Search results for: GAF domain protein
2980 De Novo Design of a Minimal Catalytic Di-Nickel Peptide Capable of Sustained Hydrogen Evolution
Authors: Saroj Poudel, Joshua Mancini, Douglas Pike, Jennifer Timm, Alexei Tyryshkin, Vikas Nanda, Paul Falkowski
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On the early Earth, protein-metal complexes likely harvested energy from a reduced environment. These complexes would have been precursors to the metabolic enzymes of ancient organisms. Hydrogenase is an essential enzyme in most anaerobic organisms for the reduction and oxidation of hydrogen in the environment and is likely one of the earliest evolved enzymes. To attempt to reinvent a precursor to modern hydrogenase, we computationally designed a short thirteen amino acid peptide that binds the often-required catalytic transition metal Nickel in hydrogenase. This simple complex can achieve hundreds of hydrogen evolution cycles using light energy in a broad range of temperature and pH. Biophysical and structural investigations strongly indicate the peptide forms a di-nickel active site analogous to Acetyl-CoA synthase, an ancient protein central to carbon reduction in the Wood-Ljungdahl pathway and capable of hydrogen evolution. This work demonstrates that prior to the complex evolution of multidomain enzymes, early peptide-metal complexes could have catalyzed energy transfer from the environment on the early Earth and enabled the evolution of modern metabolismKeywords: hydrogenase, prebiotic enzyme, metalloenzyme, computational design
Procedia PDF Downloads 2162979 Characterization of Biodiesel Produced from Cow-Tallow
Authors: Nwadike Emmanuel Chinagoron, Achebe Chukwunonso, Ezeliora Chukwuemeka Daniel, Azaka Onyemazuwa Andrew
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In this research work, the process of biodiesel production in a pilot plant was studied using cow tallow as raw material, methanol as the solvent and potassium hydroxide as catalysts. The biodiesel quality was determined by characterization. The tallow used in the production had a molecular weight of 860g. Its oil had a density value of 0.8g/ml, iodine value of 63.45, viscosity at 300C was 9.83pas, acid value was 1.96, free fatty acid (FFA) of 0.98%, saponification value of 82.75mleq/kg, specific gravity of 0.898, flash point of 1100C, cloud point of 950C and Calorific value also called Higher Heating Value (HHV) of 38.365MJ/Kg. The produced biodiesel had a density of 0.82g/ml, iodine value of 126.9, viscosity of 4.32pas at 300C, acid value of 0.561, FFA of 0.2805%, saponification value of 137.45 mleq/kg.Flash point, cloud point and centane number of the biodiesel produced are 1390C, 980C and 57.5 respectively, with fat content, protein content, ash content, moisture content, fiber content and carbohydrate content values of 10%, 2.8%, 5%, 5%, 20%, and 37.2% respectively. The biodiesel higher heating values (calorific values) when estimated from viscosity, density and flash points were 41.4MJ/Kg, 63.8MJ/Kg, and 34.6MJ/Kg respectively. The biodiesel was blended with conventional diesel. The blend B-10 had values of 1320C and 960C for flash and cloud points, with Calorific value (or HHV) of 34.6 MJ/Kg (when estimated from its Flash point) and fat content, protein content, ash content, moisture content, fiber content and carbohydrate content values of 5%, 2.1%,10%, 5%, 15%, and 62.9% respectively.Keywords: biodiesel, characterization, cow-tallow, cetane rating
Procedia PDF Downloads 5372978 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
Procedia PDF Downloads 1692977 Study Habits and Level of Difficulty Encountered by Maltese Students Studying Biology Advanced Level Topics
Authors: Marthese Azzopardi, Liberato Camilleri
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This research was performed to investigate the study habits and level of difficulty perceived by post-secondary students in Biology at Advanced-level topics after completing their first year of study. At the end of a two-year ‘sixth form’ course, Maltese students sit for the Matriculation and Secondary Education Certificate (MATSEC) Advanced-level biology exam as a requirement to pursue science-related studies at the University of Malta. The sample was composed of 23 students (16 taking Chemistry and seven taking some ‘Other’ subject at the Advanced Level). The cohort comprised seven males and 16 females. A questionnaire constructed by the authors, was answered anonymously during the last lecture at the end of the first year of study, in May 2016. The Chi square test revealed that gender plays no effect on the various study habits (c2 (6) = 5.873, p = 0.438). ‘Reading both notes and textbooks’ was the most common method adopted by males (71.4%), whereas ‘Writing notes on each topic’ was that mostly used by females (81.3%). The Mann-Whitney U test showed no significant difference in the study habits of students and the mean assessment mark obtained at the end of the first year course (p = 0.231). Statistical difference was found with the One-ANOVA test when comparing the mean assessment mark obtained at the end of the first year course when students are clustered by their Secondary Education Certificate (SEC) grade (p < 0.001). Those obtaining a SEC grade of 2 and 3 got the highest mean assessment of 68.33% and 66.9%, respectively [SEC grading is 1-7, where 1 is the highest]. The Friedman test was used to compare the mean difficulty rating scores provided for the difficulty of each topic. The mean difficulty rating score ranges from 1 to 4, where the larger the mean rating score, the higher the difficulty. When considering the whole group of students, nine topics out of 21 were perceived as significantly more difficult than the other topics. Protein synthesis, DNA Replication and Biomolecules were the most difficult, in that order. The Mann-Whitney U test revealed that the perceived level of difficulty in comprehending Biomolecules is significantly lower for students taking Chemistry compared to those not choosing the subject (p = 0.018). Protein Synthesis was claimed as the most difficult by Chemistry students and Biomolecules by those not studying Chemistry. DNA Replication was the second most difficult topic perceived by both groups. The Mann-Whitney U test was used to examine the effect of gender on the perceived level of difficulty in comprehending various topics. It was found that females have significantly more difficulty in comprehending Biomolecules than males (p=0.039). Protein synthesis was perceived as the most difficult topic by males (mean difficulty rating score = 3.14), while Biomolecules, DNA Replication and Protein synthesis were of equal difficulty for females (mean difficulty rating score = 3.00). Males and females perceived DNA Replication as equally difficult (mean difficulty rating score = 3.00). Discovering the students’ study habits and perceived level of difficulty of specific topics is vital for the lecturer to offer guidance that leads to higher academic achievement.Keywords: biology, perceived difficulty, post-secondary, study habits
Procedia PDF Downloads 1882976 Preparation of Bead-On-String Alginate/Soy Protein Isolated Nanofibers via Water-Based Electrospinning and Its Application for Drug Loading
Authors: Patcharakamon Nooeaid, Piyachat Chuysrinuan
