Search results for: recombinant protein purification
2658 Comprehending the Relationship between the Red Blood Cells of a Protein 4.1 -/- Patient and Those of Healthy Controls: A Comprehensive Analysis of Tandem Mass Spectrometry Data
Authors: Ahmed M. Hjazi, Bader M. Hjazi
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Protein 4.1 is a crucial component of complex interactions between the cytoskeleton and other junctional complex proteins. When the gene encoding this protein is altered, resulting in reduced expression, or when the protein is absent, the red cell undergoes a significant structural change. This research aims to achieve a deeper comprehension of the biochemical effects of red cell protein deficiency. A Tandem Mass Spectrometry Analysis (TMT-MS/MS) of patient cells lacking protein 4.1 compared to three healthy controls was achieved by the Proteomics Institute of the University of Bristol. The SDS-PAGE and Western blotting were utilized on the original patient sample and controls to partially confirm TMT MS/MS data analysis of the protein-4.1-deficient cells. Compared to healthy controls, protein levels in samples lacking protein 4.1 had a significantly higher concentration of proteins that probably originated from reticulocytes. This could occur if the patient has an elevated reticulocyte count. The increase in chaperone and reticulocyte-associated proteins was most notable in this study. This may result from elevated quantities of reticulocytes in patients with hereditary elliptocytosis.Keywords: hereditary elliptocytosis, protein 4.1, red cells, tandem mass spectrometry data.
Procedia PDF Downloads 792657 Computational Identification of Signalling Pathways in Protein Interaction Networks
Authors: Angela U. Makolo, Temitayo A. Olagunju
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The knowledge of signaling pathways is central to understanding the biological mechanisms of organisms since it has been identified that in eukaryotic organisms, the number of signaling pathways determines the number of ways the organism will react to external stimuli. Signaling pathways are studied using protein interaction networks constructed from protein-protein interaction data obtained using high throughput experimental procedures. However, these high throughput methods are known to produce very high rates of false positive and negative interactions. In order to construct a useful protein interaction network from this noisy data, computational methods are applied to validate the protein-protein interactions. In this study, a computational technique to identify signaling pathways from a protein interaction network constructed using validated protein-protein interaction data was designed. A weighted interaction graph of the Saccharomyces cerevisiae (Baker’s Yeast) organism using the proteins as the nodes and interactions between them as edges was constructed. The weights were obtained using Bayesian probabilistic network to estimate the posterior probability of interaction between two proteins given the gene expression measurement as biological evidence. Only interactions above a threshold were accepted for the network model. A pathway was formalized as a simple path in the interaction network from a starting protein and an ending protein of interest. We were able to identify some pathway segments, one of which is a segment of the pathway that signals the start of the process of meiosis in S. cerevisiae.Keywords: Bayesian networks, protein interaction networks, Saccharomyces cerevisiae, signalling pathways
Procedia PDF Downloads 5462656 Ingini Seeds: A Qualitative Study on Its Use in Water Purification in the Dry Zone of Sri Lanka
Authors: Iranga Weerakkody, Palitha Sri Geegana Arachchige, Dasith Tilakaratna
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The aim of this research is to study how folk wisdom can be applied to assist in the process of purification of water. This is qualitative research, and by random sampling, it is focused on to the dry zone of Sri Lanka. The research limitation has been set to the use of Ingini seeds (Strychnos potatorum) to purify water. Here the research is based on connecting traditional knowledge regarding water purification using Ingini seeds to modern times and the advantages and disadvantages of using Ingini seeds to purify water sources. Ingini seeds have been used among villagers of the dry zone to purify water for a long time by methods such as planting Ingini plants around water sources and depositing seeds covered with a cotton cloth inside wells. Crushed Ingini seeds have been put into clay water pots to reduce the hardness of water, as well as the number of impurities present in the water. This shows that Ingini seeds have a property that is successful in precipitating dissolved impurities in water. Ingini seeds are also used to precipitate solid impurities in herbal wine. The advantages of using Ingini seeds are that it can be obtained naturally from the ecology without an additional cost and that it is completely organic forest produce. Another specialty is that in practices, it is used to treat kidney stones and other water-related diseases affecting the kidneys.Keywords: folklife, Ingini seeds, Strychnos potatorum, organic forest produce, water purification
Procedia PDF Downloads 1972655 Practical Experiences in the Development of a Lab-Scale Process for the Production and Recovery of Fucoxanthin
Authors: Alma Gómez-Loredo, José González-Valdez, Jorge Benavides, Marco Rito-Palomares
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Fucoxanthin is a carotenoid that exerts multiple beneficial effects on human health, including antioxidant, anti-cancer, antidiabetic and anti-obesity activity; making the development of a whole process for its production and recovery an important contribution. In this work, the lab-scale production and purification of fucoxanthin in Isocrhysis galbana have been studied. In batch cultures, low light intensities (13.5 μmol/m2s) and bubble agitation were the best conditions for production of the carotenoid with product yields of up to 0.143 mg/g. After fucoxanthin ethanolic extraction from biomass and hexane partition, further recovery and purification of the carotenoid has been accomplished by means of alcohol – salt Aqueous Two-Phase System (ATPS) extraction followed by an ultrafiltration (UF) step. An ATPS comprised of ethanol and potassium phosphate (Volume Ratio (VR) =3; Tie-line Length (TLL) 60% w/w) presented a fucoxanthin recovery yield of 76.24 ± 1.60% among the studied