Search results for: plasma protein electrophoresis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3374

Search results for: plasma protein electrophoresis

3284 Assessment of DNA Degradation Using Comet Assay: A Versatile Technique for Forensic Application

Authors: Ritesh K. Shukla

Abstract:

Degradation of biological samples in terms of macromolecules (DNA, RNA, and protein) are the major challenges in the forensic investigation which misleads the result interpretation. Currently, there are no precise methods available to circumvent this problem. Therefore, at the preliminary level, some methods are urgently needed to solve this issue. In this order, Comet assay is one of the most versatile, rapid and sensitive molecular biology technique to assess the DNA degradation. This technique helps to assess DNA degradation even at very low amount of sample. Moreover, the expedient part of this method does not require any additional process of DNA extraction and isolation during DNA degradation assessment. Samples directly embedded on agarose pre-coated microscopic slide and electrophoresis perform on the same slide after lysis step. After electrophoresis microscopic slide stained by DNA binding dye and observed under fluorescent microscope equipped with Komet software. With the help of this technique extent of DNA degradation can be assessed which can help to screen the sample before DNA fingerprinting, whether it is appropriate for DNA analysis or not. This technique not only helps to assess degradation of DNA but many other challenges in forensic investigation such as time since deposition estimation of biological fluids, repair of genetic material from degraded biological sample and early time since death estimation could also be resolved. With the help of this study, an attempt was made to explore the application of well-known molecular biology technique that is Comet assay in the field of forensic science. This assay will open avenue in the field of forensic research and development.

Keywords: comet assay, DNA degradation, forensic, molecular biology

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3283 Learning Materials of Atmospheric Pressure Plasma Process: Turning Hydrophilic Surface to Hydrophobic

Authors: C.W. Kan

Abstract:

This paper investigates the use of atmospheric pressure plasma for improving the surface hydrophobicity of polyurethane synthetic leather with tetramethylsilane (TMS). The atmospheric pressure plasma treatment with TMS is a single-step process to enhance the hydrophobicity of polyurethane synthetic leather. The hydrophobicity of the treated surface was examined by contact angle measurement. The physical and chemical surface changes were evaluated by scanning electron microscopy (SEM) and infrared spectroscopy (FTIR). The purpose of this paper is to provide learning materials for understanding how to use atmospheric pressure plasma in the textile finishing process to transform a hydrophilic surface to hydrophobic.

Keywords: Learning materials, atmospheric pressure plasma treatment, hydrophobic, hydrophilic, surface

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3282 Effect of Plasma Radiation on Keratinocyte Cells Involved in the Wound Healing Process

Authors: B. Fazekas, I. Korolov, K. Kutasi

Abstract:

Plasma medicine, which involves the use of gas discharge plasmas for medical applications is a rapidly growing research field. The use of non-thermal atmospheric pressure plasmas in dermatology to assist tissue regeneration by improving the healing of infected and/or chronic wounds is a promising application. It is believed that plasma can activate cells, which are involved in the wound closure. Non-thermal atmospheric plasmas are rich in chemically active species (such as O and N-atoms, O2(a) molecules) and radiative species such as the NO, N2+ and N2 excited molecules, which dominantly radiate in the 200-500 nm spectral range. In order to understand the effect of plasma species, both of chemically active and radiative species on wound healing process, the interaction of physical plasma with the human skin cells is necessary. In order to clarify the effect of plasma radiation on the wound healing process we treated keratinocyte cells – that are one of the main cell types in human skin epidermis – covered with a layer of phosphate-buffered saline (PBS) with a low power atmospheric pressure plasma. For the generation of such plasma we have applied a plasma needle. Here, the plasma is ignited at the tip of the needle in flowing helium gas in contact with the ambient air. To study the effect of plasma radiation we used a plasma needle configuration, where the plasma species – chemically active radicals and charged species – could not reach the treated cells, but only the radiation. For the comparison purposes, we also irradiated the cells using a UV-B light source (FS20 lamp) with a 20 and 40 mJ cm-2 dose of 312 nm. After treatment the viability and the proliferation of the cells have been examined. The proliferation of cells has been studied with a real time monitoring system called Xcelligence. The results have indicated, that the 20 mJ cm-2 dose did not affect cell viability, whereas the 40 mJ cm-2 dose resulted a decrease in cell viability. The results have shown that the plasma radiation have no quantifiable effect on the cell proliferation as compared to the non-treated cells.

Keywords: UV radiation, non-equilibrium gas discharges (non-thermal plasmas), plasma emission, keratinocyte cells

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3281 High-Production Laser and Plasma Welding Technologies for High-Speed Vessels Production

Authors: V. M. Levshakov, N. A. Steshenkova, N. A. Nosyrev

Abstract:

Application of hulls processing technologies, based on high-concentrated energy sources (laser and plasma technologies), allow improve shipbuilding production. It is typical for high-speed vessels construction using steel and aluminum alloys with high precision hulls required. Report describes high-performance technologies for plasma welding (using direct current of reversed polarity), laser, and hybrid laser-arc welding of hulls structures developed by JSC “SSTC”.