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Electrospun natural polymers-based nanofibers are one of the most interesting materials used in tissue engineering and drug delivery applications. Bead-on-string nanofibers have gained considerable interest for sustained drug release. Vancomycin was used as the model drug and sodium alginate (SA)/soy protein isolated (SPI) as the polymer blend to fabricate the bead-on-string nanofibers by aqueous-based electrospinning. The bead-on-string SA/SPI nanofibers were successfully fabricated by the addition of poly(ethylene oxide) (PEO) as a co-blending polymer. SA-PEO with mass ratio of 70/30 showed the best spinnability with continuous nanofibers without the occurrence of beads. Bead structure formed with the addition of SPI and bead number increased with increasing SPI content. The electrospinning of 80/20 SA-PEO/SPI was obtained as a great promising bead-on-string nanofibers for drug loading, while the solution of 50/50 was not able to obtain continuous fibers. In vitro release tests showed that a more sustainable release profile up to 14 days with less initial burst release on day 1 could be obtained from the bead-on-string fibers than from smooth fibers with uniform diameter. In addition, vancomycin-loaded beaded fibers inhibited the growth of Staphylococcus aureus (S. aureus) bacteria. Therefore, the SA-PEO/SPI nanofibers showed the potential to be used as biomaterials for tissue engineering and drug delivery.Keywords: bead-on-string fibers, electrospinning, drug delivery, tissue engineering
Procedia PDF Downloads 3342975 A Mixed Thought Pattern and the Question of Justification: A Feminist Project
Authors: Angana Chatterjee
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The feminist scholars point out the various problematic issues in the traditional mainstream western thought and theories. The thought practices behind the discriminatory and oppressive social practices are based on concepts that play a pivotal role in theorisation. Therefore, many feminist philosophers take up reformation or reconceptualisation projects. Such projects have bearings on various aspects of philosophical thought, namely, ontology, epistemology, logic, ethics, social, political thought, and so on. In tune with this spirit, the present paper suggests a well-established thought pattern which is not western but has got the potential to deal with the problems of mainstream western thought culture that are identified by the feminist critics. The Indian thought pattern is theorised in the domain of Indian logic, which is a study of inference patterns. As, in the Indian context, the inference is considered as a source of knowledge, certain epistemological questions are linked with the discussion of inference. One of the key epistemological issues is one regarding justification. The study about the nature of derivation of knowledge from available evidence, and the nature of the evidence itself, are integral parts of the discipline called Indian logic. But if we contrast the western tradition of thought with the Indian one, we can find that the Indian logic has got some peculiar features which may be shown to deal with the problems identified by the feminist scholars in western thought culture more plausibly. The tradition of western logic, starting from Aristotle, has been maintaining sharp differences between two forms of reasoning, namely, deductive and inductive. These two different forms of reasoning have been theorised and dealt with separately within the domain of the study called ‘logic.’ There are various philosophical problems that are raised around concepts and issues regarding both deductive and inductive reasoning. Indian logic does not distinguish between deduction and induction as thought patterns, but their distinction is very usual to make in the western tradition. Though there can be found various interpretations about this peculiarity of Indian thought pattern, these mixed patterns were actually very close to the cross-cultural pattern in which human beings would tend to argue or infer from the available data or evidence. The feminist theories can successfully operate in the domain of lived experience if they make use of such a mixed pattern of reasoning or inference. By offering sound inferential knowledge on contextual evidences, the Indian thought pattern is potent to serve the feminist purposes in a meaningful way.Keywords: feminist thought, Indian logic, inference, justification, mixed thought pattern
Procedia PDF Downloads 1022974 X-Ray Crystallographic Studies on BPSL2418 from Burkholderia pseudomallei
Authors: Mona Alharbi
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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis
Procedia PDF Downloads 2482973 Epigenetics Regulation Play Role in the Pathogenesis of Adipose Tissue Disorder, Lipedema
Authors: Musarat Ishaq, Tara Karnezis, Ramin Shayan
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Lipedema, a poorly understood chronic disease of adipose hyper-deposition, is often mistaken for obesity and causes significant impairment to mobility and quality-of-life. To identify molecular mechanisms underpinning lipedema, we employed comprehensive omics-based comparative analyses of whole tissue, adipocyte precursors (adipose-derived stem cells (ADSCs)), and adipocytes from patients with or without lipedema. Transcriptional profiling revealed significant differences in lipedema tissue, adipocytes, and ADSCs, with altered levels of mRNAs involved inproliferation and cell adhesion. One highly up-regulated gene in lipedema adipose tissue, adipocytes and ADSCs, ZIC4, encodes Zinc Finger Protein ZIC 4, a class of transcription factor which may be involved in regulating metabolism and adipogenesis. ZIC4 inhibition impaired the adipogenesis of ADSCs into mature adipocytes. Epigenetic regulation study revealed overexpression of ZIC4 is involved in decreased promoter DNA methylation and subsequent decrease in adipogenesis. These epigenetic modifications can alter adipocytes microenvironment and adipocytes differentiation. Our study show that epigenetic events regulate the ability of ADSCs to commit and differentiate into mature adipocytes by modulating ZIC4.Keywords: lipedema, adipose-derived stem cells, adipose tisue, adipocytes, zinc finger protein, epigenetic
Procedia PDF Downloads 1752972 Cloning, Expression and N-Terminal Pegylation of Human Interferon Alpha-2b Analogs and Their Cytotoxic Evaluation against Cancer Cell Lines
Authors: Syeda Kiran Shahzadi, Nasir Mahmood, Muhammad Abdul Qadir
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In the current research, three recombinant human interferon alpha-2b proteins (two modified and one normal form) were produced and Pegylated with an aim to produce more effective drugs against viral infections and cancers. The modified recombinant human interferon alpha-2b proteins were produced by site-directed modifications of interferon alpha 2b gene, targeting the amino acids at positions ‘R23’ and ‘H34’. The resulting chemically modified and unmodified forms of human interferon alpha 2b were conjugated with methoxy-polyethylene glycol propanealdehyde (400 KDa) and methoxy-polyethylene glycol succinimidyl succinate (400 KDa). Pegylation of normal and modified forms of Interferon alpha-2b prolong their release time and enhance their efficacy. The conjugation of PEG with modified and unmodified human interferon alpha 2b protein drugs was also characterized with 1H-NMR, HPLC, and SDS-PAGE. Antiproliferative assays of modified and unmodified forms of drugs were performed in cell based bioassays using MDBK cell lines. The results indicated that experimentally produced recombinant human interferon alpha-2b proteins were biologically active and resulted in significant inhibition of cell growth.Keywords: protein refolding, antiproliferative activities, biomedical applications, human interferon alpha-2b, pegylation, mPEG-propionaldehyde, site directed mutagenesis, E. coli expression