systems and was able to remove 64.89 ± 2.64% of the carotenoid and chlorophyll pollutants. For UF, the addition of ethanol to the original recovered ethanolic ATPS stream to a final relation of 74.15% (w/w) resulted in a reduction of approximately 16% of the protein contents, increasing product purity with a recovery yield of about 63% of the compound in the permeate stream. Considering the production, extraction and primary recovery (ATPS and UF) steps, around a 45% global fucoxanthin recovery should be expected. Although other purification technologies, such as Centrifugal Partition Chromatography are able to obtain fucoxanthin recoveries of up to 83%, the process developed in the present work does not require large volumes of solvents or expensive equipment. Moreover, it has a potential for scale up to commercial scale and represents a cost-effective strategy when compared to traditional separation techniques like chromatography.Keywords: aqueous two-phase systems, fucoxanthin, Isochrysis galbana, microalgae, ultrafiltration
Procedia PDF Downloads 4242654 Experimental Research on the Effect of Activating Temperature on Combustion and Nox Emission Characteristics of Pulverized Coal in a Novel Purification-combustion Reaction System
Authors: Ziqu Ouyang, Kun Su
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A novel efficient and clean coal combustion system, namely the purification-combustion system, was designed by the Institute of Engineering Thermal Physics, Chinese Academy of Science, in 2022. Among them, the purification system was composed of a mesothermal activating unit and a hyperthermal reductive unit, and the combustion system was composed of a mild combustion system. In the purification-combustion system, the deep in-situ removal of coal-N could be realized by matching the temperature and atmosphere in each unit, and thus the NOx emission was controlled effectively. To acquire the methods for realizing the efficient and clean coal combustion, this study investigated the effect of the activating temperature (including 822 °C, 858 °C, 933 °C, 991 °C), which was the key factor affecting the system operation, on combustion and NOx emission characteristics of pulverized coal in a 30 kW purification-combustion test bench. The research result turned out that the activating temperature affected the combustion and NOx emission characteristics significantly. As the activating temperature increased, the temperature increased first and then decreased in the mild combustion unit, and the temperature change in the lower part was much higher than that in the upper part. Moreover, the main combustion region was always located at the top of the unit under different activating temperatures, and the combustion intensity along the unit was weakened gradually. Increasing the activating temperature excessively could destroy the reductive atmosphere early in the upper part of the unit, which wasn’t conducive to the full removal of coal-N in the reductive coal char. As the activating temperature increased, the combustion efficiency increased first and then decreased, while the NOx emission decreased first and then increased, illustrating that increasing the activating temperature properly promoted the efficient and clean coal combustion, but there was a limit to its growth. In this study, the optimal activating temperature was 858 °C. Hence, this research illustrated that increasing the activating temperature properly could realize the mutual matching of improving the combustion efficiency and reducing the NOx emission, and thus guaranteed the clean and efficient coal combustion well.Keywords: activating temperature, combustion characteristics, nox emission, purification-combustion system
Procedia PDF Downloads 892653 Solar-Powered Water Purification Using Ozone and Sand Filtration
Authors: Kayla Youhanaie, Kenneth Dott, Greg Gillis-Smith
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Access to clean water is a global challenge that affects nearly one-third of the world’s population. A lack of safe drinking water negatively affects a person’s health, safety, and economic status. However, many regions of the world that face this clean water challenge also have high solar energy potential. To address this worldwide issue and utilize available resources, a solar-powered water purification device was developed that could be implemented in communities around the world that lack access to potable water. The device uses ozone to destroy water-borne pathogens and sand filtration to filter out particulates from the water. To select the best method for this application, a quantitative energy efficiency comparison of three water purification methods was conducted: heat, UV light, and ozone. After constructing an initial prototype, the efficacy of the device was tested using agar petri dishes to test for bacteria growth in treated water samples at various time intervals after applying the device to contaminated water. The results demonstrated that the water purification device successfully removed all bacteria and particulates from the water within three minutes, making it safe for human consumption. These results, as well as the proposed design that utilizes widely available resources in target communities, suggest that the device is a sustainable solution to address the global water crisis and could improve the quality of life for millions of people worldwide.Keywords: clean water, solar powered water purification, ozonation, sand filtration, global water crisis
Procedia PDF Downloads 792652 Inheritance of Protein Content and Grain Yield in Half Diallel Maize (Zea mays L.) Populations
Authors: Gül Ebru Orhun
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A half diallel crossing design was carried out during 2011 and 2012 growing seasons under Çanakkale-Turkey ecological conditions. In this research, 20 F1 maize hybrids obtained by 6x6 half diallel crossing were used. Gene action for protein content and grain yield traits were explored in half set involving six elite inbred lines. According to the results diallel analysis dominance and additive gene variances were determined for protein content. Variance/Co-variance graphs revealed for grain yield and protein content traits. In this study, inheritance of grain yield and protein content demonstrated over-dominance type of gene action.Keywords: protein, maize, inheritance, gene action
Procedia PDF Downloads 5252651 Interaction of Dietary Protein and Vitamin E Supplementation on Gastrointestinal Nematode (Gnt) Parasitism of Naturally Infected Lambs
