Search results for: protein extraction
3411 Extraction of Forest Plantation Resources in Selected Forest of San Manuel, Pangasinan, Philippines Using LiDAR Data for Forest Status Assessment
Authors: Mark Joseph Quinto, Roan Beronilla, Guiller Damian, Eliza Camaso, Ronaldo Alberto
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Forest inventories are essential to assess the composition, structure and distribution of forest vegetation that can be used as baseline information for management decisions. Classical forest inventory is labor intensive and time-consuming and sometimes even dangerous. The use of Light Detection and Ranging (LiDAR) in forest inventory would improve and overcome these restrictions. This study was conducted to determine the possibility of using LiDAR derived data in extracting high accuracy forest biophysical parameters and as a non-destructive method for forest status analysis of San Manual, Pangasinan. Forest resources extraction was carried out using LAS tools, GIS, Envi and .bat scripts with the available LiDAR data. The process includes the generation of derivatives such as Digital Terrain Model (DTM), Canopy Height Model (CHM) and Canopy Cover Model (CCM) in .bat scripts followed by the generation of 17 composite bands to be used in the extraction of forest classification covers using ENVI 4.8 and GIS software. The Diameter in Breast Height (DBH), Above Ground Biomass (AGB) and Carbon Stock (CS) were estimated for each classified forest cover and Tree Count Extraction was carried out using GIS. Subsequently, field validation was conducted for accuracy assessment. Results showed that the forest of San Manuel has 73% Forest Cover, which is relatively much higher as compared to the 10% canopy cover requirement. On the extracted canopy height, 80% of the tree’s height ranges from 12 m to 17 m. CS of the three forest covers based on the AGB were: 20819.59 kg/20x20 m for closed broadleaf, 8609.82 kg/20x20 m for broadleaf plantation and 15545.57 kg/20x20m for open broadleaf. Average tree counts for the tree forest plantation was 413 trees/ha. As such, the forest of San Manuel has high percent forest cover and high CS.Keywords: carbon stock, forest inventory, LiDAR, tree count
Procedia PDF Downloads 3743410 Prospects of Milk Protein as a Potential Alternative of Natural Antibiotic
Authors: Syeda Fahria Hoque Mimmi
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Many new and promising treatments for reducing or diminishing the adverse effects of microorganisms are being discovered day by day. On the other hand, the dairy industry is accelerating the economic wheel of Bangladesh. Considering all these facts, new thoughts were developed to isolate milk proteins by the present experiment for opening up a new era of developing natural antibiotics from milk. Lactoferrin, an iron-binding glycoprotein with multifunctional properties, is crucial to strengthening the immune system and also useful for commercial applications. The protein’s iron-binding capacity makes it undoubtedly advantageous to immune system modulation and different bacterial strains. For fulfilling the purpose, 4 of raw and 17 of commercially available milk samples were collected from different farms and stores in Bangladesh (Dhaka, Chittagong, and Cox’s Bazar). Protein quantification by nanodrop technology has confirmed that raw milk samples have better quantities of protein than the commercial ones. All the samples were tested for their antimicrobial activity against 18 pathogens, where raw milk samples showed a higher percentage of antibacterial activity. In addition to this, SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis) was performed to identify lactoferrin in the milk samples. Lactoferrin was detected in 9 samples from which 4 were raw milk samples. Interestingly, Streptococcus pyogenes, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, Vibrio cholera, Staphylococcus aureus, and enterotoxigenic E. coli significantly displayed sensitivity against lactoferrin collected from raw milk. Only Bacillus cereus, Pseudomonas aeruginosa, Streptococcus pneumonia, Enterococcus faecalis, and ETEC (Enterotoxigenic Escherichia coli) were susceptible to lactoferrin obtained from a commercial one. This study suggested that lactoferrin might be used as the potential alternative of antibiotics for many diseases and also can be used to reduce microbial deterioration in the food and feed industry.Keywords: alternative of antibiotics, commercially available milk, lactoferrin, nanodrop technology, pathogens, raw milk
Procedia PDF Downloads 1633409 Separation of Mercury(Ii) from Petroleum Produced Water via Hollow Fiber Supported Liquid Membrane and Mass Transfer Modeling
Authors: Srestha Chaturabul, Wanchalerm Srirachat, Thanaporn Wannachod, Prakorn Ramakul, Ura Pancharoen, Soorathep Kheawhom
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The separation of mercury(II) from petroleum-produced water from the Gulf of Thailand was carried out using a hollow fiber supported liquid membrane system (HFSLM). Optimum parameters for feed pretreatment were 0.2 M HCl, 4% (v/v) Aliquat 336 for extractant and 0.1 M thiourea for stripping solution. The best percentage obtained for extraction was 99.73% and for recovery 90.11%, respectively. The overall separation efficiency noted was 94.92% taking account of both extraction and recovery prospects. The model for this separation developed along a combined flux principle i.e. convection–diffusion–kinetic. The results showed excellent agreement with theoretical data at an average standard deviation of 1.5% and 1.8%, respectively.Keywords: separation, mercury(ii), petroleum produced water, hollow fiber, liquid membrane
Procedia PDF Downloads 2893408 SARS-CoV-2: Prediction of Critical Charged Amino Acid Mutations
Authors: Atlal El-Assaad
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Viruses change with time through mutations and result in new variants that may persist or disappear. A Mutation refers to an actual change in the virus genetic sequence, and a variant is a viral genome that may contain one or more mutations. Critical mutations may cause the virus to be more transmissible, with high disease severity, and more vulnerable to diagnostics, therapeutics, and vaccines. Thus, variants carrying such mutations may increase the risk to human health and are considered variants of concern (VOC). