Search results for: nucleic acid binding
3599 Plasma-Assisted Nitrogen Fixation for the Elevation of Seed Germination and Plant Growth
Authors: Pradeep Lamichhane
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Plasma-assisted nitrogen fixation is a process by which atomic nitrogen generated by plasma is converted into ammonia (NH₃) or related nitrogenous compounds. Nitrogen fixation is essential to plant because fixed inorganic nitrogen compounds are required to them for the biosynthesis of all nitrogen-containing organic compounds, such as amino acids and proteins, nucleoside triphosphates and nucleic acid. Most of our atmosphere is composed of nitrogen; however, the plant cannot absorb it directly from the air ambient. As a portion of the nitrogen cycle, nitrogen fixation fundamental for agriculture and the manufacture of fertilizer. In this study, plasma-assisted nitrogen fixation was performed by exposing a non-thermal atmospheric pressure nitrogen plasma generated a sinusoidal power supply (with an applied voltage of 10 kV and frequency of 33 kHz) on a water surface. Besides this, UV excitation of water molecules at the water interface was also done in order to disassociate water. Hydrogen and hydroxyl radical obtained from this UV photolysis electrochemically combine with nitrogen atom obtained from plasma. As a result of this, nitrogen fixation on plasma-activated water (PAW) significantly enhanced. The amount of nitrogen-based products like NOₓ and ammonia (NH₃) synthesized by this combined process of UV and plasma are 1.4 and 2.8 times higher than those obtained by plasma alone. In every 48 hours, 20 ml of plasma-activated water (pH≈3.15) for 10 minutes with moderate concentrations of NOₓ, NH₃ and hydrogen peroxide (H₂O₂) was irrigated on each corn plant (Zea Mays). It was found that the PAW has shown a significant impact on seeds germination rate and improved seedling growth. The result obtained from this experiment suggested that crop yield could increase in a short duration. In the future, this experiment could open boundless opportunities in plasma agriculture to mobilize nitrogen because nitrite, nitrate, and ammonia are more suitable for plant uptake.Keywords: plasma-assisted nitrogen fixation, nitrogen plasma, UV excitation of water, ammonia synthesis
Procedia PDF Downloads 1393598 Screening of Antioxidant Activity of Exopolysaccharides Produced by Lactic Acid Bacteria From Human Origin
Authors: Piña-Ronces Laura Gabriela, Reyes-Escogido María de Lourdes
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Exist a large variability in Exopolysaccharides (EPS) produced by LAB depending on carbon source, they have multiple applications in food industry mainly, but they have become important for the health. In this study, we identified EPS-producing strains belonging to the BAL group; they were previously isolated from humans. After that, we extracted and evaluated the antioxidant activity of EPS produced by all strains. Antioxidant activity was determined by DPPH method using ascorbic acid as standard for both comparison and quantification. 31 strains (51.66 %) produced EPS at concentrations between 451 and 1.561 mg/l, 16 of EPS extracted showed antioxidant effect superior to ascorbic acid at the same concentrations. EPS-producing strains were L. plantarum, L. sp and L. fermentum corresponding to Lactobacillus genus and, E. faecium, E. durans, and E. hirae of Enterococcus genus. Antioxidant activity showed by EPS from 3 strains of L. plantarum and 3 strains of E. faecium was different into specie, while the antioxidant activity determined for EPS obtained from the other strains did not show difference at specie level, but was superior to ascorbic acid. EPS produced by L. plantarum and E. hirae had the best activity, it could be considerate for selection them as a possible new alternative for therapy or treatment of diseases related whit oxidative stress. Further studies about biological functions of EPS have to be conducted for new applications in health.Keywords: oxidative stress, lactic acid bacteria, exopolysaccharides, antioxidant activity
Procedia PDF Downloads 3623597 Effect of Chitosan and Ascorbic Acid Coating on the Refrigerated Tilapia Fish Fillet (Oreochromis niliticus)
Authors: Jau-Shya Lee, Rossita Shapawi, Vin Cent Pua
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Tilapia is a popular cultured fresh-water fish in Malaysia. The highly perishable nature of the fish and increasing demand for high-quality ready-to-cook fish has intensified the search for better fish preservation method. Chitosan edible coating has been evident to extend the shelf life of fish fillet. This work was attempted to explore the potential of ascorbic acid in enhancing the shelf life extension ability of chitosan coated Tilapia fillet under refrigeration condition (4 ± 1oC). A 3 2 Factorial Design which comprising of three concentrations of chitosan (1, 1.5 and 2%) and two concentrations of ascorbic acids (2.5 and 5%) was used. The fish fillets were analyzed for total viable count, thiobarbituric acid (TBA) value, pH, aw and colour changes at 3-day interval over 15-day storage. The shelf life of chitosan coated (1.5% and 2%) fillet was increased to 15 days as compared to uncoated fish fillet which can only last for nine days. The inhibition of microbial growth of fish fillet was enhanced with the addition of 5% of ascorbic acids in 2% of chitosan. The TBA value, pH and aw for chitosan coated samples were found lower than that of uncoated sample (p<0.05). The colour stability of the fish fillet was also improved by the composite coating. Overall, 2% of chitosan and 5% of ascorbic acid formed the most effective coating to enhance the quality and to lengthen the shelf life of refrigerated Tilapia fillet.Keywords: ascorbic acid, chitosan, edible coating, fish fillet
Procedia PDF Downloads 3943596 Analysis of Caffeic Acid from Myrica nagi Leaves by High Performance Liquid Chromatography
Authors: Preeti Panthari, Harsha Kharkwal
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Myrica nagi belongs to Myricaceae family. It is known for its therapeutic use since ancient times. The leaves were extracted with methanol and further fractioned with different solvents with increasing polarity. The n-butanol fraction of methanol extract was passed through celite, on separation through silica gel column chromatography yielded ten fractions. For the first time we report isolation of Caffeic acid from n-butanol fraction of Myrica nagi leaves in Chloroform: methanol (70:30) fraction. The mobile phase used for analysis in HPLC was Methanol: water (60:40) at the flow rate of 1 ml/min at wavelength of 280 nm. The retention time was 2.66 mins.Keywords: Myrica nagi, column chromatography, retention time, caffeic acid
Procedia PDF Downloads 5553595 Interaction Evaluation of Silver Ion and Silver Nanoparticles with Dithizone Complexes Using DFT Calculations and NMR Analysis
Authors: W. Nootcharin, S. Sujittra, K. Mayuso, K. Kornphimol, M. Rawiwan
