Search results for: plasma protein binding
3073 Covalent Binding of Cysteine to a Sol-Gel Material for Cadmium Biosorption from Aqueous Solutions
Authors: Claudiu Marcu, Cristina Paul, Adelina Andelescu, Corneliu Mircea Davidescu, Francisc Péter
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Heavy metal pollution has become a more serious environmental problem in the last several decades as a result of its toxicity and insusceptibility to the environment. Methods for removing metal ions from aqueous solution mainly consist of physical, chemical and biochemical procedures. Biosorption is defined as the removal of metal or metalloid species, compounds and particulates from solution by a biological material. Biosorption represents a very attractive method for the removal of toxic metal ions from aqueous effluents because it uses the ability of various biomass to bind the metal ions without the risk of releasing other toxic chemical compounds into the environment. The problem with using biomass or living cells as biosorbents is that their regeneration/reuse is often either impossible or very laborious. One of the most common chelating group found in biosorbents is the thiol group in cysteine. Therefore, we immobilized cysteine using covalent binding using glutaraldehyde as a linker on a synthetic sol-gel support obtained using 3-amino-propyl-trimetoxysilane and trimetoxysilane as precursors. The obtained adsorbents were used for removal of cadmium from aqueous solutions and the removal capacity was investigated in relation to the composition of the sol-gel hybrid composite, the loading of the biomolecule and the physical parameters of the biosorption process. In the same conditions, the bare sol-gel support without cysteine had no Cd removal effect, while the adsorbent with cysteine had an adsorption capacity up to 25.8 mg Cd/g adsorbent at pH 2.0 and 119 mg Cd/g adsorbent at pH 6.6, depending on cadmium concentration and adsorption conditions. We used atomic adsorption spectrometry to assess the cadmium concentration in the samples after the biosorbtion process. The parameters for the Freundlich and Langmuir adsorption isotherms where calculated from plotting the results of the adsorption experiments. The results for cysteine immobilization show a good loading capacity of the sol-gel support which indicates it could be used to immobilize metal binding proteins and by doing so boosting the heavy metal adsorption capacity of the biosorbent.Keywords: biosorbtion, cadmium, cysteine covalent binding, sol-gel
Procedia PDF Downloads 2943072 Stochastic Modeling of Secretion Dynamics in Inner Hair Cells of the Auditory Pathway
Authors: Jessica A. Soto-Bear, Virginia González-Vélez, Norma Castañeda-Villa, Amparo Gil
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Glutamate release of the cochlear inner hair cell (IHC) ribbon synapse is a fundamental step in transferring sound information in the auditory pathway. Otoferlin is the calcium sensor in the IHC and its activity has been related to many auditory disorders. In order to simulate secretion dynamics occurring in the IHC in a few milliseconds timescale and with high spatial resolution, we proposed an active-zone model solved with Monte Carlo algorithms. We included models for calcium buffered diffusion, calcium-binding schemes for vesicle fusion, and L-type voltage-gated calcium channels. Our results indicate that calcium influx and calcium binding is managing IHC secretion as a function of voltage depolarization, which in turn mean that IHC response depends on sound intensity.Keywords: inner hair cells, Monte Carlo algorithm, Otoferlin, secretion
Procedia PDF Downloads 2213071 Experimental Measurement of Equatorial Ring Current Generated by Magnetoplasma Sail in Three-Dimensional Spatial Coordinate
Authors: Masato Koizumi, Yuya Oshio, Ikkoh Funaki
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Magnetoplasma Sail (MPS) is a future spacecraft propulsion that generates high levels of thrust by inducing an artificial magnetosphere to capture and deflect solar wind charged particles in order to transfer momentum to the spacecraft. By injecting plasma in the spacecraft’s magnetic field region, the ring current azimuthally drifts on the equatorial plane about the dipole magnetic field generated by the current flowing through the solenoid attached on board the spacecraft. This ring current results in magnetosphere inflation which improves the thrust performance of MPS spacecraft. In this present study, the ring current was experimentally measured using three Rogowski Current Probes positioned in a circular array about the laboratory model of MPS spacecraft. This investigation aims to determine the detailed structure of ring current through physical experimentation performed under two different magnetic field strengths engendered by varying the applied voltage on the solenoid with 300 V and 600 V. The expected outcome was that the three current probes would detect the same current since all three probes were positioned at equal radial distance of 63 mm from the center of the solenoid. Although experimental results were numerically implausible due to probable procedural error, the trends of the results revealed three pieces of perceptive evidence of the ring current behavior. The first aspect is that the drift direction of the ring current depended on the strength of the applied magnetic field. The second aspect is that the diamagnetic current developed at a radial distance not occupied by the three current probes under the presence of solar wind. The third aspect is that the ring current distribution varied along the circumferential path about the spacecraft’s magnetic field. Although this study yielded experimental evidence that differed from the original hypothesis, the three key findings of this study have informed two critical MPS design solutions that will potentially improve thrust performance. The first design solution is the positioning of the plasma injection point. Based on the implication of the first of the three aspects of ring current behavior, the plasma injection point must be located at a distance instead of at close proximity from the MPS Solenoid for the ring current to drift in the direction that will result in magnetosphere inflation. The second design solution, predicated by the third aspect of ring current behavior, is the symmetrical configuration of plasma injection points. In this study, an asymmetrical configuration of plasma injection points using one plasma source resulted in a non-uniform distribution of ring current along the azimuthal path. This distorts the geometry of the inflated magnetosphere which minimizes the deflection area for the solar wind. Therefore, to realize a ring current that best provides the maximum possible inflated magnetosphere, multiple plasma sources must be spaced evenly apart for the plasma to be injected evenly along its azimuthal path.Keywords: Magnetoplasma Sail, magnetosphere inflation, ring current, spacecraft propulsion
Procedia PDF Downloads 3103070 Adhesion of Sputtered Copper Thin Films Deposited on Flexible Substrates
Authors: Rwei-Ching Chang, Bo-Yu Su