Procedia PDF Downloads 1772971 The Role of Piceatannol in Counteracting Glyceraldehyde-3-Phosphate Dehydrogenase Aggregation and Nuclear Translocation
Authors: Joanna Gerszon, Aleksandra Rodacka
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In the pathogenesis of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, protein and peptide aggregation processes play a vital role in contributing to the formation of intracellular and extracellular protein deposits. One of the major components of these deposits is the oxidatively modified glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Therefore, the purpose of this research was to answer the question whether piceatannol, a stilbene derivative, counteracts and/or slows down oxidative stress-induced GAPDH aggregation. The study also aimed to determine if this natural occurring compound prevents unfavorable nuclear translocation of GAPDH in hippocampal cells. The isothermal titration calorimetry (ITC) analysis indicated that one molecule of GAPDH can bind up to 8 molecules of piceatannol (7.3 ± 0.9). As a consequence of piceatannol binding to the enzyme, the loss of activity was observed. Parallel with GAPDH inactivation the changes in zeta potential, and loss of free thiol groups were noted. Nevertheless, the ligand-protein binding does not influence the secondary structure of the GAPDH. Precise molecular docking analysis of the interactions inside the active center allowed to presume that these effects are due to piceatannol ability to assemble a covalent binding with nucleophilic cysteine residue (Cys149) which is directly involved in the catalytic reaction. Molecular docking also showed that simultaneously 11 molecules of ligand can be bound to dehydrogenase. Taking into consideration obtained data, the influence of piceatannol on level of GAPDH aggregation induced by excessive oxidative stress was examined. The applied methods (thioflavin-T binding-dependent fluorescence as well as microscopy methods - transmission electron microscopy, Congo Red staining) revealed that piceatannol significantly diminishes level of GAPDH aggregation. Finally, studies involving cellular model (Western blot analyses of nuclear and cytosolic fractions and confocal microscopy) indicated that piceatannol-GAPDH binding prevents GAPDH from nuclear translocation induced by excessive oxidative stress in hippocampal cells. In consequence, it counteracts cell apoptosis. These studies demonstrate that by binding with GAPDH, piceatannol blocks cysteine residue and counteracts its oxidative modifications, that induce oligomerization and GAPDH aggregation as well as it prevents hippocampal cells from apoptosis by retaining GAPDH in the cytoplasm. All these findings provide a new insight into the role of piceatannol interaction with GAPDH and present a potential therapeutic strategy for some neurological disorders related to GAPDH aggregation. This work was supported by the by National Science Centre, Poland (grant number 2017/25/N/NZ1/02849).Keywords: glyceraldehyde-3-phosphate dehydrogenase, neurodegenerative disease, neuroprotection, piceatannol, protein aggregation
Procedia PDF Downloads 1672970 Vortex Control by a Downstream Splitter Plate in Psudoplastic Fluid Flow
Authors: Sudipto Sarkar, Anamika Paul
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Pseudoplastic (n<1, n is the power index) fluids have great importance in food, pharmaceutical and chemical process industries which require a lot of attention. Unfortunately, due to its complex flow behavior inadequate research works can be found even in laminar flow regime. A practical problem is solved in the present research work by numerical simulation where we tried to control the vortex shedding from a square cylinder using a horizontal splitter plate placed at the downstream flow region. The position of the plate is at the centerline of the cylinder with varying distance from the cylinder to calculate the critical gap-ratio. If the plate is placed inside this critical gap, the vortex shedding from the cylinder suppressed completely. The Reynolds number considered here is in unsteady laminar vortex shedding regime, Re = 100 (Re = U∞a/ν, where U∞ is the free-stream velocity of the flow, a is the side of the cylinder and ν is the maximum value of kinematic viscosity of the fluid). Flow behavior has been studied for three different gap-ratios (G/a = 2, 2.25 and 2.5, where G is the gap between cylinder and plate) and for a fluid with three different flow behavior indices (n =1, 0.8 and 0.5). The flow domain is constructed using Gambit 2.2.30 and this software is also used to generate the mesh and to impose the boundary conditions. For G/a = 2, the domain size is considered as 37.5a × 16a with 316 × 208 grid points in the streamwise and flow-normal directions respectively after a thorough grid independent study. Fine and equal grid spacing is used close to the geometry to capture the vortices shed from the cylinder and the boundary layer developed over the flat plate. Away from the geometry meshes are unequal in size and stretched out. For other gap-ratios, proportionate domain size and total grid points are used with similar kind of mesh distribution. Velocity inlet (u = U∞), pressure outlet (Neumann condition), symmetry (free-slip boundary condition) at upper and lower domain boundary conditions are used for the simulation. Wall boundary condition (u = v = 0) is considered both on the cylinder and the splitter plate surfaces. Discretized forms of fully conservative 2-D unsteady Navier Stokes equations are then solved by Ansys Fluent 14.5. SIMPLE algorithm written in finite volume method is selected for this purpose which is a default solver inculcate in Fluent. The results obtained for Newtonian fluid flow agree well with previous works supporting Fluent’s usefulness in academic research. A thorough analysis of instantaneous and time-averaged flow fields are depicted both for Newtonian and pseudoplastic fluid flow. It has been observed that as the value of n reduces the stretching of shear layers also reduce and these layers try to roll up before the plate. For flow with high pseudoplasticity (n = 0.5) the nature of vortex shedding changes and the value of critical gap-ratio reduces. These are the remarkable findings for laminar periodic vortex shedding regime in pseudoplastic flow environment.Keywords: CFD, pseudoplastic fluid flow, wake-boundary layer interactions, critical gap-ratio
Procedia PDF Downloads 1112969 The Impact of Missense Mutation in Phosphatidylinositol Glycan Class A Associated to Paroxysmal Nocturnal Hemoglobinuria and Multiple Congenital Anomalies-Hypotonia-Seizures Syndrome 2: A Computational Study
Authors: Ashish Kumar Agrahari, Amit Kumar
