Search results for: protein masked representation
3497 Interaction Effects of Dietary Ginger, Zingiber Officinale, on Plasma Protein Fractions in Rainbow Trout, Oncorhynchus Mykiss
Authors: Ali Taheri Mirghaed, Sara Ahani, Ashkan Zargar, Seyyed Morteza Hoseini
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Diseases are the major challenges in intensive aquaculture that cause significant annual losses. Antibiotic-therapy is a common way to control bacterial disease in fish, and oxytetracycline (OTC) is the only oral antibiotic in aquaculture approved FDA. OTC has been found to have negative effects on fish, such as oxidative stress and immune-suppression, thus, it is necessary to mitigate such effects. Medicinal herbs have various benefits on fish, including antioxidant, immunostimulant, and anti-microbial effects. Therefore, we hypothesized if dietary ginger meal (GM) interacts with dietary OTC by monitoring plasma protein fractions in rainbow trout. The study was conducted as a 2 × 2 factorial design, including diets containing 0 and 1% GM and 0 and 1.66 % OTC (corresponding to 100 mg/kg fish biomass per day). After ten days treating the fish (60 g individual weight) with these feeds, blood samples were taken from al treatments (n =3). Plasma was separated by centrifugation, and protein fractions were determined by electrophoresis. The results showed that OTC and GM had interaction effects on total protein (P<0.001), albumin (P<0.001), alpha-1 fraction (P=0.010), alpha-2 fraction (P=0.001), beta-2 fraction (P=0.014), and gamma fraction (P<0.001). Beta-1 fraction was significantly (P=0.030) affected by dietary GM. GM decreased plasma total protein, albumin, and beta-2 but increased beta-1 fraction. OTC significantly decreased total protein (P<0.001), albumin (P=0.001), alpha-2 fraction (P<0.001), beta-2 fraction (P=0.004), and gamma fraction (P<0.001) but had no significant effects on alpha-1 and beta-1 fractions. Dietary GM inhibited/suppressed the effects of dietary OTC on the plasma total protein and protein fractions. In conclusion, adding 1% GM to diet can mitigate the negative effects of dietary OTC on plasma proteins. Thus, GM may boost health of rainbow trout during the period of medication with OTC.Keywords: ginger, plasma protein electrophoresis, dietary additive, rainbow trout
Procedia PDF Downloads 883496 The Different Improvement of Numerical Magnitude and Spatial Representation of Numbers to Symbolic Approximate Arithmetic: A Training Study of Preschooler
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Spatial representation of numbers and numerical magnitude are important for preschoolers’ mathematical ability. Mental number line, a typical index to measure numbers spatial representation, and numerical comparison are both related to arithmetic obviously. However, they seem to rely on different mechanisms and probably influence arithmetic through different mechanisms. In line with this idea, preschool children were trained with two tasks to investigate which one is more important for approximate arithmetic. The training of numerical processing and number line estimation were proved to be effective. They both improved the ability of approximate arithmetic. When the difficulty of approximate arithmetic was taken into account, the performance in number line training group was not significantly different among three levels. However, two harder levels achieved significance in numerical comparison training group. Thus, comparing spatial representation ability, symbolic approximation arithmetic relies more on numerical magnitude. Educational implications of the study were discussed.Keywords: approximate arithmetic, mental number line, numerical magnitude, preschooler
Procedia PDF Downloads 2503495 Use RP-HPLC To Investigate Factors Influencing Sorghum Protein Extraction
Authors: Khaled Khaladi, Rafika Bibi, Hind Mokrane, Boubekeur Nadjemi
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Sorghum (Sorghum bicolor (L.) Moench) is an important cereal crop grown in the semi-arid tropics of Africa and Asia due to its drought tolerance. Sorghum grain has protein content varying from 6 to 18%, with an average of 11%, Sorghum proteins can be broadly classified into prolamin and non-prolamin proteins. Kafirins, the major storage proteins, are classified as prolamins, and as such, they contain high levels of proline and glutamine and are soluble in non-polar solvents such as aqueous alcohols. Kafirins account for 77 to 82% of the protein in the endosperm, whereas non-prolamin proteins (namely, albumins, globulins, and glutelins) make up about 30% of the proteins. To optimize the extraction of sorghum proteins, several variables were examined: detergent type and concentration, reducing agent type and concentration, and buffer pH and concentration. Samples were quantified and characterized by RP-HPLC.Keywords: sorghum, protein extraction, detergent, food science
Procedia PDF Downloads 3163494 Quantifying the Protein-Protein Interaction between the Ion-Channel-Forming Colicin A and the Tol Proteins by Potassium Efflux in E. coli Cells
Authors: Fadilah Aleanizy
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Colicins are a family of bacterial toxins that kill Escherichia coli and other closely related species. The mode of action of colicins involves binding to an outer membrane receptor and translocation across the cell envelope, leading to cytotoxicity through specific targets. The mechanism of colicin cytotoxicity includes a non-specific endonuclease activity or depolarization of the cytoplasmic membrane by pore-forming activity. For Group A colicins, translocation requires an interaction between the N-terminal domain of the colicin and a series of membrane- bound and periplasmic proteins known as the Tol system (TolB, TolR, TolA, TolQ, and Pal and the active domain must be translocated through the outer membranes. Protein-protein interactions are intrinsic to virtually every cellular process. The transient protein-protein interactions of the colicin include the interaction with much more complicated assemblies during colicin translocation across the cellular membrane to its target. The potassium release assay detects variation in the K+ content of bacterial cells (K+in). This assays is used to measure the effect of pore-forming colicins such as ColA on an indicator organism by measuring the changes of the K+ concentration in the external medium (K+out ) that are caused by cell killing with a K+ selective electrode. One of the goals of this work is to employ a quantifiable in-vivo method to spot which Tol protein are more implicated in the interaction with colicin A as it is translocated to its target.Keywords: K+ efflux, Colicin A, Tol-proteins, E. coli