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Silver has distinct antibacterial properties and has been used as a component of commercial products with many applications. An increasing number of commercial products cause risks of silver effects for human and environment such as the symptoms of Argyria and the release of silver to the environment. Therefore, the detection of silver in the aquatic environment is important. The colorimetric chemosensor is designed by the basic of ligand interactions with a metal ion, leading to the change of signals for the naked-eyes which are very useful method to this application. Dithizone ligand is considered as one of the effective chelating reagents for metal ions due to its high selectivity and sensitivity of a photochromic reaction for silver as well as the linear backbone of dithizone affords the rotation of various isomeric forms. The present study is focused on the conformation and interaction of silver ion and silver nanoparticles (AgNPs) with dithizone using density functional theory (DFT). The interaction parameters were determined in term of binding energy of complexes and the geometry optimization, frequency of the structures and calculation of binding energies using density functional approaches B3LYP and the 6-31G(d,p) basis set. Moreover, the interaction of silver–dithizone complexes was supported by UV–Vis spectroscopy, FT-IR spectrum that was simulated by using B3LYP/6-31G(d,p) and 1H NMR spectra calculation using B3LYP/6-311+G(2d,p) method compared with the experimental data. The results showed the ion exchange interaction between hydrogen of dithizone and silver atom, with minimized binding energies of silver–dithizone interaction. However, the result of AgNPs in the form of complexes with dithizone. Moreover, the AgNPs-dithizone complexes were confirmed by using transmission electron microscope (TEM). Therefore, the results can be the useful information for determination of complex interaction using the analysis of computer simulations.Keywords: silver nanoparticles, dithizone, DFT, NMR
Procedia PDF Downloads 2103594 Study of Coconut and Babassu Oils with High Acid Content and the Fatty Acids (C6 to C16) Obtained from These Oils
Authors: Flávio A. F. da Ponte, Jackson Q. Malveira, José A. S. Ramos Filho, Monica C. G. Albuquerque
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The vegetable oils have many applications in industrial processes and due to this potential have constantly increased the demand for the use of low-quality oils, mainly in the production of biofuel. This work aims to the physicochemical evaluation of babassu oil (Orbinya speciosa) and coconut (Cocos nucifera) of low quality, as well the obtaining the free fatty acids 6 to 16 carbon atoms, with intention to be used as raw material for the biofuels production. The babassu oil and coconut low quality, as well the fatty acids obtained from these oils were characterized as their physicochemical properties and fatty acid composition (using gas chromatography coupled to mass). The NMR technique was used to assess the efficiency of fractional distillation under reduced pressure to obtain the intermediate carbonic chain fatty acids. The results showed that the bad quality in terms of physicochemical evaluation of babassu oils and coconut oils interfere directly in industrial application. However the fatty acids of intermediate carbonic chain (C6 to C16) may be used in cosmetic, pharmaceutical and particularly as the biokerosene fuel. The chromatographic analysis showed that the babassu oil and coconut oil have as major fatty acids are lauric acid (57.5 and 38.6%, respectively), whereas the top phase from distillation of coconut oil showed caprylic acid (39.1%) and major fatty acid.Keywords: babassu oil (Orbinya speciosa), coconut oil (Cocos nucifera), fatty acids, biomass
Procedia PDF Downloads 3233593 Comparison of Microwave-Assisted and Conventional Leaching for Extraction of Copper from Chalcopyrite Concentrate
Authors: Ayfer Kilicarslan, Kubra Onol, Sercan Basit, Muhlis Nezihi Saridede
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Chalcopyrite (CuFeS2) is the most common primary mineral used for the commercial production of copper. The low dissolution efficiency of chalcopyrite in sulfate media has prevented an efficient industrial leaching of this mineral in sulfate media. Ferric ions, bacteria, oxygen and other oxidants have been used as oxidizing agents in the leaching of chalcopyrite in sulfate and chloride media under atmospheric or pressure leaching conditions. Two leaching methods were studied to evaluate chalcopyrite (CuFeS2) dissolution in acid media. First, the conventional oxidative acid leaching method was carried out using sulfuric acid (H2SO4) and potassium dichromate (K2Cr2O7) as oxidant at atmospheric pressure. Second, microwave-assisted acid leaching was performed using the microwave accelerated reaction system (MARS) for same reaction media. Parameters affecting the copper extraction such as leaching time, leaching temperature, concentration of H2SO4 and concentration of K2Cr2O7 were investigated. The results of conventional acid leaching experiments were compared to the microwave leaching method. It was found that the copper extraction obtained under high temperature and high concentrations of oxidant with microwave leaching is higher than those obtained conventionally. 81% copper extraction was obtained by the conventional oxidative acid leaching method in 180 min, with the concentration of 0.3 mol/L K2Cr2O7 in 0.5M H2SO4 at 50 ºC, while 93.5% copper extraction was obtained in 60 min with microwave leaching method under same conditions.Keywords: extraction, copper, microwave-assisted leaching, chalcopyrite, potassium dichromate
Procedia PDF Downloads 3703592 Surface Enhanced Raman Substrate Detection on the Structure of γ-Aminobutyric Acid(GABA) Connected with Modified Gold-Chitosan Nanoparticles by Mercaptopropionic Acid (MPA)
Authors: Bingjie Wang, Su-Yeon Kwon, Ik-Joong Kang
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A Surface-enhanced Raman Scattering (SERS) as the principle for enhancing Raman scattering by molecules adsorbed on rough metal surfaces or by nanostructures is used to detect the concentration change of γ-Aminobutyric Acid (GABA). As for the gold-chitosan nanoshell, it is made by using chitosan nanoparticles crosslinking with sodium tripolyphosphate(TPP) for the first step to form the chitosan nanoparticles, which would be covered with the gold sequentially. The size of the fabricated product was around 100nm. Based on the method that the sulfur end of the MPA linked to gold can form the very strong S–Au bond, and the carboxyl group, the other end of the MPA, can easily absorb the GABA. GABA is the mainly inhibitory neurotransmitter in the mammalian central nervous system in the human body. It plays such significant role in reducing neuronal excitability throughout the nervous system. When the system formed, it generated SERS, which made a clear difference in the intensity of Raman scattering within the range of GABA concentration. So it is obtained from the experiment that the calibration curve according to the GABA concentration relevant with the SERS scattering. In this study, DLS, SEM, FT-IR, UV, SERS were used to analyze the products to obtain the conclusion.Keywords: chitosan-gold nanoshell, mercaptopropionic acid, γ-aminobutyric acid, surface-enhanced Raman scattering
