scholarly journals Extraction and quality evaluation of exosomes from joint effusion

2020 ◽  
Author(s):  
Anmin Ruan ◽  
Yueshan Yin ◽  
Pu Chen ◽  
Jun Zhou ◽  
Xiaozhe Zhang ◽  
...  
Keyword(s):  
Knee Joint ◽  
Particle Morphology ◽  
Extraction Methods ◽  
Joint Effusion ◽  
Precipitation Method ◽  
Transmission Electron ◽  
Marker Proteins ◽  
Exclusion Chromatography ◽  
Granule Surface ◽  
Peg Precipitation Method

Abstract Objective: The aim of this study was to compare the quality and efficiency of exosomes extracted from knee joint effusion by different methods, laying a foundation for further research on exosomes from knee joint effusion. Methods:To separate and extract joint exosomes by 8% polyethylene glycol (PEG) precipitation method, ultracentrifugation method (UC) and ultrafiltration with exclusion chromatography (SECF). Transmission electron microscopy (TEM) and nanoparticle tracing technology (NTA) were used to detect particle morphology and particle size, and Western Blot (WB) was used to detect marker proteins of granule surface (CD9, CD63, Flotillin-1 and calnexin).Results: Three methods separated vesicle-like round particles from joint effusion successfully. The results of TEM show that the particles obtained by the three extraction methods are round or oval vesicles, the plasma membrane is intact, the size is different, and the diameter is distributed between 30 and 150 nm. Compared with SECF group, PEG group had more background particle impurities. and the broken vesicle fragments can be seen in the UC group; The results of NTA show that the main peaks of the three groups of particles are between 100-120nm, and the particle concentration is greater than 1×1010/ml; The results of WB show that the protein expressions of CD9, CD63 and Flotillin-1 in the suspension extracted by the three methods were higher, the expression of calnexin protein in the PEG group was higher than the UC group and SECF group.Conclusion: The three extraction methods can extract the exosomes of joint effusion successfully. The quality of exosomes obtained by the SECF method is relatively high, while the PEG precipitation method contains a small amount of impurity particles. UC method does not guarantee the integrity of exosomes. In summary, we recommend the SECF method to extract and isolate joint effusion exudates-derived exosomes when further studying the role of exosomes in the pathogenesis of knee osteoarthritis.

scholarly journals Extraction And Quality Evaluation of Exosomes From Joint Effusion

2020 ◽  
Author(s):  
Anmin Ruan ◽  
Pu Chen ◽  
Jun Zhou ◽  
Xiaozhe Zhang ◽  
Yufeng Ma ◽  
...  
Keyword(s):  
Knee Joint ◽  
Particle Morphology ◽  
Extraction Methods ◽  
Joint Effusion ◽  
Precipitation Method ◽  
Transmission Electron ◽  
Marker Proteins ◽  
Exclusion Chromatography ◽  
Peg Precipitation Method

Abstract Objective: The aim of this study was to compare the quality and efficiency of exosomes extracted from knee joint effusion by different methods, laying a foundation for further research on exosomes from knee joint effusion. Methods: Separate and extract joint exosomes by 8% polyethylene glycol (PEG) precipitation method, ultracentrifugation method (UC) and ultrafiltration with exclusion chromatography (SECF). Transmission electron microscopy (TEM) and nanoparticle tracing technology (NTA) were used to detect particle morphology and particle size, and Western Blot (WB) was used to detect granule protein surface marker proteins (CD9, CD63, Flotillin-1 and calnexin).Results: All three methods successfully separated vesicle-like round particles from joint effusion. TEM results show that the particles obtained by the three extraction methods are round or oval vesicles, the plasma membrane is intact, the size is different, and the diameter is distributed between 30 and 150 nm. Compared with SECF group, PEG group had more background particle impurities. and the broken vesicle fragments can be seen in the UC group; NTA results show that the main peaks of the three groups of particles are between 100-120nm, and the particle concentration is greater than 1×1010/ml; WB results show that the expressions of CD9, CD63 and Flotillin-1 protein in the suspension extracted by the three methods were all higher, the expression of calnexin protein was higher in the PEG group than the UC group and SECF group.Conclusion: The three extraction methods can successfully extract the exosomes of joint effusion. The quality of exosomes obtained by the SECF method is relatively high, while the PEG precipitation method contains a small amount of impurity particles. UC method does not guarantee the integrity of exosomes. In summary, when further studying the role of exosomes in the pathogenesis of knee osteoarthritis, we recommend the SECF method to extract and isolate joint exudates.

scholarly journals Extraction And Quality Evaluation of Exosomes From Joint Effusion

2020 ◽  
Author(s):  
Anmin Ruan ◽  
Yueshan Yin ◽  
Pu Chen ◽  
Jun Zhou ◽  
Xiaozhe Zhang ◽  
...  
Keyword(s):  
Knee Joint ◽  
Particle Morphology ◽  
Extraction Methods ◽  
Joint Effusion ◽  
Precipitation Method ◽  
Transmission Electron ◽  
Marker Proteins ◽  
Exclusion Chromatography ◽  
Peg Precipitation Method