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Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal blood disorder that manifests with hemolytic anemia, thrombosis, and peripheral blood cytopenias. The disease is caused by the deficiency of two glycosylphosphatidylinositols (GPI)-anchored proteins (CD55 and CD59) in the hemopoietic stem cells. The deficiency of GPI-anchored proteins has been associated with the somatic mutations in phosphatidylinositol glycan class A (PIGA). However, the mutations that do not cause PNH is associated with the multiple congenital anomalies-hypotonia-seizures syndrome 2 (MCAHS2). To best of our knowledge, no computational study has been performed to explore the atomistic level impact of PIGA mutations on the structure and dynamics of the protein. In the current work, we are mainly interested to get insights into the molecular mechanism of PIGA mutations. In the initial step, we screened the most pathogenic mutations from the pool of publicly available mutations. Further, to get a better understanding, pathogenic mutations were mapped to the modeled structure and subjected to 50ns molecular dynamics simulation. Our computational study suggests that four mutations are highly vulnerable to altering the structural conformation and stability of the PIGA protein, which illustrates its association with PNH and MCAHS2 phenotype.Keywords: homology modeling, molecular dynamics simulation, missense mutations PNH, MCAHS2, PIGA
Procedia PDF Downloads 1452968 Introduction to Two Artificial Boundary Conditions for Transient Seepage Problems and Their Application in Geotechnical Engineering
Authors: Shuang Luo, Er-Xiang Song
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Many problems in geotechnical engineering, such as foundation deformation, groundwater seepage, seismic wave propagation and geothermal transfer problems, may involve analysis in the ground which can be seen as extending to infinity. To that end, consideration has to be given regarding how to deal with the unbounded domain to be analyzed by using numerical methods, such as finite element method (FEM), finite difference method (FDM) or finite volume method (FVM). A simple artificial boundary approach derived from the analytical solutions for transient radial seepage problems, is introduced. It should be noted, however, that the analytical solutions used to derive the artificial boundary are particular solutions under certain boundary conditions, such as constant hydraulic head at the origin or constant pumping rate of the well. When dealing with unbounded domains with unsteady boundary conditions, a more sophisticated artificial boundary approach to deal with the infinity of the domain is presented. By applying Laplace transforms and introducing some specially defined auxiliary variables, the global artificial boundary conditions (ABCs) are simplified to local ones so that the computational efficiency is enhanced significantly. The introduced two local ABCs are implemented in a finite element computer program so that various seepage problems can be calculated. The two approaches are first verified by the computation of a one-dimensional radial flow problem, and then tentatively applied to more general two-dimensional cylindrical problems and plane problems. Numerical calculations show that the local ABCs can not only give good results for one-dimensional axisymmetric transient flow, but also applicable for more general problems, such as axisymmetric two-dimensional cylindrical problems, and even more general planar two-dimensional flow problems for well doublet and well groups. An important advantage of the latter local boundary is its applicability for seepage under rapidly changing unsteady boundary conditions, and even the computational results on the truncated boundary are usually quite satisfactory. In this aspect, it is superior over the former local boundary. Simulation of relatively long operational time demonstrates to certain extents the numerical stability of the local boundary. The solutions of the two local ABCs are compared with each other and with those obtained by using large element mesh, which proves the satisfactory performance and obvious superiority over the large mesh model.Keywords: transient seepage, unbounded domain, artificial boundary condition, numerical simulation
Procedia PDF Downloads 2942967 A Micro-Scale of Electromechanical System Micro-Sensor Resonator Based on UNO-Microcontroller for Low Magnetic Field Detection
Authors: Waddah Abdelbagi Talha, Mohammed Abdullah Elmaleeh, John Ojur Dennis
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This paper focuses on the simulation and implementation of a resonator micro-sensor for low magnetic field sensing based on a U-shaped cantilever and piezoresistive configuration, which works based on Lorentz force physical phenomena. The resonance frequency is an important parameter that depends upon the highest response and sensitivity through the frequency domain (frequency response) of any vibrated micro-scale of an electromechanical system (MEMS) device. And it is important to determine the direction of the detected magnetic field. The deflection of the cantilever is considered for vibrated mode with different frequencies in the range of (0 Hz to 7000 Hz); for the purpose of observing the frequency response. A simple electronic circuit-based polysilicon piezoresistors in Wheatstone's bridge configuration are used to transduce the response of the cantilever to electrical measurements at various voltages. Microcontroller-based Arduino program and PROTEUS electronic software are used to analyze the output signals from the sensor. The highest output voltage amplitude of about 4.7 mV is spotted at about 3 kHz of the frequency domain, indicating the highest sensitivity, which can be called resonant sensitivity. Based on the resonant frequency value, the mode of vibration is determined (up-down vibration), and based on that, the vector of the magnetic field is also determined.Keywords: resonant frequency, sensitivity, Wheatstone bridge, UNO-microcontroller
Procedia PDF Downloads 1272966 Novel p22-Monoclonal Antibody Based Blocking ELISA for the Detection of African Swine Fever Virus Antibodies in Serum
Authors: Ghebremedhin Tsegay, Weldu Tesfagaber, Yuanmao Zhu, Xijun He, Wan Wang, Zhenjiang Zhang, Encheng Sun, Jinya Zhang, Yuntao Guan, Fang Li, Renqiang Liu, Zhigao Bu, Dongming Zhao*
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African swine fever (ASF) is a highly infectious viral disease of pigs, resulting in significant economic loss worldwide. As there is no approved vaccines and treatments, the control of ASF entirely depends on early diagnosis and culling of infected pigs. Thus, highly specific and sensitive diagnostic assays are required for accurate and early diagnosis of ASF virus (ASFV). Currently, only a few recombinant proteins have been tested and validated for use as reagents in ASF diagnostic assays. The most promising ones for ASFV antibody detection were p72, p30, p54, and pp62. So far, three ELISA kits based on these recombinant proteins have been commercialized. Due to the complex nature of the virus and variety forms of the disease, robust serodiagnostic assays are still required. ASFV p22 protein, encoded by KP177R gene, is located in the inner membrane of viral particle and appeared transiently in the plasma membrane early after virus infection. The p22 protein interacts with numerous cellular proteins, involved in processes of phagocytosis and endocytosis through different cellular pathways. However, p22 does not seem to be involved in virus replication or swine pathogenicity. In this study, E.coli expressed recombinant p22 protein was used to generate a monoclonal antibody (mAb), and its potential use for the development of blocking ELISA (bELISA) was evaluated. A total of 806 pig serum samples were tested to evaluate the bELISA. Acording the ROC (Reciever operating chracteristic) analysis, 100% sensitivity and 98.10% of specificity was recorded when the PI cut-off value was set at 47%. The novel assay was able to detect the antibodies as early as 9 days post infection. Finaly, a highly sensitive, specific and rapid novel p22-mAb based bELISA assay was developed, and optimized for detection of antibodies against genotype I and II ASFVs. It is a promising candidate for an early and acurate detection of the antibodies and is highly expected to have a valuable role in the containment and prevention of ASF.Keywords: ASFV, blocking ELISA, diagnosis, monoclonal antibodies, sensitivity, specificity