Procedia PDF Downloads 2643591 Insight into Enhancement of CO2 Capture by Clay Minerals
Authors: Mardin Abdalqadir, Paul Adzakro, Tannaz Pak, Sina Rezaei Gomari
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Climate change and global warming recently became significant concerns due to the massive emissions of greenhouse gases into the atmosphere, predominantly CO2 gases. Therefore, it is necessary to find sustainable and inexpensive methods to capture the greenhouse gasses and protect the environment for live species. The application of naturally available and cheap adsorbents of carbon such as clay minerals became a great interest. However, the minerals prone to low storage capacity despite their high affinity to adsorb carbon. This paper aims to explore ways to improve the pore volume and surface area of two selected clay minerals, ‘montmorillonite and kaolinite’ by acid treatment to overcome their low storage capacity. Montmorillonite and kaolinite samples were treated with different sulfuric acid concentrations (0.5, 1.2 and 2.5 M) at 40 °C for 8 hours to achieve the above aim. The grain size distribution and morphology of clay minerals before and after acid treatment were explored with Scanning Electron Microscope to evaluate surface area improvement. The ImageJ software was used to find the porosity and pore volume of treated and untreated clay samples. The structure of the clay minerals was also analyzed using an X-ray Diffraction machine. The results showed that the pore volume and surface area were increased substantially through acid treatment, which speeded up the rate of carbon dioxide adsorption. XRD pattern of kaolinite did not change after sulfuric acid treatment, which indicates that acid treatment would not affect the structure of kaolinite. It was also discovered that kaolinite had a higher pore volume and porosity than montmorillonite before and after acid treatment. For example, the pore volume of untreated kaolinite was equal to 30.498 um3 with a porosity of 23.49%. Raising the concentration of acid from 0.5 M to 2.5 M in 8 hours’ time reaction led to increased pore volume from 30.498 um3 to 34.73 um3. The pore volume of raw montmorillonite was equal to 15.610 um3 with a porosity of 12.7%. When the acid concentration was raised from 0.5 M to 2.5 M for the same reaction time, pore volume also increased from 15.610 um3 to 20.538 um3. However, montmorillonite had a higher specific surface area than kaolinite. This study concludes that clay minerals are inexpensive and available material sources to model the realistic conditions and apply the results of carbon capture to prevent global warming, which is one of the most critical and urgent problems in the world.Keywords: acid treatment, kaolinite, montmorillonite, pore volume, porosity, surface area
Procedia PDF Downloads 1703590 Body Mass Index, Components of Metabolic Syndrome and Hyperuricemia among Women in Postmenopausal Period
Authors: Vladyslav Povoroznyuk, Galina Dubetska, Roksolana Povoroznyuk
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In recent years, the problem of hyperuricemia is getting a particular importance due to its increased incidence in the world population. The aim of this study was to determine uriс acid level in blood serum, incidence of hyperuricemia among women in postmenopausal period and their association with body mass index and some components of metabolic syndrome (triglyceride, cholesterol, systolic and diastolic pressure). We examined 412 women in postmenopausal period. They were divided in to the following groups: I group (BMI = 18,5-24,9), II group (BMI = 25,0-29,9), III group (BMI = 30,0-34,9), IV group (BMI > 35). We determined uric acid level among women during postmenopausal period depending on their body mass index. The higher level of uric acid was found in patients with the maximal body mass index (BMI > 35). In the I group it was 277,52 ± 8,40; in the II group – 286,81 ± 7,79; in the III group – 291,81 ± 7,56; in the IV group – 327,17 ± 12,17. Incidence of hyperuricemia among women in the I group was 10,2%, in the II group – 15,9%; in the III group – 21,2%, in the IV group – 34,2%. We found an interdependence between an uric acid level and BMI in the examined women (r = 0,21, p < 0,05). We determined that the highest level of triglyceride (F = 18,62, p < 0,05), cholesterol (F = 3,64, p < 0,05), atherogenic coefficient (F = 22,64, p < 0,05), systolic (F = 10,5, p < 0,05) and diastolic pressure (F = 4,30, p < 0,05) was among women with hyperuricemia. It was an interdependence between an uric acid level and triglyceride (r = 0,26, p < 0,05), atherogenic coefficient (r = 0,24, p < 0,05) among women in postmenopausal period.Keywords: hyperuricemia, uric acid, body mass index, women
Procedia PDF Downloads 1303589 The Effects of Ellagic Acid on Rat Heart Induced Tobacco Smoke
Authors: Nalan Kaya, D. Ozlem Dabak, Gonca Ozan, Elif Erdem, Enver Ozan
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One of the common causes of cardiovascular disease (CVD) is smoking. Moreover, tobacco smoke decreases the amount of oxygen that the blood can carry and increases the tendency for blood clots. Ellagic acid is a powerful antioxidant found especially in red fruits. It was shown to block atherosclerotic process suppressing oxidative stress and inflammation. The aim of this study was to examine the protective effects of ellagic acid against oxidative damage on heart 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. An 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. Heart tissues and blood samples were taken. Histological and biochemical analyzes were performed. Vascular congestion, hyperemic areas, inflammatory cell infiltration and increased connective tissue in the perivascular area were observed in tobacco smoke and tobacco smoke + corn oil groups. Increased connective tissue in the perivascular area, hemorrhage and inflammatory cell infiltration were decreased in tobacco smoke + EA group. Group-II GSH level was not changed (significantly), CAT, SOD, GPx activities were significantly higher than group-I. Compared to group-II, group-IV GSH, SOD, CAT, GPx activities were increased, and MDA level was decreased significantly. Group-II and Group-III levels were similar. The results indicate that ellagic acid could protect the heart tissue from the tobacco smoke harmful effects.Keywords: ellagic acid, heart, rat, tobacco smoke
Procedia PDF Downloads 2273588 Synthesis and Antibacterial Evaluation of Natural Bioactive 3,4-DihydroisocoumarinAnalogues
Authors: Hummera Rafique, Aamer Saeed
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Synthesis of structural analogues of various well known bioactive natural 3,4-dihydroisocoumarins viz. Scorzocreticin, Annulatomarin, Montroumarin, and Thunberginol B, have been carried out starting from 3,5-dimethoxy-4-methylphenyl acetic acid. 3,5-Dimethoxy-4-methylphenyl acetic acid was then condensed with various aryl acid chlorides (a-e) to afford the corresponding 6,8-dimethoxy-7-methyl-3-aryl isocoumarins (5a-e). The alkaline hydrolysis of isocoumarins yields keto-acids (3a-e), which were then reduced to hydroxyacids, followed by cyclodehydration with acetic anhydride furnish corresponding 3,4-dihydroisocoumarins (7a-e). Finally, demethylation of 3,4-dihydroisocoumarins was carried out to afford 6,8-dihydroxy-7-methyl-3-aryl-3,4-dihydroisocoumarins (7a-e). Antibacterial evaluation of all the synthesized compounds were carried out against ten bacterial strains, it was concluded that isocoumarins (5a-e) and 3,4-dihydroisocoumarins (7a-e) are more active against gram positive bacteria then gram negative. However, the 6,8-dihydroxy-3,4-dihydroisocoumarin derivatives (8a-e) are more active against gram negative then gram positive.Keywords: 3, 5-Dimethoxy-4-methylhomophthalic acid, natural 3, 4-Dihydroisocoumarin analogues, antibacterial activity, isocoumarins, demethylation