Procedia PDF Downloads 4083493 Delivery of Positively Charged Proteins Using Hyaluronic Acid Microgels
Authors: Elaheh Jooybar, Mohammad J. Abdekhodaie, Marcel Karperien, Pieter J. Dijkstra
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In this study, hyaluronic acid (HA) microgels were developed for the goal of protein delivery. First, a hyaluronic acid-tyramine conjugate (HA-TA) was synthesized with a degree of substitution of 13 TA moieties per 100 disaccharide units. Then, HA-TA microdroplets were produced using a water in oil emulsion method and crosslinked in the presence of horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). Loading capacity and the release kinetics of lysozyme and BSA, as model proteins, were investigated. It was shown that lysozyme, a cationic protein, can be incorporated efficiently in the HA microgels, while the loading efficiency for BSA, as a negatively charged protein, is low. The release profile of lysozyme showed a sustained release over a period of one month. The results demonstrated that the HA-TA microgels are a good carrier for spatial delivery of cationic proteins for biomedical applications.Keywords: microgel, inverse emulsion, protein delivery, hyaluronic acid, crosslinking
Procedia PDF Downloads 1683492 Bioinformatics and Molecular Biological Characterization of a Hypothetical Protein SAV1226 as a Potential Drug Target for Methicillin/Vancomycin-Staphylococcus aureus Infections
Authors: Nichole Haag, Kimberly Velk, Tyler McCune, Chun Wu
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Methicillin/multiple-resistant Staphylococcus aureus (MRSA) are infectious bacteria that are resistant to common antibiotics. A previous in silico study in our group has identified a hypothetical protein SAV1226 as one of the potential drug targets. In this study, we reported the bioinformatics characterization, as well as cloning, expression, purification and kinetic assays of hypothetical protein SAV1226 from methicillin/vancomycin-resistant Staphylococcus aureus Mu50 strain. MALDI-TOF/MS analysis revealed a low degree of structural similarity with known proteins. Kinetic assays demonstrated that hypothetical protein SAV1226 is neither a domain of an ATP dependent dihydroxyacetone kinase nor of a phosphotransferase system (PTS) dihydroxyacetone kinase, suggesting that the function of hypothetical protein SAV1226 might be misannotated on public databases such as UniProt and InterProScan 5.Keywords: Methicillin-resistant Staphylococcus aureus, dihydroxyacetone kinase, essential genes, drug target, phosphoryl group donor
Procedia PDF Downloads 4063491 Nutritional Potential and Functionality of Whey Powder Influenced by Different Processing Temperature and Storage
Authors: Zarmina Gillani, Nuzhat Huma, Aysha Sameen, Mulazim Hussain Bukhari
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Whey is an excellent food ingredient owing to its high nutritive value and its functional properties. However, composition of whey varies depending on composition of milk, processing conditions, processing method, and its whey protein content. The aim of this study was to prepare a whey powder from raw whey and to determine the influence of different processing temperatures (160 and 180 °C) on the physicochemical, functional properties during storage of 180 days and on whey protein denaturation. Results have shown that temperature significantly (P < 0.05) affects the pH, acidity, non-protein nitrogen (NPN), protein total soluble solids, fat and lactose contents. Significantly (p < 0.05) higher foaming capacity (FC), foam stability (FS), whey protein nitrogen index (WPNI), and a lower turbidity and solubility index (SI) were observed in whey powder processed at 160 °C compared to whey powder processed at 180 °C. During storage of 180 days, slow but progressive changes were noticed on the physicochemical and functional properties of whey powder. Reverse phase-HPLC analysis revealed a significant (P < 0.05) effect of temperature on whey protein contents. Denaturation of β-Lactoglobulin is followed by α-lacalbumin, casein glycomacropeptide (CMP/GMP), and bovine serum albumin (BSA).Keywords: whey powder, temperature, denaturation, reverse phase, HPLC
Procedia PDF Downloads 2973490 Towards the Inhibition Mechanism of Lysozyme Fibrillation by Hydrogen Sulfide
Authors: Indra Gonzalez Ojeda, Tatiana Quinones, Manuel Rosario, Igor Lednev, Juan Lopez Garriga
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Amyloid fibrils are stable aggregates of misfolded protein associated with many neurodegenerative disorders. It has been shown that hydrogen sulfide (H2S), inhibits the fibrillation of lysozyme through the formation of trisulfide (S-S-S) bonds. However, the overall mechanism remains elusive. Here, the concentration dependence of H2S effect was investigated using Atomic force microscopy (AFM), non-resonance Raman spectroscopy, Deep-UV Raman spectroscopy and circular dichroism (CD). It was found that small spherical aggregates with trisulfide bonds and a unique secondary structure were formed instead of amyloid fibrils when adding concentrations of 25 mM and 50 mM of H2S. This could indicate that H2S might serve as a protecting agent for the protein. However, further characterization of these aggregates and their trisulfide bonds is needed to fully unravel the function H2S has on protein fibrillation.Keywords: amyloid fibrils, hydrogen sulfide, protein folding, raman spectroscopy
Procedia PDF Downloads 2133489 Protein Remote Homology Detection by Using Profile-Based Matrix Transformation Approaches
Authors: Bin Liu