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Adhesion of copper thin films deposited on polyethylene terephthAdhesion of copper thin films deposited on polyethylene terephthalate substrate by direct current sputtering with different sputtering parameters is discussed in this work. The effects of plasma treatment with 0, 5, and 10 minutes on the thin film properties are investigated first. Various argon flow rates at 40, 50, 60 standard cubic centimeters per minute (sccm), deposition power at 30, 40, 50 W, and film thickness at 100, 200, 300 nm are also discussed. The 3-dimensional surface profilometer, micro scratch machine, and optical microscope are used to characterize the thin film properties. The results show that the increase of the plasma treatment time on the polyethylene terephthalate surface affects the roughness and critical load of the films. The critical load increases as the plasma treatment time increases. When the plasma treatment time was adjusted from 5 minutes to 10 minutes, the adhesion increased from 8.20 mN to 13.67 mN. When the argon flow rate is decreased from 60 sccm to 40 sccm, the adhesion increases from 8.27 mN to 13.67 mN. The adhesion is also increased by the condition of higher power, where the adhesion increased from 13.67 mN to 25.07 mN as the power increases from 30 W to 50 W. The adhesion of the film increases from 13.67 mN to 21.41mN as the film thickness increases from 100 nm to 300 nm. Comparing all the deposition parameters, it indicates the change of the power and thickness has much improvement on the film adhesion.alate substrate by direct current sputtering with different sputtering parameters is discussed in this work. The effects of plasma treatment with 0, 5, and 10 minutes on the thin film properties are investigated first. Various argon flow rates at 40, 50, 60 standard cubic centimeters per minute (sccm), deposition power at 30, 40, 50 W, and film thickness at 100, 200, 300 nm are also discussed. The 3-dimensional surface profilometer, micro scratch machine, and optical microscope are used to characterize the thin film properties. The results show that the increase of the plasma treatment time on the polyethylene terephthalate surface affects the roughness and critical load of the films. The critical load increases as the plasma treatment time increases. When the plasma treatment time was adjusted from 5 minutes to 10 minutes, the adhesion increased from 8.20 mN to 13.67 mN. When the argon flow rate is decreased from 60 sccm to 40 sccm, the adhesion increases from 8.27 mN to 13.67 mN. The adhesion is also increased by the condition of higher power, where the adhesion increased from 13.67 mN to 25.07 mN as the power increases from 30 W to 50 W. The adhesion of the film increases from 13.67 mN to 21.41mN as the film thickness increases from 100 nm to 300 nm. Comparing all the deposition parameters, it indicates the change of the power and thickness has much improvement on the film adhesion.Keywords: flexible substrate, sputtering, adhesion, copper thin film
Procedia PDF Downloads 1303069 Effect of N2-cold Plasma Treatment of Carbon Supports on the Activity of Pt3Pd3Sn2/C Towards the Dimethyl Ether Oxidation
Authors: Medhanie Gebremedhin Gebru, Alex Schechter
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Dimethyl ether (DME) possesses several advantages over other small organic molecules such as methanol, ethanol, and ammonia in terms of providing higher energy density, being less toxic, and having lower Nafion membrane crossover. However, the absence of an active and stable catalyst has been the bottleneck that hindered the commercialization of direct DME fuel cells. A Vulcan XC72 carbon-supported ternary metal catalyst, Pt₃Pd₃Sn₂/C is reported to have yielded the highest specific power density (90 mW mg-¹PGM) as compared to other catalysts tested fordirect DME fuel cell (DDMEFC). However, the micropores and sulfur groups present in Vulcan XC72 hinder the fuel utilization by causing Pt agglomeration and sulfur poisoning. Vulcan XC72 having a high carbon sp³ hybridization content, is also prone to corrosion. Therefore, carbon supports such as multi-walled carbon nanotube (MWCNT), black pearl 2000 (BP2000), and their cold N2 plasma-treated counterpartswere tested to further enhance the activity of the catalyst, and the outputs with these carbons were compared with the originally used support. Detailed characterization of the pristine and carbon supports was conducted. Electrochemical measurements in three-electrode cells and laboratory prototype fuel cells were conducted.Pt₃Pd₃Sn₂/BP2000 exhibited excellent performance in terms of electrochemical active surface area (ECSA), peak current density (jp), and DME oxidation charge (Qoxi). The effect of the plasma activation on the activity improvement was observed only in the case of MWCNT while having little or no effect on the other carbons. A Pt₃Pd₃Sn₂ supported on the optimized mixture of carbons containing 75% plasma-activated MWCNT and 25% BP2000 (Pt₃Pd₃Sn₂/75M25B) provided the highest reported power density of 117 mW mg-1PGM using an anode loading of1.55 mgPGMcm⁻².Keywords: DME, DDMEFC, ternary metal catalyst, carbon support, plasma activation
Procedia PDF Downloads 1443068 The Study of Platelet-Rich Plasma(PRP) on Wounds of OLEFT Rats Using Expression of MMP-2, MMP-9 mRNA
Authors: Ho Seong Shin
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Introduction: A research in relation to wound healing also showed that platelet-rich plasma (PRP) was effective on normal tissue regeneration. Nonetheless, there is no evidence that when platelet-rich plasma was applied on diabetic wound, it normalize diabetic wound healing process. In this study, we have analyzed matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) expression to know the effect of PRP on diabetic wounds using Reverse transcription-polymerase chain reaction (RT-PCR) of MMP-2, MMP-9 mRNA. Materials and Methods: Platelet-rich plasma (PRP) was prepared from blood of 6 rats. The whole 120-mL was added immediately to an anticoagulant. Citrate phosphonate dextrose(CPD) buffer (0.15 mg CPDmL) in a ratio of 1 mL of CPD buffer to 5 mL of blood. The blood was then centrifuged at 220g for 20minutes. The supernatant was saved to produce fibrin glue. The participate containing PRP was used for second centrifugation at 480g for 20 minutes. The pellet from the second centrifugation was saved and diluted with supernatant until the platelet concentration became 900,000/μL. Twenty male, 4week-old OLETF rats were underwent operation; each rat had two wounds created on left and right sides. The each wound of left side was treated with PRP gel, the wound of right side was treated with physiologic saline gauze. Results: RT-PCR analysis; The levels of MMP-2 mRNA in PRP applied tissues were positively related to postwounding days, whereas MMP-2 mRNA expression in saline-applied tissues remained in 5day after treatment. MMP-9 mRNA was undetectable in saline-applied tissues for either tissue, except 3day after treatment. Following PRP-applied tissues, MMP-9 mRNA expression was detected, with maximal expression being seen at third day. The levels of MMP-9 mRNA in PRP applied tissues were reported high intensity of optical density related to saline applied tissues.Keywords: diabetes, MMP-2, MMP-9, OLETF, PRP, wound healing MMP-9
Procedia PDF Downloads 2733067 Homology Modelling of Beta Defensin 3 of Bos taurus and Its Docking Studies with Molecules Responsible for Formation of Biofilm
Authors: Ravinder Singh, Ankita Gurao, Saroj Bandhan, Sudhir Kumar Kashyap