Authors: Ayobami Adeyemo, Michael Chimonyo, Munyaradzi Marufu
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Gastrointestinal nematode (GNT) infection significantly hinder sustainable and profitable sheep production on rangelands. While vitamin E and protein supplementation have individually proven to improve host immunity to parasitism in lambs, to our knowledge, there is no information on the interaction of dietary vitamin E and protein supplementation on lamb growth and GIN faecal egg counts in naturally infected lambs. Therefore, the current study investigated the interaction of dietary protein and vitamin E supplementation on faecal egg counts (FEC) and growth performance of lambs. Twenty four Dohne Merino lambs aged 12 months were allocated equally to each of four treatment combinations, with six lambs in each treatment group for a period of eight weeks. Treatment one lambs received dietary protein and vitamin E (PE), treatment two lambs received dietary protein and no vitamin E (PNE), treatment three received dietary vitamin E and no protein (NPE), and treatment four received no dietary protein and vitamin E supplementation (NPNE). The lambs were allowed to graze on Pennisetum clandestinum contaminated with a heavy load of nematodes. Dietary protein supplementation increased (P < 0.01) average daily gain (ADG) and body condition scores (BCS). Dietary vitamin E supplementation had no effect (P > 0.05) on ADG and BCS. There was no interaction (P > 0.05) between dietary protein and vitamin E supplementation on ADG and BCS. Combined supplementation of dietary protein and vitamin E supplementation significantly reduced (P < 0.01) faecal egg counts and larval counts, respectively. Also, dietary protein and vitamin E supplementation reduced GNT faecal egg counts over the exposure period. The current findings support the hypothesis that the interaction of dietary protein and vitamin E supplementation reduced faecal egg counts and larval counts in lambs. This necessitates future findings on the interaction of dietary protein and vitamin E supplementation on blood associated profiles.Keywords: gastrointestinal nematodes, nematode eggs, Haemonchus, Trichostrongylus
Procedia PDF Downloads 2102650 Effects of Dietary Protein and Lipid Levels on Growth and Body Composition of Juvenile Fancy Carp, Cyprinus carpio var. Koi
Authors: Jin Choi, Zahra Aminikhoei, Yi-Oh Kim, Sang-Min Lee
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A 4 × 2 factorial experiment was conducted to determine the optimum dietary protein and lipid levels for juvenile fancy carp, Cyprinus carpio var. koi. Eight experimental diets were formulated to contain four protein levels (200, 300, 400, and 500 g kg-1) with two lipid levels (70 and 140 g kg-1). Triplicate groups of fish (initial weight, 12.1±0.2 g fish-1) were hand-fed the diets to apparent satiation for 8 weeks. Weight gain, daily feed intake, feed efficiency ratio and protein efficiency ratio were significantly (P < 0.0001) affected by dietary protein level, but not by dietary lipid level (P > 0.05). Weight gain and feed efficiency ratio tended to increase as dietary protein level increased up to 400 and 500 g kg-1, respectively. Daily feed intake of fish decreased with increasing dietary protein level and that of fish fed diet contained 500 g kg-1 protein was significantly lower than other fish groups. The protein efficiency ratio of fish fed 400 and 500 g kg-1 protein was lower than that of fish fed 200 and 300 g kg-1 protein. Moisture, crude protein and crude lipid contents of muscle and liver were significantly affected by dietary protein, but not by dietary lipid level (P > 0.05). The increase in dietary lipid level resulted in an increase in linoleic acid in liver and muscle paralleled with a decrease in n-3 highly unsaturated fatty acids content in muscle of fish. In considering these results, it was concluded that the diet containing 400 g kg-1 protein with 70 g kg-1 lipid level is optimal for growth and efficient feed utilization of juvenile fancy carp.Keywords: fancy carp, dietary protein, dietary lipid, Cyprinus carpio, fatty acid
Procedia PDF Downloads 4052649 Different Methods of Producing Bioemulsifier by Bacillus licheniformis Strains
Authors: Saba Pajuhan, Afshin Farahbakhsh, S. M. M. Dastgheib
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Biosurfactants and bioemulsifiers are a structurally diverse group of surface-active molecules synthesized by microorganisms, they are amphipathic molecules which reduce surface and interfacial tensions and widely used in pharmaceutical, cosmetic, food and petroleum industries. In this paper, several methods of bioemulsifer synthesis and purification by Bacillus licheniformis strains (namely ACO1, PTCC 1595 and ACO4) were investigated. Strains were grown in nutrient broth with different conditions in order to get maximum production of bioemulsifer. The purification of bio emulsifier and the quality evaluation of the product was done by adding sulfuric acid (H₂SO₄) (98%), Ethanol or HCl to the solution followed by centrifuging. To determine the optimal conditions yielding the highest bioemulsifier production, the effect of various carbon and nitrogen sources, temperature, NaCl concentration, pH, O₂ levels, incubation time are indispensable and all of them were highly effective in bioemulsifiers production.Keywords: biosurfactant, bioemulsifier, purification, surface tension, interfacial tension
Procedia PDF Downloads 2712648 Isolation, Preparation and Biological Properties of Soybean-Flaxseed Protein Co-Precipitates
Authors: Muhammad H. Alu’datt, Inteaz Alli
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This study was conducted to prepare and evaluate the biological properties of protein co-precipitates from flaxseed and soybean. Protein was prepared by NaOH extraction through the mixing of soybean flour (Sf) and flaxseed flour (Ff) or mixtures of soybean extract (Se) and flaxseed extract (Fe). The protein co-precipitates were precipitated by isoelectric (IEP) and isoelectric-heating (IEPH) co-precipitation techniques. Effects of extraction and co-precipitation techniques on co-precipitate yield were investigated. Native-PAGE, SDS-PAGE were used to study the molecular characterization. Content and antioxidant activity of extracted free and bound phenolic compounds were evaluated for protein co-precipitates. Removal of free and bound phenolic compounds from protein co-precipitates showed little effects on the electrophoretic behavior of the proteins or the protein subunits of protein co-precipitates. Results showed that he highest protein contents and yield were obtained in for Sf-Ff/IEP co-precipitate with values of 53.28 and 25.58% respectively as compared to protein isolates and other co-precipitates. Results revealed that the Sf-Ff/IEP showed a higher content of bound phenolic compounds (53.49% from total phenolic content) as compared to free phenolic compounds (46.51% from total phenolic content). Antioxidant activities of extracted bound phenolic compounds with and without heat treatment from Sf-Ff/IEHP were higher as compared to free phenolic compounds extracted from other protein co-precipitates (29.68 and 22.84%, respectively).Keywords: antioxidant, phenol, protein co-precipitate, yield