Keywords: flat sections, hybrid laser-arc welding, plasma welding, plasmatron

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3280 Effect of Electromagnetic Fields on Protein Extraction from Shrimp By-Products for Electrospinning Process

Authors: Guido Trautmann-Sáez, Mario Pérez-Won, Vilbett Briones, María José Bugueño, Gipsy Tabilo-Munizaga, Luis Gonzáles-Cavieres

Abstract:

Shrimp by-products are a valuable source of protein. However, traditional protein extraction methods have limitations in terms of their efficiency. Protein extraction from shrimp (Pleuroncodes monodon) industrial by-products assisted with ohmic heating (OH), microwave (MW) and pulsed electric field (PEF). It was performed by chemical method (using NaOH and HCl 2M) assisted with OH, MW and PEF in a continuous flow system (5 ml/s). Protein determination, differential scanning calorimetry (DSC) and Fourier-transform infrared (FTIR). Results indicate a 19.25% (PEF) 3.65% (OH) and 28.19% (MW) improvement in protein extraction efficiency. The most efficient method was selected for the electrospinning process and obtaining fiber.

Keywords: electrospinning process, emerging technology, protein extraction, shrimp by-products

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3279 Physicochemical Properties of Soy Protein Isolate (SPI): Starch Conjugates Treated by Sonication

Authors: Gulcin Yildiz, Hao Feng

Abstract:

In recent years there is growing interested in using soy protein because of several advantages compared to other protein sources, such as high nutritional value, steady supply, and low cost. Soy protein isolate (SPI) is the most refined soy protein product. It contains 90% protein in a moisture-free form and has some desirable functionalities. Creating a protein-polysaccharide conjugate to be the emulsifying agent rather than the protein alone can markedly enhance its stability. This study was undertaken to examine the effects of ultrasound treatments on the physicochemical properties of SPI-starch conjugates. The soy protein isolate (SPI, Pro-Fam® 955) samples were obtained from the Archer Daniels Midland Company. Protein concentrations were analyzed by the Bardford method using BSA as the standard. The volume-weighted mean diameters D [4,3] of protein–polysaccharide conjugates were measured by dynamic light scattering (DLS). Surface hydrophobicity of the conjugates was measured by using 1-anilino-8-naphthalenesulfonate (ANS) (Sigma-Aldrich, St. Louis, MO, USA). Increasing the pH from 2 to 12 resulted in increased protein solubility. The highest solubility was 69.2% for the sample treated with ultrasonication at pH 12, while the lowest (9.13%) was observed in the Control. For the other pH conditions, the protein solubility values ranged from 40.53 to 49.65%. The ultrasound treatment significantly decreased the particle sizes of the SPI-modified starch conjugates. While the D [4,3] for the Control was 731.6 nm, it was 293.7 nm for the samples treated by sonication at pH 12. The surface hydrophobicity (H0) of SPI-starch at all pH conditions were significantly higher than those in the Control. Ultrasonication was proven to be effective in improving the solubility and emulsifying properties of soy protein isolate-starch conjugates.

Keywords: particle size, solubility, soy protein isolate, ultrasonication

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3278 Resistive Instability in a Multi Ions Hall Thrusters Plasma

Authors: Sukhmander Singh

Abstract:

Hall thrusters are preferred over chemical thrusters because of its high exhaust velocity (around 10 times higher) and high specific impulse. The propellant Xenon is ionized inside the channel and controlled by the magnetic field. The strength of the magnetic field is such that only electrons get magnetized and ions remain unmagnetized because of larger Larmor radius as compared with the length of the channel of the device. There is quite a possibility of the existence of multi ions in a Hall thruster plasma because of dust contribution or another process which take place in the chamber. In this paper, we have derived the dispersion relation for multi ions resistive instability in a hall plasma. The analytical approach is also used to find out the propagating speed and the growth rate of the instability. In addition, some growing waves are also found to exist in the plasma. The dispersion relation is solved numerically to see the behavior of the instability with the plasma parameters viz, the temperature of plasma species, wave number, drift velocity, collision frequency, magnetic field.

Keywords: instability, resisitive, thrusters, waves

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3277 Effect of Removing Hub Domain on Human CaMKII Isoforms Sensitivity to Calcium/Calmodulin

Authors: Ravid Inbar

Abstract:

CaMKII (calcium-calmodulin dependent protein kinase II) makes up 2% of the protein in our brain and has a critical role in memory formation and long-term potentiation of neurons. Despite this, research has yet to uncover the role of one of the domains on the activation of this kinase. The following proposes to express the protein without the hub domain in E. coli, leaving only the kinase and regulatory segment of the protein. Next, a series of kinase assays will be conducted to elucidate the role the hub domain plays on CaMKII sensitivity to calcium/calmodulin activation. The hub domain may be important for activation; however, it may also be a variety of domains working together to influence protein activation and not the hub alone. Characterization of a protein is critical to the future understanding of the protein's function, as well as for producing pharmacological targets in cases of patients with diseases.

