish words. Immediate feedback was implemented by incorrect bricks recoiling, while correct brick moved to form part of the finished phrase. In the delayed feedback condition, the user did not know if the bricks they pressed were correct until the phrase was complete. The level of Flow experienced by the participants was measured after each session using the Flow Short Scale. The results showed that higher levels of Flow were experienced in the immediate feedback session. It was also found that 14 of the participants indicated that the demands of the task were ‘just right’ in the immediate feedback session, while only one did in the delayed feedback session. These results have implications for how to design educational technology and opens up questions for how Flow experience can be used to increase performance and engagement.

Keywords: feedback timing, flow experience, L2 language learning, mobile learning

Procedia PDF Downloads 104