Procedia PDF Downloads 2402647 Study of Environmental Impact
Authors: Houmame Benbouali
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The risks, in general, exist in any project; one can hardly carry out a project without taking risks. The hydraulic works are rather complex projects in their design, realization and exploitation, and are often subjected at the multiple risks being able to influence with their good performance, and can have an negative impact on their environment. The present study was carried out to quote the impacts caused by purification plant STEP Chlef on the environment, it aims has studies the environmental impacts during construction and when designing this STEP, it is divided into two parts: The first part results from a research task bibliographer which contain three chapters (-cleansing of water worn-general information on water worn-proceed of purification of waste water). The second part is an experimental part which is divided into four chapters (detailed state initial-description of the station of purification-evaluation of the impacts of the project analyzes measurements and recommendations).Keywords: treatment plant, waste water, waste water treatment, environmental impact
Procedia PDF Downloads 5112646 Lucilia Sericata Netrin-A: Secreted by Salivary Gland Larvae as a Potential to Neuroregeneration
Authors: Hamzeh Alipour, Masoumeh Bagheri, Tahereh Karamzadeh, Abbasali Raz, Kourosh Azizi
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Netrin-A, a protein identified for conducting commissural axons, has a similar role in angiogenesis. In addition, studies have shown that one of the netrin-A receptors is expressed in the growing cells of small capillaries. It will be interesting to study this new group of molecules because their role in wound healing will become clearer in the future due to angiogenesis. The greenbottle blowfly Luciliasericata (L. sericata) larvae are increasingly used in maggot therapy of chronic wounds. This aim of this was the identification of moleculareatures of Netrin-A in L. sericata larvae. Larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericataNetrin-A (LSN-A) was then identified using Rapid Amplification of cDNA Ends (RACE) and Rapid Amplification of Genomic Ends (RAGE). Parts of the Netrin-A gene, including the middle, 3′-, and 5′-ends were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its molecular weight is estimated to be 78.6 kDa. The 3-D structure ofNetrin-A drawn by SWISS-MODEL revealed its similarity to the Netrin-1 of humans with 66.8% identity. The LSN-A protein conduces to repair the myelin membrane in neuronal cells. Ultimately, it can be an effective candidate in neural regeneration and wound healing. Furthermore, our next attempt is to deplore recombinant proteins for use in medical sciences.Keywords: maggot therapy, netrin-A, RACE, RAGE, lucilia sericata
Procedia PDF Downloads 1112645 Hermeneutical Understanding of 2 Cor. 7:1 in the Light of Igbo Cultural Concept of Purification
Authors: H. E. Amolo
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The concepts of pollution or contamination and purification or ritual cleansing are very important concepts among traditional Africans. This is because in relation to human behaviors and attitudes, they constitute on the one hand what could be referred to as moral demands and on the other, what results in the default of such demands. The many taboos which a man has to observe are not to be regarded as things mechanical which do not touch the heart, but that the avoidance is a sacred law respected by the community. In breaking it, you offend the divine power’. Researches have shown that, Africans tenaciously hold the belief that, moral values are based upon the recognition of the divine will and that sin in the community must be expelled if perfect peace is to be enjoyed. Sadly enough, these moral values are gradually eroding in contemporary times. Thus, this study proposal calls for a survey of the passage from an African cultural context; how it can enhance the understanding of the text, as well as how it can complement its scholarly interpretation with the view of institutionalizing the concept of holiness as a means of bringing the people closer to God, and also instilling ethical purity and righteousness.Keywords: cultural practices, Igbo ideology, purification, rituals
Procedia PDF Downloads 3092644 Wastewater Treatment Using Sodom Apple Tree in Arid Regions
Authors: D. Oulhaci, M. Zehah, S. Meguellati
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Collected by the sewerage network, the wastewater contains many polluting elements, coming from the population, commercial, industrial and agricultural activities. These waters are collected and discharged into the natural environment and pollute it. Hence the need to transport them before discharge to a treatment plant to undergo several treatment phases. The objective of this study is to highlight the purification performance of the "Sodom apple tree" which is a very common shrub in the region of Djanet and Illizi in Algeria. As material, we used small buckets filled with sand with a gravel substrate. We sowed seeds that we let grow a few weeks. The water supply is under a horizontal flow regime under-ground. The urban wastewater used is preceded by preliminary treatment. The water obtained after purification is collected using a tap in a container placed under the seal. The comparison between the inlet and the outlet waters showed that the presence of the Sodom apple tree contributes to reducing their pollutant parameters with significant rates: 81% for COD, 84%, for BOD , 95% for SM , 82% for NO⁻² , and 85% for NO⁻³ and can be released into the environment without risk of pollutionKeywords: arid zone, pollution, purification, re-use, wastewater.