Procedia PDF Downloads 772965 Nutritional Management of Polycystic Ovary Syndrome Using a Mediterranean Diet
Authors: Mohamed Radwan
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Introduction: Polycystic ovary syndrome is becoming very common among girls from ages 14 to 21 years old, alongside women in their childbearing period. A combination of menstrual irregularities, overweight or obesity, insulin resistance (type 2 diabetes), and symptoms of virilization are characteristics of these cases. Background: A multidisciplinary team should be involved in managing these cases and may consist of A gynecologist, endocrinologist, nutritionist, and psychologist. The role of nutritionists is crucial in these cases in weight reduction and nutritional management. It is mandatory to present some questions that may help in expanding further our understanding of the nutritional management of this syndrome, such as it is not an easy process to reduce the weight of PCO patients (2) we need to know what suitable nutritional plans for these cases are. Methodology: It is not successful to use one diet plan for all patients or a specific plan, but we need to give tailored plans for each patient. We will summarize the medical, nutritional therapy and weight management in polycystic ovary patients and highlight the best eating plan and dietary composition in the treatment of these women. We will also discuss the role of dieticians in treating polycystic ovary cases and overcoming the challenges these women face. Conclusion: All diet plans would decrease the weight of Polycystic ovary cases as low calory diet, low glycemic index diet, high protein diet, as well as Dash diet, but the most suitable diet plan to improve hormones and lead to spontaneous pregnancy is the Mediterranean diet.Keywords: obesity, PCO mediteranian diet, dash diet, high protein diet
Procedia PDF Downloads 762964 Molecular Characterization of Functional Domain (LRR) of TLR9 Genes in Malnad Gidda Cattle and Their Comparison to Cross Breed Cattle
Authors: Ananthakrishna L. R., Ramesh D., Kumar Wodeyar, Kotresh A. M., Gururaj P. M.
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Malnad Gidda is the indigenous recognized cattle breed of Shivamogga District of Karnataka state, India is known for its disease resistance to many of the infectious diseases. There are 25 LRR (Leucine Rich Repeats) identified in bovine (Bos indicus) TLR9. The amino acid sequence of LRR is deduced to nucleotide sequence in BLASTx bioinformatic online tools. LRR2 to LRR10 are involved in pathogen recognition and binding in human TLR9 which showed a higher degree of nucleotide variations with respect to disease resistance to various pathogens. Hence, primers were designed to amplify the flanking sequences of LRR2 to LRR10, to discover the nucleotide variations if any, in Malnad Gidda breed of Cattle which is associated with disease resistance. The DNA isolated from peripheral blood mononuclear cells of ten Malnad Gidda cattle. A desired and specific amplification product of 0.8 kb was obtained at an annealing temperature of 56.6ᵒC. All the PCR products were sequenced on both sides by gene-specific primers. The sequences were compared with TLR9 sequence of cross breed cattle obtained from NCBI data bank. The sequence analysis between Malnad Gidda and crossbreed cattle revealed no nucleotide variations in the region LRR2 to LRR9 which shows the conserved in pathogen binding domain (LRR) of TLR9.Keywords: leucine rich repeats, Malnad Gidda, cross breed, TLR9
Procedia PDF Downloads 2262963 Expression of Metallothionein Gen and Protein on Hepatopancreas, Gill and Muscle of Perna viridis Caused by Biotoxicity Hg, Pb and Cd
Authors: Yulia Irnidayanti , J. J. Josua, A. Sugianto
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Jakarta Bay with 13 rivers that flow into, the environment has deteriorated and is the most polluted bays in Asia. The entry of waste into the waters of the Bay of Jakarta has caused pollution. Heavy metal contamination has led to pollution levels and may cause toxicity to organisms that live in the sea, down to the cellular level and may affect the ecological balance. Various ways have been conducted to measure the impact of environmental degradation, such as by measuring the levels of contaminants in the environment, including measuring the accumulation of toxic compounds in the tissues of organisms. Biological responses or biomarkers known as a sensitive indicator but need relevant predictions. In heavy metal pollution monitoring, analysis of aquatic biota is very important from the analysis of the water itself. The content of metals in aquatic biota will usually always be increased from time to time due to the nature of metal bioaccumulation, so the aquatic biota is best used as an indicator of metal pollution in aquatic environments. The results of the content analysis results of sea water in coastal estuaries Angke, Kaliadem and Panimbang detected heavy metals cadmium, mercury, lead, but did not find zinc metal. Based on the results of protein electrophoresis methallotionein found heavy metals in the tissues hepatopancreas, gills and muscles, and also the mRNA expression of has detected.Keywords: gills, heavy metal, hepatopancreas, metallothionein, muscle
Procedia PDF Downloads 3892962 Ontology Expansion via Synthetic Dataset Generation and Transformer-Based Concept Extraction
Authors: Andrey Khalov
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The rapid proliferation of unstructured data in IT infrastructure management demands innovative approaches for extracting actionable knowledge. This paper presents a framework for ontology-based knowledge extraction that combines relational graph neural networks (R-GNN) with large language models (LLMs). The proposed method leverages the DOLCE framework as the foundational ontology, extending it with concepts from ITSMO for domain-specific applications in IT service management and outsourcing. A key component of this research is the use of transformer-based models, such as DeBERTa-v3-large, for automatic entity and relationship extraction from unstructured texts. Furthermore, the paper explores how transfer learning techniques can be applied to fine-tune large language models (LLaMA) for using to generate synthetic datasets to improve precision in BERT-based entity recognition and ontology alignment. The resulting IT Ontology (ITO) serves as a comprehensive knowledge base that integrates domain-specific insights from ITIL processes, enabling more efficient decision-making. Experimental results demonstrate significant improvements in knowledge extraction and relationship mapping, offering a cutting-edge solution for enhancing cognitive computing in IT service environments.Keywords: ontology expansion, synthetic dataset, transformer fine-tuning, concept extraction, DOLCE, BERT, taxonomy, LLM, NER