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) - the contagious in humans, positive-sense single-stranded RNA virus that caused coronavirus disease 2019 (COVID-19) - has been studied thoroughly, and several variants were revealed across the world with their corresponding mutations. SARS-CoV-2 has four structural proteins, known as the S (spike), E (envelope), M (membrane), and N (nucleocapsid) proteins, but prior study and vaccines development focused on genetic mutations in the S protein due to its vital role in allowing the virus to attach and fuse with the membrane of a host cell. Specifically, subunit S1 catalyzes attachment, whereas subunit S2 mediates fusion. In this perspective, we studied all charged amino acid mutations of the SARS-CoV-2 viral spike protein S1 when bound to Antibody CC12.1 in a crystal structure and assessed the effect of different mutations. We generated all missense mutants of SARS-CoV-2 protein amino acids (AAs) within the SARS-CoV-2:CC12.1 complex model. To generate the family of mutants in each complex, we mutated every charged amino acid with all other charged amino acids (Lysine (K), Arginine (R), Glutamic Acid (E), and Aspartic Acid (D)) and studied the new binding of the complex after each mutation. We applied Poisson-Boltzmann electrostatic calculations feeding into free energy calculations to determine the effect of each mutation on binding. After analyzing our data, we identified charged amino acids keys for binding. Furthermore, we validated those findings against published experimental genetic data. Our results are the first to propose in silico potential life-threatening mutations of SARS-CoV-2 beyond the present mutations found in the five common variants found worldwide.Keywords: SARS-CoV-2, variant, ionic amino acid, protein-protein interactions, missense mutation, AESOP
Procedia PDF Downloads 1023407 GraphNPP: A Graphormer-Based Architecture for Network Performance Prediction in Software-Defined Networking
Authors: Hanlin Liu, Hua Li, Yintan AI
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Network performance prediction (NPP) is essential for the management and optimization of software-defined networking (SDN) and contributes to improving the quality of service (QoS) in SDN to meet the requirements of users. Although current deep learning-based methods can achieve high effectiveness, they still suffer from some problems, such as difficulty in capturing global information of the network, inefficiency in modeling end-to-end network performance, and inadequate graph feature extraction. To cope with these issues, our proposed Graphormer-based architecture for NPP leverages the powerful graph representation ability of Graphormer to effectively model the graph structure data, and a node-edge transformation algorithm is designed to transfer the feature extraction object from nodes to edges, thereby effectively extracting the end-to-end performance characteristics of the network. Moreover, routing oriented centrality measure coefficient for nodes and edges is proposed respectively to assess their importance and influence within the graph. Based on this coefficient, an enhanced feature extraction method and an advanced centrality encoding strategy are derived to fully extract the structural information of the graph. Experimental results on three public datasets demonstrate that the proposed GraphNPP architecture can achieve state-of-the-art results compared to current NPP methods.Keywords: software-defined networking, network performance prediction, Graphormer, graph neural network
Procedia PDF Downloads 333406 On Exploring Search Heuristics for improving the efficiency in Web Information Extraction
Authors: Patricia Jiménez, Rafael Corchuelo
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Nowadays the World Wide Web is the most popular source of information that relies on billions of on-line documents. Web mining is used to crawl through these documents, collect the information of interest and process it by applying data mining tools in order to use the gathered information in the best interest of a business, what enables companies to promote theirs. Unfortunately, it is not easy to extract the information a web site provides automatically when it lacks an API that allows to transform the user-friendly data provided in web documents into a structured format that is machine-readable. Rule-based information extractors are the tools intended to extract the information of interest automatically and offer it in a structured format that allow mining tools to process it. However, the performance of an information extractor strongly depends on the search heuristic employed since bad choices regarding how to learn a rule may easily result in loss of effectiveness and/or efficiency. Improving search heuristics regarding efficiency is of uttermost importance in the field of Web Information Extraction since typical datasets are very large. In this paper, we employ an information extractor based on a classical top-down algorithm that uses the so-called Information Gain heuristic introduced by Quinlan and Cameron-Jones. Unfortunately, the Information Gain relies on some well-known problems so we analyse an intuitive alternative, Termini, that is clearly more efficient; we also analyse other proposals in the literature and conclude that none of them outperforms the previous alternative.Keywords: information extraction, search heuristics, semi-structured documents, web mining.
Procedia PDF Downloads 3293405 Characteristics of Sorghum (Sorghum bicolor L. Moench) Flour on the Soaking Time of Peeled Grains and Particle Size Treatment
Authors: Sri Satya Antarlina, Elok Zubaidah, Teti Istiana, Harijono
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Sorghum bicolor (Sorghum bicolor L. Moench) has the potential as a flour for gluten-free food products. Sorghum flour production needs grain soaking treatment. Soaking can reduce the tannin content which is an anti-nutrient, so it can increase the protein digestibility. Fine particle size decreases the yield of flour, so it is necessary to study various particle sizes to increase the yield. This study aims to determine the characteristics of sorghum flour in the treatment of soaking peeled grain and particle size. The material of white sorghum varieties KD-4 from farmers in East Java, Indonesia. Factorial randomized factorial design (two factors), repeated three times, factor I were the time of grain soaking (five levels) that were 0, 12, 24, 36, and 48 hours, factor II was the size of the starch particles sifted with a fineness level of 40, 60, 80, and 100 mesh. The method of making sorghum flour is grain peeling, soaking peeled grain, drying using the oven at 60ᵒC, milling, and sieving. Physico-chemical analysis of sorghum flour. The results show that there is an interaction between soaking time of grain with the size of sorghum flour particles. Interaction in yield of flour, L* color (brightness level), whiteness index, paste properties, amylose content, protein content, bulk density, and protein digestibility. The method of making sorghum flour through the soaking of peeled grain and the difference in particle size has an important role in producing the physicochemical properties of the specific flour. Based on the characteristics of sorghum flour produced, it is determined the method of making sorghum flour through sorghum grain soaking for 24 hours, the particle size of flour 80 mesh. The sorghum flour with characteristic were 24.88% yield of flour, 88.60 color L* (brightness level), 69.95 whiteness index, 3615 Cp viscosity, 584.10 g/l of bulk density, 24.27% db protein digestibility, 90.02% db starch content, 23.4% db amylose content, 67.45% db amylopectin content, 0.22% db crude fiber content, 0.037% db tannin content, 5.30% db protein content, ash content 0.18% db, carbohydrate content 92.88 % db, and 1.94% db fat content. The sorghum flour is recommended for cookies products.Keywords: characteristic, sorghum (Sorghum bicolor L. Moench) flour, grain soaking, particle size, physicochemical properties