Keywords: CaMKII, hub domain, kinase assays, kinase + reg seg

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3276 Comparison of Structure and Corrosion Properties of Titanium Oxide Films Prepared by Thermal Oxidation, DC Plasma Oxidation, and by the Sol-Gel

Authors: O. Çomaklı, M. Yazıcı, T. Yetim, A. F. Yetim, A. Çelik

Abstract:

In this work, TiO₂ films were deposited on Cp-Ti substrates by thermal oxidation, DC plasma oxidation, and by the sol-gel method. Microstructures of uncoated and TiO₂ film coated samples were examined by X-ray diffraction and SEM. Thin oxide film consisting of anatase (A) and rutile (R) TiO₂ structures was observed on the surface of CP-Ti by under three different treatments. Also, the more intense anatase and rutile peaks appeared at samples plasma oxidized at 700˚C. The thicknesses of films were about 1.8 μm at the TiO₂ film coated samples by sol-gel and about 2.7 μm at thermal oxidated samples, while it was measured as 3.9 μm at the plasma oxidated samples. Electrochemical corrosion behaviour of uncoated and coated specimens was mainly carried out by potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) in simulated body fluid (SBF) solution. Results showed that at the plasma oxidated samples exhibited a better resistance property to corrosion than that of other treatments.

Keywords: TiO₂, CP-Ti, corrosion properties, thermal oxidation, plasma oxidation, sol-gel

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3275 Testing Serum Proteome between Elite Sprinters and Long-Distance Runners

Authors: Hung-Chieh Chen, Kuo-Hui Wang, Tsu-Lin Yeh

Abstract:

Proteomics represent the performance of genomic complement proteins and the protein level on functional genomics. This study adopted proteomic strategies for comparing serum proteins among three groups: elite sprinter (sprint runner group, SR), long-distance runners (long-distance runner group, LDR), and the untrained control group (control group, CON). Purposes: This study aims to identify elite sprinters and long-distance runners’ serum protein and to provide a comparison of their serum proteome’ composition. Methods: Serum protein fractionations that separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by a quantitative nano-LC-MS/MS-based proteomic profiling. The one-way analysis of variance (ANOVA) and Scheffe post hoc comparison (α= 0.05) was used to determine whether there is any significant difference in each protein level among the three groups. Results: (1) After analyzing the 307 identified proteins, there were 26 unique proteins in the SR group, and 18 unique proteins in the LDR group. (2) For the LDR group, 7 coagulation function-associated proteins’ expression levels were investigated: vitronectin, serum paraoxonase/arylesterase 1, fibulin-1, complement C3, vitamin K-dependent protein, inter-alpha-trypsin inhibitor heavy chain H3 and von Willebrand factor, and the findings show the seven coagulation function-associated proteins were significantly lower than the group of SR. (3) Comparing to the group of SR, this study found that the LDR group’s expression levels of the 2 antioxidant proteins (afamin and glutathione peroxidase 3) were also significantly lower. (4) The LDR group’s expression levels of seven immune function-related proteins (Ig gamma-3 chain C region, Ig lambda-like polypeptide 5, clusterin, complement C1s subcomponent, complement factor B, complement C4-A, complement C1q subcomponent subunit A) were also significantly lower than the group of SR. Conclusion: This study identified the potential serum protein markers for elite sprinters and long-distance runners. The changes in the regulation of coagulation, antioxidant, or immune function-specific proteins may also provide further clinical applications for these two different track athletes.

Keywords: biomarkers, coagulation, immune response, oxidative stress

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3274 Biocompatibility and Electrochemical Assessment of Biomedical Ti-24Nb-4Zr-8Sn Produced by Spark Plasma Sintering

Authors: Jerman Madonsela, Wallace Matizamhuka, Akiko Yamamoto, Ronald Machaka, Brendon Shongwe

Abstract:

In this study, biocompatibility evaluation of nanostructured near beta Ti-24Nb-4Zr-8Sn (Ti2448) alloy with non-toxic elements produced utilizing Spark plasma sintering (SPS) of very fine microsized powders attained through mechanical alloying was performed. The results were compared with pure titanium and Ti-6Al-4V (Ti64) alloy. Cell proliferation test was performed using murine osteoblastic cells, MC3T3-E1 at two cell densities; 400 and 4000 cells/mL for 7 days incubation. Pure titanium took a lead under both conditions suggesting that the presence of other oxide layers influence cell proliferation. No significant difference in cell proliferation was observed between Ti64 and Ti2448. Potentiodynamic measurement in Hanks, 0.9% NaCl and cell culture medium showed no distinct difference on the anodic polarization curves of the three alloys, indicating that the same anodic reaction occurred on their surface but with different rates. However, Ti2448 showed better corrosion resistance in cell culture medium with a slightly lower corrosion rate of 2.96 nA/cm2 compared to 4.86 nA/cm2 and 5.62 nA/cm2 of Ti and Ti64 respectively. Ti2448 adsorbed less protein as compared to Ti and Ti64 though no notable difference in surface wettability was observed.

