Search results for: retinoic acid signaling pathway
3637 Kinetic Study on Extracting Lignin from Black Liquor Using Deep Eutectic Solvents
Authors: Fatemeh Saadat Ghareh Bagh, Srimanta Ray, Jerald Lalman
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Lignin, the largest inventory of organic carbon with a high caloric energy value is a major component in woody and non-woody biomass. In pulping mills, a large amount of the lignin is burned for energy. At the same time, the phenolic structure of lignin enables it to be converted to value-added compounds.This study has focused on extracting lignin from black liquor using deep eutectic solvents (DESs). Therefore, three choline chloride (ChCl)-DESs paired with lactic acid (LA) (1:11), oxalic acid.2H₂O (OX) (1:4), and malic acid (MA) (1:3) were synthesized at 90oC and atmospheric pressure. The kinetics of lignin recovery from black liquor using DES was investigated at three moderate temperatures (338, 353, and 368 K) at time intervals from 30 to 210 min. The extracted lignin (acid soluble lignin plus Klason lignin) was characterized by Fourier transform infrared spectroscopy (FTIR). The FTIR studies included comparing the extracted lignin with a model Kraft lignin. The extracted lignin was characterized spectrophotometrically to determine the acid soluble lignin (ASL) [TAPPI UM 250] fraction and Klason lignin was determined gravimetrically using TAPPI T 222 om02. The lignin extraction reaction using DESs was modeled by first-order reaction kinetics and the activation energy of the process was determined. The ChCl:LA-DES recovered lignin was 79.7±2.1% at 368K and a DES:BL ratio of 4:1 (v/v). The quantity of lignin extracted for the control solvent, [emim][OAc], was 77.5+2.2%. The activation energy measured for the LA-DES system was 22.7 KJ mol⁻¹, while the activation energy for the OX-DES and MA-DES systems were 7.16 KJ·mol⁻¹ and 8.66 KJ·mol⁻¹ when the total lignin recovery was 75.4 ±0.9% and 62.4 ±1.4, % respectively.Keywords: black liquor, deep eutectic solvents, kinetics, lignin
Procedia PDF Downloads 1513636 Evaluation of Phytochemical and Fatty Acids Content and Composition in Iranian Borage (Echium amoenum) in Different Habitate of Iran
Authors: Esmaeil Babakhanzadeh Sajirani, Mohamadjavad Shakouri
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Iranian Gole GavZaban (Echium amoenum fich & mey), is one of the most important medicinal plant in north of iran . is dry petals used for tonic, tranquillizer, diaphoretic, cough suppressant and a remedy for sore throat in treditional Iranian medicine. This study is the report about the analysis of phytochemical and seeds oil of Echium amoenum's in different habitates and accessions of Iran. The results showed that the oil content of seeds was 36% and eleven fatty acids were identified and quantified by gas chromatography (GC). The major fatty acids wereα-Linolenicacid (39.99), Linoleic acid (20.86), linolenic acid (20%) and Oleic acid (15.36) respectively. The amount of phenols, tannins, flavonoids and anthocyanins with increasing height, increased amount of these compounds. So that the highest rates of these compounds were observed at an altitude of 2125 meters in ciposht accession.Keywords: accession, phytochemical, oil components, Iranian borage
Procedia PDF Downloads 2573635 Investigation of Parameters Affecting Copper Recovery from Brass Melting Dross
Authors: Sercan Basit, Muhlis N. Sarıdede
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Metal amounts of copper based compounds in the various wastes have been recovered successfully by hydrometallurgical treatment methods in the literature. X-ray diffraction pattern of the brass melting slag demonstrates that it contains sufficient amount of recoverable copper. Recovery of copper from brass melting dross by sulfuric acid leaching and the effect of temperature and acid and oxidant concentration on recovery rate of copper have been investigated in this study. Experiments were performed in a temperature-controlled reactor in sulfuric acid solution in different molarities using solid liquid ratio of 100 g/L, with leaching time of 300 min. Temperature was changed between 25 °C and 80 °C and molarity was between 0.5 and 3M. The results obtained showed that temperature has important positive effect on recovery whereas it decreases with time. Also copper was recovered in larger amounts from brass dross in the presence of H2O2 as an oxidant according to the case that oxidant was not used.Keywords: brass dross, copper recovery, hydrogen peroxide, leaching
Procedia PDF Downloads 3363634 Treatment of Acid Mine Drainage with Modified Fly Ash
Authors: Sukla Saha, Alok Sinha
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Acid mine drainage (AMD) is the generation of acidic water from active as well as abandoned mines. AMD generates due to the oxidation of pyrites present in the rock in mining areas. Sulfur oxidizing bacteria such as Thiobacillus ferrooxidans acts as a catalyst in this oxidation process. The characteristics of AMD is extreme low pH (2-3) with elevated concentration of different heavy metals such as Fe, Al, Zn, Mn, Cu and Co and anions such sulfate and chloride. AMD contaminate the ground water as well as surface water which leads to the degradation of water quality. Moreover, it carries detrimental effect for aquatic organism and degrade the environment. In the present study, AMD is treated with fly ash, modified with alkaline agent (NaOH). This modified fly ash (MFA) was experimentally proven as a very effective neutralizing agent for the treatment of AMD. It was observed that pH of treated AMD raised to 9.22 from 1.51 with 100g/L of MFA dose. Approximately, 99% removal of Fe, Al, Mn, Cu and Co took place with the same MFA dose. The treated water comply with the effluent discharge standard of (IS: 2490-1981).Keywords: acid mine drainage, heavy metals, modified fly ash, neutralization
Procedia PDF Downloads 1563633 Investigation on the Capacitive Deionization of Functionalized Carbon Nanotubes (F-CNTs) and Silver-Decorated F-CNTs for Water Softening
Authors: Khrizelle Angelique Sablan, Rizalinda De Leon, Jaeyoung Lee, Joey Ocon
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The impending water shortage drives us to find alternative sources of water. One of the possible solutions is desalination of seawater. There are numerous processes by which it can be done and one if which is capacitive deionization. Capacitive deionization is a relatively new technique for water desalination. It utilizes the electric double layer for ion adsorption. Carbon-based materials are commonly used as electrodes for capacitive deionization. In this study, carbon nanotubes (CNTs) were treated in a mixture of nitric and sulfuric acid. The silver addition was also facilitated to incorporate antimicrobial action. The acid-treated carbon nanotubes (f-CNTs) and silver-decorated f-CNTs (Ag@f-CNTs) were used as electrode materials for seawater deionization and compared with CNT and acid-treated CNT. The synthesized materials were characterized using TEM, EDS, XRD, XPS and BET. The electrochemical performance was evaluated using cyclic voltammetry, and the deionization performance was tested on a single cell with water containing 64mg/L NaCl. The results showed that the synthesized Ag@f-CNT-10 H could have better performance than CNT and a-CNT with a maximum ion removal efficiency of 50.22% and a corresponding adsorption capacity of 3.21 mg/g. It also showed antimicrobial activity against E. coli. However, the said material lacks stability as the efficiency decreases with repeated usage of the electrode.Keywords: capacitive deionization, carbon nanotubes, desalination, acid functionalization, silver