Procedia PDF Downloads 1523404 Inclusion Body Refolding at High Concentration for Large-Scale Applications
Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening
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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.Keywords: dialysis, inclusion body, refolding, solubilization
Procedia PDF Downloads 2863403 Volarization of Sugarcane Bagasse: The Effect of Alkali Concentration, Soaking Time and Temperature on Fibre Yield
Authors: Tamrat Tesfaye, Tilahun Seyoum, K. Shabaridharan
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The objective of this paper was to determine the effect of NaOH concentration, soaking time, soaking temperature and their interaction on percentage yield of fibre extract using Response Surface Methodology (RSM). A Box-Behnken design was employed to optimize the extraction process of cellulosic fibre from sugar cane by-product bagasse using low alkaline extraction technique. The quadratic model with the optimal technological conditions resulted in a maximum fibre yield of 56.80% at 0.55N NaOH concentration, 4 h steeping time and 60ᵒC soaking temperature. Among the independent variables concentration was found to be the most significant (P < 0.005) variable and the interaction effect of concentration and soaking time leads to securing the optimized processes.Keywords: sugarcane bagasse, low alkaline, Box-Behnken, fibre
Procedia PDF Downloads 2403402 Production and Evaluation of Jam Made from Pineapple (Ananas comosus) and Grape (Vitis vinifera)
Authors: Z. O. Apotiola, J. F. Fashakin
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This project studied the production and evaluation of jam produced from pineapple and grape at different level of ratio (90:10, 80:20, 70:30, 60:40, 50:50, and 100%). The proximate and sensory properties were determined using standard methods. The (GDZ) was the highest for protein, moisture, fat and ash, (KFJ) was the highest for carbohydrate. There were significant differences (p<0.05) in samples (PAB, GDZ, BEN) for moisture. Also, there were significant differences (p<0.05) in samples (PAB, BBL, GDZ, KFJ) for protein. There were significant differences (p<0.05) in samples (PAB, BBL, BEN) for carbohydrate. Also, there were significant differences (p<0.05) in samples (PAB, BBL, QCM, GDZ, BEN) for fat and there were significant differences (p<0.05) in samples (PAB, BBL, GDZ) for ash. (KFJ) was the highest for pH, (BBL and QCM) was the highest for Vitamin C; (GDZ) was the highest for titratable acidity. For sensory properties, for aroma, colour, flavour, and overall acceptability were tested using panellists; the result showed that (KFJ) had the highest for all samples. From the results of chemical and sensory characteristics sample BBL was the best combination.Keywords: chemical, characteristic, combination, titratable, sensory, significant
Procedia PDF Downloads 2673401 The Effects of Highly Active Antiretroviral Therapy (HAART) on the Expression of Muc1 and P65 in a Cervical Cancer Cell Line, HCS-2
Authors: K. R. Thabethe, G. A. Adefolaju, M. J. Hosie
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Cervical cancer is the third most commonly diagnosed cancer globally and it is one of three AIDS defining malignancies. Highly active antiretroviral therapy (HAART) is a combination of three or more antiretroviral drugs and has been shown to play a significant role in reducing the incidence of some AIDS defining malignancies, although its effect on cervical cancer is still unclear. The aim of this study was to investigate the relationship between cervical cancer and HAART. This was achieved by studying the expression of two signalling molecules expressed in cervical cancer; MUC1 and P65. Following the 24 hour treatment of a cervical cancer cell line, HCS-2, with drugs which are commonly used as part of HAART at their clinical plasma concentrations, real-time qPCR and immunofluorescence were used in order to study gene and protein expression. A one way ANOVA followed by a Tukey Kramer Post Hoc test was conducted using JMP 11 software on both sets of data. The drug classified as a protease inhibitor (PI) (i.e. LPV/r) reduced MUC1 and P65 gene and protein expression more than the other drug tested. PIs are known to play a significant role in cell death, therefore the cells were thought to be more susceptible to cell death following treatment with PIs. In conclusion, the drugs used, especially the PI showed some anticancer effects by facilitating cell death through decreased gene and protein expression of MUC1 and P65 and present promising agents for cancer treatment.Keywords: cervical cancer, haart, MUC1, P65
Procedia PDF Downloads 3253400 Producing Lutein Powder from Algae by Extraction and Drying
Authors: Zexin Lei, Timothy Langrish
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Lutein is a type of carotene believed to be beneficial to the eyes. This study aims to explore the possibility of using a closed cycle spray drying system to produce lutein. The system contains a spray dryer, a condenser, a heater, and a pressure seal. Hexane, ethanol, and isopropanol will be used as organic solvents to compare the extraction effects. Several physical and chemical methods of cell disruption will be compared. By continuously sweeping the system with nitrogen, the oxygen content will be controlled below 2%, reducing the concentration of organic solvent below the explosion limit and preventing lutein from being oxidized. Lutein powder will be recovered in the collection device. The volatile organic solvent will be cooled in the condenser and deposited in the bottom until it is discharged from the bottom of the condenser.Keywords: closed cycle spray drying system, Chlorella vulgaris, organic solvent, solvent recovery
Procedia PDF Downloads 1273399 Exploring Penicillin Resistance in Gonococcal Penicillin Binding Protein-2: Molecular Docking and Ligand Interaction Analysis
Authors: Sinethemba Yakobi, Lindiwe Zuma, Ofentse Pooe