Keywords: biocompatibility, osteoblast, corrosion, surface wettability, protein adsorption

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3273 Fortification of Concentrated Milk Protein Beverages with Soy Proteins: Impact of Divalent Cations and Heating Treatment on the Physical Stability

Authors: Yichao Liang, Biye Chen, Xiang Li, Steven R. Dimler

Abstract:

This study investigated the effects of adding calcium and magnesium chloride on heat and storage stability of milk protein concentrate-soy protein isolate (8:2 respectively) mixtures containing 10% w/w total protein subjected to the in-container sterilization (115 °C x 15 min). The particle size does not change when emulsions are heated at pH between 6.7 and 7.3 irrespective of the mixed protein ratio. Increasing concentration of divalent cation salts resulted in an increase in protein particle size, dry sediment formation and sediment height and a decrease in pH, heat stability and hydration in milk protein concentrate-soy protein isolate mixtures solutions on sterilization at 115°C. Fortification of divalent cation salts in milk protein concentrate-soy protein isolate mixture solutions resulted in an accelerated protein sedimentation and two unique sediment regions during accelerated storage stability testing. Moreover, the heat stability decreased upon sterilization at 115°C, with addition of MgCl₂ causing a greater increase in sedimentation velocity and compressibility than CaCl₂. Increasing pH value of protein milk concentrate-soy protein isolate mixtures solutions from 6.7 to 7.2 resulted in an increase in viscosity following the heat treatment. The study demonstrated that the type and concentration of divalent cation salts used strongly impact heat and storage stability of milk protein concentrate-soy protein isolate mixture nutritional beverages.

Keywords: divalent cation salts, heat stability, milk protein concentrate, soy protein isolate, storage stability

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3272 Plasma Treatment of a Lignite Using Water-Stabilized Plasma Torch at Atmospheric Pressure

Authors: Anton Serov, Alan Maslani, Michal Hlina, Vladimir Kopecky, Milan Hrabovsky

Abstract:

Recycling of organic waste is an increasingly hot topic in recent years. This issue becomes even more interesting if the raw material for the fuel production can be obtained as the result of that recycling. A process of high-temperature decomposition of a lignite (a non-hydrolysable complex organic compound) was studied on the plasma gasification reactor PLASGAS, where water-stabilized plasma torch was used as a source of high enthalpy plasma. The plasma torch power was 120 kW and allowed heating of the reactor to more than 1000 °C. The material feeding rate in the gasification reactor was selected 30 and 60 kg per hour that could be compared with small industrial production. An efficiency estimation of the thermal decomposition process was done. A balance of the torch energy distribution was studied as well as an influence of the lignite particle size and an addition of methane (CH4) in a reaction volume on the syngas composition (H2+CO). It was found that the ratio H2:CO had values in the range of 1,5 to 2,5 depending on the experimental conditions. The recycling process occurred at atmospheric pressure that was one of the important benefits because of the lack of expensive vacuum pump systems. The work was supported by the Grant Agency of the Czech Republic under the project GA15-19444S.

Keywords: atmospheric pressure, lignite, plasma treatment, water-stabilized plasma torch

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3271 BTEX (Benzene, Toluene, Ethylbenzene and Xylene) Degradation by Cold Plasma

Authors: Anelise Leal Vieira Cubas, Marina de Medeiros Machado, Marília de Medeiros Machado

Abstract:

The volatile organic compounds - BTEX (Benzene, Toluene, Ethylbenzene, and Xylene) petroleum derivatives, have high rates of toxicity, which may carry consequences for human health, biota and environment. In this direction, this paper proposes a method of treatment of these compounds by using corona discharge plasma technology. The efficiency of the method was tested by analyzing samples of BTEX after going through a plasma reactor by gas chromatography method. The results show that the optimal residence time of the sample in the reactor was 8 minutes.

Keywords: BTEX, degradation, cold plasma, ecological sciences

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3270 The Haemoglobin, Transferrin, Ceruloplasmin and Glutathione Polymorphism of Native Goat Breeds of Turkey, I-Angora and Hair

Authors: Ayse Ozge Demir, Nihat Mert

Abstract:

This study has been carried out in order to determine the polymorphic traits of various biochemical parameters in goat breeds which are native to Turkey. For this purpose, Angora and Hair goats breeds were chosen as live materials. Two different herds for each breed were selected from Ankara and Antalya, respectively. Blood samples were taken from a total of 120 goats aged between 2 and 4 which was made up of 60 Angora goats and 60 Hair goats. All which derived equally from 4 lots of herds. Analyses were performed for the polymorphic determination of the Haemoglobin (Hb), Transferrine (Tf), Ceruloplasmin (Cp) and Glutathione (GSH). Hb types were determined by starch gel electrophoresis and Tf types were detected by SDS-PAGE electrophoresis. Furthermore, Cp and GSH analyses were performed by spectrophotometrically. Following the analysis, Hb types were found as 3 genotypes (AA, AB, BB) controlled by 2 allel genes. Tf types were found as 6 genotypes (AA, AB, AC, BB, BC, CC) controlled by 3 allele genes. Findings for Hb was in line with the Hardy-Weinberg Equilibrium (HWE) in Angora goats while the Hair goat was not found to be in line. Moreover, Tf was found in line with the HWE for 2 separate goat breeds. The levels of Cp and GSH of two breeds were significantly different from other (P<0.0001). The findings are recorded as a source of reference for prospective polymorphism studies.

Keywords: electrophoresis, genetic resources, goats, spectrophotometer

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3269 Particleboard Production from Atmospheric Plasma Treated Wheat Straw Particles

Authors: Štěpán Hýsek, Milan Podlena, Miloš Pavelek, Matěj Hodoušek, Martin Böhm, Petra Gajdačová

Abstract:

Particle boards have being used in the civil engineering as a decking for load bearing and non-load bearing vertical walls and horizontal panels (e. g. floors, ceiling, roofs) in a large scale. When the straw is used as non-wood material for manufacturing of lignocellulosic panels, problems with wax layer on the surface of the material can occur. Higher percentage of silica and wax cause the problems with the adhesion of the adhesive and this is the reason why it is necessary to break the surface layer for the better bonding effect. Surface treatment of the particles cause better mechanical properties, physical properties and the overall better results of the composite material are reached. Plasma application is one possibility how to modify the surface layer. The aim of this research is to modify the surface of straw particles by using cold plasma treatment. Surface properties of lignocellulosic materials were observed before and after cold plasma treatment. Cold plasma does not cause any structural changes deeply in the material. There are only changes in surface layers, which are required. Results proved that the plasma application influenced the properties of surface layers and the properties of composite material.

Keywords: composite, lignocellulosic materials, straw, cold plasma, surface treatment

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3268 Simulation for the Magnetized Plasma Compression Study

Authors: Victor V. Kuzenov, Sergei V. Ryzhkov

Abstract:

Ongoing experimental and theoretical studies on magneto-inertial confinement fusion (Angara, C-2, CJS-100, General Fusion, MagLIF, MAGPIE, MC-1, YG-1, Omega) and new constructing facilities (Baikal, C-2W, Z300 and Z800) require adequate modeling and description of the physical processes occurring in high-temperature dense plasma in a strong magnetic field. This paper presents a mathematical model, numerical method, and results of the computer analysis of the compression process and the energy transfer in the target plasma, used in magneto-inertial fusion (MIF). The computer simulation of the compression process of the magnetized target by the high-power laser pulse and the high-speed plasma jets is presented. The characteristic patterns of the two methods of the target compression are being analysed.

Keywords: magnetized target, magneto-inertial fusion, mathematical model, plasma and laser beams

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3267 The Relation Between Protein-Protein and Polysaccharide-Protein Interaction on Aroma Release from Brined Cheese Model

Authors: Mehrnaz Aminifar

Abstract:

The relation between textural parameters and casein network on release of aromatic compounds was investigated over 90-days of ripening. Low DE maltodextrin and WPI were used to modify the textural properties of low fat brined cheese. Hardness, brittleness and compaction of casein network were affected by addition of maltodextrin and WPI. Textural properties and aroma release from cheese texture were affected by interaction of WPI protein-cheese protein and maltodexterin-cheese protein.

Keywords: aroma release, brined cheese, maltodexterin, WPI

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3266 Water-Repellent Finishing on Cotton Fabric by SF₆ Plasma

Authors: We'aam Alali, Ziad Saffour, Saker Saloum

Abstract:

Low-pressure, sulfur hexafluoride (SF₆) remote radio-frequency (RF) plasma, ignited in a hollow cathode discharge (HCD-L300) plasma system, has been shown to be a powerful method in cotton fabric finishing to achieve water-repellent property. This plasma was ignited at an SF6 flow rate of (200 cm), low pressure (0.5 mbar), and radio frequency (13.56 MHz) with a power of (300 W). The contact angle has been measured as a function of the plasma exposure period using the water contact angle measuring device (WCA), and the changes in the morphology, chemical structure, and mechanical properties as tensile strength and elongation at the break of the fabric have also been investigated using the scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy (EDX), attenuated total reflectance Fourier transform Infrared spectroscopy (ATR-FTIR), and tensile test device, respectively. In addition, weight loss of the fabric and the fastness of washing have been studied. It was found that the exposure period of the fabric to the plasma is an important parameter. Moreover, a good water-repellent cotton fabric can be obtained by treating it with SF₆ plasma for a short time (1 min) without degrading its mechanical properties. Regarding the modified morphology of the cotton fabric, it was found that grooves were formed on the surface of the fibers after treatment. Chemically, the fluorine atoms were attached to the surface of the fibers.