Procedia PDF Downloads 2343632 Identification of Genes Regulating Differentiation and Stemness of Human Mesenchymal Stem Cells for Gene Therapy in Regenerative Medicine
Authors: Tong Ming Liu
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Human mesenchymal stem cells (MSCs) represent the most used stem cells for clinical application, which have been used in over 1000 clinical trials to treat over 30 diseases due to multilineage differentiation potential, secretome and immunosuppression. Gene therapies of MSCs hold great promise in the treatment of many diseases due to enhanced MSC-based clinical outcomes. To identify genes for gene therapy of MSCs, by comparing gene expression profile before and after MSC differentiation following by functional screening, we have identified ZNF145 that regulated MSC differentiation. Forced expression of ZNF145 resulted in enhanced in vitro chondrogenesis of MSCs as an upstream factor of SOX9 and improved osteochondral repair upon implant into osteochondral defects in rodents. By comparing gene expression profile during differentiation of iPSCs toward MSCs, we also identified gene HOX regulating MSC stemness, which was much downregulated in late-passaged MSCs. Knockdown of this gene greatly compromised MSC stemness including abolished proliferation, decreased CFU-F, promoted senescence and reduced expression of cell surface antigens linked to the MSC phenotype. In addition, multi-linage differentiation was also greatly impaired. Notably, HOX overexpression resulted in improved multi-lineage differentiation. In the mechanism, HOX expression significantly deceased in late passage of MSCs compared with early passage of MSCs, correlating with MSC important genes. ChIP-seq data shown that HOX binds to genes related to MSC self-renewal and differentiation. Most importantly, most HOX binding sites are lost in late passage of MSCs. HOX exerts its effects by directing binding Twist1, one important gene of MSCs. The identification of the genes regulating MSC differentiation and stemness will provide and promising strategies for gene therapy of MSCs in regenerative medicine.Keywords: mesenchymal stem cell, novel transcription factor, stemness, gene therapy, cartilage repair, signaling pathway
Procedia PDF Downloads 603631 Development of Flipped Classroom in Chemistry on 'Acid-Base' for Enrichment Science Classroom Students
Authors: Waratthaya Maneerattana, Piyarat Dornbundit
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The study aimed to develop flipped classroom in Chemistry on ‘acid-base’ for high school students and study efficiency of students on academic achievement and problem-solving skills. The evaluating result from the experts showed that developed flipped classroom was ranked in high score level. The flipped classroom efficiency E1/E2 was higher than the criteria of 70/70. The flipped classroom was used by 24 students in grade 11 in the second semester of the academic year 2016 at Bodindecha (Sing Singhaseni) School. Sampling group was chosen using a purposive sampling approach. The results revealed that academic achievement and problem solving skills of students after studying flipped classroom was significantly higher at .05 level.Keywords: flipped classroom, acid-base, academic achievement, problem solving skill
Procedia PDF Downloads 3153630 The Influence of Lactic Acid Bacteria Combinations on Wheat Bread Quality
Authors: Vita Lele, Vadims Bartkevics, Iveta Pugajeva, Paulina Zavistanaviciute, Daiva Zadeike, Grazina Juodeikiene, Elena Bartkiene
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Different combinations of appropriate technological properties showing lactic acid bacteria (Pediococcus pentosaceus VLGL183 and Enterococcus pseudoavium VLGL 234, Lactobacillus plantarum VLGL135 and Pediococcus pentosaceus VLGL183, Pediococcus pentosaceus VLGL183 and Lactobacillus brevis VLGL173, Pediococcus pentosaceus VLGL183 and Leuconostoc mesenteroides VLGL242, Pediococcus pentosaceus VLGL183 and Lactobacillus curvatus VLGL51, Lactobacillus plantarum VLGL135 and Lactobacillus curvatus VLGL51) for wheat sourdough production were used, and the influence of different sourdoughs on wheat bread quality parameters was evaluated. The highest overall acceptability (135.8 mm in 140 mm hedonic scale) of the bread produced with L. plantarum VLGL135 and P. pentosaceus VLGL183 sourdough was established. Also, bread produced with above mention sourdough, has the highest specific volume, shape coefficient, moisture content, and porosity, 3.40 ml /g; 2.59, 33.7 %, and 76.6 %, respectively. It was found, that the used sourdoughs reduce acrylamide content in bread (from 29.5 to 67.2%), just, the isolated lactic acid bacteria strains could be recommended for higher quality and safer bread production.Keywords: acrylamide, lactic acid bacteria, quality, sourdough, wheat bread
Procedia PDF Downloads 1783629 Comparison of Fatty Acids Composition of Three Commercial Fish Species Farmed in the Adriatic Sea
Authors: Jelka Pleadin, Greta Krešić, Tina Lešić, Ana Vulić, Renata Barić, Tanja Bogdanović, Dražen Oraić, Ana Legac, Snježana Zrnčić
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Fish has been acknowledged as an integral component of a well-balanced diet, providing a healthy source of energy, high-quality proteins, vitamins, essential minerals and, especially, n-3 long-chain polyunsaturated fatty acids (n-3 LC PUFA), mainly eicosapentaenoic acid (20:5 n-3 EPA), and docosahexaenoicacid, (22:6 n-3 DHA), whose pleiotropic effects in terms of health promotion and disease prevention have been increasingly recognised. In this study, the fatty acids composition of three commercially important farmed fish species: sea bream (Sparus aurata), sea bass (Dicentrarchus labrax) and dentex (Dentex dentex) was investigated. In total, 60 fish samples were retrieved during 2015 (n = 30) and 2016 (n = 30) from different locations in the Adriatic Sea. Methyl esters of fatty acids were analysed using gas chromatography (GC) with flame ionization detection (FID). The results show that the most represented fatty acid in all three analysed species is oleic acid (C18:1n-9, OA), followed by linoleic acid (C18:2n-6, LA) and palmitic acid (C16:0, PA). Dentex was shown to have two to four times higher eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid content as compared to sea bream and sea bass. The recommended n-6/n-3 ratio was determined in all fish species but obtained results pointed to statistically significant differences (p < 0.05) in fatty acid composition among the analysed fish species and their potential as a dietary source of valuable fatty acids. Sea bass and sea bream had a significantly higher proportion of n-6 fatty acids, while dentex had a significantly higher proportion of n-3 (C18:4n-3, C20:4n-3, EPA, DHA) fatty acids. A higher hypocholesterolaemic and hypercholesterolaemic fatty acids (HH) ratio was determined for sea bass and sea bream, which comes as the consequence of a lower share of SFA determined in these two species in comparison to dentex. Since the analysed fish species vary in their fatty acids composition consumption of diverse fish species would be advisable. Based on the established lipid quality indicators, dentex, a fish species underutilised by the aquaculture, seems to be a highly recommendable and important source of fatty acids recommended to be included into the human diet.Keywords: dentex, fatty acids, farmed fish, sea bass, sea bream