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As one of the most important tasks in protein sequence analysis, protein remote homology detection has been studied for decades. Currently, the profile-based methods show state-of-the-art performance. Position-Specific Frequency Matrix (PSFM) is widely used profile. However, there exists noise information in the profiles introduced by the amino acids with low frequencies. In this study, we propose a method to remove the noise information in the PSFM by removing the amino acids with low frequencies called Top frequency profile (TFP). Three new matrix transformation methods, including Autocross covariance (ACC) transformation, Tri-gram, and K-separated bigram (KSB), are performed on these profiles to convert them into fixed length feature vectors. Combined with Support Vector Machines (SVMs), the predictors are constructed. Evaluated on two benchmark datasets, and experimental results show that these proposed methods outperform other state-of-the-art predictors.Keywords: protein remote homology detection, protein fold recognition, top frequency profile, support vector machines
Procedia PDF Downloads 1243488 C-Reactive Protein in Patients with Type 2 Diabetes Mellitus
Authors: Athar Hussain Memon
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Objectives: We tried to determine the frequency of raised C-reactive protein (CRP) in patients with type 2 diabetes mellitus. Patients and Methods: This cross-sectional descriptive study of six months study was conducted at Liaquat University Hospital Hyderabad from March 2013 to August 2013. All diabetic patients of ≥35 years age of either gender for >01 year duration visited at OPD were evaluated for C-reactive protein and their glycemic status by hemoglobin A1c. The data was analyzed in SPSS and the frequency and percentage were calculated. Results: During six month study period, total 100 diabetic patients were evaluated for C-reactive protein. The majority of patients were from urban areas 75/100 (75%). The mean ±SD for age of patients with diabetes mellitus was 51.63±7.82. The mean age ±SD of patient with raised CRP was 53±7.21. The mean ±SD for HbA1c in patients with raised CRP is 9.55±1.73. The mean random blood sugar level in patients with raised CRP was 247.42 ± 6.62. The majority of subjects were of 50-69 years of age group with female predominance (p=0.01) while the CRP was raised in 70 (70%) patients in relation to age (p=0.02) and gender (p=0.01), respectively. Both HbA1c and CRP were raised in 64.9% (p=0.04) in patients with type 2 diabetes mellitus. The mean ±SD of CRP was 5.8±1.21 while for male and female individuals with raised CRP was 3.52±1.22 and 5.7±1.63, respectively. Conclusions: The raised CRP was observed in patients with type 2 diabetes mellitus.Keywords: diabetes mellitus, C-reactive protein, hemoglobin A1c, diabetes and metabolism
Procedia PDF Downloads 4123487 Arabic Text Classification: Review Study
Authors: M. Hijazi, A. Zeki, A. Ismail
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An enormous amount of valuable human knowledge is preserved in documents. The rapid growth in the number of machine-readable documents for public or private access requires the use of automatic text classification. Text classification can be defined as assigning or structuring documents into a defined set of classes known in advance. Arabic text classification methods have emerged as a natural result of the existence of a massive amount of varied textual information written in the Arabic language on the web. This paper presents a review on the published researches of Arabic Text Classification using classical data representation, Bag of words (BoW), and using conceptual data representation based on semantic resources such as Arabic WordNet and Wikipedia.Keywords: Arabic text classification, Arabic WordNet, bag of words, conceptual representation, semantic relations
Procedia PDF Downloads 4253486 Production of Recombinant VP2 Protein of Canine Parvovirus Type 2c Using Baculovirus Expression System
Authors: Jae Young Song, In-Ohk Ouh, Seyeon Park, Byeong Sul Kang, Soo Dong Cho, In-Soo Cho
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Canine parvovirus (CPV) is a major pathogen of diarrhea disease in dogs. CPV type 2 has three of antigenic variants such as 2a, 2b, and 2c. CPV constructs a small non-enveloped, icosahedral capsid that contains single-stranded DNA. It has capsids that two largely overlapping virion proteins (VP), VP1 (82 kDa), and VP2 (65 kDa). Baculoviruses are insect pathogens that regulate insect populations in nature and are being successfully used to control insect pests. The proteins produced in the baculovirus-expression system are used for instance for functional studies, vaccine preparations, or diagnostics. The vaccines produced by baculovirus-expression system showed elicitation of antibodies. The recombinant baculovirus infected SF9 cells showed broken shape. The recombinant VP2 proteins from cell pellet or supernatant were confirmed by western blotting. The result showed that the recombinant VP2 protein bands were appeared at 65 kDa molecular weight in both cell pellet and supernatant of infected SF9 cell. These results indicated that the recombinant baculovirus infected SF9 cell express the recombinant VP2 protein successfully. In addition, the expressed recombinant VP2 protein is secreted from cell to supernatant. The baculovirus expression system can be used to produce the VP2 protein of CPV 2c. In addition, the secretion property of the expression of VP2 protein may decrease the cost of production, because it can be skipped the cell breaking step. The produced VP2 protein could be used for vaccine and the agent of diagnostic tests. This study provides the foundation of the production of CPV 2c vaccine and the diagnostic agent.Keywords: baculovirus, canine parvovirus 2c, dog, Korea
Procedia PDF Downloads 1493485 Phase Transition in Iron Storage Protein Ferritin
Authors: Navneet Kaur, S. D. Tiwari