Keywords: cotton fabric, SEM, SF₆ plasma, water-repellency

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3265 Enhanced Functional Production of a Crucial Biomolecule Human Serum Albumin in Escherichia coli

Authors: Ashima Sharma

Abstract:

Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

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3264 Amino Acid Profile, Protein Digestibility, Antioxidant and Functional Properties of Protein Concentrate of Local Varieties (Kwandala, Yardass, Jeep, and Jamila) of Rice Brands from Nigeria

Authors: C. E. Chinma, S. O. Azeez, J. C. Anuonye, O. B. Ocheme, C. M. Yakubu, S. James, E. U. Ohuoba, I. A. Baba

Abstract:

There is growing interest in the use of rice bran protein in food formulation due to its hypoallergenic protein, high nutritional value and health promoting potentials. For the first time, the amino acid profile, protein digestibility, antioxidant, and functional properties of protein concentrate from some local varieties of rice bran from Nigeria were studied for possible food applications. Protein concentrates were prepared from rice bran and analysed using standard methods. Results showed that protein content of Kwandala, Yardass, Jeep, and Jamila were 69.24%, 69.97%, 68.73%, and 71.62%, respectively while total essential amino acid were 52.71, 53.03, 51.86, and 55.75g/100g protein, respectively. In vitro protein digestibility of protein concentrate from Kwandala, Yardass, Jeep and Jamila were 90.70%, 91.39%, 90.57% and 91.63% respectively. DPPH radical inhibition of protein from Kwandala, Yardass, Jeep, and Jamila were 48.15%, 48.90%, 47.56%, and 53.29%, respectively while ferric reducing ability power were 0.52, 0.55, 0.47 and 0.67mmol TE per gram, respectively. Protein concentrate from Jamila had higher onset (92.57oC) and denaturation temperature (102.13oC), and enthalpy (0.72J/g) than Jeep (91.46oC, 101.76oC, and 0.68J/g, respectively), Kwandala (90.32oC, 100.54oC and 0.57J/g, respectively), and Yardass (88.94oC, 99.45oC, and 0.51J/g, respectively). In vitro digestibility of protein from Kwandala, Yardas, Jeep, and Jamila were 90.70%, 91.39%, 90.57% and 91.63% respectively. Oil absorption capacity of Kwandala, Yardass, Jeep, and Jamila were 3.61, 3.73, 3.40, and 4.23g oil/g sample respectively, while water absorption capacity were 4.19, 4.32, 3.55 and 4.48g water/g sample, respectively. Protein concentrates had low bulk density (0.37-0.43g/ml). Protein concentrate from Jamila rice bran had the highest foam capacity (37.25%), followed by Yardass (34.20%), Kwandala (30.14%) and Jeep (28.90%). Protein concentrates showed low emulsifying and gelling capacities. In conclusion, protein concentrate prepared from these local rice bran varieties could serve as functional ingredients in food formulations and for enriching low protein foods.

Keywords: rice bran protein, amino acid profile, protein digestibility, antioxidant and functional properties

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3263 Analysis of Formyl Peptide Receptor 1 Protein Value as an Indicator of Neutrophil Chemotaxis Dysfunction in Aggressive Periodontitis

Authors: Prajna Metta, Yanti Rusyanti, Nunung Rusminah, Bremmy Laksono

Abstract:

The decrease of neutrophil chemotaxis function may cause increased susceptibility to aggressive periodontitis (AP). Neutrophil chemotaxis is affected by formyl peptide receptor 1 (FPR1), which when activated will respond to bacterial chemotactic peptide formyl methionyl leusyl phenylalanine (FMLP). FPR1 protein value is decreased in response to a wide number of inflammatory stimuli in AP patients. This study was aimed to assess the alteration of FPR1 protein value in AP patients and if FPR1 protein value could be used as an indicator of neutrophil chemotaxis dysfunction in AP. This is a case control study with 20 AP patients and 20 control subjects. Three milliliters of peripheral blood were drawn and analyzed for FPR1 protein value with ELISA. The data were statistically analyzed with Mann-Whitney test (p>0,05). Results showed that the mean value of FPR1 protein value in AP group is 0,353 pg/mL (0,11 to 1,18 pg/mL) and the mean value of FPR1 protein value in control group is 0,296 pg/mL (0,05 to 0,88 pg/mL). P value 0,787 > 0,05 suggested that there is no significant difference of FPR1 protein value in both groups. The present study suggests that FPR1 protein value has no significance alteration in AP patients and could not be used as an indicator of neutrophil chemotaxis dysfunction.