Procedia PDF Downloads 4053587 The Role of Metallic Mordant in Natural Dyeing Process: Experimental and Quantum Study on Color Fastness
Authors: Bo-Gaun Chen, Chiung-Hui Huang, Mei-Ching Chiang, Kuo-Hsing Lee, Chia-Chen Ho, Chin-Ping Huang, Chin-Heng Tien
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It is known that the natural dyeing of cloth results moderate color, but with poor color fastness. This study points out the correlation between the macroscopic color fastness of natural dye to the cotton fiber and the microscopic binding energy of dye molecule to the cellulose. With the additive metallic mordant, the new-formed coordination bond bridges the dye to the fiber surface and thus affects the color fastness as well as the color appearance. The density functional theory (DFT) calculation is therefore used to explore the most possible mechanism during the dyeing process. Finally, the experimental results reflect the strong effect of three different metal ions on the natural dyeing clothes.Keywords: binding energy, color fastness, density functional theory (DFT), natural dyeing, metallic mordant
Procedia PDF Downloads 5593586 The Role of Piceatannol in Counteracting Glyceraldehyde-3-Phosphate Dehydrogenase Aggregation and Nuclear Translocation
Authors: Joanna Gerszon, Aleksandra Rodacka
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In the pathogenesis of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, protein and peptide aggregation processes play a vital role in contributing to the formation of intracellular and extracellular protein deposits. One of the major components of these deposits is the oxidatively modified glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Therefore, the purpose of this research was to answer the question whether piceatannol, a stilbene derivative, counteracts and/or slows down oxidative stress-induced GAPDH aggregation. The study also aimed to determine if this natural occurring compound prevents unfavorable nuclear translocation of GAPDH in hippocampal cells. The isothermal titration calorimetry (ITC) analysis indicated that one molecule of GAPDH can bind up to 8 molecules of piceatannol (7.3 ± 0.9). As a consequence of piceatannol binding to the enzyme, the loss of activity was observed. Parallel with GAPDH inactivation the changes in zeta potential, and loss of free thiol groups were noted. Nevertheless, the ligand-protein binding does not influence the secondary structure of the GAPDH. Precise molecular docking analysis of the interactions inside the active center allowed to presume that these effects are due to piceatannol ability to assemble a covalent binding with nucleophilic cysteine residue (Cys149) which is directly involved in the catalytic reaction. Molecular docking also showed that simultaneously 11 molecules of ligand can be bound to dehydrogenase. Taking into consideration obtained data, the influence of piceatannol on level of GAPDH aggregation induced by excessive oxidative stress was examined. The applied methods (thioflavin-T binding-dependent fluorescence as well as microscopy methods - transmission electron microscopy, Congo Red staining) revealed that piceatannol significantly diminishes level of GAPDH aggregation. Finally, studies involving cellular model (Western blot analyses of nuclear and cytosolic fractions and confocal microscopy) indicated that piceatannol-GAPDH binding prevents GAPDH from nuclear translocation induced by excessive oxidative stress in hippocampal cells. In consequence, it counteracts cell apoptosis. These studies demonstrate that by binding with GAPDH, piceatannol blocks cysteine residue and counteracts its oxidative modifications, that induce oligomerization and GAPDH aggregation as well as it prevents hippocampal cells from apoptosis by retaining GAPDH in the cytoplasm. All these findings provide a new insight into the role of piceatannol interaction with GAPDH and present a potential therapeutic strategy for some neurological disorders related to GAPDH aggregation. This work was supported by the by National Science Centre, Poland (grant number 2017/25/N/NZ1/02849).Keywords: glyceraldehyde-3-phosphate dehydrogenase, neurodegenerative disease, neuroprotection, piceatannol, protein aggregation
Procedia PDF Downloads 1673585 Effect of Slag Application to Soil Chemical Properties and Rice Yield on Acid Sulphate Soils with Different Pyrite Depth
Authors: Richardo Y. E. Sihotang, Atang Sutandi, Joshua Ginting
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The expansion of marginal soil such as acid sulphate soils for the development of staple crops, including rice was unavoidable. However, acid sulphate soils were less suitable for rice field due to the low fertility and the threats of pyrite oxidation. An experiment using Randomized Complete Block Design was designed to investigate the effect of slag in stabilizing soil reaction (pH), improving soil fertility and rice yield. Experiments were conducted in two locations with different pyrite depth. The results showed that slag application was able to decrease the exchangeable Al and available iron (Fe) as well as increase the soil pH, available-P, soil exchangeable Ca2+, Mg2+, and K+. Furthermore, the slag application increased the plant nutrient uptakes, particularly N, P, K, followed by the increasing of rice yield significantly. Nutrients availability, nutrient uptake, and rice yield were higher in the shallow pyrite soil instead of the deep pyrite soil. In addition, slag application was economically feasible due to the ability to reduce standard fertilizer requirements.Keywords: acid sulphate soils, available nutrients, pyrite, slag
Procedia PDF Downloads 3033584 Microwave Assisted Thermal Cracking of Castor Oil Zeolite ZSM-5 as Catalyst for Biofuel Production
Authors: Ghazi Faisal Najmuldeen, Ali Abdul Rahman–Al Ezzi, Tharmathas A/L Alagappan