Procedia PDF Downloads 142961 Visualizing Matrix Metalloproteinase-2 Activity Using Extracellular Matrix-Immobilized Fluorescence Resonance Energy Transfer Bioprobe in Cancer Cells
Authors: Hawon Lee, Young-Pil Kim
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Visualizing matrix metalloproteinases (MMPs) activity is necessary for understanding cancer metastasis because they are implicated in cell migration and invasion by degrading the extracellular matrix (ECM). While much effort has been made to sense the MMP activity, but extracellularly long-term monitoring of MMP activity still remains challenging. Here, we report a collagen-bound fluorescent bioprobe for the detection of MMP-2 activity in the extracellular environment. This bioprobe consists of ECM-immobilized part (including collagen-bound protein) and MMP-sensing part (including peptide substrate linked with fluorescence resonance energy transfer (FRET) coupler between donor green fluorescent protein (GFP) and acceptor TAMRA dye), which was constructed through intein-mediated self-splicing conjugation. Upon being immobilized on the collagen-coated surface, this bioprobe enabled efficient long-lasting observation of MMP-2 activity in the cultured cells without affecting cell growth and viability. As a result, the FRET ratio (acceptor/donor) decreased as the MMP2 activity increased in cultured cancer cells. Furthermore, unlike wild-type MMP-2, mutated MMP-2 expression (Y580A in the hemopexin region) gave rise to lowering the secretion of MMP-2 in HeLa. Conclusively, our method is anticipated to find applications for tracing and visualizing enzyme activity.Keywords: collagen, ECM, FRET, MMP
Procedia PDF Downloads 2022960 Toward Understanding the Glucocorticoid Receptor Network in Cancer
Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden
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The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor
Procedia PDF Downloads 2272959 Development of Ferric Citrate Complex Draw Solute and Its Application for Liquid Product Enrichment through Forward Osmosis
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Forward osmosis is an emerging technology for separation and has great potential in the concentration of liquid products such as protein, pharmaceutical, and natural products. In pharmacy industry, one of the very tough talks is to concentrate the product in a gentle way since some of the key components may lose bioactivity when exposed to heating or pressurization. Therefore, forward osmosis (FO), which uses inherently existed osmosis pressure instead of externally applied hydraulic pressure, is attractive for pharmaceutical enrichments in a much efficient and energy-saving way. Recently, coordination complexes have been explored as the new class of draw solutes in FO processes due to their bulky configuration and excellent performance in terms of high water flux and low reverse solute flux. Among these coordination complexes, ferric citrate complex with lots of hydrophilic groups and ionic species which make them good solubility and high osmotic pressure in aqueous solution, as well as its low toxicity, has received much attention. However, the chemistry of ferric complexation by citrate is complicated, and disagreement prevails in the literature, especially for the structure of the ferric citrate. In this study, we investigated the chemical reaction with various molar ratio of iron and citrate. It was observed that the ferric citrate complex (Fe-CA2) with molar ratio of 1:1 for iron and citrate formed at the beginning of the reaction, then Fecit would convert to ferric citrate complex at the molar ratio of 1:2 with the proper excess of citrate in the base solution. The structures of the ferric citrate complexes synthesized were systematically characterized by X-ray diffraction (XRD), UV-vis spectroscopy, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR) and Thermogravimetric analysis (TGA). Fe-CA2 solutions exhibit osmotic pressures more than twice of that for NaCl solutions at the same concentrations. Higher osmotic pressure means higher driving force, and this is preferable for the FO process. Fe-CA2 and NaCl draw solutions were prepared with the same osmotic pressure and used in FO process for BSA protein concentration. Within 180 min, BSA concentration was enriched from 0.2 to 0.27 L using Fe-CA draw solutions. However, it was only increased from 0.20 to 0.22 g/L using NaCl draw solutions. A reverse flux of 11 g/m²h was observed for NaCl draw solutes while it was only 0.1 g/m²h for Fe-CA2 draw solutes. It is safe to conclude that Fe-CA2 is much better than NaCl as draw solute and it is suitable for the enrichment of liquid product.Keywords: draw solutes, ferric citrate complex, forward osmosis, protein enrichment
Procedia PDF Downloads 1532958 Functional Ingredients from Potato By-Products: Innovative Biocatalytic Processes
Authors: Salwa Karboune, Amanda Waglay
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Recent studies indicate that health-promoting functional ingredients and nutraceuticals can help support and improve the overall public health, which is timely given the aging of the population and the increasing cost of health care. The development of novel ‘natural’ functional ingredients is increasingly challenging. Biocatalysis offers powerful approaches to achieve this goal. Our recent research has been focusing on the development of innovative biocatalytic approaches towards the isolation of protein isolates from potato by-products and the generation of peptides. Potato is a vegetable whose high-quality proteins are underestimated. In addition to their high proportion in the essential amino acids, potato proteins possess angiotensin-converting enzyme-inhibitory potency, an ability to reduce plasma triglycerides associated with a reduced risk of atherosclerosis, and stimulate the release of the appetite regulating hormone CCK. Potato proteins have long been considered not economically feasible due to the low protein content (27% dry matter) found in tuber (Solanum tuberosum). However, potatoes rank the second largest protein supplying crop grown per hectare following wheat. Potato proteins include patatin (40-45 kDa), protease inhibitors (5-25 kDa), and various high MW proteins. Non-destructive techniques for the extraction of proteins from potato pulp and for the generation of peptides are needed in order to minimize functional losses and enhance quality. A promising approach for isolating the potato proteins was developed, which involves the use of multi-enzymatic systems containing selected glycosyl hydrolase enzymes that synergistically work to open the plant cell wall network. This enzymatic approach is advantageous due to: (1) the use of milder reaction conditions, (2) the high selectivity and specificity of enzymes, (3) the low cost and (4) the ability to market natural ingredients. Another major benefit to this enzymatic approach is the elimination of a costly purification step; indeed, these multi-enzymatic systems have the ability to isolate proteins, while fractionating them due to their specificity and selectivity with minimal proteolytic activities. The isolated proteins were used for the enzymatic generation of active peptides. In addition, they were applied into a reduced gluten cookie formulation as consumers are putting a high demand for easy ready to eat snack foods, with high nutritional quality and limited to no gluten incorporation. The addition of potato protein significantly improved the textural hardness of reduced gluten cookies, more comparable to wheat flour alone. The presentation will focus on our recent ‘proof-of principle’ results illustrating the feasibility and the efficiency of new biocatalytic processes for the production of innovative functional food ingredients, from potato by-products, whose potential health benefits are increasingly being recognized.Keywords: biocatalytic approaches, functional ingredients, potato proteins, peptides