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The Bos taurus Beta defensin 3 is a defensin peptide secreted by neutrophils and epithelial that exhibits anti-microbial activity. It is one of the crucial components forming an innate defense against intra mammary infections in livestock. The beta defensin 3 by virtue of its anti-microbial activity inhibits major mastitis pathogens including Staphylococcus aureus and Pseudomonas aeruginosa etc, which are also responsible for biofilm formation leading to antibiotic resistance phenomenon. Therefore, the defensin may prove as a non-conventional option to treat mastitis. In this study, computational analysis has been performed including sequence comparison among species and homology modeling of Bos taurus beta defensin 3 protein. The assessments of protein structure were done using the protein structure and model assessment tools integrated in Swiss Model server, which employs various local and global quality evaluation parameters. Further, molecular docking was also carried out between the defensin peptide and the components of biofilm to gain insight into various interactions and structural differences crucial for functionality of this protein.Keywords: beta defensin 3, bos taurus, docking, homology modeling
Procedia PDF Downloads 2913066 Magnetic Bio-Nano-Fluids for Hyperthermia
Authors: Z. Kolacinski, L. Szymanski. G. Raniszewski, D. Koza, L. Pietrzak
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Magnetic Bio-Nano-Fluid (BNF) can be composed of a buffer fluid such as plasma and magnetic nanoparticles such as iron, nickel, cobalt and their oxides. However iron is one of the best elements for magnetization by electromagnetic radiation. It can be used as a tool for medical diagnosis and treatment. Radio frequency (RF) radiation is able to heat iron nanoparticles due to magnetic hysteresis. Electromagnetic heating of iron nanoparticles and ferro-fluids BNF can be successfully used for non-invasive thermal ablation of cancer cells. Moreover iron atoms can be carried by carbon nanotubes (CNTs) if iron is used as catalyst for CNTs synthesis. Then CNTs became the iron containers and they screen the iron content against oxidation. We will present a method of CNTs addressing to the required cells. For thermal ablation of cancer cells we use radio frequencies for which the interaction with human body should be limited to minimum. Generally, the application of RF energy fields for medical treatment is justified by deep tissue penetration. The highly iron doped CNTs as the carriers creating magnetic fluid will be presented. An excessive catalyst injection method using electrical furnace and microwave plasma reactor will be presented. This way it is possible to grow the Fe filled CNTs on a moving surface in continuous synthesis process. This also allows producing uniform carpet of the Fe filled CNTs carriers. For the experimental work targeted to cell ablation we used RF generator to measure the increase in temperature for some samples like: solution of Fe2O3 in BNF which can be plasma-like buffer, solutions of pure iron of different concentrations in plasma-like buffer and in buffer used for a cell culture, solutions of carbon nanotubes (MWCNTs) of different concentrations in plasma-like buffer and in buffer used for a cell culture. Then the targeted therapies which can be effective if the carriers are able to distinguish the difference between cancerous and healthy cell’s physiology are considered. We have developed an approach based on ligand-receptor or antibody-antigen interactions for the case of colon cancer.Keywords: cancer treatment, carbon nano tubes, drag delivery, hyperthermia, iron
Procedia PDF Downloads 4133065 Analysis of Nutritional Value for Soybean Genotypes Grown in Lesotho
Authors: Motlatsi Eric Morojele, Moleboheng Patricia Lekota, Pulane Nkhabutlane, Motanyane Stanley Motake
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Soybean was introduced in Lesotho to increase the spectrum of nutritious foods, especially protein, oil and carbohydrates. However, since then, determination of nutritional value has not been performed, hence this study. The objective of the study was to distinguish soybean genotypes on the basis of nutritive value. The experiment was laid out using a Randomized Complete Block Design with 27 treatments (genotypes) and three replications. Compound fertilizer 2:3:2 (22) was broadcasted over the experimental plot at the rate of 250kg ha-1. Dimensions of the main experimental plot were 135m long and 10m wide, with each sub-plot being 4m and 3.6m. Inter-row and intra-row spacing were 0.9m and 0.20m, respectively. Samples of seeds from each plot were taken to the laboratory to analyze protein content, ash, ca, mg, fiber, starch and ether extract. There were significant differences (P>0.05) among 28 soybean genotypes for protein content, acid detergent fiber, calcium, magnesium and ash. The soybean cultivars with the highest amount of protein were P48T48R, PAN 1663 and PAN 155R. High ADF content was expressed by PAN 1521R. LS 6868 exhibited the highest value of 0.788mg calcium, and the cultivars with the highest magnesium were NA 5509 with 1.306mg. PAN 1663, LCD 5.9, DM5302 RS and NS 6448R revealed higher nutritional values than other genotypes.Keywords: genotypes, Lesotho, nutritive value, proximate analysis, soya-bean
Procedia PDF Downloads 253064 Antimicrobial Agents Produced by Yeasts
Authors: T. Büyüksırıt, H. Kuleaşan
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Natural antimicrobials are used to preserve foods that can be found in plants, animals, and microorganisms. Antimicrobial substances are natural or artificial agents that produced by microorganisms or obtained semi/total chemical synthesis are used at low concentrations to inhibit the growth of other microorganisms. Food borne pathogens and spoilage microorganisms are inactivated by the use of antagonistic microorganisms and their metabolites. Yeasts can produce toxic proteins or glycoproteins (toxins) that cause inhibition of sensitive bacteria and yeast species. Antimicrobial substance producing phenotypes belonging different yeast genus were isolated from different sources. Toxins secreted by many yeast strains inhibiting the growth of other yeast strains. These strains show antimicrobial activity, inhibiting the growth of mold and bacteria. The effect of antimicrobial agents produced by yeasts can be extremely fast, and therefore may be used in various treatment procedures. Rapid inhibition of microorganisms is possibly caused by microbial cell membrane lipopolysaccharide binding and in activation (neutralization) effect. Antimicrobial agents inhibit the target cells via different mechanisms of action.Keywords: antimicrobial agents, yeast, toxic protein, glycoprotein
Procedia PDF Downloads 3623063 PARP1 Links Transcription of a Subset of RBL2-Dependent Genes with Cell Cycle Progression
Authors: Ewelina Wisnik, Zsolt Regdon, Kinga Chmielewska, Laszlo Virag, Agnieszka Robaszkiewicz