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The aim of this investigation was to produce biofuel from castor oil through microwave assisted thermal cracking with zeolite ZSM-5 as catalyst. The obtained results showed that microwave assisted thermal cracking of castor oil with Zeolite ZSM-5 as catalyst generates products consisting of alcohol, methyl esters and fatty acids. The products obtained from this experimental procedure by the cracking of castor oil are components of biodiesel. Samples of cracked castor oil containing 1, 3 and 5wt % catalyst was analyzed, however, only the sample containing the 5wt % catalyst showed significant presence of condensate. FTIR and GCMS studies show that the condensate obtained is an unsaturated fatty acid, is 9, 12-octadecadienoic acid, suitable for biofuel use. 9, 12-octadecadienoic acid is an unsaturated fatty acid with a molecular weight of 280.445 g/mol. Characterization of the sample demonstrates that functional group for the products from the three samples display a similar peak in the FTIR graph analysis at 1700 cm-1 and 3600 cm-1. The result obtained from GCMS shows that there are 16 peaks obtained from the sample. The compound with the highest peak area is 9, 12-octadecadienoic acid with a retention time of 9.941 and 24.65 peak areas. All these compounds are organic material and can be characterized as biofuel and biodiesel.Keywords: castor oil, biofuel, biodiesel, thermal cracking, microwave
Procedia PDF Downloads 2333583 Chemiluminescent Detection of Microorganisms in Food/Drug Product Using Reducing Agents and Gold Nanoplates
Authors: Minh-Phuong Ngoc Bui, Abdennour Abbas
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Microbial spoilage of food/drug has been a constant nuisance and an unavoidable problem throughout history that affects food/drug quality and safety in a variety of ways. A simple and rapid test of fungi and bacteria in food/drugs and environmental clinical samples is essential for proper management of contamination. A number of different techniques have been developed for detection and enumeration of foodborne microorganism including plate counting, enzyme-linked immunosorbent assay (ELISA), polymer chain reaction (PCR), nucleic acid sensor, electrical and microscopy methods. However, the significant drawbacks of these techniques are highly demand of operation skills and the time and cost involved. In this report, we introduce a rapid method for detection of bacteria and fungi in food/drug products using a specific interaction between a reducing agent (tris(2-carboxylethyl)phosphine (TCEP)) and the microbial surface proteins. The chemical reaction was transferred to a transduction system using gold nanoplates-enhanced chemiluminescence. We have optimized our nanoplates synthetic conditions, characterized the chemiluminescence parameters and optimized conditions for the microbial assay. The new detection method was applied for rapid detection of bacteria (E.coli sp. and Lactobacillus sp.) and fungi (Mucor sp.), with limit of detection as low as single digit cells per mL within 10 min using a portable luminometer. We expect our simple and rapid detection method to be a powerful alternative to the conventional plate counting and immunoassay methods for rapid screening of microorganisms in food/drug products.Keywords: microorganism testing, gold nanoplates, chemiluminescence, reducing agents, luminol
Procedia PDF Downloads 2993582 Structural and Functional Comparison of Untagged and Tagged EmrE Protein
Authors: S. Junaid S. Qazi, Denice C. Bay, Raymond Chew, Raymond J. Turner
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EmrE, a member of the small multidrug resistance protein family in bacteria is considered to be the archetypical member of its family. It confers host resistance to a wide variety of quaternary cation compounds (QCCs) driven by proton motive force. Generally, purification yield is a challenge in all membrane proteins because of the difficulties in their expression, isolation and solubilization. EmrE is extremely hydrophobic which make the purification yield challenging. We have purified EmrE protein using two different approaches: organic solvent membrane extraction and hexahistidine (his6) tagged Ni-affinity chromatographic methods. We have characterized changes present between ligand affinity of untagged and his6-tagged EmrE proteins in similar membrane mimetic environments using biophysical experimental techniques. Purified proteins were solubilized in a buffer containing n-dodecyl-β-D-maltopyranoside (DDM) and the conformations in the proteins were explored in the presence of four QCCs, methyl viologen (MV), ethidium bromide (EB), cetylpyridinium chloride (CTP) and tetraphenyl phosphonium (TPP). SDS-Tricine PAGE and dynamic light scattering (DLS) analysis revealed that the addition of QCCs did not induce higher multimeric forms of either proteins at all QCC:EmrE molar ratios examined under the solubilization conditions applied. QCC binding curves obtained from the Trp fluorescence quenching spectra, gave the values of dissociation constant (Kd) and maximum specific one-site binding (Bmax). Lower Bmax values to QCCs for his6-tagged EmrE shows that the binding sites remained unoccupied. This lower saturation suggests that the his6-tagged versions provide a conformation that prevents saturated binding. Our data demonstrate that tagging an integral membrane protein can significantly influence the protein.Keywords: small multidrug resistance (SMR) protein, EmrE, integral membrane protein folding, quaternary ammonium compounds (QAC), quaternary cation compounds (QCC), nickel affinity chromatography, hexahistidine (His6) tag
Procedia PDF Downloads 3793581 The Multiple Sclerosis condition and the Role of Varicella-zoster virus in its Progression
Authors: Sina Mahdavi, Mahdi Asghari Ozma
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Multiple sclerosis (MS) is the most common inflammatory autoimmune disease of the CNS that affects the myelination process in the central nervous system (CNS). Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially human Varicella-zoster virus (VZV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on VZV retrovirus infection in MS disease progression. For this study, the keywords "Multiple sclerosis", " Human Varicella-zoster virus ", and "central nervous system" in the databases PubMed, Google Scholar, Sid, and MagIran between 2016 and 2022 were searched and 14 articles were chosen, studied, and analyzed. Analysis of the amino acid sequences of HNRNPA1 with VZV proteins has shown a 62% amino acid sequence similarity between VZV gE and the PrLD/M9 epitope region (TNPO1 binding domain) of mutant HNRNPA1. A heterogeneous nuclear ribonucleoprotein (hnRNP), which is produced by HNRNPA1, is involved in the processing and transfer of mRNA and pre-mRNA. Mutant HNRNPA1 mimics gE of VZV as an antigen that leads to autoantibody production. Mutant HnRNPA1 translocates to the cytoplasm, after aggregation is presented by MHC class I, followed by CD8 + cells. Of these, antibodies and immune cells against the gE epitopes of VZV remain due to the memory immune response, causing neurodegeneration and the development of MS in genetically predisposed individuals. VZV expression during the course of MS is present in genetically predisposed individuals with HNRNPA1 mutation, suggesting a link between VZV and MS, and that this virus may play a role in the development of MS by inducing an inflammatory state. Therefore, measures to modulate VZV expression may be effective in reducing inflammatory processes in demyelinated areas of MS patients in genetically predisposed individuals.Keywords: multiple sclerosis, varicella-zoster virus, central nervous system, autoimmunity
Procedia PDF Downloads 763580 Production of Soy Yoghurt Using Soymilk-Based Lactic Acid Bacteria as Starter Culture
Authors: Ayobami Solomon Popoola, Victor N. Enujiugha