Procedia PDF Downloads 3973628 Innovative Preparation Techniques: Boosting Oral Bioavailability of Phenylbutyric Acid Through Choline Salt-Based API-Ionic Liquids and Therapeutic Deep Eutectic Systems
Authors: Lin Po-Hsi, Sheu Ming-Thau
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Urea cycle disorders (UCD) are rare genetic metabolic disorders that compromise the body's urea cycle. Sodium phenylbutyrate (SPB) is a medication commonly administered in tablet or powder form to lower ammonia levels. Nonetheless, its high sodium content poses risks to sodium-sensitive UCD patients. This necessitates the creation of an alternative drug formulation to mitigate sodium load and optimize drug delivery for UCD patients. This study focused on crafting a novel oral drug formulation for UCD, leveraging choline bicarbonate and phenylbutyric acid. The active pharmaceutical ingredient-ionic liquids (API-ILs) and therapeutic deep eutectic systems (THEDES) were formed by combining these with choline chloride. These systems display characteristics like maintaining a liquid state at room temperature and exhibiting enhanced solubility. This in turn amplifies drug dissolution rate, permeability, and ultimately oral bioavailability. Incorporating choline-based phenylbutyric acid as a substitute for traditional SPB can effectively curtail the sodium load in UCD patients. Our in vitro dissolution experiments revealed that the ILs and DESs, synthesized using choline bicarbonate and choline chloride with phenylbutyric acid, surpassed commercial tablets in dissolution speed. Pharmacokinetic evaluations in SD rats indicated a notable uptick in the oral bioavailability of phenylbutyric acid, underscoring the efficacy of choline salt ILs in augmenting its bioavailability. Additional in vitro intestinal permeability tests on SD rats authenticated that the ILs, formulated with choline bicarbonate and phenylbutyric acid, demonstrate superior permeability compared to their sodium and acid counterparts. To conclude, choline salt ILs developed from choline bicarbonate and phenylbutyric acid present a promising avenue for UCD treatment, with the added benefit of reduced sodium load. They also hold merit in formulation engineering. The sustained-release capabilities of DESs position them favorably for drug delivery, while the low toxicity and cost-effectiveness of choline chloride signal potential in formulation engineering. Overall, this drug formulation heralds a prospective therapeutic avenue for UCD patients.Keywords: phenylbutyric acid, sodium phenylbutyrate, choline salt, ionic liquids, deep eutectic systems, oral bioavailability
Procedia PDF Downloads 1213627 Hydroponic Cultivation Enhances the Morpho-Physiological Traits and Quality Flower Production in Tagetes patula L
Authors: Ujala, Diksha Sharma, Mahinder Partap, Ashish R. Warghat, Bhavya Bhargava
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In soil-less agriculture, hydroponic is considered a potential farming system for the production of uniform quality plant material in significantly less time. Therefore, for the first time, the current investigation corroborates the effect of different cultivation conditions (open-field, poly-house, and hydroponic) on morpho-physiological traits, phenolic content, and essential oil components analysis in three flower color variants (yellow, scarlet red, and orange) of Tagetes patula. The results revealed that the maximum plant height, number of secondary branches, number of flowers, photosynthesis, stomatal conductance, and transpiration rate were observed under the hydroponic system as compared to other conditions. However, the maximum content of gallic acid (0.82 mg/g DW), syringic acid (3.98 mg/g DW), epicatechin (0.48 mg/g DW), p-coumaric acid (7.28 mg/g DW), protocatechuic acid (0.59 mg/g DW), ferulic acid (2.58 mg/g DW), and luteolin (8.24 mg/g DW) were quantified maximally under open-field conditions. However, under hydroponic conditions, the higher content of vanillic acid (0.43 mg/g DW), caffeic acid (0.49 mg/g DW), and quercetin (0.92 mg/g DW) were quantified. Moreover, a total of nineteen volatile components were identified in the essential oil of different flower color variants of T. patula cultivated under different conditions. The major reported volatile components in essential oil were (-)-caryophyllene oxide, trans-β-caryophyllene, trans-geraniol, 3 methyl-benzyl alcohol, and 2,2’:5’,2”-terthiophene. It has also been observed that the volatile component percentage range in all variants was observed in open-field (70.85 % to 90.54 %), poly-house (59.03 % to 77.93 %), and hydroponic (68.78 % to 89.41 %). In conclusion, the research highlighted that morpho-physiological performance with flower production was enhanced in the hydroponic system. However, phenolic content and volatile components were maximally observed in open-field conditions. However, significant results have been reported under hydroponic conditions in all studied parameters, so it could be a potential strategy for quality biomass production in T. patula.Keywords: Tagetes patula, cultivation conditions, hydroponic, morpho-physiology
Procedia PDF Downloads 793626 Opportunity Integrated Assessment Facilitating Critical Thinking and Science Process Skills Measurement on Acid Base Matter
Authors: Anggi Ristiyana Puspita Sari, Suyanta
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To recognize the importance of the development of critical thinking and science process skills, the instrument should give attention to the characteristics of chemistry. Therefore, constructing an accurate instrument for measuring those skills is important. However, the integrated instrument assessment is limited in number. The purpose of this study is to validate an integrated assessment instrument for measuring students’ critical thinking and science process skills on acid base matter. The development model of the test instrument adapted McIntire model. The sample consisted of 392 second grade high school students in the academic year of 2015/2016 in Yogyakarta. Exploratory factor analysis (EFA) was conducted to explore construct validity, whereas content validity was substantiated by Aiken’s formula. The result shows that the KMO test is 0.714 which indicates sufficient items for each factor and the Bartlett test is significant (a significance value of less than 0.05). Furthermore, content validity coefficient which is based on 8 expert judgments is obtained at 0.85. The findings support the integrated assessment instrument to measure critical thinking and science process skills on acid base matter.Keywords: acid base matter, critical thinking skills, integrated assessment instrument, science process skills, validity