Keywords: aggressive periodontitis, chemotaxis dysfunction, FPR1 protein value, neutrophil

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3262 Selection of Pichia kudriavzevii Strain for the Production of Single-Cell Protein from Cassava Processing Waste

Authors: Phakamas Rachamontree, Theerawut Phusantisampan, Natthakorn Woravutthikul, Peerapong Pornwongthong, Malinee Sriariyanun

Abstract:

A total of 115 yeast strains isolated from local cassava processing wastes were measured for crude protein content. Among these strains, the strain MSY-2 possessed the highest protein concentration (>3.5 mg protein/mL). By using molecular identification tools, it was identified to be a strain of Pichia kudriavzevii based on similarity of D1/D2 domain of 26S rDNA region. In this study, to optimize the protein production by MSY-2 strain, Response Surface Methodology (RSM) was applied. The tested parameters were the carbon content, nitrogen content, and incubation time. Here, the value of regression coefficient (R2) = 0.7194 could be explained by the model, which is high to support the significance of the model. Under the optimal condition, the protein content was produced up to 3.77 g per L of the culture and MSY-2 strain contain 66.8 g protein per 100 g of cell dry weight. These results revealed the plausibility of applying the novel strain of yeast in single-cell protein production.

Keywords: single cell protein, response surface methodology, yeast, cassava processing waste

Procedia PDF Downloads 403
3261 Effect of Different Irrigation Intervals on Protein and Gel Production of Aloe Vera (Aloe Barbadensis M.) in Iran

Authors: Seyed Mohammad Hosein Al Omrani Nejad, Ali Rezvani Aghdam

Abstract:

This study was done in order to evaluation different irrigation intervals on amount of protein, and gel production in Aloe vera, a traditional medicinal plant. Plants was plnted in Greenhouse and irrigated according to Accumulative Pan Evaporation(APE). The treatments were included 20, 40, 60, 80, 100, 120, 140, 160, 180, and 200 mm APE which has been showed W1,W2, W3, W4, W5, W6, W7, W8,W9 and W10 respectively.The amount of protein and gel production was measured seperately. Results showed that highest protein and fresh weight of gel obtained plants which irrigated W6 and W7 respectively. According to these results can recomend which if plant irrigatedwhen APE reached 120 and 140 mm by Class A Evaporation Pan method gel production and protein would besuitable in north of khozestan province in limited irrigation conditions.

Keywords: irrigation, protein, gel, aloe vera, Iran

Procedia PDF Downloads 389
3260 Role of Biomaterial Surface Nanotopography on Protein Unfolding and Immune Response

Authors: Rahul Madathiparambil Visalakshan, Alex Cavallaro, John Hayball, Krasimir Vasilev

Abstract:

The role of biomaterial surface nanotopograhy on fibrinogen adsorption and unfolding, and the subsequent immune response were studied. Inconsistent topography and varying chemical functionalities along with a lack of reproducibility pose a challenge in determining the specific effects of nanotopography or chemistry on proteins and cells. It is important to have a well-defined nanotopography with a homogeneous chemistry to study the real effect of nanotopography on biological systems. Therefore, we developed a technique that can produce well-defined and highly reproducible topography to identify the role of specific roughness, size, height and density with the presence of homogeneous chemical functionality. Using plasma polymerisation of oxazoline monomers and immobilized gold nanoparticles we created surfaces with an equal number density of nanoparticles of different sizes. This surface was used to study the role of surface nanotopography and the interplay of surface chemistry on proteins and immune cells. The effect of nanotopography on fibrinogen adsorption was investigated using Quartz Cristal Microbalance with Dissipation and micro BCA. The mass of fibrinogen adsorbed on the surface increased with increasing size of nano-topography. Protein structural changes up on adsorption to the nano rough surface was studied using circular dichroism spectroscopy. Fibrinogen unfolding varied depending on the specific nanotopography of the surfaces. It was revealed that the in vitro immune response to the nanotopography surfaces changed due to this protein unfolding.

Keywords: biomaterial inflammation, protein and cell responses, protein unfolding, surface nanotopography

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3259 Bio-Functional Polymeric Protein Based Materials Utilized for Soft Tissue Engineering Application

Authors: Er-Yuan Chuang

Abstract:

Bio-mimetic matters have biological functionalities. This might be valuable in the development of versatile biomaterials. At biological fields, protein-based materials might be components to form a 3D network of extracellular biomolecules, containing growth factors. Also, the protein-based biomaterial provides biochemical and structural assistance of adjacent cells. In this study, we try to prepare protein based biomaterial, which was harvested from living animal. We analyzed it’s chemical, physical and biological property in vitro. Besides, in vivo bio-interaction of the prepared biomimetic matrix was tested in an animal model. The protein-based biomaterial has degradability and biocompatibility. This development could be used for tissue regenerations and be served as platform technologies.