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Production of soy-yogurt by fermentation of soymilk with lactic acid bacteria isolated from soymilk was studied. Soymilk was extracted from dehulled soybean seeds and pasteurized at 95 °C for 15 min. The soymilk was left to naturally ferment (temperature 40 °C; time 8 h) and lactic acid bacteria were isolated, screened and selected for yogurt production. Freshly prepared soymilk was pasteurized (95 °C, 15 min), inoculated with the lactic acid bacteria isolated (3% w/v starter culture) and incubated at 40 °C for 8 h. The yogurt produced was stored at 4 °C. Investigations were carried out with the aim of improving the sensory qualities and acceptability of soy yogurt. Commercial yogurt was used as a control. The percentage of soymilk inoculated was 70% of the broth. Soy-yoghurt samples produced were subsequently subjected to biochemical and microbiological assays which included total viable counts of fresh milk and soy-based yoghurt; proximate composition of functional soy-based yoghurt fermented with Lactobacillus plantarum; changes in pH, Titratable acidity, and lactic acid bacteria during a 14 day period of storage; as well as morphological and biochemical characteristics of lactic acid bacteria isolated. The results demonstrated that using Lactobacillus plantarum to inoculate soy milk for yogurt production takes about 8 h. The overall acceptability of the soy-based yogurt produced was not significantly different from that of the control sample. The use of isolate from soymilk had the added advantage of reducing the cost of yogurt starter culture, thereby making soy-yogurt, a good source of much desired good quality protein. However, more experiments are needed to improve the sensory qualities such as beany or astringent flavor and color.Keywords: soy, soymilk, yoghurt, starter culture
Procedia PDF Downloads 2643579 Polyacrylates in Poly (Lactic Acid) Matrix, New Biobased Polymer Material
Authors: Irena Vuković-Kwiatkowska, Halina Kaczmarek
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Poly (lactic acid) is well known polymer, often called green material because of its origin (renewable resources) and biodegradability. This biopolymer can be used in the packaging industry very often. Poor resistance to permeation of gases is the disadvantage of poly (lactic acid). The permeability of gases and vapor through the films applied for packages and bottles generally should be very low to prolong products shelf-life. We propose innovation method of PLA gas barrier modification using electromagnetic radiation in ultraviolet range. Poly (lactic acid) (PLA) and multifunctional acrylate monomers were mixed in different composition. Final films were obtained by photochemical reaction (photocrosslinking). We tested permeability to water vapor and carbon dioxide through these films. Also their resistance to UV radiation was also studied. The samples were conditioned in the activated sludge and in the natural soil to test their biodegradability. An innovative method of PLA modification allows to expand its usage, and can reduce the future costs of waste management what is the result of consuming such materials like PET and HDPE. Implementation of our material for packaging will contribute to the protection of the environment from the harmful effects of extremely difficult to biodegrade materials made from PET or other plasticKeywords: interpenetrating polymer network, packaging films, photocrosslinking, polyacrylates dipentaerythritol pentaacrylate DPEPA, poly (lactic acid), polymer biodegradation
Procedia PDF Downloads 4793578 Metabolic Engineering of Yarrowia Lipolytica for the Simultaneous Production of Succinic Acid (SA) and Polyhydroxyalkanoates (PHAs)
Authors: Qingsheng Qi, Cuijuan Gao, Carol Sze Ki Lin
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Food waste can be defined as a by-product of food processing by industries and consumers, which has not been recycled or used for other purposes. Stringent waste regulations worldwide are pushing local companies and sectors towards higher sustainability standards. The development of novel strategies for food waste re-use is economically and environmentally sound, as it solves a waste management issue and represents an inexpensive nutrient source for biotechnological processes. For example, Yarrowia lipolytica is a yeast which can utilize hydrophobic substrates, such as fatty acids, lipids, and alkanes and simple carbon sources, such as glucose and glycerol, which can all be found in food waste. This broad substrate range makes Y. lipolytica a promising candidate for the degradation and valorisation of food waste, and for the production of organic acids, such as citric and α-ketoglutaric acids. Current research conducted in our group demonstrated that Y. lipolytica was shown to be able to produce succinic acid. In this talk, we will focus on the application of genetically modified yeast Y. lipolytica for fermentative succinic acid production with an aim to increase productivity and yield.Keywords: food waste, succinic acid, Yarrowia lipolytica, bioplastic
Procedia PDF Downloads 2933577 Structural and Functional Characterization of the Transcriptional Regulator Rv1176 of Mycobacterium tuberculosis H37Rv
Authors: Vikash Yadav, Ashish Arora
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Microorganisms have self-defense mechanisms to protect themselves from toxic environments. Phenolic acid decarboxylase(pad) is responsible for the defense against toxicity caused by phenolic acids, converting them into less toxic vinyl derivatives. The transcription of the pad gene is regulated by a negative transcription factor, phenolic acid decarboxylase regulators (PadR), in a substrate-inducible manner. The PadR family members share the conserved DNA-binding features and interact with the operator DNA using a winged helix-turn-helix (wHTH) motif, which contains a three-helix motif and a β-stranded wing. The members of this family function as transcriptional regulators that are involved in various cellular survival processes, such as toxin production, detoxification, multidrug resistance, antibiotic biosynthesis, and carbon catabolism. Rv1176 of Mycobacterium tuberculosis H37Rv has been assigned to the PadR family protein that remains to be structurally and functionally uncharacterized. To reveal the structural mechanism by which Rv1176 could regulates effector-responsive transcription, several experiments were performed, including Electrophoretic Mobility Shift Assay (EMSA) for DNA protein interaction, differential scanning calorimetry (DSC) and Differential Scanning Fluorimetry (DSF) for temperature and ligand-dependent protein stability, Circular Dichroism (CD) spectroscopy for secondary structure analysis. Further, to evaluate the functional role of Rv1176, the intracellular survival of recombinant M. smegmatis was examined in murine macrophage cell line J774A.1 and different stressed conditions like oxidative, pH, and nutritive stress. All these studies demonstrated that Rv1176 could behave as a transcription regulator and its expression in recombinant M. smegmatis increases intracellular survival.Keywords: EMSA, Mycobacterium tuberculosis, PadR family protein, transcriptional regulator