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Gonococcal infections present a notable public health issue, and the major approach for treatment involves using β-lactam antibiotics that specifically target penicillin-binding protein 2 (PBP2) in Neisseria gonorrhoeae. This study examines the influence of flavonoids, namely rutin, on the structural changes of PBP2 in both penicillin-resistant (FA6140) and penicillin-susceptible (FA19) strains. The research clarifies the structural effects of particular mutations, such as inserting an aspartate residue at position 345 (Asp-345a) in the PBP2 protein. The strain FA6140, which is resistant to penicillin, shows specific changes that lead to a decrease in penicillin binding. These mutations, namely P551S and F504L, significantly impact the pace at which acylation occurs and the stability of the strain under high temperatures. Molecular docking analyses investigate the antibacterial activities of rutin and other phytocompounds, emphasizing its exceptional binding affinity and potential as an inhibitor of PBP2. Quercetin and protocatechuic acid have encouraging antibacterial effectiveness, with quercetin displaying characteristics similar to those of drugs. Molecular dynamics simulations offer a detailed comprehension of the interactions between flavonoids and PBP2, highlighting rutin's exceptional antioxidant effects and strong affinity for the substrate binding site. The study's wider ramifications pertain to the pressing requirement for antiviral treatments in the context of the ongoing COVID-19 epidemic. Flavonoids have a strong affinity for binding to PBP2, indicating their potential as inhibitors to impair cell wall formation in N. gonorrhoeae. Ultimately, this study provides extensive knowledge on the interactions between proteins and ligands, the dynamics of the structure, and the ability of flavonoids to combat penicillin-resistant N. gonorrhoeae bacteria. The verified simulation outcomes establish a basis for creating potent inhibitors and medicinal therapies to combat infectious illnesses.Keywords: phytochemicals, penicillin-binding protein 2, gonococcal infection, ligand-protein interaction, binding energy, neisseria gonorrhoeae FA19, neisseria gonorrhoeae FA6140, flavonoids
Procedia PDF Downloads 583398 DNAJB6 Chaperone Prevents the Aggregation of Intracellular but not Extracellular Aβ Peptides Associated with Alzheimer’s Disease
Authors: Rasha M. Hussein, Reem M. Hashem, Laila A. Rashed
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Alzheimer’s disease is the most common dementia disease in the elderly. It is characterized by the accumulation of extracellular amyloid β (Aβ) peptides and intracellular hyper-phosphorylated tau protein. In addition, recent evidence indicates that accumulation of intracellular amyloid β peptides may play a role in Alzheimer’s disease pathogenesis. This suggests that intracellular Heat Shock Proteins (HSP) that maintain the protein quality control in the cell might be potential candidates for disease amelioration. DNAJB6, a member of DNAJ family of HSP, effectively prevented the aggregation of poly glutamines stretches associated with Huntington’s disease both in vitro and in cells. In addition, DNAJB6 was found recently to delay the aggregation of Aβ42 peptides in vitro. In the present study, we investigated the ability of DNAJB6 to prevent the aggregation of both intracellular and extracellular Aβ peptides using transfection of HEK293 cells with Aβ-GFP and recombinant Aβ42 peptides respectively. We performed western blotting and immunofluorescence techniques. We found that DNAJB6 can prevent Aβ-GFP aggregation, but not the seeded aggregation initiated by extracellular Aβ peptides. Moreover, DNAJB6 required interaction with HSP70 to prevent the aggregation of Aβ-GFP protein and its J-domain was essential for this anti-aggregation activity. Interestingly, overexpression of other DNAJ proteins as well as HSPB1 suppressed Aβ-GFP aggregation efficiently. Our findings suggest that DNAJB6 is a promising candidate for the inhibition of Aβ-GFP mediated aggregation through a canonical HSP70 dependent mechanism.Keywords: Aβ, Alzheimer’s disease, chaperone, DNAJB6, aggregation
Procedia PDF Downloads 5033397 Improving the Quality of Casava Peel-Leaf Mixture through Fermentation with Rhizopus oligosporusas Poultry Ration
Authors: Mirnawati, G. Ciptaan, Ferawati
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This study aims to improve the quality of the cassava peel-leaf mixture (CPLM) through fermentation with Rhizopus oligosporusas poultry ration. This research is an experimental study using a completely randomized design (CRD) with four treatments and five replications. The treatments were cassava peel-leaf mixture (CPLM) fermented with Rhizopus oligosporus. The treatments were a combination of cassava peel and leaves with the ratio of; A (9:1), B (8:2), C (7:3), and D (6:4). The observed variables were protease enzyme activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and metabolic energy. The results of the diversity analysis showed that there was a very significant (p < 0.01) effect on protease activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and energy metabolism of fermented CPLM. Based on the results of the study, it can be concluded that CPLM (6:4) fermented with Rhizopus oligosporus gave the best results seen from protease activity 7,25 U/ml, 21.23% crude protein, 19.80% crude fiber, 59.65% nitrogen retention, 62.99% crude fiber digestibility and metabolic energy 2671 Kcal/kg.Keywords: quality, Casava peel-leaf mixture, fermentation, Rhizopus oligosporus
Procedia PDF Downloads 1723396 Transfer of Constraints or Constraints on Transfer? Syntactic Islands in Danish L2 English
Authors: Anne Mette Nyvad, Ken Ramshøj Christensen
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In the syntax literature, it has standardly been assumed that relative clauses and complement wh-clauses are islands for extraction in English, and that constraints on extraction from syntactic islands are universal. However, the Mainland Scandinavian languages has been known to provide counterexamples. Previous research on Danish has shown that neither relative clauses nor embedded questions are strong islands in Danish. Instead, extraction from this type of syntactic environment is degraded due to structural complexity and it interacts with nonstructural factors such as the frequency of occurrence of the matrix verb, the possibility of temporary misanalysis leading to semantic incongruity and exposure over time. We argue that these facts can be accounted for with parametric variation in the availability of CP-recursion, resulting in the patterns observed, as Danish would then “suspend” the ban on movement out of relative clauses and embedded questions. Given that Danish does not seem to adhere to allegedly universal syntactic constraints, such as the Complex NP Constraint and the Wh-Island Constraint, what happens in L2 English? We present results from a study investigating how native Danish speakers judge extractions from island structures in L2 English. Our findings suggest that Danes transfer their native language parameter setting when asked to judge island constructions in English. This is compatible with the Full Transfer Full Access Hypothesis, as the latter predicts that Danish would have difficulties resetting their [+/- CP-recursion] parameter in English because they are not exposed to negative evidence.Keywords: syntax, islands, second language acquisition, danish