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Apart from protecting genome, PARP1 has been documented to regulate many intracellular processes inter alia gene transcription by physically interacting with chromatin bound proteins and by their ADP-ribosylation. Our recent findings indicate that expression of PARP1 decreases during the differentiation of human CD34+ hematopoietic stem cells to monocytes as a consequence of differentiation-associated cell growth arrest and formation of E2F4-RBL2-HDAC1-SWI/SNF repressive complex at the promoter of this gene. Since the RBL2 complexes repress genes in a E2F-dependent manner and are widespread in the genome in G0 arrested cells, we asked (a) if RBL2 directly contributes to defining monocyte phenotype and function by targeting gene promoters and (b) if RBL2 controls gene transcription indirectly by repressing PARP1. For identification of genes controlled by RBL2 and/or PARP1,we used primer libraries for surface receptors and TLR signaling mediators, genes were silenced by siRNA or shRNA, analysis of gene promoter occupation by selected proteins was carried out by ChIP-qPCR, while statistical analysis in GraphPad Prism 5 and STATISTICA, ChIP-Seq data were analysed in Galaxy 2.5.0.0. On the list of 28 genes regulated by RBL2, we identified only four solely repressed by RBL2-E2F4-HDAC1-BRM complex. Surprisingly, 24 out of 28 emerged genes controlled by RBL2 were co-regulated by PARP1 in six different manners. In one mode of RBL2/PARP1 co-operation, represented by MAP2K6 and MAPK3, PARP1 was found to associate with gene promoters upon RBL2 silencing, which was previously shown to restore PARP1 expression in monocytes. PARP1 effect on gene transcription was observed only in the presence of active EP300, which acetylated gene promoters and activated transcription. Further analysis revealed that PARP1 binding to MA2K6 and MAPK3 promoters enabled recruitment of EP300 in monocytes, while in proliferating cancer cell lines, which actively transcribe PARP1, this protein maintained EP300 at the promoters of MA2K6 and MAPK3. Genome-wide analysis revealed a similar distribution of PARP1 and EP300 around transcription start sites and the co-occupancy of some gene promoters by PARP1 and EP300 in cancer cells. Here, we described a new RBL2/PARP1/EP300 axis which controls gene transcription regardless of the cell type. In this model cell, cycle-dependent transcription of PARP1 regulates expression of some genes repressed by RBL2 upon cell cycle limitation. Thus, RBL2 may indirectly regulate transcription of some genes by controlling the expression of EP300-recruiting PARP1. Acknowledgement: This work was financed by Polish National Science Centre grants nr DEC-2013/11/D/NZ2/00033 and DEC-2015/19/N/NZ2/01735. L.V. is funded by the National Research, Development and Innovation Office grants GINOP-2.3.2-15-2016-00020 TUMORDNS, GINOP-2.3.2-15-2016-00048-STAYALIVE and OTKA K112336. AR is supported by Polish Ministry of Science and Higher Education 776/STYP/11/2016.Keywords: retinoblastoma transcriptional co-repressor like 2 (RBL2), poly(ADP-ribose) polymerase 1 (PARP1), E1A binding protein p300 (EP300), monocytes
Procedia PDF Downloads 2093062 A Recombinant Group a Streptococcus (GAS-2W) Strain Elicits Protective Immunity in Mice through Induction of an IFN-γ Dependent Humoral Response
Authors: Shiva Emami, Jenny Persson, Bengt Johansson Lindbom
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Group A streptococcus (GAS) is a prevalent human pathogen, causing a wide range of infections and diseases. One of the most well-known virulence factors in GAS is M protein, a surface protein that facilitates bacterial invasion. In this study, we used a recombinant GAS strain (GAS-2W) expressing M protein containing a hyper immunogenic peptide (2W). Mice were immunized three times with heat-killed-GAS subcutaneously at three weeks intervals. Three weeks post last immunization, mice were challenged intraperitoneally with a lethal dose of live GAS. In order to investigate the impact of IFN-ƴ and antibodies in protection against GAS infection, we used a mouse model knock-out for IFN-ƴ (IFN-ƴ KO). We observed immunization with GAS-2W strain can increase protection against GAS infection in mice compared with the original GAS strain. Higher levels of antibodies against M1 protein were measured in GAS-2W-immunized mice. There was also a significant increase in IgG2c response in mice immunized with GAS2W. By using IFN-ƴ KO mice, we showed that not a high level of total IgG, but IgG2c was correlated with protection through the i.p challenge. It also emphasizes the importance of IFN-ƴ cytokine to combat GAS by isotype switching to IgG2c (which is opsonic for phagocytosis). Our data indicate the crucial role of IFN-ƴ in the protective immune response that, together with IgG2c, can induce protection against GAS.Keywords: Group A streptococcus, IgG2c, IFN-γ, protection
Procedia PDF Downloads 903061 Insights of Interaction Studies between HSP-60, HSP-70 Proteins and HSF-1 in Bubalus bubalis
Authors: Ravinder Singh, C Rajesh, Saroj Badhan, Shailendra Mishra, Ranjit Singh Kataria
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Heat shock protein 60 and 70 are crucial chaperones that guide appropriate folding of denatured proteins under heat stress conditions. HSP60 and HSP70 provide assistance in correct folding of a multitude of denatured proteins. The heat shock factors are the family of some transcription factors which controls the regulation of gene expression of proteins involved in folding of damaged or improper folded proteins during stress conditions. Under normal condition heat shock proteins bind with HSF-1 and act as its repressor as well as aids in maintaining the HSF-1’s nonactive and monomeric confirmation. The experimental protein structure for all these proteins in Bubalus bubalis is not known till date. Therefore computational approach was explored to identify three-dimensional structure analysis of all these proteins. In this study, an extensive in silico analysis has been performed including sequence comparison among species to comparative modeling of Bubalus bubalis HSP60, HSP70 and HSF-1 protein. The stereochemical properties of proteins were assessed by utilizing several scrutiny bioinformatics tools to ensure model accuracy. Further docking approach was used to study interactions between Heat shock proteins and HSF-1.Keywords: Bubalus bubalis, comparative modelling, docking, heat shock protein
Procedia PDF Downloads 3223060 Molecular Docking Study of Quinazoline and Quinoline Derivatives against EGFR
Authors: Asli Faiza, Khamouli Saida
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With the development of computer tools over the past 20 years. Molecular modeling and, more precisely, molecular docking has very quickly entered field of pharmaceutical research. EGFR enzyme involved in cancer disease.Our work consists of studying the inhibition of EGFR (1M17) with deferent inhibitors derived from quinazoline and quinoline by molecular docking. The values of ligands L148 and L177 are the best ligands for inhibit the activity of 1M17 since it forms a stable complex with this enzyme by better binding to the active site. The results obtained show that the ligands L148 and L177 give weak interactions with the active site residues EGFR (1M17), which stabilize the complexes formed of this ligands, which gives a better binding at the level of the active site, and an RMSD of L148 [1,9563 Å] and of L177 [ 1,2483 Å]. [1, 9563, 1.2483] ÅKeywords: docking, EGFR, quinazoline, quinoliène, MOE
Procedia PDF Downloads 683059 A Multi-Arm Randomized Trial Comparing the Weight Gain of Very Low Birth Weight Neonates: High Glucose versus High Protein Intake
Authors: Farnaz Firuzian, Farhad Choobdar, Ali Mazouri