Keywords: protein based, in vitro study, in vivo study, biomaterials

Procedia PDF Downloads 189
3258 Investigation of Fusion Zone Microstructures in Plasma Arc Welding of Austenitic Stainless Steel (SS-304L) with Low Carbon Steel (A-36) with or without Filler Alloy

Authors: Shan-e-Fatima, Mushtaq Khan, Syed Imran Hussian

Abstract:

Plasma arc welding technology is used for welding SS-304L with A-36. Two different optimize butt welded joints were produced by using austenitic filler alloy E-309L and with direct fusion at 45 A, 2mm/sec by keeping plasma gas flow rate at 0.5LPM. Microstructure analysis of the weld bead was carried out. The results reveal complex heterogeneous microstructure in austenitic base filler alloy sample where as full martensite was found in directly fused sample.

Keywords: fusion zone microstructure, stainless steel, low carbon steel, plasma arc welding

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3257 A Study on the Etching Characteristics of High aspect ratio Oxide Etching Using C4F6 Plasma in Inductively Coupled Plasma with Low Frequency Bias

Authors: ByungJun Woo

Abstract:

In this study, high-aspect-ratio (HAR) oxide etching characteristics in inductively coupled plasma were investigated using low frequency (2 MHz) bias power with C4F6 gas. An experiment was conducted using CF4/C4F6/He as the mixed gas. A 100 nm (etch area)/500 nm (mask area) line patterns were used, and the etch cross-section and etch selectivity of the amorphous carbon layer thin film were derived using a scanning electron microscope. Ion density was extracted using a double Langmuir probe, and CFx and F neutral species were observed via optical emission spectroscopy. Based on these results, the possibility for HAR oxide etching using C4F6 gas chemistry was suggested in this work. These etching results also indicate that the use of C4F6 gas can significantly contribute to the development of next-generation HAR oxide etching.

Keywords: plasma, etching, C4F6, high aspect ratio, inductively coupled plasma

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3256 Reforming of CO₂-Containing Natural Gas by Using an AC Gliding Arc Discharge Plasma System

Authors: Krittiya Pornmai, Sumaeth Chavadej

Abstract:

The increasing in global energy demand has affected the climate change caused by the generation of greenhouse gases. Therefore, the objective of this work was to investigate a direct production of synthesis gas from a CO₂-containing natural gas by using gliding arc discharge plasma technology. In this research, the effects of steam reforming, combined steam reforming and partial oxidation, and using multistage gliding arc discharge system on the process performance have been discussed. The simulated natural gas used in this study contains 70% methane, 5% ethane, 5% propane, and 20% carbon dioxide. In comparison with different plasma reforming processes (under their optimum conditions), the steam reforming provides the highest H₂ selectivity resulting from the cracking reaction of steam. In addition, the combined steam reforming and partial oxidation process gives a very high CO production implying that the addition of both oxygen and steam can offer the acceptably highest synthesis gas production. The stage number of plasma reactor plays an important role in the improvement of CO₂ conversion. Moreover, 3 stage number of plasma reactor is considered as an optimum stage number for the reforming of CO₂-containing natural gas with steam and partial oxidation in term of providing low energy consumption as compared with other plasma reforming processes.

Keywords: natural gas, reforming process, gliding arc discharge, plasma technology

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3255 Prospects of Milk Protein as a Potential Alternative of Natural Antibiotic

Authors: Syeda Fahria Hoque Mimmi

Abstract:

Many new and promising treatments for reducing or diminishing the adverse effects of microorganisms are being discovered day by day. On the other hand, the dairy industry is accelerating the economic wheel of Bangladesh. Considering all these facts, new thoughts were developed to isolate milk proteins by the present experiment for opening up a new era of developing natural antibiotics from milk. Lactoferrin, an iron-binding glycoprotein with multifunctional properties, is crucial to strengthening the immune system and also useful for commercial applications. The protein’s iron-binding capacity makes it undoubtedly advantageous to immune system modulation and different bacterial strains. For fulfilling the purpose, 4 of raw and 17 of commercially available milk samples were collected from different farms and stores in Bangladesh (Dhaka, Chittagong, and Cox’s Bazar). Protein quantification by nanodrop technology has confirmed that raw milk samples have better quantities of protein than the commercial ones. All the samples were tested for their antimicrobial activity against 18 pathogens, where raw milk samples showed a higher percentage of antibacterial activity. In addition to this, SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis) was performed to identify lactoferrin in the milk samples. Lactoferrin was detected in 9 samples from which 4 were raw milk samples. Interestingly, Streptococcus pyogenes, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, Vibrio cholera, Staphylococcus aureus, and enterotoxigenic E. coli significantly displayed sensitivity against lactoferrin collected from raw milk. Only Bacillus cereus, Pseudomonas aeruginosa, Streptococcus pneumonia, Enterococcus faecalis, and ETEC (Enterotoxigenic Escherichia coli) were susceptible to lactoferrin obtained from a commercial one. This study suggested that lactoferrin might be used as the potential alternative of antibiotics for many diseases and also can be used to reduce microbial deterioration in the food and feed industry.

Keywords: alternative of antibiotics, commercially available milk, lactoferrin, nanodrop technology, pathogens, raw milk

Procedia PDF Downloads 180