Procedia PDF Downloads 1143395 A Galectin from Rock Bream Oplegnathus fasciatus: Molecular Characterization and Immunological Properties
Authors: W. S. Thulasitha, N. Umasuthan, G. I. Godahewa, Jehee Lee
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In fish, innate immune defense is the first immune response against microbial pathogens which consists of several antimicrobial components. Galectins are one of the carbohydrate binding lectins that have the ability to identify pathogen by recognition of pathogen associated molecular patterns. Galectins play a vital role in the regulation of innate and adaptive immune responses. Rock bream Oplegnathus fasciatus is one of the most important cultured species in Korea and Japan. Considering the losses due to microbial pathogens, present study was carried out to understand the molecular and functional characteristics of a galectin in normal and pathogenic conditions, which could help to establish an understanding about immunological components of rock bream. Complete cDNA of rock bream galectin like protein B (rbGal like B) was identified from the cDNA library, and the in silico analysis was carried out using bioinformatic tools. Genomic structure was derived from the BAC library by sequencing a specific clone and using Spidey. Full length of rbGal like B (contig14775) cDNA containing 517 nucleotides was identified from the cDNA library which comprised of 435 bp in the open reading frame encoding a deduced protein composed of 145 amino acids. The molecular mass of putative protein was predicted as 16.14 kDa with an isoelectric point of 8.55. A characteristic conserved galactose binding domain was located from 12 to 145 amino acids. Genomic structure of rbGal like B consisted of 4 exons and 3 introns. Moreover, pairwise alignment showed that rock bream rbGal like B shares highest similarity (95.9 %) and identity (91 %) with Takifugu rubripes galectin related protein B like and lowest similarity (55.5 %) and identity (32.4 %) with Homo sapiens. Multiple sequence alignment demonstrated that the galectin related protein B was conserved among vertebrates. A phylogenetic analysis revealed that rbGal like B protein clustered together with other fish homologs in fish clade. It showed closer evolutionary link with Takifugu rubripes. Tissue distribution and expression patterns of rbGal like B upon immune challenges were performed using qRT-PCR assays. Among all tested tissues, level of rbGal like B expression was significantly high in gill tissue followed by kidney, intestine, heart and spleen. Upon immune challenges, it showed an up-regulated pattern of expression with Edwardsiella tarda, rock bream irido virus and poly I:C up to 6 h post injection and up to 24 h with LPS. However, In the presence of Streptococcus iniae rbGal like B showed an up and down pattern of expression with the peak at 6 - 12 h. Results from the present study revealed the phylogenetic position and role of rbGal like B in response to microbial infection in rock bream.Keywords: galectin like protein B, immune response, Oplegnathus fasciatus, molecular characterization
Procedia PDF Downloads 3453394 Comparative Analysis of Oil Extracts from Cotton and Watermelon Seeds
Authors: S. A. Jumare, A. O. Tijani, M. F. Siraj, B. V. Babatunde
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This research investigated the comparative analysis of oil extracted from cotton and watermelon seeds using solvent extraction process. Normal ethyl-ether was used as solvent in the extraction process. The AOAC method of Analysis was employed in the determination of the physiochemical properties of the oil. The chemical properties of the oil determined include the saponification value, free fatty acid, iodine value, peroxide value and acid value. The physical properties of the oil determined include specific gravity, refractive index, colour, odour, taste and pH. The value obtained for cottonseed oil are saponification value (187mgKOH/g), free fatty acid (5.64mgKOH/g), iodine value (95.2g/100), peroxide value (9.33meq/kg), acid value (11.22mg/KOH/g), pH value (4.62), refractive index (1.46), and specific gravity (0.9) respectively, it has a bland odour, a reddish brown colour and a mild taste. The values obtained for watermelon seed oil are saponification value (83.3mgKOH/g), free fatty acid (6.58mg/KOH/g), iodine value (122.6g/100), peroxide value (5.3meq/kg), acid value (3.74mgKOH/g), pH value (6.3), refractive index (1.47), and specific gravity (0.9) respectively, it has a nutty flavour, a golden yellow colour and a mild taste. From the result obtained, it shows that cottonseed oil has high acid value which shows the stability of the oil and its stability to rancidity. Consequently, watermelon seed oil is order wise.Keywords: extraction, solvent, cotton seeds, watermelon seeds
Procedia PDF Downloads 3493393 Competition in Petroleum Extraction and the Challenges of Climate Change
Authors: Saeid Rabiei Majd, Motahareh Alvandi, Bahareh Asefi
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Extraction of maximum natural resources is one of the common policies of governments, especially petroleum resources that have high economic and strategic value. The incentive to access and maintain profitable oil markets for governments or international oil companies, causing neglects them to pay attention to environmental principles and sustainable development, which in turn drives up environmental and climate change. Significant damage to the environment can cause severe damage to citizens and indigenous people, such as the compulsory evacuation of their zone due to contamination of water and air resources, destruction of animals and plants. Hawizeh Marshes is a common aquatic and environmental ecosystem along the Iran-Iraq border that also has oil resources. This marsh has been very rich in animal, vegetative, and oil resources. Since 1990, the political motives, the strategic importance of oil extraction, and the disregard for the environmental rights of the Iraqi and Iranian governments in the region have caused 90% of the marshes and forced migration of indigenous people. In this paper, we examine the environmental degradation factors resulting from the adoption of policies and practices of governments in this region based on the principles of environmental rights and sustainable development. Revision of the implementation of the government’s policies and natural resource utilization systems can prevent the spread of climate change, which is a serious international challenge today.Keywords: climate change, indigenous rights, petroleum operation, sustainable development principles, sovereignty on resources
Procedia PDF Downloads 1033392 Reorientation of Sustainable Livestock Management: A Case Study Applied to Wastes Management in Faculty of Animal Husbandry, Padjadjaran University, Indonesia
Authors: Raka Rahmatulloh, Mohammad Ilham Nugraha, Muhammad Ifan Fathurrahman