Procedia PDF Downloads 3792957 Biomass and Biogas Yield of Maize as Affected by Nitrogen Rates with Varying Harvesting under Semi-Arid Condition of Pakistan
Authors: Athar Mahmood, Asad Ali
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Management considerations including harvesting time and nitrogen application considerably influence the biomass yield, quality and biogas production. Therefore, a field study was conducted to determine the effect of various harvesting times and nitrogen rates on the biomass yield, quality and biogas yield of maize crop. This experiment was consisted of various harvesting times i.e., harvesting after 45, 55 and 65 days of sowing (DAS) and nitrogen rates i.e., 0, 100, 150 and 200 kg ha-1 respectively. The data indicated that maximum plant height, leaf area, dry matter (DM) yield, protein, acid detergent fiber, neutral detergent fiber, crude fiber contents and biogas yield were recorded 65 days after sowing while lowest was recorded 45 days after sowing. In contrary to that significantly higher chlorophyll contents were observed at 45 DAS. In case of nitrogen rates maximum plant height, leaf area, and DM yield, protein contents, ash contents, acid detergent fiber, neutral detergent fiber, crude fiber contents and chlorophyll contents were determined with nitrogen at the rate of 200 kg ha-1, while minimum was observed when no N was applied. Therefore, harvesting 65 DAS and N application @ 200 kg ha-1 can be suitable for getting the higher biomass and biogas production.Keywords: chemical composition, fiber contents, biogas, nitrogen, harvesting time
Procedia PDF Downloads 1602956 Protein Stabilized Foam Structures as Protective Carrier Systems during Microwave Drying of Probiotics
Authors: Jannika Dombrowski, Sabine Ambros, Ulrich Kulozik
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Due to the increasing popularity of healthy products, probiotics are still of rising importance in food manufacturing. With the aim to amplify the field of probiotic application to non-chilled products, the cultures have to be preserved by drying. Microwave drying has proved to be a suitable technique to achieve relatively high survival rates, resulting from drying at gentle temperatures, among others. However, diffusion limitation due to compaction of cell suspension during drying can prolong drying times as well as deteriorate product properties (grindability, rehydration performance). Therefore, we aimed to embed probiotics in an aerated matrix of whey proteins (surfactants) and di-/polysaccharides (foam stabilization, probiotic protection) during drying. As a result of the manifold increased inner surface of the cell suspension, drying performance was enhanced significantly as compared to non-foamed suspensions. This work comprises investigations on suitable foam matrices, being stable under vacuum (variation of protein concentration, type and concentration of di-/polysaccharide) as well as development of an applicable microwave drying process in terms of microwave power, chamber pressure and maximum product temperatures. Performed analyses included foam characteristics (overrun, drainage, firmness, bubble sizes), and properties of the dried cultures (survival, activity). In addition, efficiency of the drying process was evaluated.Keywords: foam structure, microwave drying, polysaccharides, probiotics
Procedia PDF Downloads 2622955 Application of Lattice Boltzmann Method to Different Boundary Conditions in a Two Dimensional Enclosure
Authors: Jean Yves Trepanier, Sami Ammar, Sagnik Banik
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Lattice Boltzmann Method has been advantageous in simulating complex boundary conditions and solving for fluid flow parameters by streaming and collision processes. This paper includes the study of three different test cases in a confined domain using the method of the Lattice Boltzmann model. 1. An SRT (Single Relaxation Time) approach in the Lattice Boltzmann model is used to simulate Lid Driven Cavity flow for different Reynolds Number (100, 400 and 1000) with a domain aspect ratio of 1, i.e., square cavity. A moment-based boundary condition is used for more accurate results. 2. A Thermal Lattice BGK (Bhatnagar-Gross-Krook) Model is developed for the Rayleigh Benard convection for both test cases - Horizontal and Vertical Temperature difference, considered separately for a Boussinesq incompressible fluid. The Rayleigh number is varied for both the test cases (10^3 ≤ Ra ≤ 10^6) keeping the Prandtl number at 0.71. A stability criteria with a precise forcing scheme is used for a greater level of accuracy. 3. The phase change problem governed by the heat-conduction equation is studied using the enthalpy based Lattice Boltzmann Model with a single iteration for each time step, thus reducing the computational time. A double distribution function approach with D2Q9 (density) model and D2Q5 (temperature) model are used for two different test cases-the conduction dominated melting and the convection dominated melting. The solidification process is also simulated using the enthalpy based method with a single distribution function using the D2Q5 model to provide a better understanding of the heat transport phenomenon. The domain for the test cases has an aspect ratio of 2 with some exceptions for a square cavity. An approximate velocity scale is chosen to ensure that the simulations are within the incompressible regime. Different parameters like velocities, temperature, Nusselt number, etc. are calculated for a comparative study with the existing works of literature. The simulated results demonstrate excellent agreement with the existing benchmark solution within an error limit of ± 0.05 implicates the viability of this method for complex fluid flow problems.Keywords: BGK, Nusselt, Prandtl, Rayleigh, SRT
Procedia PDF Downloads 1282954 Expression of Fused Plasmodium falciparum Orotate Phosphoribosyltransferase and Orotidine 5'-Monophosphate Decarboxylase in Escherichia coli
Authors: Waranya Imprasittichai, Patsarawadee Paojinda, Sudaratana R. Krungkrai, Nirianne Marie Q. Palacpac, Toshihiro Horii, Jerapan Krungkrai
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Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. In this study, we constructed gene fusions of Plasmodium falciparum OMPDC-OPRT (1,836 bp) in pTrcHisA vector and expressed as an 6xHis-tag bifunctional protein in three Escherichia coli strains (BL21, Rosetta, TOP10) at 18 °C, 25 °C and 37 °C. The recombinant bifunctional protein was partially purified by Ni-Nitrilotriacetic acid-affinity chromatography. Specific activities of OPRT and OMPDC domains in the bifunctional enzyme expressed in E. coli TOP10 cells were approximately 3-4-fold higher than those in BL21 cells. There were no enzymatic activities when the construct vector expressed in Rosetta cells. Maximal expression of the fused gene was observed at 18 °C and the bifunctional enzyme had specific activities of OPRT and OMPDC domains in a ratio of 1:2. These results provide greater yields and better catalytic activities of the bifunctional OMPDC-OPRT enzyme for further purification and kinetic study.Keywords: bifunctional enzyme, orotate phosphoribosyltransferase, orotidine 5'-monophosphate decarboxylase, plasmodium falciparum