Procedia PDF Downloads 3243625 Development of Broad Spectrum Nitrilase Biocatalysts and Bioprocesses for Nitrile Biotransformation
Authors: Avinash Vellore Sunder, Shikha Shah, Pramod P. Wangikar
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The enzymatic conversion of nitriles to carboxylic acids by nitrilases has gained significance in the green synthesis of several pharmaceutical precursors and fine chemicals. While nitrilases have been characterized from different sources, the industrial application requires the identification of nitrilases that possess higher substrate tolerance, wider specificity and better thermostability, along with the development of an efficient bioprocess for producing large amounts of nitrilase. To produce large amounts of nitrilase, we developed a fed-batch fermentation process on defined media for the high cell density cultivation of E. coli cells expressing the well-studied nitrilase from Alcaligenes fecalis. A DO-stat feeding approach was employed combined with an optimized post-induction strategy to achieve nitrilase titer of 2.5*105 U/l and 78 g/l dry cell weight. We also identified 16 novel nitrilase sequences from genome mining and analysis of substrate binding residues. The nitrilases were expressed in E. coli and their biocatalytic potential was evaluated on a panel of 22 industrially relevant nitrile substrates using high-throughput screening and HPLC analysis. Nine nitrilases were identified to exhibit high activity on structurally diverse nitriles including aliphatic and aromatic dinitriles, heterocyclic, -hydroxy and -keto nitriles. With fed-batch biotransformation, whole-cell Zobelia galactanivorans nitrilase achieved yields of 2.4 M nicotinic acid and 1.8 M isonicotinic acid from 3-cyanopyridine and 4-cyanopyridine respectively within 5 h, while Cupravidus necator nitrilase enantioselectively converted 740 mM mandelonitrile to (R)–mandelic acid. The nitrilase from Achromobacter insolitus could hydrolyze 542 mM iminodiacetonitrile in 1 h. The availability of highly active nitrilases along with bioprocesses for enzyme production expands the toolbox for industrial biocatalysis.Keywords: biocatalysis, isonicotinic acid, iminodiacetic acid, mandelic acid, nitrilase
Procedia PDF Downloads 2383624 Glycyrrhizic Acid Inhibits Lipopolysaccharide-Stimulated Bovine Fibroblast-Like Synoviocyte, Invasion through Suppression of TLR4/NF-κB-Mediated Matrix Metalloproteinase-9 Expression
Authors: Hosein Maghsoudi
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Rheumatois arthritis (RA) is progressive inflammatory autoimmune diseases that primarily affect the joints, characterized by synovial hyperplasia and inflammatory cell infiltration, deformed and painful joints, which can lead tissue destruction, functional disability systemic complications, and early dead and socioeconomic costs. The cause of rheumatoid arthritis is unknown, but genetic and environmental factors are contributory and the prognosis is guarded. However, advances in understanding the pathogenesis of the disease have fostered the development of new therapeutics, with improved outcomes. The current treatment strategy, which reflects this progress, is to initiate aggressive therapy soon after diagnosis and to escalate the therapy, guided by an assessment of disease activity, in pursuit of clinical remission. The pathobiology of RA is multifaceted and involves T cells, B cells, fibroblast-like synoviocyte (FLSc) and the complex interaction of many pro-inflammatory cytokine. Novel biologic agents that target tumor necrosis or interlukin (IL)-1 and Il-6, in addition T- and B-cells inhibitors, have resulted in favorable clinical outcomes in patients with RA. Despite this, at least 30% of RA patients are résistance to available therapies, suggesting novel mediators should be identified that can target other disease-specific pathway or cell lineage. Among the inflammatory cell population that might participated in RA pathogenesis, FLSc are crucial in initiaing and driving RA in concert of cartilage and bone by secreting metalloproteinase (MMPs) into the synovial fluid and by direct invasion into extracellular matrix (ECM), further exacerbating joint damage. Invasion of fibroblast-like synoviocytes (FLSc) is critical in the pathogenesis of rheumatoid-arthritis. The metalloproteinase (MMPs) and activator of Toll-like receptor 4 (TLR4)/nuclear factor- κB pthway play a critical role in RA-FLS invasion induced by lipopolysaccharide (LPS). The present study aimed to explore the anti-invasion activity of Glycyrrhizic Acid as a pharmacologically safe phytochemical agent with potent anti-inflammatory properties on IL-1beta and TNF-alpha signalling pathways in Bovine fibroblast-like synoviocyte ex- vitro, on LPS-stimulated bovine FLS migration and invasion as well as MMP expression and explored the upstream signal transduction. Results showed that Glycyrrhizic Acid suppressed LPS-stimulated bovine FLS migration and invasion by inhibition MMP-9 expression and activity. In addition our results revealed that Glycyrrhizic Acid inhibited the transcriptional activity of MMP-9 by suppression the nbinding activity of NF- κB in the MMP-9 promoter pathway. The extract of licorice (Glycyrrhiza glabra L.) has been widely used for many centuries in the traditional Chinese medicine as native anti-allergic agent. Glycyrrhizin (GL), a triterpenoidsaponin, extracted from the roots of licorice is the most effective compound for inflammation and allergic diseases in human body. The biological and pharmacological studies revealed that GL possesses many pharmacological effects, such as anti-inflammatory, anti-viral and liver protective effects, and the biological effects, such as induction of cytokines (interferon-γ and IL-12), chemokines as well as extrathymic T and anti-type 2 T cells. GL is known in the traditional Chinese medicine for its anti-inflammatory effect, which is originally described by Finney in 1959. The mechanism of the GL-induced anti-inflammatory effect is based on different pathways of the GL-induced selective inhibition of the prostaglandin E2 production, the CK-II- mediated activation of both GL-binding lipoxygenas (gbLOX; 17) and PLA2, an anti-thrombin action of GL and production of the reactive oxygen species (ROS; GL exerts liver protection properties by inhibiting PLA2 or by the hydroxyl radical trapping action, leading to the lowering of serum alanine and aspartate transaminase levels. The present study was undertaken to examine the possible mechanism of anti-inflammatory properties GL on IL-1beta and TNF-alpha signalling pathways in bovine fibroblast-like synoviocyte ex-vivo, on LPS-stimulated bovine FLS migration and invasion as well as MMP expression and explored the upstream signal transduction. Our results clearly showed that treatment of bovine fibroblast-like synoviocyte with GL suppressed LPS-induced cell migration and invasion. Furthermore, it