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Ferritin is a protein which present in the blood of mammals. It maintains the need of iron inside the body. It has an antiferromagnetic iron core, 7-8 nm in size, which is encapsulated inside a protein cage. The thickness of this protein shell is about 2-3 nm. This protein shell reduces the interaction among particles and make ferritin a model superparamagnet. The major composition of ferritin core is mineral ferrihydrite. The molecular formula of ferritin core is (FeOOH)8[FeOOPO3H2]. In this study, we discuss the phase transition of ferritin. We characterized ferritin using x-ray diffractometer, transmission electron micrograph, thermogravimetric analyzer and vibrating sample magnetometer. It is found that ferritin core is amorphous in nature with average particle size of 8 nm. The thermogravimetric and differential thermogravimetric analysis curves shows mass loss at different temperatures. We heated ferritin at these temperatures. It is found that ferritin core starts decomposing after 390^o C. At 1020^o C, the ferritin core is finally converted to alpha phase of iron oxide. Magnetization behavior of final sample clearly shows the iron oxyhydroxide core is completely converted to alpha iron oxide.Keywords: Antiferromagnetic, Ferritin, Phase, Superparamagnetic
Procedia PDF Downloads 1173484 Evaluation Methods for Question Decomposition Formalism
Authors: Aviv Yaniv, Ron Ben Arosh, Nadav Gasner, Michael Konviser, Arbel Yaniv
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This paper introduces two methods for the evaluation of Question Decomposition Meaning Representation (QDMR) as predicted by sequence-to-sequence model and COPYNET parser for natural language questions processing, motivated by the fact that previous evaluation metrics used for this task do not take into account some characteristics of the representation, such as partial ordering structure. To this end, several heuristics to extract such partial dependencies are formulated, followed by the hereby proposed evaluation methods denoted as Proportional Graph Matcher (PGM) and Conversion to Normal String Representation (Nor-Str), designed to better capture the accuracy level of QDMR predictions. Experiments are conducted to demonstrate the efficacy of the proposed evaluation methods and show the added value suggested by one of them- the Nor-Str, for better distinguishing between high and low-quality QDMR when predicted by models such as COPYNET. This work represents an important step forward in the development of better evaluation methods for QDMR predictions, which will be critical for improving the accuracy and reliability of natural language question-answering systems.Keywords: NLP, question answering, question decomposition meaning representation, QDMR evaluation metrics
Procedia PDF Downloads 763483 Interaction of Histone H1 with Chromatin-associated Protein HMGB1 Studied by Microscale Thermophoresis
Authors: Michal Štros, Eva Polanská, Šárka Pospíšilová
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HMGB1 is an architectural protein in chromatin, acting also as a signaling molecule outside the cell. Recent reports from several laboratories provided evidence that a number of both the intracellular and extracellular functions of HMGB1 may depend on redox-sensitive cysteine residues of the protein. MALDI-TOF analysis revealed that mild oxidization of HMGB1 resulted in a conformational change of the protein due to formation of an intramolecular disulphide bond by opposing Cys23 and Cys45 residues. We have demonstrated that redox state of HMGB1 could significantly modulate the ability of the protein to bind and bend DNA. We have also shown that reduced HMGB1 could easily displace histone H1 from DNA, while oxidized HMGB1 had limited capacity for H1 displacement. Using microscale thermophoresis (MST) we have further studied mechanism of HMGB1 interaction with histone H1 in free solution or when histone H1 was bound to DNA. Our MST analysis indicated that reduced HMGB1 exhibited in free solution > 1000 higher affinity of for H1 (KD ~ 4.5 nM) than oxidized HMGB1 (KD <10 M). Finally, we present a novel mechanism for the HMGB1-mediated modulation of histone H1 binding to DNA.Keywords: HMGB1, histone H1, redox state, interaction, cross-linking, DNA bending, DNA end-joining, microscale thermophoresis
Procedia PDF Downloads 3333482 Evidence of Paternal Protein Provisioning During Male Pregnancy in the Seahorse, Hippocampus Abdominalis
Authors: Zoe M. G. Skalkos, Sam N. Dowland, James U. Van Dyke, Camilla. M. Whittington
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Syngnathid fishes (seahorses, pipefishes, and seadragons) are unique because embryos develop on or in the male in a specialised brooding structure. Many seahorse species are endangered or vulnerable, while others are popular in the ornamental fish trade. Seahorses are capable of nutrient provisioning (patrotrophy) of lipids during pregnancy via their fully enclosed brood pouch. Protein is vital for gene regulation and tissue growth during embryogenesis. We tested the hypothesis that protein is paternally transported to developing embryos during pregnancy in the Australian Pot-bellied seahorse, Hippocampus abdominalis. We compared the dry masses and nitrogen content in recently fertilised H. abdominalis embryos and newborns. We calculated an updated patrotrophy index, 1.34, but without a significant difference in dry mass between the two developmental stages. There was, however, a significant increase in total protein content from recently fertilised embryos to neonates. This suggests paternal protein transport is essential for H. abdominalis embryogenesis because protein yolk reserves are depleted by embryonic metabolism, and supplementation is required. This study is the first to provide evidence for paternal protein transport during pregnancy in seahorses. It furthers our understanding of the paternal influence on embryonic development in male pregnancy and how a protein-deficient diet during pregnancy may limit the allocation of resources to embryos, reducing offspring fitness. This research contributes to a deeper understanding of the fundamental reproductive biology of seahorses, which can help improve conservation and farming production outcomes.Keywords: brood pouch, embryonic provisioning, nitrogen, parentotrophy, paternal investment, reproduction
Procedia PDF Downloads 1033481 Insight into Structure and Functions of of Acyl CoA Binding Protein of Leishmania major
Authors: Rohit Singh Dangi, Ravi Kant Pal, Monica Sundd