Procedia PDF Downloads 3542953 Hotel Deposit Contract and Coverage of Risks Resulting, through Insurance Contracts, in Tourism within the HoReCa Domain: Alternative Dispute Resolution Methods on These Contracts
Authors: Laura Ramona Nae
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The issue of risks faced by companies providing tourist and hotel services in the HoReCa field, related to the goods belonging to consumer tourists left in hotel storage, has acquired a new dimension in the context of the economic and geo-political influences that have recently intervened at the global level. Thus, hoteliers and not only had to create contractual mechanisms regarding the risks and to protect the businesses in this field of activity. This situation has led to a reassessment of the importance of insurance, in particular with regard to hotel liability insurance-premises liability, safety, and security of goods. Interpretation of clauses in contracts concluded between hoteliers and tourists consuming hotel services and products, all the more so in the current pandemic context of Covid 19, stressed the increase in the number of disputes generated by them. This article presents a general picture of the significance of the risks related to the activity carried out in the hospitality industry, tourism, respectively within the HoReCa field. The study mainly marks the specificities of the hotel deposit contract, as well as the related insurance specific to the field, as a way to cover these risks. The article also refers to alternative methods of out-of-court settlement of disputes (ADR) in the HoReCa domain, generally used in both Romania and the European Union.Keywords: consumer tourist, disputes and ADR methods, deposit contract, hotel warehouse and hotelier insurance, hotel services and tourist products, HoReCa
Procedia PDF Downloads 572952 Interethnic Communication in Multicultural Areas: A Case Study of Intercultural Sensitivity Between Baloch and Persians in Iran
Authors: Mehraveh Taghizadeh
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Iran is home to a diverse range of ethnic groups such as Baloch, Kurds, Persians, Lors, Arabs, and Turks. The Persian ethnicity is the largest group, while Baloch people are considered a minority residing on the southeastern border of the country with different language and religion. As a consequence, Political discussions have often prioritized national identity and national security over Baloch ethnic identity. However, to improve intercultural understanding and reduce cultural schemas, it's crucial to decrease ethnocentrism and increase intercultural communication. In the meantime, Kerman, a multicultural province that borders Sistan and Baluchistan, has become a destination for Baloch immigrants. By recognizing the current status of intercultural competence, we can develop effective policies for expanding intercultural communication and creating a more inclusive and peaceful society. As a result, this research aims to study the domain of intercultural sensitivity between Persians and Baloch in Kerman. Therefore, the question is how do Persians and Baloch ethnicities perceive each other? This study represents the first exploration of communication dynamics between Persians and Baloch individuals. Utilizing a qualitative approach, this study employs thematic analysis in conjunction with Bennett's intercultural sensitivities model. The model comprises two components: ethnocentrism, which spans from denial and defense to minimization, and ethno-relativism, which ranges from acceptance and adaptation to integration. To attain this objective, 30 individuals from Persian and Baloch ethnicities were interviewed using a semi-structured format. it analysis suggests that the Baluch and Persians exhibit a range of intercultural sensitivities characterized by defensive and minimizing attitudes in the ethnocentrism domain, and accepting attitudes in the ethno-relativism domain. The concept of minimization involves recognizing the shared humanity and positive schemas of both groups. Furthermore, in the adaptation domain, Persians' efforts to assimilate into Baloch culture at an acceptance level are primarily focused on the civilizational dimension, including using traditional Balochi clothing designs on their clothes. The Persians hold intercultural schemas about the Baloch people, including notions of religious fanaticism, tribalism, poverty, smuggling, and a nomadic way of life. Conversely, the Baloch people have intercultural schemas about Persians including religious fanaticism, disdain towards the Baloch, and ethnocentrism. Both groups tend to tie ethnicity to religion and judge each other accordingly. Also, the origin of these schemas is in the representation of the media and the encounter without interaction between the two ethnic groups. These findings indicate that they have not received adaptation and integration levels in ethno-relativism. Furthermore, the results indicate that developing personal communication in multicultural environments reduces intercultural sensitivity, and increases positive interactions and civilizational dialogues. People can understand each other better and perform better in their daily lives.Keywords: intercultural communication, intercultural sensitivity, interethnic communication, Iran, Baloch, Persians
Procedia PDF Downloads 512951 A Novel Gene Encoding Ankyrin-Repeat Protein, SHG1, Is Indispensable for Seed Germination under Moderate Salt Stress
Authors: H. Sakamoto, J. Tochimoto, S. Kurosawa, M. Suzuki, S. Oguri
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Salt stress adversely affects plant growth at various stages of development including seed germination, seedling establishment, vegetative growth and finally reproduction. Because of their immobile nature, plants have evolved mechanisms to sense and respond to salt stress. Seed dormancy is an adaptive trait that enables seed germination to coincide with favorable environmental conditions. We identified a novel locus of Arabidopsis, designated SHG1 (salt hypersensitive germination 1), whose disruption leads to reduced germination rate under moderate salt stress conditions. SHG1 encodes a transmembrane protein with an ankyrin repeat motif that has been implicated in diverse cellular processes such as signal transduction. The SGH1-disrupted Arabidopsis mutant died at the cotyledon stage when sown on salt-containing medium, although wild type plants could form true leaves under the same conditions. On the other hand, this mutant showed similar phenotypes to wild type plants when sown on medium without salt and transferred to salt-containing medium at the vegetative stage. These results suggested that SHG1 played indispensable role in the seed germination and seedling establishment under moderate salt stress conditions. SHG1 may be involved in the release of seed dormancy.Keywords: germination, ankyrin repeat, arabidopsis, salt tolerance
Procedia PDF Downloads 398