revealed that GL inhibited the transcription activity of MMP-9 by suppressing the binding activity of NF-κB in the MM-9 promoter. MMP-9 is an important ECM-degrading enzyme and overexpression of MMPs in important of RA-FLSs. LPS can stimulate bovine FLS to secret MMPs, and this induction is regulated at the transcription and translational levels. In this study, LPS treatment of bovine FLS caused an increase in MMP-2 and MMP-9 levels. The increase in MMP-9 expression and secretion was inhibited by ex- vitro. Furthermore, these effects were mimicked by MMP-9 siRNA. These result therefore indicate the the inhibition of LPS-induced bovine FLS invasion by GL occurs primarily by inhibiting MMP-9 expression and activity. Next we analyzed the functional significance of NF-κB transcription of MMP-9 activation in Bovine FLSs. Results from EMSA showed that GL suppressed LPS-induced NF-κB binding to the MMP-9 promotor, as NF-κB regulates transcriptional activation of multiple inflammatory cytokines, we predicted that GL might target NF-κB to suppress MMP-9 transcription by LPS. Myeloid differentiation-factor 88 (MyD88) and TIR-domain containing adaptor protein (TIRAP) are critical proteins in the LPS-induced NF-κB and apoptotic signaling pathways, GL inhibited the expression of TLR4 and MYD88. These results demonstrated that GL suppress LPS-induced MMP-9 expression through the inhibition of the induced TLR4/NFκB signaling pathway. Taken together, our results provide evidence that GL exerts anti-inflammatory effects by inhibition LPS-induced bovine FLSs migration and invasion, and the mechanisms may involve the suppression of TLR4/NFκB –mediated MMP-9 expression. Although further work is needed to clarify the complicated mechanism of GL-induced anti-invasion of bovine FLSs, GL might be used as a further anti-invasion drug with therapeutic efficacy in the treatment of immune-mediated inflammatory disease such as RA.Keywords: glycyrrhizic acid, bovine fibroblast-like synoviocyte, tlr4/nf-κb, metalloproteinase-9
Procedia PDF Downloads 3923623 Conjugated Linoleic Acid (CLA) Health Benefits and Sources
Authors: Hilal Ahmad Punoo
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Conjugated linoleic acid (CLA) is a mixture of positional and geometric isomers of octadecadienoic acid with two conjugated double bonds. Of more than a dozen isomers of CLA found naturally in dairy and meat products from ruminants, c-9, t-11 and t-10, c-12 are the two isomers with known physiological importance, including anticarcinogenic, antidiabetic, antilipogenic, and antiatherosclerotic effects. Conjugated linoleic acids (CLA) may influence the onset and severity of several chronic diseases, including various cancers, atherosclerosis, obesity, bone density loss, and diabetes. These findings are of special interest to the agriculture community, because dietary sources of CLA are almost exclusively beef and dairy products. Thus, a better understanding of the specific isomers and mechanisms responsible for these positive effects of CLA on human health would be both prudent and economically beneficial. To date, research related to the advantages of CLA consumption on human health has been conducted using experimental laboratory animals and cell culture systems. These data consistently show that relatively low levels of CLA can influence risk of cancer. Further, very recent investigations suggest that the predominate CLA isoform (cis-9, trans-11 CLA or rumenic acid) found in beef and milk fat possesses anticarcinogenic effects but does not alter body composition; the trans-10, cis-12 CLA has been shown to inhibit lipogenesis. Clearly, further work, especially using human subjects, will be required to characterize the potential benefits of CLA consumption on human health. Moreover, we suggest that foods naturally containing high amounts of CLA (e.g., beef and dairy products) be considered as meeting the definition of functional foods.Keywords: conjugated linoleic acid, potential health benefits, fats, animals, humans
Procedia PDF Downloads 3123622 Agro-Industrial Waste as a Source of Catalyst Production
Authors: Brenda Cecilia Ledesma, Andrea Beltramone
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This work deals with the bio-waste valorization approach for catalyst development, the use of products derived from biomass as raw material and the obtaining of biofuels. In this research, activated carbons were synthesized from the orange peel using different synthesis conditions. With the activated carbons obtained with the best structure and texture, PtIr bimetallic catalysts were prepared. Carbon activation was carried out through a chemical process with phosphoric acid as an activating agent, varying the acid concentration, the ratio substrate/activating agent and time of contact between them. The best support was obtained using a carbonization time of 1 h, the temperature of carbonization of 470oC, the phosphoric acid concentration of 50 wt.% and a BET area of 1429 m2/g. Subsequently, the metallic nanoparticles were deposited in the activated carbon to use the solid as a catalytic material for the hydrogenation of HMF to 2,5-DMF. The catalyst presented an excellent performance for biofuels generation.Keywords: orange peel, bio-waste valorization, platinum, iridium, 5-hydroxymethylfurfural
Procedia PDF Downloads 2033621 Caspase-11 and AIM2 Inflammasome are Involved in Smoking-Induced COPD and Lung Adenocarcinoma
Authors: Chiara Colarusso, Michela Terlizzi, Aldo Pinto, Rosalinda Sorrentino
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Cigarette smoking is the main cause and the most common risk factor for both COPD and lung cancer. In our previous studies, we proved that caspase-11 in mice and its human analogue, caspase-4, are involved in lung carcinogenesis and that AIM2 inflammasome might play a pro-cancerous role in lung cancer. Therefore, the aim of this study was to investigate potential crosstalk between COPD and lung cancer, focusing on AIM2 and caspase-11-dependent inflammasome signaling pathway. To mimic COPD, we took advantage of an experimental first-hand smoking mouse model and, to confirm what was observed in mice, we used human samples of lung adenocarcinoma patients stratified according to the smoking and COPD status. We demonstrated that smoke exposure led to emphysema-like features, bronchial tone impairment, and release of IL-1-like cytokines (IL-1α, IL-1β, IL-33, IL-18) in a caspase-1 independent manner in C57Bl/6N. Rather, a dysfunctional caspase-11 in smoke-exposed 129Sv mice was associated to lower bronchial inflammation, collagen deposition, and IL-1-like inflammation. In addition, for the first time, we found that AIM2 inflammasome is involved in lung inflammation in smoking and COPD, in that its expression was higher in smoke-exposed C57Bl/6N compared to 129Sv smoking mice, who instead did not show any alteration of AIM2 in both macrophages and dendritic cells. Moreover, we found that AIM2 expression in the cancerous tissue, albeit higher than non-cancerous tissue, was not statistically different according to the COPD and smoking status. Instead, the higher expression of AIM2 in non-cancerous tissue of smoker COPD patients than smokers who did not have COPD was correlated to a higher hazard ratio of poor survival rate than patients who presented lower levels of AIM2. In conclusion, our data highlight that caspase-11 in mice is associated to smoke-induced lung latent inflammation which could drive the establishment of lung cancer, and that AIM2 inflammasome plays a role at the crosstalk between smoking/COPD and lung adenocarcinoma in that its higher presence is correlated to lower survival rate of smoker COPD adenocarcinoma.Keywords: COPD, inflammasome, lung cancer, lung inflammation, smoke