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Acyl-CoA binding protein (ACBP) is a housekeeping protein which functions as an intracellular carrier of acyl-CoA esters. Given the fact that the amastigote stage (blood stage) of Leishmania depends largely on fatty acids as the energy source, of which a large part is derived from its host, these proteins might have an important role in its survival. In Leishmania major, genome sequencing suggests the presence of six ACBPs, whose function remains largely unknown. For functional and structural characterization, one of the ACBP genes was cloned, and the protein was expressed and purified heterologously. Acyl-CoA ester binding and stoichiometry were analyzed by isothermal titration calorimetry and Dynamic light scattering. Our results shed light on high affinity of ACBP towards longer acyl-CoA esters, such as myristoyl-CoA to arachidonoyl-CoA with single binding site. To understand the binding mechanism & dynamics, Nuclear magnetic resonance assignments of this protein are being done. The protein's crystal structure was determined at 1.5Å resolution and revealed a classical topology for ACBP, containing four alpha-helical bundles. In the binding pocket, the loop between the first and the second helix (16 – 26AA) is four residues longer from other extensively studied ACBPs (PfACBP) and it curls upwards towards the pantothenate moiety of CoA to provide a large tunnel space for long acyl chain insertion.Keywords: acyl-coa binding protein (ACBP), acyl-coa esters, crystal structure, isothermal titration, calorimetry, Leishmania
Procedia PDF Downloads 4473480 The Effect of Dendrobium nobile Lindl. Alkaloids on the Blood Glucose and Amyloid Precursor Protein Metabolic Pathways in Db/Db Mice
Authors: Juan Huang, Nanqu Huang, Jingshan Shi, Yu Qiu
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Objectives: There are pathophysiological connections between type 2 diabetes mellitus (T2DM) and Alzheimer's disease (AD), and research on drugs with hypoglycemic and beta-amyloid (Aβ)-clearing effects have great therapeutic potential for AD. Dendrobium nobile Lindl. Alkaloids (DNLA) as one of the active compounds of Dendrobium nobile Lindl. In this study, we attempted to verify the hypoglycemic effect and investigate the effects of DNLA on the amyloid precursor protein (APP) metabolic pathway of the hippocampus in db/db mice. Methods: 4-weeks-old male C57BL/KsJ mice were the control group. And the same age and sexuality db/db mice were: model, DNLA-L (20 mg/kg), DNLA-M (40 mg/kg), and DNLA-H (80 mg/kg). After, mice were treated with different concentrations of DNLA for 17 weeks. The fasting blood glucose (FBG) was detected by glucose oxidase assay every week from the 4th to last week. The protein expression of β-amyloid 1-42 (Aβ1-42), β-site amyloid precursor protein-cleaving enzyme 1 (BACE1), and APP were examined by Western blotting. Results: The concentration of FBG and the protein expression of Aβ1-42, BACE1, and APP were increased in the hippocampus of the model group. Moreover, DNLA not only significantly decreased the concentration of FBG but also reduced the protein expressions of Aβ1-42, BACE1 and APP in the hippocampus of db/db mice in a dose-dependent manner. Conclusions: DNLA can decrease the protein expressions of Aβ1-42 in the hippocampus of db/db mice, and the mechanism may be involved in the APP metabolic pathway.Keywords: Alzheimer's disease, type 2 diabetes mellitus, β-site amyloid precursor protein-cleaving enzyme 1, traditional Chinese medicines, beta-amyloid
Procedia PDF Downloads 2483479 Persistent Ribosomal In-Frame Mis-Translation of Stop Codons as Amino Acids in Multiple Open Reading Frames of a Human Long Non-Coding RNA
Authors: Leonard Lipovich, Pattaraporn Thepsuwan, Anton-Scott Goustin, Juan Cai, Donghong Ju, James B. Brown
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Two-thirds of human genes do not encode any known proteins. Aside from long non-coding RNA (lncRNA) genes with recently-discovered functions, the ~40,000 non-protein-coding human genes remain poorly understood, and a role for their transcripts as de-facto unconventional messenger RNAs has not been formally excluded. Ribosome profiling (Riboseq) predicts translational potential, but without independent evidence of proteins from lncRNA open reading frames (ORFs), ribosome binding of lncRNAs does not prove translation. Previously, we mass-spectrometrically documented translation of specific lncRNAs in human K562 and GM12878 cells. We now examined lncRNA translation in human MCF7 cells, integrating strand-specific Illumina RNAseq, Riboseq, and deep mass spectrometry in biological quadruplicates performed at two core facilities (BGI, China; City of Hope, USA). We excluded known-protein matches. UCSC Genome Browser-assisted manual annotation of imperfect (tryptic-digest-peptides)-to-(lncRNA-three-frame-translations) alignments revealed three peptides hypothetically explicable by 'stop-to-nonstop' in-frame replacement of stop codons by amino acids in two ORFs of the lncRNA MMP24-AS1. To search for this phenomenon genomewide, we designed and implemented a novel pipeline, matching tryptic-digest spectra to wildcard-instead-of-stop versions of repeat-masked, six-frame, whole-genome translations. Along with singleton putative stop-to-nonstop events affecting four other lncRNAs, we identified 24 additional peptides with stop-to-nonstop in-frame substitutions from multiple positive-strand MMP24-AS1 ORFs. Only UAG and UGA, never UAA, stop codons were impacted. All MMP24-AS1-matching spectra met the same significance thresholds as high-confidence known-protein signatures. Targeted resequencing of MMP24-AS1 genomic DNA and cDNA from the same samples did not reveal any mutations, polymorphisms, or sequencing-detectable RNA editing. This unprecedented apparent gene-specific violation of the genetic code highlights the importance of matching peptides to whole-genome, not known-genes-only, ORFs in mass-spectrometry workflows, and suggests a new mechanism enhancing the combinatorial complexity of the proteome. Funding: NIH Director’s New Innovator Award 1DP2-CA196375 to LL.Keywords: genetic code, lncRNA, long non-coding RNA, mass spectrometry, proteogenomics, ribo-seq, ribosome, RNAseq
Procedia PDF Downloads 2333478 Fluorescence Spectroscopy of Lysozyme-Silver Nanoparticles Complex