Procedia PDF Downloads 803576 The Effect of Supercritical Fluid on the Extraction Efficiency of Heavy Metal from Soil
Authors: Haifa El-Sadi, Maria Elektorowicz, Reed Rushing, Ammar Badawieh, Asif Chaudry
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Clay soils have particular properties that affect the assessment and remediation of contaminated sites. In clay soils, electro-kinetic transport of heavy metals has been carried out. The transport of these metals is predicated on maintaining a low pH throughout the cell, which, in turn, keeps the metals in the pore water phase where they are accessible to electro-kinetic transport. Supercritical fluid extraction and acid digestion were used for the analysis of heavy metals concentrations after the completion of electro-kinetic experimentation. Supercritical fluid (carbon dioxide) extraction is a new technique used to extract the heavy metal (lead, nickel, calcium and potassium) from clayey soil. The comparison between supercritical extraction and acid digestion of different metals was carried out. Supercritical fluid extraction, using ethylenediaminetetraacetic acid (EDTA) as a modifier, proved to be efficient and a safer technique than acid digestion technique in extracting metals from clayey soil. Mixing time of soil with EDTA before extracting heavy metals from clayey soil was investigated. The optimum and most practical shaking time for the extraction of lead, nickel, calcium and potassium was two hours.Keywords: clay soil, heavy metals, supercritical fluid extraction, acid digestion
Procedia PDF Downloads 4693575 Improved Signal-To-Noise Ratio by the 3D-Functionalization of Fully Zwitterionic Surface Coatings
Authors: Esther Van Andel, Stefanie C. Lange, Maarten M. J. Smulders, Han Zuilhof
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False outcomes of diagnostic tests are a major concern in medical health care. To improve the reliability of surface-based diagnostic tests, it is of crucial importance to diminish background signals that arise from the non-specific binding of biomolecules, a process called fouling. The aim is to create surfaces that repel all biomolecules except the molecule of interest. This can be achieved by incorporating antifouling protein repellent coatings in between the sensor surface and it’s recognition elements (e.g. antibodies, sugars, aptamers). Zwitterionic polymer brushes are considered excellent antifouling materials, however, to be able to bind the molecule of interest, the polymer brushes have to be functionalized and so far this was only achieved at the expense of either antifouling or binding capacity. To overcome this limitation, we combined both features into one single monomer: a zwitterionic sulfobetaine, ensuring antifouling capabilities, equipped with a clickable azide moiety which allows for further functionalization. By copolymerizing this monomer together with a standard sulfobetaine, the number of azides (and with that the number of recognition elements) can be tuned depending on the application. First, the clickable azido-monomer was synthesized and characterized, followed by copolymerizing this monomer to yield functionalizable antifouling brushes. The brushes were fully characterized using surface characterization techniques like XPS, contact angle measurements, G-ATR-FTIR and XRR. As a proof of principle, the brushes were subsequently functionalized with biotin via strain-promoted alkyne azide click reactions, which yielded a fully zwitterionic biotin-containing 3D-functionalized coating. The sensing capacity was evaluated by reflectometry using avidin and fibrinogen containing protein solutions. The surfaces showed excellent antifouling properties as illustrated by the complete absence of non-specific fibrinogen binding, while at the same time clear responses were seen for the specific binding of avidin. A great increase in signal-to-noise ratio was observed, even when the amount of functional groups was lowered to 1%, compared to traditional modification of sulfobetaine brushes that rely on a 2D-approach in which only the top-layer can be functionalized. This study was performed on stoichiometric silicon nitride surfaces for future microring resonator based assays, however, this methodology can be transferred to other biosensor platforms which are currently being investigated. The approach presented herein enables a highly efficient strategy for selective binding with retained antifouling properties for improved signal-to-noise ratios in binding assays. The number of recognition units can be adjusted to a specific need, e.g. depending on the size of the analyte to be bound, widening the scope of these functionalizable surface coatings.Keywords: antifouling, signal-to-noise ratio, surface functionalization, zwitterionic polymer brushes
Procedia PDF Downloads 3073574 Drought Alters the Expression of a Candidate Zea Mays P-Coumarate 3-Hydroxylase Gene and Caffeic Acid Biosynthesis
Authors: Zintle Kolo, Ndiko Ludidi
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The enzymatic activity of p-coumarate 3-hydroxylase (C3H) synthesize caffeic acid from p-coumaric acid. We recently showed that exogenously applied caffeic acid confers salinity tolerance in soybean (Glycine max) by inducing antioxidant enzymatic activity to promote enhanced scavenging or reactive oxygen species, thus limiting salinity-induced oxidative stress. Recent evidence also establishes that pre-treatment of plants with exogenously supplied caffeic acid improves plant tolerance to osmotic stress by improving plant antioxidant capacity and enhancing biosynthesis of compatible solutes. We aimed to identify a C3H in maize (Zea mays) and evaluate the effect of drought on the spatial and temporal expression of the gene encoding the candidate maize C3H (ZmC3H). Primary sequence analysis shows that ZmC3H shares 71% identity with an Arabidopsis thaliana C3H that is implicated in the control of Arabidopsis cell expansion, growth, and responses to stress. In silico ZmC3H promoter analysis reveals the presence of cis-acting elements that interact with transcription factors implicated in plant responses to drought. Spatial expression analysis by semi-quantitative RT-PCR shows that ZmC3H is expressed in both leaves and roots under normal conditions. However, drought represses the expression of ZmC3H in leaves whereas it up-regulates its expression in roots. These changes in ZmC3H expression correlate with the changes in the content of caffeic acid in maize in response to drought. We illustrate the implications of these changes in the expression of the gene in relation to maize responses to drought and discuss the potential of regulating caffeic acid biosynthesis towards genetic improvement of maize tolerance to drought stress. These findings have implications for food security because of the potential of the implications of the study for drought tolerance in maize.Keywords: caffeic acid, drought-responsive expression, maize drought tolerance, p-coumarate 3-hydroxylase
Procedia PDF Downloads 4753573 Interaction of Low-Energy Positrons with Mg Atoms: Elastic Scattering, Bound States, and Annihilation
Authors: Mahasen M. Abdel Mageed, H. S. Zaghloul