Procedia PDF Downloads 1593620 Effect of Solvents in the Extraction and Stability of Anthocyanin from the Petals of Caesalpinia pulcherrima for Natural Dye-Sensitized Solar Cell
Authors: N. Prabavathy, R. Balasundaraprabhu, S. Shalini, Dhayalan Velauthapillai, S. Prasanna, N. Muthukumarasamy
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Dye sensitized solar cell (DSSC) has become a significant research area due to their fundamental and scientific importance in the area of energy conversion. Synthetic dyes as sensitizer in DSSC are efficient and durable but they are costlier, toxic and have the tendency to degrade. Natural sensitizers contain plant pigments such as anthocyanin, carotenoid, flavonoid, and chlorophyll which promote light absorption as well as injection of charges to the conduction band of TiO2 through the sensitizer. But, the efficiency of natural dyes is not up to the mark mainly due to instability of the pigment such as anthocyanin. The stability issues in vitro are mainly due to the effect of solvents on extraction of anthocyanins and their respective pH. Taking this factor into consideration, in the present work, the anthocyanins were extracted from the flower Caesalpinia pulcherrima (C. pulcherrimma) with various solvents and their respective stability and pH values are discussed. The usage of citric acid as solvent to extract anthocyanin has shown good stability than other solvents. It also helps in enhancing the sensitization properties of anthocyanins with Titanium dioxide (TiO2) nanorods. The IPCE spectra show higher photovoltaic performance for dye sensitized TiO2nanorods using citric acid as solvent. The natural DSSC using citric acid as solvent shows a higher efficiency compared to other solvents. Hence citric acid performs to be a safe solvent for natural DSSC in boosting the photovoltaic performance and maintaining the stability of anthocyanins.Keywords: Caesalpinia pulcherrima, citric acid, dye sensitized solar cells, TiO₂ nanorods
Procedia PDF Downloads 2963619 Service Life Modelling of Concrete Deterioration Due to Biogenic Sulphuric Acid (BSA) Attack-State-of-an-Art-Review
Authors: Ankur Bansal, Shashank Bishnoi
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Degradation of Sewage pipes, sewage pumping station and Sewage treatment plants(STP) is of major concern due to difficulty in their maintenance and the high cost of replacement. Most of these systems undergo degradation due to Biogenic sulphuric acid (BSA) attack. Since most of Waste water treatment system are underground, detection of this deterioration remains hidden. This paper presents a literature review, outlining the mechanism of this attack focusing on critical parameters of BSA attack, along with available models and software to predict the deterioration due to this attack. This paper critically examines the various steps and equation in various Models of BSA degradation, detail on assumptions and working of different softwares are also highlighted in this paper. The paper also focuses on the service life design technique available through various codes and method to integrate the servile life design with BSA degradation on concrete. In the end, various methods enhancing the resistance of concrete against Biogenic sulphuric acid attack are highlighted. It may be concluded that the effective modelling for degradation phenomena may bring positive economical and environmental impacts. With current computing capabilities integrated degradation models combining the various durability aspects can bring positive change for sustainable society.Keywords: concrete degradation, modelling, service life, sulphuric acid attack
Procedia PDF Downloads 3153618 Effect of Resveratrol and Ascorbic Acid on the Stability of Alfa-Tocopherol in Whey Protein Isolate Stabilized O/W Emulsions
Authors: Lei Wang, Yingzhou Ni, Amr M. Bakry, Hao Cheng, Li Liang
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Food proteins have been widely used as carrier materials because of their multiple functional properties. In this study, alfa-tocopherol was encapsulated in the oil phase of an oil-in-water emulsion stabilized with whey protein isolate (WPI). The influence of WPI concentration and resveratrol or ascorbic acid on the decomposition of alfa-tocopherol in the emulsion during storage is discussed. Decomposition decreased as WPI concentrations increased. Decomposition was delayed at ascorbic acid/WPI molar ratios lower than 5 but was promoted at higher ratios. Resveratrol partitioned into the oil-water interface by binding to WPI and its cis-isomer is believed to have contributed most of the protective effect of this polyphenol. These results suggest the possibility of using the emulsifying and ligand-binging properties of WPI to produce carriers for simultaneous encapsulation of alfa-tocopherol and resveratrol in a single emulsion system.Keywords: stability, alfa-tocopherol, resveratrol, whey protein isolate
Procedia PDF Downloads 5353617 Comparison of Nucleic Acid Extraction Platforms On Tissue Samples
Authors: Siti Rafeah Md Rafei, Karen Wang Yanping, Park Mi Kyoung
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Tissue samples are precious supply for molecular studies or disease identification diagnosed using molecular assays, namely real-time PCR (qPCR). It is critical to establish the most favorable nucleic acid extraction that gives the PCR-amplifiable genomic DNA. Furthermore, automated nucleic acid extraction is an appealing alternative to labor-intensive manual methods. Operational complexity, defined as the number of steps required to obtain an extracted sample, is one of the criteria in the comparison. Here we are comparing the One BioMed’s automated X8 platform with the commercially available manual-operated kits from QIAGEN Mini Kit and Roche. We extracted DNA from rat fresh-frozen tissue (from different type of organs) in the matrices. After tissue pre-treatment, it is added to the One BioMed’s X8 pre-filled cartridge, and the QIAGEN QIAmp column respectively. We found that the results after subjecting the eluates to the Real Time PCR using BIORAD CFX are comparable.Keywords: DNA extraction, frozen tissue, PCR, qPCR, rat
Procedia PDF Downloads 1673616 The Effects of Ellagic Acid on Rat Lungs Induced Tobacco Smoke
Authors: Nalan Kaya, Gonca Ozan, Elif Erdem, Neriman Colakoglu, Enver Ozan