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The agricultural sector covers a wide area, one of them is livestock subsector that supply needs of the food source of animal protein. Animal protein is produced by the main livestock production such as meat, milk, eggs, etc. Besides the main production, livestock would produce metabolic residue, so called livestock wastes. Characteristics of livestock wastes can be either solid (feces), liquid (urine), and gas (methane) which turned out to be useful and has economical value when well-processed and well-controlled. Nowadays, this livestock wastes is considered as a source of pollutants, especially water pollution. If the source of pollutants used in an integrated way, it will have a positive impact on organic farming and a healthy environment. Management of livestock wastes can be integrated with the farming sector to the planting and caring that rely on fertilizers. Most Indonesian farmers still use chemical fertilizers, where the use of it in the long term will disturb the ecological balance of the environment. One of the main efforts is to use organic fertilizers instead of chemical fertilizer that conducted by the Faculty of Animal Husbandry, Padjadjaran University. The method is to use the solid waste of livestock and agricultural wastes into liquid organic fertilizer, feed additive, biogas and vermicompost through decomposition. The decomposition takes as long as 14 days including aeration and extraction process using water as a nutrients solvent media which contained in decomposes and disinfection media to release pathogenic microorganisms in decomposes. Liquid organic fertilizer has highly efficient for the farmers to have a ratio of carbon/nitrogen (C/N) 25/1 to 30/1 and neutral pH (6.5-7.5) which is good for plant growth. Feed additive may be given to improve the digestibility of feed so that substances can be easily absorbed by the body for production. Biogas contains methane (CH4), which has a high enough heat to produce electricity. Vermicompost is an overhaul of waste organic material that has excellent structure, porosity, aeration, drainage, and moisture holding capacity. Based on the case study above, an integrated livestock wastes management program strongly supports the Indonesian government in the achievement of sustainable livestock development.Keywords: integrated, livestock wastes, organic fertilizer, sustainable livestock development
Procedia PDF Downloads 4283391 Effect of Texturised Soy Protein and Yeast on the Instrumental and Sensory Quality of Hybrid Beef Meatballs
Authors: Simona Grasso, Gabrielle Smith, Sophie Bowers, Oluseyi Moses Ajayi, Mark Swainson
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Hybrid meat analogues are meat products whereby a proportion of meat has been partially replaced by more sustainable protein sources. These products could bridge the gap between meat and meat-free products, providing convenience, and allowing consumers to continue using meat products as they conventionally would, while lowering their overall meat intake. The study aimed to investigate the effect of introducing texturized soy protein (TSP) at different levels (15% and 30%) with and without nutritional yeast as flavour enhancer on the sensory and instrumental quality of beef meatballs, compared to a soy and yeast-free control. Proximate analysis, yield, colour, instrumental texture, and sensory quality were investigated. The addition of soy and yeast did not have significant effects on the overall protein content, but the total fat and moisture content went down with increasing soy substitution. Samples with 30% TSP had significantly higher yield than the other recipes. In terms of colour, a* redness values tended to go down and b* yellowness values tended to go up with increasing soy addition. The addition of increasing levels of soy and yeast modified the structure of meatballs resulting in a progressive decrease in hardness and chewiness compared to control. Sixty participants assessed the samples using Check-all-that-apply (CATA) questions and hedonic scales. The texture of all TSP-containing samples received significantly higher acceptability scores than control, while 15% TSP with yeast received significantly higher flavour and overall acceptability scores than control. Control samples were significantly more often associated than the other recipes to the term 'hard' and the least associated to 'soft' and 'crumbly and easy to cut'. All recipes were similarly associated to the terms 'weak meaty', 'strong meaty', 'characteristic' and 'unusual'. Correspondence analysis separated the meatballs in three distinct groups: 1) control; 2) 30%TSP with yeast; and 3) 15%TSP, 15%TSP with yeast and 30%TSP located together on the sensory map, showing similarity. Adding 15-30% TSP with or without yeast inclusion could be beneficial for the development of future meat hybrids with acceptable sensory quality. These results can provide encouragement for the use of the hybrid concept by the meat industry to promote the partial substitution of meat in flexitarians’ diets.Keywords: CATA, hybrid meat products, texturised soy protein, yeast
Procedia PDF Downloads 1573390 Modeling of Gas Extraction from a Partially Gas-Saturated Porous Gas Hydrate Reservoir with Respect to Thermal Interactions with Surrounding Rocks
Authors: Angelina Chiglintseva, Vladislav Shagapov
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We know from the geological data that quite sufficient gas reserves are concentrated in hydrates that occur on the Earth and on the ocean floor. Therefore, the development of these sources of energy and the storage of large reserves of gas hydrates is an acute global problem. An advanced technology for utilizing gas is to store it in a gas-hydrate state. Under natural conditions, storage facilities can be established, e.g., in underground reservoirs, where quite large volumes of gas can be conserved compared with reservoirs of pure gas. An analysis of the available experimental data of the kinetics and the mechanism of the gas-hydrate formation process shows the self-conservation effect that allows gas to be stored at negative temperatures and low values of pressures of up to several atmospheres. A theoretical model has been constructed for the gas-hydrate reservoir that represents a unique natural chemical reactor, and the principal possibility of the full extraction of gas from a hydrate due to the thermal reserves of the reservoirs themselves and the surrounding rocks has been analyzed. The influence exerted on the evolution of a gas hydrate reservoir by the reservoir thicknesses and the parameters that determine its initial state (a temperature, pressure, hydrate saturation) has been studied. It has been established that the shortest time of exploitation required by the reservoirs with a thickness of a few meters for the total hydrate decomposition is recorded in the cyclic regime when gas extraction alternated with the subsequent conservation of the gas hydrate deposit. The study was performed by a grant from the Russian Science Foundation (project No.15-11-20022).Keywords: conservation, equilibrium state, gas hydrate reservoir, rocks