Procedia PDF Downloads 802643 Antifungal Protein ~35kDa Produced by Bacillus cereus Inhibits the Growth of Some Molds and Yeasts
Authors: Saleh H. Salmen, Sulaiman Ali Alharbi, Hany M. Yehia, Mohammad A. Khiyami, Milton Wainwright, Naiyf S. Alharbi, Arunachalam Chinnathambi
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An antifungal protein synthesized by Bacillus cereus has been partially purified by the use of ammonium sulfate precipitation and Sephadex-G-200 column chromatography. The protein was produced from Bacillus cereus grown in potato Dextrose Broth Medium (PDB) at 30 ºC for 3 days at 100 rpm. The protein showed antagonistic effect against some fungi and yeasts. Crude extract from medium and semi-purified protein were tested in vitro against both fungi and yeasts using the disc diffusion method in order to detect the inhibitory effect of the protein. Zones of inhibition of the following diameter were found (mm) were Alternaria alternate (28), Rhodotorula glutinis (20), Fusarium sp. (16), Rhizopus sp. (15), Penicillium digitatum (13), Mucor sp. (13) and Aspergillus niger (10). The isolated protein was found to have a molecular weight of ~35kDa by sodium deodecyl sulfate-poly acrylamide gel electrophoresis. The data showed that the protein of Bacillus cereus has antifungal activity, a fact which points to the possibility of using it as a bio-control agent against some fungi, findings which emphasize the potential role of B. cereus as an important bio-control agent.Keywords: bacillus cereus, ~35kDa protein, molds, yeasts
Procedia PDF Downloads 2912642 Experimental Design in Extraction of Pseudomonas sp. Protease from Fermented Broth by Polyethylene Glycol/Citrate Aqueous Two-Phase System
Authors: Omar Pillaca-Pullo, Arturo Alejandro-Paredes, Carol Flores-Fernandez, Marijuly Sayuri Kina, Amparo Iris Zavaleta
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Aqueous two-phase system (ATPS) is an interesting alternative for separating industrial enzymes due to it is easy to scale-up and low cost. Polyethylene glycol (PEG) mixed with potassium phosphate or magnesium sulfate is one of the most frequently polymer/salt ATPS used, but the consequences of its use is a high concentration of phosphates and sulfates in wastewater causing environmental issues. Citrate could replace these inorganic salts due to it is biodegradable and does not produce toxic compounds. On the other hand, statistical design of experiments is widely used for ATPS optimization and it allows to study the effects of the involved variables in the purification, and to estimate their significant effects on selected responses and interactions. The 24 factorial design with four central points (20 experiments) was employed to study the partition and purification of proteases produced by Pseudomonas sp. in PEG/citrate ATPS system. ATPS was prepared with different sodium citrate concentrations [14, 16 and 18% (w/w)], pH values (7, 8 and 9), PEG molecular weight (2,000; 4,000 and 6,000 g/mol) and PEG concentrations [18, 20 and 22 % (w/w)]. All system components were mixed with 15% (w/w) of the fermented broth and deionized water was added to a final weight of 12.5 g. Then, the systems were mixed and kept at room temperature until to reach two-phases separation. Volumes of the top and bottom phases were measured, and aliquots from both phases were collected for subsequent proteolytic activity and total protein determination. Influence of variables such as PEG molar mass (MPEG), PEG concentration (CPEG), citrate concentration (CSal) and pH were evaluated on the following responses: purification factor (PF), activity yield (Y), partition coefficient (K) and selectivity (S). STATISTICA program version 10 was used for the analysis. According to the obtained results, higher levels of CPEG and MPEG had a positive effect on extraction, while pH did not influence on the process. On the other hand, the CSal could be related with low values of Y because of the citrate ions have a negative effect on solubility and enzymatic structure. The optimum values of Y (66.4 %), PF (1.8), K (5.5) and S (4.3) were obtained at CSal (18%), MPEG (6,000 g/mol), CPEG (22%) and pH 9. These results indicated that the PEG/citrate system is accurate to purify these Pseudomonas sp. proteases from fermented broth as a first purification step.Keywords: citrate, polyethylene glycol, protease, Pseudomonas sp
Procedia PDF Downloads 1952641 Aqueous Two Phase Extraction of Jonesia denitrificans Xylanase 6 in PEG 1000/Phosphate System
Authors: Nawel Boucherba, Azzedine Bettache, Abdelaziz Messis, Francis Duchiron, Said Benallaoua
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The impetus for research in the field of bioseparation has been sparked by the difficulty and complexity in the downstream processing of biological products. Indeed, 50% to 90% of the production cost for a typical biological product resides in the purification strategy. There is a need for efficient and economical large scale bioseparation techniques which will achieve high purity and high recovery while maintaining the biological activity of the molecule. One such purification technique which meets these criteria involves the partitioning of biomolecules between two immiscible phases in an aqueous system (ATPS). The Production of xylanases is carried out in 500ml of a liquid medium based on birchwood xylan. In each ATPS, PEG 1000 is added to a mixture consisting of dipotassium phosphate, sodium chloride and the culture medium inoculated with the strain Jonesia denitrificans, the mixture was adjusted to different pH. The concentration of PEG 1000 was varied: 8 to 16 % and the NaCl percentages are also varied from 2 to 4% while maintaining the other parameters constant. The results showed that the best ATPS for purification of xylanases is composed of PEG 1000 at 8.33%, 13.14 % of K2HPO4, 1.62% NaCl at pH 7. We obtained a yield of 96.62 %, a partition coefficient of 86.66 and a purification factor of 2.9. The zymogram showed that the activity is mainly detected in the top phase.Keywords: Jonesia denitrificans BN13, xylanase, aqueous two phases system, zymogram
Procedia PDF Downloads 3992640 Valorization of Sawdust for the Treatment of Purified Water for Irrigation
Authors: Dalila Oulhaci, Mohammed Zahaf
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The watering technique is essential to maintain a moist perimeter around the roots of the crop. This is the case with topical watering, where the soil around the root system can be kept permanently moist between the two extremes of water content. Moreover, one of the oldest methods used since Roman times throughout North Africa and the Near East was based on the repeated pouring of water into porous earthen vessels buried in the ground. In this context, these two techniques have been combined by replacing the earthen vase with plastic bottles filled with sand which release water through their perforated walls into the surrounding soil. The objective of this work is to first determine the purifying power of the activated sludge treatment plant of Toggourt and then that of the bottled Sawdust filter. For the station, the BOD purification rate was (96.5%), the COD purification rate was (87%) and suspended solids (90%). For the bottle, the BOD removal rate was (35%), and COD removal rate was (12.58%). This work falls within the framework of water saving, sustainable development and environmental protection, and also within the framework of agriculture.Keywords: wasterwater, sawdust, purification, irrigation, touggourt (Algeria)