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As Very Low Birth Weight (VLBW) neonates cannot tolerate enteral feeding, parenteral nutrition (PN) must be administered shortly after birth. To find an optimal combination of nutrition, in this study, we compare administering high glucose versus high protein intake as a component of total parenteral nutrition (TPN) to test their effect on birth weight (BW) regain in VLBW. This study employs a multi-arm randomized trial: 145 newborns with BW < 1500 g were randomized to control (C) or experimental groups: high glucose (G) or high protein (P). All samples in each group received the same TPN regimens except glucose and protein intake: Glocuse was provided by dextrose water (DW) serum: 7-15 g/kg/d (10% DW) in groups C and P versus 8.75-18.75 g/kg/d (12.5% DW) in group G. Protein provided by amino acids 3 g/kg/d for groups C and G versus 4 g/kg/d for group P. Outcomes (weight, height, and head circumference) was monitored on a daily basis until the BW was regained. Data has been gathered recently and is being processed. We hypothesize that neonates with higher amino acid intake will result in sooner BW regain than other groups. The result will be presented at the conference. The findings of this study not only can help optimize nutrition, cost reduction, and shorter NICU admission of VLBW neonates at the hospital level but eventually contribute to reduced healthcare-associated infections (HAIs) and an improved health economy.Keywords: very low birth weight neonates, weight gain, parenteral nutrition, glucose, amino acids
Procedia PDF Downloads 833058 Large-Scale Screening for Membrane Protein Interactions Involved in Platelet-Monocyte Interactions
Authors: Yi Sun, George Ed Rainger, Steve P. Watson
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Background: Beyond the classical roles in haemostasis and thrombosis, platelets are important in the initiation and development of various thrombo-inflammatory diseases. In atherosclerosis and deep vein thrombosis, for example, platelets bridge monocytes with endothelium and form heterotypic aggregates with monocytes in the circulation. This can alter monocyte phenotype by inducing their activation, stimulating adhesion and migration. These interactions involve cell surface receptor-ligand pairs on both cells. This list is likely incomplete as new interactions of importance to platelet biology are continuing to be discovered as illustrated by our discovery of PEAR-1 binding to FcεR1α. Results: We have developed a highly sensitive avidity-based assay to identify novel extracellular interactions among 126 recombinantly-expressed platelet cell surface and secreted proteins involved in platelet aggregation. In this study, we will use this method to identify novel platelet-monocyte interactions. We aim to identify ligands for orphan receptors and novel partners of well-known proteins. Identified interactions will be studied in preliminary functional assays to demonstrate relevance to the inflammatory processes supporting atherogenesis. Conclusions: Platelet-monocyte interactions are essential for the development of thromboinflammatory disease. Up until relatively recently, technologies only allow us to limit our studies on each individual protein interaction at a single time. These studies propose for the first time to study the cell surface platelet-monocyte interactions in a systematic large-scale approach using a reliable screening method we have developed. If successful, this will likely to identify previously unknown ligands for important receptors that will be investigated in details and also provide a list of novel interactions for the field. This should stimulate studies on developing alternative therapeutic strategies to treat vascular inflammatory disorders such as atherosclerosis, DVT and sepsis and other clinically important inflammatory conditions.Keywords: membrane proteins, large-scale screening, platelets, recombinant expression
Procedia PDF Downloads 1513057 In Silico Studies on Selected Drug Targets for Combating Drug Resistance in Plasmodium Falcifarum
Authors: Deepika Bhaskar, Neena Wadehra, Megha Gulati, Aruna Narula, R. Vishnu, Gunjan Katyal
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With drug resistance becoming widespread in Plasmodium falciparum infections, development of the alternative drugs is the desired strategy for prevention and cure of malaria. Three drug targets were selected to screen promising drug molecules from the GSK library of around 14000 molecules. Using an in silico structure-based drug designing approach, the differences in binding energies of the substrate and inhibitor were exploited between target sites of parasite and human to design a drug molecule against Plasmodium. The docking studies have shown several promising molecules from GSK library with more effective binding as compared to the already known inhibitors for the drug targets. Though stronger interaction has been shown by several molecules as compare to reference, few molecules have shown the potential as drug candidates though in vitro studies are required to validate the results.Keywords: plasmodium, malaria, drug targets, in silico studies
Procedia PDF Downloads 4473056 Metabolic Profiling in Breast Cancer Applying Micro-Sampling of Biological Fluids and Analysis by Gas Chromatography – Mass Spectrometry
Authors: Mónica P. Cala, Juan S. Carreño, Roland J.W. Meesters
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Recently, collection of biological fluids on special filter papers has become a popular micro-sampling technique. Especially, the dried blood spot (DBS) micro-sampling technique has gained much attention and is momently applied in various life sciences reserach areas. As a result of this popularity, DBS are not only intensively competing with the venous blood sampling method but are at this moment widely applied in numerous bioanalytical assays. In particular, in the screening of inherited metabolic diseases, pharmacokinetic modeling and in therapeutic drug monitoring. Recently, microsampling techniques were also introduced in “omics” areas, whereunder metabolomics. For a metabolic profiling study we applied micro-sampling of biological fluids (blood and plasma) from healthy controls and from women with breast cancer. From blood samples, dried blood and plasma samples were prepared by spotting 8uL sample onto pre-cutted 5-mm paper disks followed by drying of the disks for 100 minutes. Dried disks were then extracted by 100 uL of methanol. From liquid blood and plasma samples 40 uL were deproteinized with methanol followed by centrifugation and collection of supernatants. Supernatants and extracts were evaporated until dryness by nitrogen gas and residues derivated by O-methyxyamine and MSTFA. As internal standard C17:0-methylester in heptane (10 ppm) was used. Deconvolution and alignment of and full scan (m/z 50-500) MS data were done by AMDIS and SpectConnect (http://spectconnect.mit.edu) software, respectively. Statistical Data analysis was done by Principal Component Analysis (PCA) using R software. The results obtained from our preliminary study indicate that the use of dried blood/plasma on paper disks could be a powerful new tool in metabolic profiling. Many of the metabolites observed in plasma (liquid/dried) were also positively identified in whole blood samples (liquid/dried). Whole blood could be a potential substitute matrix for plasma in Metabolomic profiling studies as well also micro-sampling techniques for the collection of samples in clinical studies. It was concluded that the separation of the different sample methodologies (liquid vs. dried) as observed by PCA was due to different sample treatment protocols applied. More experiments need to be done to confirm obtained observations as well also a more rigorous validation .of these micro-sampling techniques is needed. The novelty of our approach can be found in the application of different biological fluid micro-sampling techniques for metabolic profiling.Keywords: biofluids, breast cancer, metabolic profiling, micro-sampling
Procedia PDF Downloads 4113055 Artificial Cells Capable of Communication by Using Polymer Hydrogel
Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng
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The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.Keywords: artificial cell, cell-free system, gene circuit, synthetic biology