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Annihilations, phase shifts, scattering lengths, and elastic cross sections of low energy positrons scattering from magnesium atoms were studied using the least-squares variational method (LSVM). The possibility of positron binding to the magnesium atoms is investigated. A trial wavefunction is suggested to represent e+-Mg elastic scattering and scattering parameters were derived to estimate the binding energy and annihilation rates. The trial function is taken to depend on several adjustable parameters and is improved iteratively by increasing the number of terms. The present results have the same behavior as reported semi-empirical, theoretical, and experimental results. Especially, the estimated positive scattering length supports the possibility of positron-magnesium bound state system that was confirmed in previous experimental and theoretical work.Keywords: bound wavefunction, positron annihilation, scattering phase shift, scattering length
Procedia PDF Downloads 5573572 Carbothermic Reduction of Phosphoric Acid Extracted from Dephosphorization Slags to Produce Yellow Phosphorus
Authors: Ryoko Yoshida, Jyunpei Yoshida, Hua Fang Yu, Yasushi Sasaki, Tetsuya Nagasaka
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Phosphorous is an important element for agriculture and industry and is a non-renewable resource. Especially, yellow phosphorus is an essential material in advanced industrial technology, but phosphorus resources were not produced in Japan at all, and all depend on imports. It has been suggested, however, that the remaining accessible reserves of phosphate ore will be depleted within 50 years. Therefore, alternative resources for phosphate ore must be found. In this research, we have developed a process that enables the production of high-purity yellow phosphorus from domestic unused phosphorus resources such as steelmaking slags. The process consists of two parts: (1) the production of crude phosphoric acid from wastes such as steelmaking slag; (2) producing high-purity yellow phosphorus by low-temperature carbothermic reduction of phosphoric acid (H3PO4). The details of the carbothermic reduction of phosphoric acid are presented in this paper. Yellow phosphorus is commercially produced by carbothermic reduction of phosphate ore in an electric arc furnace at more than 1673K. In the newly developed system, gaseous P4O10 evaporated from H3PO4 is successfully reduced to yellow phosphorus by using carbon packed bed at less than 1273K. To meet the depletion of phosphate ore, the proposed process in this study to produce yellow phosphorus by carbothermic reduction of H3PO4 that are extracted from dephosphorization slags will be one of the effective and economical solutions.Keywords: carbothermic reduction, phosphoric acid, dephosphorization slags, yellow phosphorus
Procedia PDF Downloads 1213571 Benefits of Monitoring Acid Sulfate Potential of Coffee Rock (Indurated Sand) across Entire Dredge Cycle in South East Queensland
Authors: S. Albert, R. Cossu, A. Grinham, C. Heatherington, C. Wilson
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Shipping trends suggest increasing vessel size and draught visiting Australian ports highlighting potential challenges to port infrastructure and requiring optimization of shipping channels to ensure safe passage for vessels. The Port of Brisbane in Queensland, Australia has an 80 km long access shipping channel which vessels must transit 15 km of relatively shallow coffee rock (generic class of indurated sands where sand grains are bound within an organic clay matrix) outcrops towards the northern passage in Moreton Bay. This represents a risk to shipping channel deepening and maintenance programs as the dredgeability of this material is more challenging due to its high cohesive strength compared with the surrounding marine sands and potential higher acid sulfate risk. In situ assessment of acid sulfate sediment for dredge spoil control is an important tool in mitigating ecological harm. The coffee rock in an anoxic undisturbed state does not pose any acid sulfate risk, however when disturbed via dredging it’s vital to ensure that any present iron sulfides are either insignificant or neutralized. To better understand the potential risk we examined the reduction potential of coffee rock across the entire dredge cycle in order to accurately portray the true outcome of disturbed acid sulfate sediment in dredging operations in Moreton Bay. In December 2014 a dredge trial was undertaken with a trailing suction hopper dredger. In situ samples were collected prior to dredging revealed acid sulfate potential above threshold guidelines which could lead to expensive dredge spoil management. However, potential acid sulfate risk was then monitored in the hopper and subsequent discharge, both showing a significant reduction in acid sulfate potential had occurred. Additionally, the acid neutralizing capacity significantly increased due to the inclusion of shell fragments (calcium carbonate) from the dredge target areas. This clearly demonstrates the importance of assessing potential acid sulfate risk across the entire dredging cycle and highlights the need to carefully evaluate sources of acidity.Keywords: acid sulfate, coffee rock, indurated sand, dredging, maintenance dredging
Procedia PDF Downloads 3683570 Computational Investigation of V599 Mutations of BRAF Protein and Its Control over the Therapeutic Outcome under the Malignant Condition
Authors: Mayank, Navneet Kaur, Narinder Singh
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The V599 mutations in the BRAF protein are extremely oncogenic, responsible for countless of malignant conditions. Along with wild type, V599E, V599D, and V599R are the important mutated variants of the BRAF proteins. The BRAF inhibitory anticancer agents are continuously developing, and sorafenib is a BRAF inhibitor that is under clinical use. The crystal structure of sorafenib bounded to wild type, and V599 is known, showing a similar interaction pattern in both the case. The mutated 599th residue, in both the case, is also found not interacting directly with the co-crystallized sorafenib molecule. However, the IC50 value of sorafenib was found extremely different in both the case, i.e., 22 nmol/L for wild and 38 nmol/L for V599E protein. Molecular docking study and MMGBSA binding energy results also revealed a significant difference in the binding pattern of sorafenib in both the case. Therefore, to explore the role of distinctively situated 599th residue, we have further conducted comprehensive computational studies. The molecular dynamics simulation, residue interaction network (RIN) analysis, and residue correlation study results revealed the importance of the 599th residue on the therapeutic outcome and overall dynamic of the BRAF protein. Therefore, although the position of 599th residue is very much distinctive from the ligand-binding cavity of BRAF, still it has exceptional control over the overall functional outcome of the protein. The insight obtained here may seem extremely important and guide us while designing ideal BRAF inhibitory anticancer molecules.Keywords: BRAF, oncogenic, sorafenib, computational studies
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