Authors: Shahnaz Ashrafpour, Tahereh Tohidi Moghadam, Bijan Ranjbar
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Identifying the nature of protein-nanoparticle interactions and favored binding sites is an important issue in functional characterization of biomolecules and their physiological responses. Herein, interaction of silver nanoparticles with lysozyme as a model protein has been monitored via fluorescence spectroscopy. Formation of complex between the biomolecule and silver nanoparticles (AgNPs) induced a steady state reduction in the fluorescence intensity of protein at different concentrations of nanoparticles. Tryptophan fluorescence quenching spectra suggested that silver nanoparticles act as a foreign quencher, approaching the protein via this residue. Analysis of the Stern-Volmer plot showed quenching constant of 3.73 µM−1. Moreover, a single binding site in lysozyme is suggested to play role during interaction with AgNPs, having low affinity of binding compared to gold nanoparticles. Unfolding studies of lysozyme showed that complex of lysozyme-AgNPs has not undergone structural perturbations compared to the bare protein. Results of this effort will pave the way for utilization of sensitive spectroscopic techniques for rational design of nanobiomaterials in biomedical applications.Keywords: nanocarrier, nanoparticles, surface plasmon resonance, quenching fluorescence
Procedia PDF Downloads 3293477 Spectrofluorometric Studies on the Interactions of Bovine Serum Albumin with Dimeric Cationic Surfactants
Authors: Srishti Sinha, Deepti Tikariha, Kallol K. Ghosh
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Over the past few decades protein-surfactant interactions have been a subject of extensive studies as they are of great importance in wide variety of industries, biological, pharmaceutical and cosmetic systems. Protein-surfactant interactions have been explored the effect of surfactants on structure of protein in the form of solubilization and denaturing or renaturing of protein. Globular proteins are frequently used as functional ingredients in healthcare and pharmaceutical products, due to their ability to catalyze biochemical reactions, to be adsorbed on the surface of some substance and to bind other moieties and form molecular aggregates. One of the most widely used globular protein is bovine serum albumin (BSA), since it has a well-known primary structure and been associated with the binding of many different categories of molecules, such as dyes, drugs and toxic chemicals. Protein−surfactant interactions are usually dependent on the surfactant features. Most of the research has been focused on single-chain surfactants. More recently, the binding between proteins and dimeric surfactants has been discussed. In present study interactions of one dimeric surfactant Butanediyl-1,4-bis (dimethylhexadecylammonium bromide) (16-4-16, 2Br-) and the corresponding single-chain surfactant cetyl trimethylammonium bromide (CTAB) with bovine serum albumin (BSA) have been investigated by surface tension and spectrofluoremetric methods. It has been found that the bindings of all gemini surfactant to BSA were cooperatively driven by electrostatic and hydrophobic interactions. The gemini surfactant carrying more charges and hydrophobic tails, showed stronger interactions with BSA than the single-chain surfactant.Keywords: bovine serum albumin, gemini surfactants, hydrophobic interactions, protein surfactant interaction
Procedia PDF Downloads 5063476 Alterations of Malondialdehyde and Heat Shock Protein-27 in Sheep with Naturally Infected Liver Cystic Echinococcosis
Authors: K. Azimzadeh, S. Rasouli
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The present study investigates whether malondialdehyde (MDA) and heat shock protein-27 (HSP-27) are altered in sheep with cystic echinococcosis (CE). For this purpose, forty parasitized and thirty healthy sheep were selected based on severe cystic form observation in liver and lack of blood parasite along with no cystic conformation in carcass respectively. The results revealed a significant decrease (p<0.01) in albumin (Alb) and total plasma protein (TPP) and a significant increase (p<0.01) in HSP-27, MDA, total bilirubin and unconjugated bilirubin in the infected group compared with healthy ones.The results indicate low levels of TPP and Alb reveal liver damage in suffered sheep and MDA elevation demonstrates oxidative stress in infected group. In addition, HSP-27 enhancement may attribute to disease-induced stress conditions.Keywords: malondialdehyde, heat shock protein-27, Echinococcosis, blood parasites
Procedia PDF Downloads 6073475 A Sparse Representation Speech Denoising Method Based on Adapted Stopping Residue Error
Authors: Qianhua He, Weili Zhou, Aiwu Chen
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A sparse representation speech denoising method based on adapted stopping residue error was presented in this paper. Firstly, the cross-correlation between the clean speech spectrum and the noise spectrum was analyzed, and an estimation method was proposed. In the denoising method, an over-complete dictionary of the clean speech power spectrum was learned with the K-singular value decomposition (K-SVD) algorithm. In the sparse representation stage, the stopping residue error was adaptively achieved according to the estimated cross-correlation and the adjusted noise spectrum, and the orthogonal matching pursuit (OMP) approach was applied to reconstruct the clean speech spectrum from the noisy speech. Finally, the clean speech was re-synthesised via the inverse Fourier transform with the reconstructed speech spectrum and the noisy speech phase. The experiment results show that the proposed method outperforms the conventional methods in terms of subjective and objective measure.Keywords: speech denoising, sparse representation, k-singular value decomposition, orthogonal matching pursuit
Procedia PDF Downloads 4983474 In vitro and invivo Antioxidant Studies of Grewia crenata Leaves Extract in Albino Rats
Authors: A. N.Ukwuani, A. K. Abdulfatah