Procedia PDF Downloads 1523054 Human LACE1 Functions Pro-Apoptotic and Interacts with Mitochondrial YME1L Protease
Authors: Lukas Stiburek, Jana Cesnekova, Josef Houstek, Jiri Zeman
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Cellular function depends on mitochondrial function and integrity that is therefore maintained by several classes of proteins possessing chaperone and/or proteolytic activities. In this work, we focused on characterization of LACE1 (lactation elevated 1) function in mitochondrial protein homeostasis maintenance. LACE1 is the human homologue of yeast mitochondrial Afg1 ATPase, a member of SEC18-NSF, PAS1, CDC48-VCP, TBP family. Yeast Afg1 was shown to be involved in mitochondrial complex IV biogenesis, and based on its similarity with CDC48 (p97/VCP) it was suggested to facilitate extraction of polytopic membrane proteins. Here we show that LACE1, which is a mitochondrial integral membrane protein, exists as part of three complexes of approx. 140, 400 and 500 kDa and is essential for maintenance of fused mitochondrial reticulum and lamellar cristae morphology. Using affinity purification of LACE1-FLAG expressed in LACE1 knockdown background we show that the protein physically interacts with mitochondrial inner membrane protease YME1L. We further show that human LACE1 exhibits significant pro-apoptotic activity and that the protein is required for normal function of the mitochondrial respiratory chain. Thus, our work establishes LACE1 as a novel factor with the crucial role in mitochondrial homeostasis maintenance.Keywords: LACE1, mitochondria, apoptosis, protease
Procedia PDF Downloads 3133053 Differentially Expressed Genes in Atopic Dermatitis: Bioinformatics Analysis Of Pooled Microarray Gene Expression Datasets In Gene Expression Omnibus
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Background: Atopic dermatitis (AD) is a chronic and refractory inflammatory skin disease characterized by relapsing eczematous and pruritic skin lesions. The global prevalence of AD ranges from 1~ 20%, and its incidence rates are increasing. It affects individuals from infancy to adulthood, significantly impacting their daily lives and social activities. Despite its major health burden, the precise mechanisms underlying AD remain unknown. Understanding the genetic differences associated with AD is crucial for advancing diagnosis and targeted treatment development. This study aims to identify candidate genes of AD by using bioinformatics analysis. Methods: We conducted a comprehensive analysis of four pooled transcriptomic datasets (GSE16161, GSE32924, GSE130588, and GSE120721) obtained from the Gene Expression Omnibus (GEO) database. Differential gene expression analysis was performed using the R statistical language. The differentially expressed genes (DEGs) between AD patients and normal individuals were functionally analyzed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Furthermore, a protein-protein interaction (PPI) network was constructed to identify candidate genes. Results: Among the patient-level gene expression datasets, we identified 114 shared DEGs, consisting of 53 upregulated genes and 61 downregulated genes. Functional analysis using GO and KEGG revealed that the DEGs were mainly associated with the negative regulation of transcription from RNA polymerase II promoter, membrane-related functions, protein binding, and the Human papillomavirus infection pathway. Through the PPI network analysis, we identified eight core genes: CD44, STAT1, HMMR, AURKA, MKI67, and SMARCA4. Conclusion: This study elucidates key genes associated with AD, providing potential targets for diagnosis and treatment. The identified genes have the potential to contribute to the understanding and management of AD. The bioinformatics analysis conducted in this study offers new insights and directions for further research on AD. Future studies can focus on validating the functional roles of these genes and exploring their therapeutic potential in AD. While these findings will require further verification as achieved with experiments involving in vivo and in vitro models, these results provided some initial insights into dysfunctional inflammatory and immune responses associated with AD. Such information offers the potential to develop novel therapeutic targets for use in preventing and treating AD.Keywords: atopic dermatitis, bioinformatics, biomarkers, genes
Procedia PDF Downloads 823052 Role of Estrogen Receptor-alpha in Mammary Carcinoma by Single Nucleotide Polymorphisms and Molecular Docking: An In-silico Analysis
Authors: Asif Bilal, Fouzia Tanvir, Sibtain Ahmad
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Estrogen receptor alpha, also known as estrogen receptor-1, is highly involved in risk of mammary carcinoma. The objectives of this study were to identify non-synonymous SNPs of estrogen receptor and their association with breast cancer and to identify the chemotherapeutic responses of phytochemicals against it via in-silico study design. For this purpose, different online tools. to identify pathogenic SNPs the tools were SIFT, Polyphen, Polyphen-2, fuNTRp, SNAP2, for finding disease associated SNPs the tools SNP&GO, PhD-SNP, PredictSNP, MAPP, SNAP, MetaSNP, PANTHER, and to check protein stability Mu-Pro, I-Mutant, and CONSURF were used. Post-translational modifications (PTMs) were detected by Musitedeep, Protein secondary structure by SOPMA, protein to protein interaction by STRING, molecular docking by PyRx. Seven SNPs having rsIDs (rs760766066, rs779180038, rs956399300, rs773683317, rs397509428, rs755020320, and rs1131692059) showing mutations on I229T, R243C, Y246H, P336R, Q375H, R394S, and R394H, respectively found to be completely deleterious. The PTMs found were 96 times Glycosylation; 30 times Ubiquitination, a single time Acetylation; and no Hydroxylation and Phosphorylation were found. The protein secondary structure consisted of Alpha helix (Hh) is (28%), Extended strand (Ee) is (21%), Beta turn (Tt) is 7.89% and Random coil (Cc) is (44.11%). Protein-protein interaction analysis revealed that it has strong interaction with Myeloperoxidase, Xanthine dehydrogenase, carboxylesterase 1, Glutathione S-transferase Mu 1, and with estrogen receptors. For molecular docking we used Asiaticoside, Ilekudinuside, Robustoflavone, Irinoticane, Withanolides, and 9-amin0-5 as ligands that extract from phytochemicals and docked with this protein. We found that there was great interaction (from -8.6 to -9.7) of these ligands of phytochemicals at ESR1 wild and two mutants (I229T and R394S). It is concluded that these SNPs found in ESR1 are involved in breast cancer and given phytochemicals are highly helpful against breast cancer as chemotherapeutic agents. Further in vitro and in vivo analysis should be performed to conduct these interactions.Keywords: breast cancer, ESR1, phytochemicals, molecular docking
Procedia PDF Downloads 693051 Predicting Aggregation Propensity from Low-Temperature Conformational Fluctuations
Authors: Hamza Javar Magnier, Robin Curtis