Procedia PDF Downloads 2883389 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 703388 Terrain Classification for Ground Robots Based on Acoustic Features
Authors: Bernd Kiefer, Abraham Gebru Tesfay, Dietrich Klakow
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The motivation of our work is to detect different terrain types traversed by a robot based on acoustic data from the robot-terrain interaction. Different acoustic features and classifiers were investigated, such as Mel-frequency cepstral coefficient and Gamma-tone frequency cepstral coefficient for the feature extraction, and Gaussian mixture model and Feed forward neural network for the classification. We analyze the system’s performance by comparing our proposed techniques with some other features surveyed from distinct related works. We achieve precision and recall values between 87% and 100% per class, and an average accuracy at 95.2%. We also study the effect of varying audio chunk size in the application phase of the models and find only a mild impact on performance.Keywords: acoustic features, autonomous robots, feature extraction, terrain classification
Procedia PDF Downloads 3533387 Extraction of Inulin from Cichorium Intybus and Its Application as Fat Replacer in Yoghurt
Authors: Hafiz Khuram Wasim Aslam, Muhammad Saeed, Azam Shakeel, Muhammad Inam Ur Raheem, Moazzam Rafiq Khan, Muhammad Atif Randhawa
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Inulin is significant ingredient used in food industry that functions technologically as a fat replacer often without compromising taste and texture. In this study inulin was extracted from the chicory roots and the effect of inulin addition as a fat replacer on the physiochemical, microbiological and sensory properties of non-fat yogurt was investigated. The supplementation of chicory inulin reduced the magnitude of firmness in comparison with non-inulin ¬supplemented non-fat yoghurt. Higher values of acidity were observed due to the more microbial fermentation in the inulin containing yogurt as compared to non-inulin yogurt and were in the range of 0.56 to 0.75 during storage days. Syneresis in control sample increased from 43.9% to 47.9% during the storage study. However inulin addition at different treatment enhanced syneresis from 44.5% to 47.6%. Inulin addition at various concentrations caused an increase in the TPC due to its probiotic effect. No effects of inuline addition on fat and protein contents were observed. Non-fat yoghurt supplemented with inulin demonstrated sensory behavior better than that of the control yoghurt. The most important effect of the addition of inulin to non-fat yoghurt is an increase in the sensory attributes appearance, body and texture, taste and mouth feel, overall acceptability. On an average, yoghurt supplemented with 1 to 2% inulin was better in overall acceptance as compared to control yoghurt.Keywords: inulin, fat replacer, yoghurt, sensory evaluation, low fat
Procedia PDF Downloads 5813386 Engineering C₃ Plants with SbtA, a Cyanobacterial Transporter, for Enhancing CO₂ Fixation
Authors: Vandana Deopanée Tomar, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra, Sandhya Mehrotra
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The cyanobacterial CO₂ concentrating mechanism (CCM) operates to raise the levels of CO₂ in the vicinity of the main carboxylation enzyme Rubisco which is encapsulated in protein micro compartments called carboxysomes. Thus, due to the presence of CCM, cyanobacterial cells are able to work with high photosynthetic efficiency even at low Ci conditions and can accumulate 1000 folds high internal concentrations of Ci than external environment. Engineering of some useful CCM components into higher plants is one of the plausible approaches to improve their photosynthetic performance. The first step and the simplest approach for attaining this objective would be the transfer of cyanobacterial bicarbonate transporter such as SbtA to inner chloroplast envelope of C₃ plants. For this, SbtA transporter gene from Synechococcus elongatus PCC 7942 was fused to a transit peptide element to generate chimeric constructs in order to direct it to chloroplast inner envelope. Two transit peptides namely, TnaXTP (transit peptide from AT3G56160) and TMDTP (transit peptide from AT2G02590) were shortlisted from Arabidopsis thaliana genome and cloned in plant expression vector pCAMBIA1302 having mgfp5 as a reporter gene. Plant transformation was done by agro infiltration and Agrobacterium mediated co-culture. DNA, RNA, and protein were isolated from the leaves four days post infiltration, and the presence of transgene was confirmed by gene specific PCR (Polymerase Chain Reaction) analysis and by RT-PCR (Reverse Transcription Polymerase Chain Reaction). The expression was confirmed at the protein level by western blotting using anti-GFP primary antibody and horseradish peroxidase (HRP) conjugated secondary antibody. The localization of the protein was detected by confocal microscopy of isolated protoplasts. We observed chloroplastic expression for both the fusion constructs which suggest that the transit peptide sequences are capable of taking the cargo protein to the chloroplasts. These constructs are now being used to generate stable transgenic plants by Agrobacterium mediated transformation. The stability of transgene expression will be analyzed from T₀ to T₂ generation.Keywords: agro infiltration, bicarbonate transporter, carbon concentrating mechanisms, cyanobacteria, SbtA
Procedia PDF Downloads 2083385 Nanoparticles-Protein Hybrid-Based Magnetic Liposome
Authors: Amlan Kumar Das, Avinash Marwal, Vikram Pareek
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Liposome plays an important role in medical and pharmaceutical science as e.g. nano scale drug carriers. Liposomes are vesicles of varying size consisting of a spherical lipid bilayer and an aqueous inner compartment. Magnet-driven liposome used for the targeted delivery of drugs to organs and tissues1. These liposome preparations contain encapsulated drug components and finely dispersed magnetic particles. Liposomes are vesicles of varying size consisting of a spherical lipid bilayer and an aqueous inner compartment that are generated in vitro. These are useful in terms of biocompatibility, biodegradability, and low toxicity, and can control biodistribution by changing the size, lipid composition, and physical characteristics2. Furthermore, liposomes can entrap both hydrophobic and hydrophilic drugs and are able to continuously release the entrapped substrate, thus being useful drug carriers. Magnetic liposomes (MLs) are phospholipid vesicles that encapsulate magneticor paramagnetic nanoparticles. They are applied as contrast agents for magnetic resonance imaging (MRI)3. The biological synthesis of nanoparticles