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G. crenata is used locally for the treatment of fractured bones, wound healing and inflammatory conditions. In vitro and in vivo antioxidant activity of hydromethanolic extracts of the leaves of G. crenata were assessed. The phytochemical analysis shows the presence of phenols, flavonoids, saponins, cardiac glycosides and tannins. An in vitro quantitative analysis of phenols, flavonoids and tannins respectively were (164±1.20, 199±0.88 and 88.67±0.88 mg/100g FW). In vivo studies of hydromethanolic extract demonstrated a dose dependent increase in hepatic superoxide dismutase (1.14±0.14, 2.13±0.11, 2.55±0.11 U/mg Protein) with improvement in hepatic glutathione (6.98±0.42, 8.91±0.37, 11.07±0.46 µM/mg Protein) and Catalase (4.47±0.05, 6.24±0.02, 7.17±0.04 U/mg Protein) and Total protein (6.18±0.08, 6.69±0.18, 7.27±0.16 mg/ml) respectively at 100-300mg/kg body weight Grewia crenata leaves when compared to the control and standard drug. It can be concluded from the present findings of that G. crenata leaves possess antioxidant potential.Keywords: Grewia crenata, antioxidant, hydromethanolic extract, in vivo, in vitro
Procedia PDF Downloads 5523473 Functional Cell Surface Display Using Ice Nucleation Protein from Erwina ananas on Escherischia coli
Authors: Mei Yuin Joanne Wee, Rosli Md. Illias
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Cell surface display is the expression of a protein with an anchoring motif on the surface of the cell. This approach offers advantages when used in bioconversion in terms of easier purification steps and more efficient enzymatic reaction. A surface display system using ice nucleation protein (InaA) from Erwina ananas as an anchoring motif has been constructed to display xylanase (xyl) on the surface of Escherischia coli. The InaA was truncated so that it is made up of the N- and C-terminal domain (INPANC-xyl) and it has successfully directed xylanase to the surface of the cell. A study was also done on xylanase fused to two other ice nucleation proteins, InaK (INPKNC-xyl) and InaZ (INPZNC-xyl) from Pseudomonas syringae KCTC 1832 and Pseudomonas syringae S203 respectively. Surface localization of the fusion protein was verified using SDS-PAGE and Western blot on the cell fractions and all anchoring motifs were successfully displayed on the outer membrane of E. coli. Upon comparison, whole-cell activity of INPANC-xyl was more than six and five times higher than INPKNC-xyl and INPZNC-xyl respectively. Furthermore, the expression of INPANC-xyl on the surface of E. coli did not inhibit the growth of the cell. This is the first report of surface display system using ice nucleation protein, InaA from E. ananas. From this study, this anchoring motif offers an attractive alternative to the current surface display systems.Keywords: cell surface display, Escherischia coli, ice nucleation protein, xylanase
Procedia PDF Downloads 3893472 Representation of Pashtuns in the Context of Terrorism: A Comparative Study of Bollywood and Lollywood Movies After 9/11
Authors: Aamir Ayub, Yasir Shehzad, Shakeel Ahmad
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This research paper aims to understand how the Pashtuns have been represented in relationship to terrorism in post-9/11 Bollywood and Lollywood movies. It focuses particularly on ‘Torbaaz’ from Bollywood and ‘Waar’ from Lollywood in order to define the nature of Pashtun characterization, the functioning of intelligence agencies, as well as the socio-political side of the represented narratives. In this research, the analytical approach developed is applied to contemplate how these films represent or fail to represent Pashtun identity, taking into consideration the cultural, historical and social dimensions. The study also aims to examine the effects of the media, particularly on the different ethnic groups’ perceptions of terrorism. In this case, it covers how the movie relates actual events in society – specifically, socio-political – to the messages in the film regarding the Pashtun people and their portrayal. Such elements may constitute the portrayal of intelligence agencies and their fight against terrorism, state-security dynamics, and the Pashtun society. In conclusion, this research paper focuses on the representation of Pashtuns in films after 9/11 and addresses the issue concerning the representation of ethnic groups in the method of the theme of terrorism. It provides ideas about the role of media in influencing the mind of the society and their attitude towards certain communities after geopolitics upheavals.Keywords: pashtun representation, terrorism, 9/11 attacks, socio-political implications, ethnic representation in media
Procedia PDF Downloads 203471 Extraction of Rice Bran Protein Using Enzymes and Polysaccharide Precipitation
Authors: Sudarat Jiamyangyuen, Tipawan Thongsook, Riantong Singanusong, Chanida Saengtubtim
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Rice is a staple food as well as exported commodity of Thailand. Rice bran, a 10.5% constituent of rice grain, is a by-product of rice milling process. Rice bran is normally used as a raw material for rice bran oil production or sold as feed with a low price. Therefore, this study aimed to increase value of defatted rice bran as obtained after extracting of rice bran oil. Conventionally, the protein in defatted rice bran was extracted using alkaline extraction and acid precipitation, which results in reduction of nutritious components in rice bran. Rice bran protein concentrate is suitable for those who are allergenic of protein from other sources eg. milk, wheat. In addition to its hypoallergenic property, rice bran protein also contains good quantity of lysine. Thus it may act as a suitable ingredient for infant food formulations while adding variety to the restricted diets of children with food allergies. The objectives of this study were to compare properties of rice bran protein concentrate (RBPC) extracted from defatted rice bran using enzymes together with precipitation step using polysaccharides (alginate and carrageenan) to those of a control sample extracted using a conventional method. The results showed that extraction of protein from rice bran using enzymes exhibited the higher protein recovery compared to that extraction with alkaline. The extraction conditions using alcalase 