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There have been rapid advances in the upstream processing of protein therapeutics, which has shifted the bottleneck to downstream purification and formulation. Finding liquid formulations with shelf lives of up to two years is increasingly difficult for some of the newer therapeutics, which have been engineered for activity, but their formulations are often viscous, can phase separate, and have a high propensity for irreversible aggregation1. We explore means to develop improved predictive ability from a better understanding of how protein-protein interactions on formulation conditions (pH, ionic strength, buffer type, presence of excipients) and how these impact upon the initial steps in protein self-association and aggregation. In this work, we study the initial steps in the aggregation pathways using a minimal protein model based on square-well potentials and discontinuous molecular dynamics. The effect of model parameters, including range of interaction, stiffness, chain length, and chain sequence, implies that protein models fold according to various pathways. By reducing the range of interactions, the folding- and collapse- transition come together, and follow a single-step folding pathway from the denatured to the native state2. After parameterizing the model interaction-parameters, we developed an understanding of low-temperature conformational properties and fluctuations, and the correlation to the folding transition of proteins in isolation. The model fluctuations increase with temperature. We observe a low-temperature point, below which large fluctuations are frozen out. This implies that fluctuations at low-temperature can be correlated to the folding transition at the melting temperature. Because proteins “breath” at low temperatures, defining a native-state as a single structure with conserved contacts and a fixed three-dimensional structure is misleading. Rather, we introduce a new definition of a native-state ensemble based on our understanding of the core conservation, which takes into account the native fluctuations at low temperatures. This approach permits the study of a large range of length and time scales needed to link the molecular interactions to the macroscopically observed behaviour. In addition, these models studied are parameterized by fitting to experimentally observed protein-protein interactions characterized in terms of osmotic second virial coefficients.Keywords: protein folding, native-ensemble, conformational fluctuation, aggregation
Procedia PDF Downloads 3613050 Development of Adhesive from Prosopis african Seed Endosperm (OKPEYI)
Authors: Florence Chinyere Nwangwu, Rosemary Ene
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An experimental study was carried out to develop an adhesive from Prosopis africana seed endosperm. The Prosopis seeds for this work were obtained from Enugu State in the South East part of Nigeria. The Prosopis seeds were prepared by separating the Prosopis endosperm from the seed coat and cotyledon. The dry adhesive gotten from the endosperm was later dissolved to get the adhesive solution. Confirmatory tests like viscosity, density, pH, and binding strength were carried out. The effect of time, temperature, concentration on the yield and properties of the adhesive were investigated. The results obtained showed that increase in concentration, time, temperature decreases the viscosity of the Prosopis adhesive and yield of Prosopis endosperm. It was also deduced that increase in viscosity increases the binding strength of the Prosopis adhesive. The percentage of the adhesive yield from Prosopis endosperm showed that the commercialization of the seed in Nigeria will be possible and profitable.Keywords: adhesive, Prosopis, viscosity, endosperm
Procedia PDF Downloads 3093049 The Effects of Inoculation and N Fertilization on Soybean (Glycine max (L.) Merr.) Seed Yield and Protein Concentration under Drought Stress
Authors: Oqba Basal, Andras Szabo
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Using mineral fertilization is increasing worldwide, as it is claimed to be majorly responsible for achieving high yields; however, the negative impacts of mineral fertilization on soil and environment are becoming more obvious, with alternative methods being more necessary and applicable, especially with the current climatic changes which have imposed serious abiotic stresses, such as drought. An experiment was made during 2017 growing season in Debrecen, Hungary to investigate the effects of inoculation and N fertilization on the seed yield and protein concentration of the soybean (Glycine max (L.) Merr.) cultivar (Panonia Kincse) under three different irrigation regimes: severe drought stress (SD), moderate drought stress (MD) and control with no drought stress (ND). Three N fertilizer rates were applied: no N fertilizer (0 N), 35 kg ha⁻¹ of N fertilizer (35 N) and 105 kg ha⁻¹ of N fertilizer (105 N). Half of the seeds in each treatment was inoculated with Bradyrhizobium japonicum inoculant, and the other half was not inoculated. The results showed significant differences in the seed yield associated with inoculation, irrigation and the interaction between them, whereas there were no significant differences in the seed yield associated with fertilization alone or in interaction with inoculation or irrigation or both. When seeds were inoculated, yield was increased when (35 N) was applied compared to (0 N) but not significantly; however, the high rate of N fertilizer (105 N) reduced the yield to a level even less than (0 N). When seeds were not inoculated, the highest rate of N increased the yield the most compared to the other two N fertilizer rates whenever the drought was present (moderate or severe). Under severe drought stress, inoculation was positively and significantly correlated with yield; however, adding N fertilizer increased the yield of uninoculated plants compared to the inoculated ones, regardless of the rate of N fertilizer. Protein concentration in the seeds was significantly affected by irrigation and by fertilization, but not by inoculation. Protein concentration increased as the N fertilization rate increased, regardless of the inoculation or irrigation treatments; moreover, increasing the N rate reduced the correlation coefficient of protein concentration with the irrigation. It was concluded that adding N fertilizer is not always recommended, especially when seeds are inoculated before being sown; however, it is very important under severe drought stress to sustain yield. Enhanced protein concentrations could be achieved by applying N fertilization, whether the seeds were pre-inoculated or not.Keywords: drought stress, N fertilization, protein concentration, soybean
Procedia PDF Downloads 1543048 Research of the Rotation Magnetic Field Current Driven Effect on Pulsed Plasmoid Acceleration of Electric Propulsion
Authors: X. F. Sun, X. D. Wen, L. J. Liu, C. C. Wu, Y. H. Jia
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The field reversed closed magnetic field configuration plasmoid has a potential for large thrust and high power propulsion missions such as deep space exploration due to its high plasma density and larger azimuthal current, which will be a most competitive program for the next generation electric propulsion technology. Moreover, without the electrodes, it also has a long lifetime. Thus, the research on this electric propulsion technology is quite necessary. The plasmoid will be formatted and accelerated by applying a rotation magnetic field (RMF) method. And, the essence of this technology lies on the generation of the azimuthal electron currents driven by RMF. Therefore, the effect of RMF current on the plasmoid acceleration efficiency is a concerned problem. In the paper, the influences of the penetration process of RMF in plasma, the relations of frequency and amplitude of input RF power with current strength and the RMF antenna configuration on the plasmoid acceleration efficiency will be given by a two-fluid numerical simulation method. The results show that the radio-frequency and input power have remarkable influence on the formation and acceleration of plasmoid. These results will provide useful advice for the development, and optimized designing of field reversed configuration plasmoid thruster.Keywords: rotation magnetic field, current driven, plasma penetration, electric propulsion