using plant extracts plays an important role in the field of nanotechnology4. Green-synthesized magnetite nanoparticles-protein hybrid has been produced by treating Iron (III)/Iron(II) chloride with the leaf extract of Dhatura Inoxia. The phytochemicals present in the leaf extracts act as a reducing as well stabilizing agents preventing agglomeration, which include flavonoids, phenolic compounds, cardiac glycosides, proteins and sugars. The magnetite nanoparticles-protein hybrid has been trapped inside the aqueous core of the liposome prepared by reversed phase evaporation (REV) method using oleic and linoleic acid which has been shown to be driven under magnetic field confirming the formation magnetic liposome (ML). Chemical characterization of stealth magnetic liposome has been performed by breaking the liposome and release of magnetic nanoparticles. The presence iron has been confirmed by colour complex formation with KSCN and UV-Vis study using spectrophotometer Cary 60, Agilent. This magnet driven liposome using nanoparticles-protein hybrid can be a smart vesicles for the targeted drug delivery.Keywords: nanoparticles-protein hybrid, magnetic liposome, medical, pharmaceutical science
Procedia PDF Downloads 2433384 Liquid-Liquid Equilibrium Study in Solvent Extraction of o-Cresol from Coal Tar
Authors: Dewi Selvia Fardhyanti, Astrilia Damayanti
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Coal tar is a liquid by-product of the process of coal gasification and carbonation, also in some industries such as steel, power plant, cement, and others. This liquid oil mixture contains various kinds of useful compounds such as aromatic compounds and phenolic compounds. These compounds are widely used as raw material for insecticides, dyes, medicines, perfumes, coloring matters, and many others. This research investigates thermodynamic modelling of liquid-liquid equilibria (LLE) in solvent extraction of o-Cresol from the coal tar. The equilibria are modeled by ternary components of Wohl, Van Laar, and Three-Suffix Margules models. The values of the parameters involved are obtained by curve-fitting to the experimental data. Based on the comparison between calculated and experimental data, it turns out that among the three models studied, the Three-Suffix Margules seems to be the best to predict the LLE of o-Cresol for those system.Keywords: coal tar, o-Cresol, Wohl, Van Laar, three-suffix margules
Procedia PDF Downloads 2703383 Development and Investigation of Efficient Substrate Feeding and Dissolved Oxygen Control Algorithms for Scale-Up of Recombinant E. coli Cultivation Process
Authors: Vytautas Galvanauskas, Rimvydas Simutis, Donatas Levisauskas, Vykantas Grincas, Renaldas Urniezius
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The paper deals with model-based development and implementation of efficient control strategies for recombinant protein synthesis in fed-batch E.coli cultivation processes. Based on experimental data, a kinetic dynamic model for cultivation process was developed. This model was used to determine substrate feeding strategies during the cultivation. The proposed feeding strategy consists of two phases – biomass growth phase and recombinant protein production phase. In the first process phase, substrate-limited process is recommended when the specific growth rate of biomass is about 90-95% of its maximum value. This ensures reduction of glucose concentration in the medium, improves process repeatability, reduces the development of secondary metabolites and other unwanted by-products. The substrate limitation can be enhanced to satisfy restriction on maximum oxygen transfer rate in the bioreactor and to guarantee necessary dissolved carbon dioxide concentration in culture media. In the recombinant protein production phase, the level of substrate limitation and specific growth rate are selected within the range to enable optimal target protein synthesis rate. To account for complex process dynamics, to efficiently exploit the oxygen transfer capability of the bioreactor, and to maintain the required dissolved oxygen concentration, adaptive control algorithms for dissolved oxygen control have been proposed. The developed model-based control strategies are useful in scale-up of cultivation processes and accelerate implementation of innovative biotechnological processes for industrial applications.Keywords: adaptive algorithms, model-based control, recombinant E. coli, scale-up of bioprocesses
Procedia PDF Downloads 2503382 In Silico Analysis of Deleterious nsSNPs (Missense) of Dihydrolipoamide Branched-Chain Transacylase E2 Gene Associated with Maple Syrup Urine Disease Type II
Authors: Zainab S. Ahmed, Mohammed S. Ali, Nadia A. Elshiekh, Sami Adam Ibrahim, Ghada M. El-Tayeb, Ahmed H. Elsadig, Rihab A. Omer, Sofia B. Mohamed
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Maple syrup urine (MSUD) is an autosomal recessive disease that causes a deficiency in the enzyme branched-chain alpha-keto acid (BCKA) dehydrogenase. The development of disease has been associated with SNPs in the DBT gene. Despite that, the computational analysis of SNPs in coding and noncoding and their functional impacts on protein level still remains unknown. Hence, in this study, we carried out a comprehensive in silico analysis of missense that was predicted to have a harmful influence on DBT structure and function. In this study, eight different in silico prediction algorithms; SIFT, PROVEAN, MutPred, SNP&GO, PhD-SNP, PANTHER, I-Mutant 2.0 and MUpo were used for screening nsSNPs in DBT including. Additionally, to understand the effect of mutations in the strength of the interactions that bind protein together the ELASPIC servers were used. Finally, the 3D structure of DBT was formed using Mutation3D and Chimera servers respectively. Our result showed that a total of 15 nsSNPs confirmed by 4 software (R301C, R376H, W84R, S268F, W84C, F276C, H452R, R178H, I355T, V191G, M444T, T174A, I200T, R113H, and R178C) were found damaging and can lead to a shift in DBT gene structure. Moreover, we found 7 nsSNPs located on the 2-oxoacid_dh catalytic domain, 5 nsSNPs on the E_3 binding domain and 3 nsSNPs on the Biotin Domain. So these nsSNPs may alter the putative structure of DBT’s domain. Furthermore, we detected all these nsSNPs are on the core residues of the protein and have the ability to change the stability of the protein. Additionally, we found W84R, S268F, and M444T have high significance, and they affected Leucine, Isoleucine, and Valine, which reduces or disrupt the function of BCKD complex, E2-subunit which the DBT gene encodes. In conclusion, based on our extensive in-silico analysis, we report 15 nsSNPs that have possible association with protein deteriorating and disease-causing abilities. These candidate SNPs can aid in future studies on Maple Syrup Urine Disease type II base in the genetic level.Keywords: DBT gene, ELASPIC, in silico analysis, UCSF chimer
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