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The toxic effects of tobacco smoke exposure have been detected in numerous studies. Ellagic acid (EA), (2,3,7,8-tetrahydroxy [1]-benzopyranol [5,4,3-cde] benzopyran 5,10-dione), a natural phenolic lactone compound, is found in various plant species including pomegranate, grape, strawberries, blackberries and raspberries. Similar to the other effective antioxidants, EA can safely interact with the free radicals and reduces oxidative stress through the phenolic ring and hydroxyl components in its structure. The aim of the present study was to examine the protective effects of ellagic acid against oxidative damage on lung tissues of rats induced by tobacco smoke. Twenty-four male adult (8 weeks old) Spraque-Dawley rats were divided randomly into 4 equal groups: group I (Control), group II (Tobacco smoke), group III (Tobacco smoke + corn oil) and group IV (Tobacco smoke + ellagic acid). The rats in group II, III and IV, were exposed to tobacco smoke 1 hour twice a day for 12 weeks. In addition to tobacco smoke exposure, 12 mg/kg ellagic acid (dissolved in corn oil), was applied to the rats in group IV by oral gavage. Equal amount of corn oil used in solving ellagic acid was applied to the rats by oral gavage in group III. At the end of the experimental period, rats were decapitated. Lung tissues and blood samples were taken. The lung slides were stained by H&E and Masson’s Trichrome methods. Also, galactin-3 stain was applied. Biochemical analyzes were performed. Vascular congestion and inflammatory cell infiltration in pulmonary interstitium, thickness in interalveolar septum, cytoplasmic vacuolation in some macrophages and galactin-3 positive cells were observed in histological examination of tobacco smoke group. In addition to these findings, hemorrhage in pulmonary interstitium and bronchial lumen was detected in tobacco smoke + corn oil group. Reduced vascular congestion and hemorrhage in pulmoner interstitium and rarely thickness in interalveolar septum were shown in tobacco smoke + EA group. Compared to group-I, group-II GSH level was decreased and MDA level was increased significantly. Nevertheless group-IV GSH level was higher and MDA level was lower than group-II. The results indicate that ellagic acid could protect the lung tissue from the tobacco smoke harmful effects.Keywords: ellagic acid, lung, rat, tobacco smoke
Procedia PDF Downloads 2203615 Study of Self-Assembled Photocatalyst by Metal-Terpyridine Interactions in Polymer Network
Authors: Dong-Cheol Jeong, Jookyung Lee, Yu Hyeon Ro, Changsik Song
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The design and synthesis of photo-active polymeric systems are important in regard to solar energy harvesting and utilization. In this study, we synthesized photo-active polymer, thin films, and polymer gel via iterative self-assembly using reversible metal-terpyridine (M-tpy) interactions. The photocurrent generated in the polymeric thin films with Zn(II) was much higher than those of other films. Apparent diffusion rate constant (kapp) was measured for the electron hopping process via potential-step chronoamperometry. As a result, the kapp for the polymeric thin films with Zn(II) was almost two times larger than those with other metal ions. We found that the anodic photocurrents increased with the inclusion of the multi-walled carbon nanotube (MWNT) layer. Inclusion of MWNTs can provide efficient electron transfer pathways. In addition, polymer gel based on interactions between terpyridine and metal ions was shown the photocatalytic activity. Interestingly, in the Mg-terpyridine gel, the reaction rate of benzylamine to imine photo-oxidative coupling was faster than Fe-terpyridine gel because the Mg-terpyridine gel has two steps electron transfer pathway but Fe-terpyridine gel has three steps electron transfer pathway.Keywords: terpyridine, photocatalyst, self-assebly, metal-ligand
Procedia PDF Downloads 3133614 Neuroprotective Effect of Chrysin on Thioacetamide-Induced Hepatic Encephalopathy in Rats: Role of Oxidative Stress and TLR-4/NF-κB Pathway
Authors: S. A. El-Marasy, S. A. El Awdan, R. M. Abd-Elsalam
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This study aimed to investigate the possible neuroprotective effect of chrysin on thioacetamide (TAA)-induced hepatic encephalopathy in rats. Also, the effect of chrysin on motor impairment, cognitive deficits, oxidative stress, neuroinflammation, apoptosis and histopathological damage was assessed. Male Wistar rats were randomly allocated into five groups. The first group received the vehicle (distilled water) for 21 days and is considered as normal group. While the second one received intraperitoneal dose of TAA (200 mg/kg) at three alternative days during the third week of the experiment to induce HE and is considered as control group. The other three groups were orally administered chrysin for 21 days (25, 50, 100 mg/kg) and starting from day 17; rats received intraperitoneal dose of TAA (200 mg/kg) at three alternative days. Then behavioral, biochemical, histopathological and immunohistochemical analyses were assessed. Then behavioral, biochemical, histopathological and immunohistochemical analyses were assessed. Chrysin reversed TAA-induced motor coordination in rotarod test, cognitive deficits in object recognition test (ORT) and attenuated serum ammonia, hepatic liver enzymes, reduced malondialdehyde (MDA), elevated reduced glutathione (GSH), reduced nuclear factor kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α) and Interleukin-6 (IL-6) brain contents. Chrysin administration also reduced Toll-4 receptor (TLR-4) gene expression, caspase-3 protein expression, hepatic necrosis and astrocyte swelling. This study depicts that chrysin exerted neuroprotective effect in TAA-induced HE rats, evidenced by improvement of cognitive deficits, motor incoordination and histopathological changes such as astrocyte swelling and vacuolization; hallmarks in HE, via reducing hyperammonemia, ameliorating hepatic function, in addition to its anti-oxidant, inactivation of TLR-4/NF-κB inflammatory pathway, and anti-apoptotic effects.Keywords: chrysin, hepatic encephalopathy, oxidative stress, rats, thioacetamide, TLR4/NF-κB pathway
Procedia PDF Downloads 1633613 Synthesis of Bismuth-Hyaluronic Acid Nanoparticles Containing Melittin Coated with Chitosan for Treating Eye Cancer Cells with Radiotherapy
Authors: Akbar Esmaeili, Fateme Dadashi
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Bismuth can increase radiation and reduce the dose of radiotherapy. On the other hand, hyaluronic acid plays a role in healing damaged cells, and melittin has been used to destroy cancer cells. This research aims to destroy eye cancer cells and accelerate the recovery of damaged healthy cells during treatment. In this research, we used this nanoparticle, the sol-gel method. According to the optimization process that was carried out, we obtained the optimal value of the desired variables for the manufacture of nanoparticles. The advantage of doing this is reducing the amount of medicine used, as a result of reducing the number of side effects during the treatment and using melittin as an anti-eye cancer drug and the presence of hyaluronic acid to accelerate the recovery of cells, as well as coating the bismuth nanoparticle with chitosan to increase the half-life of the nanoparticle and prevent its adhesion.Keywords: synthesis, nanoparticles, coated, cancer