Procedia PDF Downloads 1163047 Ramadan as a Model of Intermittent Fasting: Effects on Gut Hormones, Appetite and Body Composition in Diabetes vs. Controls
Authors: Turki J. Alharbi, Jencia Wong, Dennis Yue, Tania P. Markovic, Julie Hetherington, Ted Wu, Belinda Brooks, Radhika Seimon, Alice Gibson, Stephanie L. Silviera, Amanda Sainsbury, Tanya J. Little
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Fasting has been practiced for centuries and is incorporated into the practices of different religions including Islam, whose followers intermittently fast throughout the month of Ramadan. Thus, Ramadan presents a unique model of prolonged intermittent fasting (IF). Despite a growing body of evidence for a cardio-metabolic and endocrine benefit of IF, detailed studies of the effects of IF on these indices in type 2 diabetes are scarce. We studied 5 subjects with type 2 diabetes (T2DM) and 7 healthy controls (C) at baseline (pre), and in the last week of Ramadan (post). Fasting circulating levels of glucose, HbA1c and lipids, as well as body composition (with DXA) and resting energy expenditure (REE) were measured. Plasma gut hormone levels and appetite responses to a mixed meal were also studied. Data are means±SEM. Ramadan decreased total fat mass (-907±92 g, p=0.001) and trunk fat (-778±190 g, p=0.014) in T2DM but not in controls, without any reductions in lean mass or REE. There was a trend towards a decline in plasma FFA in both groups. Ramadan had no effect on body weight, glycemia, blood pressure, or plasma lipids in either group. In T2DM only, the area under the curve for post-meal plasma ghrelin concentrations increased after Ramadan (pre:6632±1737 vs. post:9025±2518 pg/ml.min-1, p=0.045). Despite this increase in orexigenic ghrelin, subjective appetite scores were not altered by Ramadan. Meal-induced plasma concentrations of the satiety hormone pancreatic polypeptide did not change during Ramadan, but were higher in T2DM compared to controls (post: C: 23486±6677 vs. T2DM: 62193±6880 pg/ml.min-1, p=0.003. In conclusion, Ramadan, as a model for IF appears to have more favourable effects on body composition in T2DM, without adverse effects on metabolic control or subjective appetite. These data suggest that IF may be particularly beneficial in T2DM as a nutritional intervention. Larger studies are warranted.Keywords: type 2 diabetes, obesity, intermittent fasting, appetite regulating hormones
Procedia PDF Downloads 3123046 Folding Pathway and Thermodynamic Stability of Monomeric GroEL
Authors: Sarita Puri, Tapan K. Chaudhuri
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Chaperonin GroEL is a tetradecameric Escherichia coli protein having identical subunits of 57 kDa. The elucidation of thermodynamic parameters related to stability for the native GroEL is not feasible as it undergoes irreversible unfolding because of its large size (800kDa) and multimeric nature. Nevertheless, it is important to determine the thermodynamic stability parameters for the highly stable GroEL protein as it helps in folding and holding of many substrate proteins during many cellular stresses. Properly folded monomers work as building-block for the formation of native tetradecameric GroEL. Spontaneous refolding behavior of monomeric GroEL makes it suitable for protein-denaturant interactions and thermodynamic stability based studies. The urea mediated unfolding is a three state process which means there is the formation of one intermediate state along with native and unfolded states. The heat mediated denaturation is a two-state process. The unfolding process is reversible as observed by the spontaneous refolding of denatured protein in both urea and head mediated refolding processes. Analysis of folding/unfolding data provides a measure of various thermodynamic stability parameters for the monomeric GroEL. The proposed mechanism of unfolding of monomeric GroEL is a three state process which involves formation of one stable intermediate having folded apical domain and unfolded equatorial, intermediate domains. Research in progress is to demonstrate the importance of specific residues in stability and oligomerization of GroEL protein. Several mutant versions of GroEL are under investigation to resolve the above mentioned issue.Keywords: equilibrium unfolding, monomeric GroEl, spontaneous refolding, thermodynamic stability
Procedia PDF Downloads 2823045 Characterization of Calcium-Signalling Mediated by Human GPR55 Expressed in HEK293 Cells
Authors: Yousuf M. Al Suleimani, Robin Hiley
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The endogenous phospholipid lysophosphatidylinositol (LPI) was recently identified as a novel ligand for the G protein-coupled receptor 55 (GPR55) and an inducer of intracellular Ca2+ [Ca2+]i release. This study attempts to characterize Ca2+ signals provoked by LPI in HEK293 cells engineered to stably express human GPR55 and to test cannabinoid ligand activity at GPR55. The study shows that treatment with LPI stimulates a sustained, oscillatory Ca2+ release. The response is characterized by an initial rapid rise, which is mediated by the Gαq-PLC-IP3 pathway, and this is followed by prolonged oscillations that require RhoA activation. Ca2+ oscillations are initiated by intracellular mechanisms and extracellular Ca2+ is only required to replenish Ca2+ lost from the cytoplasm. Analysis of cannabinoid ligand activity at GPR55 revealed no clear effect of the endocannabinoid anandamide, however, rimonabant and the CB1 receptor antagonist AM251 evoked GPR55-mediated [Ca2+]i. Thus, LPI is likely to be a key plasma membrane mediator of signaling events and changes in gene expression through GPR55 activation.Keywords: lysophosphatidylinositol, calcium, GPR55, cannabinoid
Procedia PDF Downloads 3593044 Effect of Whey Protein Based Edible Coating on the Moisture Loss and Sensory Attributes of Fresh Mutton
Authors: Saba Belgheisi
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Food packaging, is an important discipline in the area of food technology, concerns preservation and protection of foods. The objective of this research was to determine of the effect of whey protein based edible coating on the moisture loss and sensory attributes of fresh mutton after 0, 1, 3 and 5 days at 5° C. The moisture content, moisture loss and sensory attributes (juiciness, color and odor) of the coated and uncoated samples were analyzed. The results showed that, moisture content, moisture loss, juiciness and color of the coated and uncoated samples have significant differences (p < 0.05) at the intervals of 0 to 1 and 1 to 3 days of storage. But no significant difference was observed at interval time 3 to 5 days of storage (p > 0.05). Also, there was no significant differences in the odor values of the coated and uncoated samples (p > 0.05). Therefore, the coated samples had consistently more moisture, juiciness and colored values than uncoated samples after 3 days at 5° C. So, whey protein edible coating could enhance product presentation and eliminate the need for placing absorbent pads at the bottom of the trays.Keywords: coating, whey protein, mutton, moisture, sensory
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