2% (v/w) at 50 C, pH 9.5 gave the highest protein (2.44%) and yield (32.09%) in extracted solution compared to other enzymes. Rice bran protein concentrate powder prepared by a precipitation step using alginate (protein in solution: alginate 1:0.006) exhibited the highest protein (27.55%) and yield (6.62%). Precipitation using alginate was better than that of acid. RBPC extracted with alkaline (ALK) or enzyme alcalase (ALC), then precipitated with alginate (AL) (samples RBP-ALK-AL and RBP-ALC-AL) yielded the precipitation rate of 75% and 91.30%, respectively. Therefore, protein precipitation using alginate was then selected. Amino acid profile of control sample, and sample precipitated with alginate, as compared to casein and soy protein isolated, showed that control sample showed the highest content among all sample. Functional property study of RBP showed that the highest nitrogen solubility occurred in pH 8-10. There was no statically significant between emulsion capacity and emulsion stability of control and sample precipitated by alginate. However, control sample showed a higher of foaming and lower foam stability compared to those of sample precipitated with alginate. The finding was successful in terms of minimizing chemicals used in extraction and precipitation steps in preparation of rice bran protein concentrate. This research involves in a production of value-added product in which the double amount of protein (28%) compared to original amount (14%) contained in rice bran could be beneficial in terms of adding to food products eg. healthy drink with high protein and fiber. In addition, the basic knowledge of functional property of rice bran protein concentrate was obtained, which can be used to appropriately select the application of this value-added product from rice bran.Keywords: alginate, carrageenan, rice bran, rice bran protein
Procedia PDF Downloads 2933470 Life Expansion: Visual Autobiography, Identity, Representation and the Degrees of Fictionalization of the Self on Instagram
Authors: Pablo De Macedo Silveira Vallejos
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This article aims to observe autobiographical and visual narrative practices among users on Instagram. In this way, the work proposes to reflect on how image resources are used to develop edited representations of the self in that social network. The research aims to explore the uses of editing and the degrees of fictionalization present on Instagram.Keywords: autobiography, visual narratives, representation, fiction, social media
Procedia PDF Downloads 723469 Beauty Representation and Body Politic of Women Writers in Magdalene
Authors: Putri Alya Ramadhani
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This research analysed how women writers represent their beauty in a platform called Magdalene. With the vision “Supporting diversity, empowering minds,” Magdalene is a new media that seeks to represent women's voices rarely heard in mainstream media. This research elaborates further on how women writers, through their writing, use their body politic to subvert patriarchal values. This research used a qualitative method with an explorative design by using text analysis based on the representation theory of Stuart Hall and in-dept-interview with Women Writers in Magdalene. The result illustrated that women writers represent their beauty in Magdalene to subvert body and beauty-representation in mainstream discourse. Furthermore, the authors have identified an identity negotiation as tension from inevitable oppression and power towards and from women’s bodies. In addition, Women Writers showed the power of their bodies through the redefinition of beauty practices and self. Hence, they subvert body dichotomy to redefine body values in society. In conclusion, this study shows various representations of beauty and body that are underrepresented in the mainstream media through the innovative new medium, Magdalena.Keywords: women writers, beauty-representation, body politic, new media, identity negotiation
Procedia PDF Downloads 1743468 “Lightyear” – The Battle for LGBTQIA+ Representation Behind Disney/Pixar’s Failed Blockbuster
Authors: Ema Vitória Fonseca Lavrador
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In this work, we intend to explore the impact that the film "Lightyear" (2022) had on the social context of its production, distribution, and reception. This film, produced by Walt Disney Animation Studios and Pixar Animation Studios, depicts the story of Buzz Lightyear, a Space Ranger from which the character of the same name in the "Toy Story" film franchise is based. This prequel was predicted to be the blockbuster of the year, but it was a financial fiasco and the subject of numerous controversies, which also caused it to be drowned out by the film "Minions: The Rise of Gru" (2022). The reason for its failure is not based on the film's narrative or quality but on its controversial context for being a commitment to LGBTQIA+ representation in an unexpected way, by featuring a same-sex couple and showing a kiss shared by them. This representation cost Disney distribution in countries against LGBTQIA+ representation in media and involved Disney in major disagreements with fans and politicians, especially for being a direct opposition to the Florida House Bill 1557, also called the “Don't Say Gay” bill. Many major companies have taken a stand against this law because it jeopardizes the safety of the LGBTQIA+ community, and, although Disney initially cut the kiss off the film, pressure from the staff and audience resulted in unprecedented progress. For featuring a brief homosexual kiss, its exhibition was banned in several countries and discouraged by the same public that was previously the focus of Disney's attention, as this is a conservative “family-friendly” branded company. We believe it is relevant to study the case of "Lightyear" because it is a work that raises awareness and promotes representation of communities affected during the dark times while less legislation is being approved to protect the rights and safety of queer people.Keywords: Don’t Say Gay” bill, gender stereotypes, LGBTQIA+ representation, lightyear, Disney/Pixar
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