Procedia PDF Downloads 733612 Control of Spoilage Fungi by Lactobacilli
Authors: Laref Nora, Guessas Bettache
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Lactic acid bacteria (LAB) have a major potential to be used in biopreservation methods because they are safe to consume (GRAS: generally regarded as safe) and they naturally occurring microflora of many foods. The preservative action of LAB is due to several antimicrobial metabolites, including lactic acid, acetic acid, hydrogen peroxide, bacteriocins, carbon dioxide, diacetyl, and reuterin. Several studies have focused on the antifungal activity compounds from natural sources for biopreservation in alternatives to chemical use. LAB has an antifungal activity which may inhibit food spoilage fungi. Lactobacillus strains isolated from silage prepared in our laboratory by fermentation of grass in anaerobic condition were screened for antifungal activity with overlay assay against Aspergillus spp. The antifungal compounds were originated from organic acids; inhibitory activity did not change after treatment with proteolytic enzymes. Lactobacillus strains were able also to inhibit Trichoderma spp, Penicillium spp, Fusarium roseum, and Stemphylim spp by confrontation assay. The inhibitory activity could be detected against the mould Aspergillus spp in the apricot juice but not in a bakery product. These antifungal compounds have the potential to be used as food biopreservation to inhibit conidia germination, and mycelia growth of spoilage fungi depending on food type, pH of food especially in heat, and cold processed foods.Keywords: lactic acid bacteria, Lactobacillus, Aspergillus, antifungal activity
Procedia PDF Downloads 3353611 Identification of EEG Attention Level Using Empirical Mode Decompositions for BCI Applications
Authors: Chia-Ju Peng, Shih-Jui Chen
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This paper proposes a method to discriminate electroencephalogram (EEG) signals between different concentration states using empirical mode decomposition (EMD). Brain-computer interface (BCI), also called brain-machine interface, is a direct communication pathway between the brain and an external device without the inherent pathway such as the peripheral nervous system or skeletal muscles. Attention level is a common index as a control signal of BCI systems. The EEG signals acquired from people paying attention or in relaxation, respectively, are decomposed into a set of intrinsic mode functions (IMF) by EMD. Fast Fourier transform (FFT) analysis is then applied to each IMF to obtain the frequency spectrums. By observing power spectrums of IMFs, the proposed method has the better identification of EEG attention level than the original EEG signals between different concentration states. The band power of IMF3 is the most obvious especially in β wave, which corresponds to fully awake and generally alert. The signal processing method and results of this experiment paves a new way for BCI robotic system using the attention-level control strategy. The integrated signal processing method reveals appropriate information for discrimination of the attention and relaxation, contributing to a more enhanced BCI performance.Keywords: biomedical engineering, brain computer interface, electroencephalography, rehabilitation
Procedia PDF Downloads 3943610 Genome Analysis of Lactobacillus Plantarum and Lactobacillus Brevis Isolated From Traditionally Fermented Ethiopian Kocho and Their Probiotic Properties
Authors: Guesh Mulaw, Haile Beruhulay, Anteneh Tesfaye, Tesfaye Sisay Diriba Muleta
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Probiotics are live microorganisms that, when administered in adequate amounts, promote the health of a consumer. The present work aims to study the whole genome sequence of probiotic strains of lactic acid bacteria (LAB) isolated from traditional Ethiopian fermented kocho for bacteriocin production and to evaluate their probiotic properties. LAB were isolated from traditionally fermented kocho samples and characterized following standard methods. Accordingly, a total of 150 LAB were isolated, of which 7 (4.67%) isolates showed 50.52-74.05% and 33.33-62.40% survival rates at pH 2 for 3 and 6 h, respectively. The 7 acid-tolerant isolates were also tolerated 0.3% bile salt for 24 h with 88.96 to 98.10% survival. The acid and bile salt-tolerant LAB isolates also inhibited some reference foodborne pathogenic bacteria to varying degrees. All 7 acid- and bile salt-tolerant isolates were susceptible to ampicillin, tetracycline and erythromycin. However, the potent isolates showed remarkable resistance to kanamycin. Likewise, four of the 7 isolates were resistant to streptomycin, but three of the 7 isolates were sensitive to streptomycin. The identification of the seven selected probiotic LAB isolates and their genetic relatedness was performed based on whole-genome sequence comparisons. Consequently, these isolates belonged to Lactobacillus species, including 6 Lb. plantarum, 1 Lb. brevis. Among the 7 potential probiotic LAB strains, BAGEL predicted 2 bacteriocin for class II in the genome of 7 strains. The 7 Lactobacillus strains were found to be potentially useful for producing functional products and could be suitable probiotic candidates for food processing industriesKeywords: ferneted foods, kocho, probiotics, lactic acid bacteria
Procedia PDF Downloads 373609 Deciphering Tumor Stroma Interactions in Retinoblastoma
Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar
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Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment
Procedia PDF Downloads 3863608 Effect of Nitrogen and Gibberellic Acid at Different Level and their Interaction on Calendula
Authors: Pragnyashree Mishra, Shradhanjali Mohapatra
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The present investigation is carried out to know the effect of foliar feeding of nitrogen and gibberellic acid on vegetative growth, flowering behaviour and yield of calendula variety ‘Golden Emporer’. The experiment was laid out in RBD in rabi season of 2013-14. There are 16 treatments are taken at different level such as nitrogen (at 0%,1%,2%,3%) and GA3 (at 50 ppm,100ppm,150 ppm). Among them maximum height at bud initiation stage was obtained at 3% nitrogen (27.00 cm) and at 150 ppm GA3 (26.5 cm), fist flowering was obtained at 3% nitrogen(60.00 days) and at 150 ppm GA3 (63.75 days), maximum flower stalk length was obtained at 3% nitrogen(3.50 cm) and at 150 ppm GA3 (5.42 cm),maximum duration of flowering was obtained at 3% nitrogen(46.00 days) and at 150 ppm GA3 (46.50days), maximum number of flower was obtained at 3% nitrogen (89.00per plant) and at 150 ppm GA3 (83.50 per plant), maximum flower weight was obtained at 3% nitrogen(1.25 gm per flower) and at 150 ppm GA3 (1.50 gm per flower), maximum yield was was obtained at 3% nitrogen (110.00 gm per plant) and at 150 ppm GA3 (105.00gm per plant) and minimum of all character was obtained when 0% nitrogen0 ppm GA3. All interaction between nitrogen and GA3 was found in significant except the yield .Keywords: calendula, golden emporer, GA3, nitrogen and gibberellic acid
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