Procedia PDF Downloads 862639 Sustainable Membranes Based on 2D Materials for H₂ Separation and Purification
Authors: Juan A. G. Carrio, Prasad Talluri, Sergio G. Echeverrigaray, Antonio H. Castro Neto
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Hydrogen as a fuel and environmentally pleasant energy carrier is part of this transition towards low-carbon systems. The extensive deployment of hydrogen production, purification and transport infrastructures still represents significant challenges. Independent of the production process, the hydrogen generally is mixed with light hydrocarbons and other undesirable gases that need to be removed to obtain H₂ with the required purity for end applications. In this context, membranes are one of the simplest, most attractive, sustainable, and performant technologies enabling hydrogen separation and purification. They demonstrate high separation efficiencies and low energy consumption levels in operation, which is a significant leap compared to current energy-intensive options technologies. The unique characteristics of 2D laminates have given rise to a diversity of research on their potential applications in separation systems. Specifically, it is already known in the scientific literature that graphene oxide-based membranes present the highest reported selectivity of H₂ over other gases. This work explores the potential of a new type of 2D materials-based membranes in separating H₂ from CO₂ and CH₄. We have developed nanostructured composites based on 2D materials that have been applied in the fabrication of membranes to maximise H₂ selectivity and permeability, for different gas mixtures, by adjusting the membranes' characteristics. Our proprietary technology does not depend on specific porous substrates, which allows its integration in diverse separation modules with different geometries and configurations, looking to address the technical performance required for industrial applications and economic viability. The tuning and precise control of the processing parameters allowed us to control the thicknesses of the membranes below 100 nanometres to provide high permeabilities. Our results for the selectivity of new nanostructured 2D materials-based membranes are in the range of the performance reported in the available literature around 2D materials (such as graphene oxide) applied to hydrogen purification, which validates their use as one of the most promising next-generation hydrogen separation and purification solutions.Keywords: membranes, 2D materials, hydrogen purification, nanocomposites
Procedia PDF Downloads 1342638 The Effect of Sorafenibe on Soat1 Protein by Using Molecular Docking Method
Authors: Mahdiyeh Gholaminezhad
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Context: The study focuses on the potential impact of Sorafenib on SOAT1 protein in liver cancer treatment, addressing the need for more effective therapeutic options. Research aim: To explore the effects of Sorafenib on the activity of SOAT1 protein in liver cancer cells. Methodology: Molecular docking was employed to analyze the interaction between Sorafenib and SOAT1 protein. Findings: The study revealed a significant effect of Sorafenib on the stability and activity of SOAT1 protein, suggesting its potential as a treatment for liver cancer. Theoretical importance: This research highlights the molecular mechanism underlying Sorafenib's anti-cancer properties, contributing to the understanding of its therapeutic effects. Data collection: Data on the molecular structure of Sorafenib and SOAT1 protein were obtained from computational simulations and databases. Analysis procedures: Molecular docking simulations were performed to predict the binding interactions between Sorafenib and SOAT1 protein. Question addressed: How does Sorafenib influence the activity of SOAT1 protein and what are the implications for liver cancer treatment? Conclusion: The study demonstrates the potential of Sorafenib as a targeted therapy for liver cancer by affecting the activity of SOAT1 protein. Reviewers' Comments: The study provides valuable insights into the molecular basis of Sorafenib's action on SOAT1 protein, suggesting its therapeutic potential. To enhance the methodology, the authors could consider validating the docking results with experimental data for further validation.Keywords: liver cancer, sorafenib, SOAT1, molecular docking
Procedia PDF Downloads 282637 Protein and Lipid Extraction from Microalgae with Ultrasound Assisted Osmotic Shock Method
Authors: Nais Pinta Adetya, H. Hadiyanto
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Microalgae has a potential to be utilized as food and natural colorant. The microalgae components consists of three main parts, these are lipid, protein, and carbohydrate. Crucial step in producing lipid and protein from microalgae is extraction. Microalgae has high water level (70-90%), it causes drying process of biomass needs much more energy and also has potential to distract lipid and protein from microalgae. Extraction of lipid from wet biomass is able to take place efficiently with cell disruption of microalgae by osmotic shock method. In this study, osmotic shock method was going to be integrated with ultrasound to maximalize the extraction yield of lipid and protein from wet biomass Spirulina sp. with osmotic shock method assisted ultrasound. This study consisted of two steps, these were osmotic shock process toward wet biomass and ultrasound extraction assisted. NaCl solution was used as osmotic agent, with the variation of concentrations were 10%, 20%, and 30%. Extraction was conducted in 40°C for 20 minutes with frequency of ultrasound wave was 40kHz. The optimal yield of protein (2.7%) and (lipid 38%) were achieved at 20% osmotic agent concentration.Keywords: extraction, lipid, osmotic shock, protein, ultrasound
Procedia PDF Downloads 3592636 Study of Treatment Plant of The City Chlef Study of Environmental Impact
Authors: Houmame Benbouali, Aboubakr Gribi
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The risks, in general, exist in any project, one can hardly carry out a project without taking risks. The hydraulic works are rather complex projects in their design, realization and exploitation and are often subjected at the multiple risks being able to influence with their good performance and can have a negative impact on their environment. The present study was carried out to quote the impacts caused by purification plant STEP Chlef on the environment, it aims has studied the environmental impacts during construction and when designing this STEP, it is divided into two parts: The first part results from a research task bibliographer which contain three chapters (- cleansing of water-worn- general information on water worn-proceed of purification of waste water). The second part is an experimental part which is divided into four chapters (detailed state initial description of the station of purification-evaluation of the impacts of the project analyzes measurements and recommendations).Keywords: treatment plant, waste water, waste water treatment, Chlef
Procedia PDF Downloads 3342635 Myeloid Zinc Finger 1/Ets-Like Protein-1/Protein Kinase C Alpha Associated with Poor Prognosis in Patients with Hepatocellular Carcinoma
Authors: Jer-Yuh Liu, Je-Chiuan Ye, Jin-Ming Hwang
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Protein kinase C alpha (PKCα) is a key signaling molecule in human cancer development. As a therapeutic strategy, targeting PKCα is difficult because the molecule is ubiquitously expressed in non-malignant cells. PKCα is regulated by the cooperative interaction of the transcription factors myeloid zinc finger 1 (MZF-1) and Ets-like protein-1 (Elk-1) in human cancer cells. By conducting tissue array analysis, herein, we determined the protein expression of MZF-1/Elk-1/PKCα in various cancers. The data show that the expression of MZF-1/Elk-1 is correlated with that of PKCα in hepatocellular carcinoma (HCC), but not in bladder and lung cancers. In addition, the PKCα down-regulation by shRNA Elk-1 was only observed in the HCC SK-Hep-1 cells. Blocking the interaction between MZF-1 and Elk-1 through the transfection of their binding domain MZF-160–72 decreased PKCα expression. This step ultimately depressed the epithelial-mesenchymal transition potential of the HCC cells. These findings could be used to develop an alternative therapeutic strategy for patients with the PKCα-derived HCC.Keywords: protein kinase C alpha, myeloid zinc finger 1, ets-like protein-1, hepatocellular carcinoma
Procedia PDF Downloads 2272634 Development of Monoclonal Antibodies against the Acute Hepatopancreatic Necrosis Disease Toxins
Authors: Naveen Kumar B. T., Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, Shanthanagouda A. H., Orawan Boodde, Shankar K. M., Prakash Patil, Shubhkaramjeet Kaur
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Since 2009, Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks have increased rapidly, and these have led to the major economic losses to the global shrimp industry. In comparison to other treatments, passive immunity and monoclonal antibody (MAb) based farmer level kit have proved their importance in controlling and treating the diseases in the shrimp industry. In the present study, MAbs were produced against the recombinant PirB protein Vibrio parahaemolyticus strain causing AHPND. Briefly, Balb/C mice were immunized with rPirB at 15 days interval, and antibody titer was determined by ELISA. Spleen cells from mice showing high antibody titer were fused with SP2O myeloma cells for hybridoma production. Among 130 hybridomas, four showed high antibody titer and positive reactivity in an immunoblot assay. In Western blot assay, three out of four MAbs (4C4, 2C2 and 4G3) showed reactivity to rPirB protein. However, in the natural host, only Mab clone 4G3 show strong reactivity (with a strain of V. parahemolyticus causing EMS/AHPND). These clones also showed reactivity with less than 20 kDa proteins in AHPND free V. parahaemolyticus (Thailand stain). Further, on from MAb 4G3 clone, four panels of single cell MAbs clones (G3F5, G3B8, G3H2, and G3D6) were produced of which three showed strong positive reactivity to rPirB protein in the Western blot. These MAbs have potential for controlling and prevention of the AHPND through passive immunity and development of filed level rapid diagnostic kits.Keywords: shrimp, economic loss, AHPND, MAb
Procedia PDF Downloads 2532633 Investigation on Porcine Follicular Fluid Protein Pattern of Medium and Large Follicles
Authors: Hatairuk Tungkasen, Somrudee Phetchrid, Suwapat Jaidee, Supinya Yoomak, Chantana Kankamol, Mayuree Pumipaiboon, Mayuva Areekijseree
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Ovaries of reproductive female pigs were obtained from local slaughterhouses in Nakorn Pathom Province, Thailand. Follicular fluid of medium follicle (5-6 diameters) and large follicles (7-8 mm and 10 mm in diameter) were aspirated and collected by sterile technique and analyzed protein pattern. The follicular fluid protein bands were found by SDS-PAGE which has no protein band in difference compared to standard protein band. So we chose protein band molecular weight 50, 62-65, 75-80, 90, 120-160, and >220 kDa were analyzed by LC/MS/MS. The result was found immunoglobulin gamma chain, keratin, transferrin, heat shock protein, and plasminogen precursor, ceruloplasmin, and hemopexin, and protease, respectively. All proteins play important roles in promotion and regulation on growth and development of reproductive cells. The result of this study found many proteins which were useful and important for in vitro oocyte maturation and embryonic development of cell technology in animals. The further study will be use porcine follicular fluid protein of medium and large follicles as feeder cells in in vitro condition to promote oocyte and embryo maturation.Keywords: follicular fluid protein, LC/MS/MS, porcine oocyte, SDS-PAGE
Procedia PDF Downloads 5852632 Purification, Extraction and Visualization of Lipopolysaccharide of Escherichia coli from Urine Samples of Patients with Urinary Tract Infection
Authors: Fariha Akhter Chowdhury, Mohammad Nurul Islam, Anamika Saha, Sabrina Mahboob, Abu Syed Md. Mosaddek, Md. Omar Faruque, Most. Fahmida Begum, Rajib Bhattacharjee
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Urinary tract infection (UTI) is one of the most common infectious diseases in Bangladesh where Escherichia coli is the prevalent organism and responsible for most of the infections. Lipopolysaccharide (LPS) is known to act as a major virulence factor of E. coli. The present study aimed to purify, extract and visualize LPS of E. coli clinical isolates from urine samples of patients with UTI. The E. coli strain was isolated from the urine samples of 10 patients with UTI and then the antibiotic sensitivity pattern of the isolates was determined. The purification of LPS was carried out using the hot aqueous-phenol method and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, which was directly stained using the modified silver staining method and Coomassie blue. The silver-stained gel demonstrated both smooth and rough type LPS by showing trail-like band patterns with the presence and lacking O-antigen region, respectively. Coomassie blue staining showed no band assuring the absence of any contaminating protein. Our successful extraction of purified LPS from E. coli isolates of UTI patients’ urine samples can be an important step to understand the UTI disease conditions.Keywords: Escherichia coli, electrophoresis, polyacrylamide gel, silver staining, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
Procedia PDF Downloads 3912631 Thrombophilic Risk Factors and Pregnancy Complications
Authors: Hanan Azzam1, Nashwa Abousamra1, Amany Mansour1, Yaser Abd El-dayem2, , Solafa Elsharawy1
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Background: Inherited thrombophilias are a heterogenous group of conditions which have been implicated in a variety of pregnancy complications. More recently, deficiency of protein Z (PZ) has been liked to pregnancy complications, including preterm delivery. Aim: We designed this study to evaluate the association of inherited thrombophilias including [Protein C (PC), Protein S (PS), Anti thrombin III (ATIII) deficiency and activated protein C (APC) resistance] and protein Z deficiency with a variety of pregnancy complications. Patients and Methods: 60 women with different pregnancy complications, including 20 patients with preeclampsia, 20 patients with intrauterine growth resistance (IUGR), and 20 patients with intrauterine fetal death (IUFD), in addition to 30 healthy pregnant women were recruited for the present study. PC and free PS antigen, ATIII activity, modified functional APC-resistance, and PZ levels were determined. Results: There was no significant association between inherited thrombophilias and complicated pregnancies as regards PC deficiency (p=1.0), AT III and PS deficiency (p=0.312), and APC-resistance (P=0.083). PZ was significantly associated with complicated pregnancies (p=0.012). Patients with protein Z levels below 1.5 µg/ml were considered deficient. Accordingly, we demonstrated protein Z deficiency in 30% of complicated pregnancies (RR 6.0, 95% CI 1.29-27.90;p=0.022), 20% of preeclampsia (RR 3.5, 95% CI 0.57 – 21.28; P = 0.174), 40% of IUGR (RR 9.3 95% CI 1.72-50.61; P = 0.010) and 30% of IUFD (RR 6, 95% CI 1.07 – 33.64; P = 0.042). Conclusions: These findings indicate the absence of association of inherited thrombophilias, including PC, PS, AT III deficiency, and APC resistance with pregnancy complications. However, PZ deficiency is associated with increased risk of pregnancy complications, especially intrauterine growth restriction and intrauterine fetal death.Keywords: protein C, protein S, thrombophelia, pregnancy, protein Z
Procedia PDF Downloads 2342630 Unequal Contributions of Parental Isolates in Somatic Recombination of the Stripe Rust Fungus
Authors: Xianming Chen, Yu Lei, Meinan Wang
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The dikaryotic basidiomycete fungus, Puccinia striiformis, causes stripe rust, one of the most important diseases of wheat and barley worldwide. The pathogen is largely reproduced asexually, and asexual recombination has been hypothesized to be one of the mechanisms for the pathogen variations. To test the hypothesis and understand the genetic process of asexual recombination, somatic recombinant isolates were obtained under controlled conditions by inoculating susceptible host plants with a mixture of equal quantity of urediniospores of isolates with different virulence patterns and selecting through a series of inoculation on host plants with different genes for resistance to one of the parental isolates. The potential recombinant isolates were phenotypically characterized by virulence testing on the set of 18 wheat lines used to differentiate races of the wheat stripe rust pathogen, P. striiformis f. sp. tritici (Pst), for the combinations of Pst isolates; or on both sets of the wheat differentials and 12 barley differentials for identifying races of the barley stripe rust pathogen, P. striiformis f. sp. hordei (Psh) for combinations of a Pst isolate and a Psh isolate. The progeny and parental isolates were also genotypically characterized with 51 simple sequence repeat and 90 single-nucleotide polymorphism markers. From nine combinations of parental isolates, 68 potential recombinant isolates were obtained, of which 33 (48.5%) had similar virulence patterns to one of the parental isolates, and 35 (51.5%) had virulence patterns distinct from either of the parental isolates. Of the 35 isolates of distinct virulence patterns, 11 were identified as races that had been previously detected from natural collections and 24 were identified as new races. The molecular marker data confirmed 66 of the 68 isolates as recombinants. The percentages of parental marker alleles ranged from 0.9% to 98.9% and were significantly different from equal proportions in the recombinant isolates. Except for a couple of combinations, the greater or less contribution was not specific to any particular parental isolates as the same parental isolates contributed more to some of the progeny isolates but less to the other progeny isolates in the same combination. The unequal contributions by parental isolates appear to be a general role in somatic recombination for the stripe rust fungus, which may be used to distinguish asexual recombination from sexual recombination in studying the evolutionary mechanisms of the highly variable fungal pathogen.Keywords: molecular markers, Puccinia striiformis, somatic recombination, stripe rust
Procedia PDF Downloads 2442629 Novel Recombinant Betasatellite Associated with Vein Thickening Symptoms on Okra Plants in Saudi Arabia
Authors: Adel M. Zakri, Mohammed A. Al-Saleh, Judith. K. Brown, Ali M. Idris
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Betasatellites are small circular single stranded DNA molecules found associated with begomoviruses on field symptomatic plants. Their genome size is about half that of the helper begomovirus, ranging between 1.3 and 1.4 kb. The helper begomoviruses are usually members of the family Geminiviridae. Okra leaves showing vein thickening were collected from okra plants growing in Jazan, Saudi Arabia. Total DNA was extracted from leaves and used as a template to amplify circular DNA using rolling circle amplification (RCA) technology. Products were digested with PstI to linearize the helper viral genome(s), and associated DNA satellite(s), yielding a 2.8kbp and 1.4kbp fragment, respectively. The linearized fragments were cloned into the pGEM-5Zf (+) vector and subjected to DNA sequencing. The 2.8 kb fragment was identified as Cotton leaf curl Gezira virus genome, at 2780bp, an isolate closely related to strains reported previously from Saudi Arabia. A clone obtained from the 1.4 kb fragments he 1.4kb was blasted to GeneBank database found to be a betasatellite. The genome of betasatellite was 1357-bp in size. It was found to be a recombinant containing one fragment (877-bp) that shared 91% nt identity with Cotton leaf curl Gezira betasatellite [KM279620], and a smaller fragment [133--bp) that shared 86% nt identity with Tomato leaf curl Sudan virus [JX483708]. This satellite is thus a recombinant between a malvaceous-infecting satellite and a solanaceous-infecting begomovirus.Keywords: begomovirus, betasatellites, cotton leaf curl Gezira virus, okra plants
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