Abstract Objective: The aim of this study was to compare the quality and efficiency of exosomes extracted from knee joint effusion by different methods, laying a foundation for further research on exosomes from knee joint effusion. Methods: Separate and extract joint exosomes by 8% polyethylene glycol (PEG) precipitation method, ultracentrifugation method (UC) and ultrafiltration with exclusion chromatography (SECF). Transmission electron microscopy (TEM) and nanoparticle tracing technology (NTA) were used to detect particle morphology and particle size, and Western Blot (WB) was used to detect granule protein surface marker proteins (CD9, CD63, Flotillin-1 and calnexin).Results: All three methods successfully separated vesicle-like round particles from joint effusion. TEM results show that the particles obtained by the three extraction methods are round or oval vesicles, the plasma membrane is intact, the size is different, and the diameter is distributed between 30 and 150 nm. Compared with SECF group, PEG group had more background particle impurities. and the broken vesicle fragments can be seen in the UC group; NTA results show that the main peaks of the three groups of particles are between 100-120nm, and the particle concentration is greater than 1×1010/ml; WB results show that the expressions of CD9, CD63 and Flotillin-1 protein in the suspension extracted by the three methods were all higher, the expression of calnexin protein was higher in the PEG group than the UC group and SECF group.Conclusion: The three extraction methods can successfully extract the exosomes of joint effusion. The quality of exosomes obtained by the SECF method is relatively high, while the PEG precipitation method contains a small amount of impurity particles. UC method does not guarantee the integrity of exosomes. In summary, when further studying the role of exosomes in the pathogenesis of knee osteoarthritis, we recommend the SECF method to extract and isolate joint exudates.

Cryomicroscopy of multiphase latices

1991 ◽  
Vol 49 ◽  
pp. 1060-1061
Author(s):  
O. L. Shaffer ◽  
M.S. El-Aasser ◽  
C. L. Zhao ◽  
M. A. Winnik ◽  
R. R. Shivers
Keyword(s):  
Electron Microscopy ◽  
Radiation Damage ◽  
Freeze Fracture ◽  
Particle Morphology ◽  
Second Stage ◽  
Transmission Electron ◽  
Important Approach ◽  
Carbon Coated ◽  
Preparation Techniques

Transmission electron microscopy is an important approach to the characterization of the morphology of multiphase latices. Various sample preparation techniques have been applied to multiphase latices such as OsO4, RuO4 and CsOH stains to distinguish the polymer phases or domains. Radiation damage by an electron beam of latices imbedded in ice has also been used as a technique to study particle morphology. Further studies have been developed in the use of freeze-fracture and the effect of differential radiation damage at liquid nitrogen temperatures of the latex particles embedded in ice and not embedded.Two different series of two-stage latices were prepared with (1) a poly(methyl methacrylate) (PMMA) seed and poly(styrene) (PS) second stage; (2) a PS seed and PMMA second stage. Both series have varying amounts of second-stage monomer which was added to the seed latex semicontinuously. A drop of diluted latex was placed on a 200-mesh Formvar-carbon coated copper grid.

Magnetic and dielectric properties of BiFeO3 nanoparticles

2015 ◽  
Vol 7 (2) ◽  
pp. 1393-1403
Author(s):  
Dr R.P VIJAYALAKSHMI ◽  
N. Manjula ◽  
S. Ramu ◽  
Amaranatha Reddy
Keyword(s):  
Diffuse Reflectance Spectrum ◽  
Precipitation Method ◽  
Secondary Phases ◽  
X Ray Diffraction ◽  
Bifeo3 Nanoparticles ◽  
Transmission Electron ◽  
Multiferroic Bifeo3 ◽  
Pure Phase ◽  
Simple Chemical ◽  
Co Precipitation

Single crystalline nano-sized multiferroic BiFeO3 (BFO) powders were synthesized through simple chemical co-precipitation method using polyethylene glycol (PEG) as capping agent. We obtained pure phase BiFeO3 powder by controlling pHand calcination temperature. From X-ray diffraction studies the nanoparticles were unambiguously identified to have a rhombohedrally distorted perovskite structure belonging to the space group of R3c. No secondary phases were detected. It indicates single phase structure. EDX spectra indicated the appearance of three elements Bi, Fe, O in 1:1:3. From the UV-Vis diffuse reflectance spectrum, the absorption cut-off wavelength of the BFO sample is around 558nm corresponding to the energy band gap of 2.2 eV. The size (60-70 nm) and morphology of the nanoparticles have been analyzed using transmission electron microscopy (TEM).   Linear M−H behaviour and slight hysteresis at lower magnetic field is observed for BiFeO3 nanoparticles from Vibrating sample magnetometer studies. It indicates weak ferromagnetic behaviour at room temperature. From dielectric studies, the conductivity value is calculated from the relation s = L/RbA Sm-1 and it is around 7.2 x 10-9 S/m.

Effect of synthesis and activation methods on the character of CoMo/ultrastable Y-zeolite catalysts

2021 ◽  
Vol 19 (1) ◽  
pp. 745-754
Author(s):  
Khoirina Dwi Nugrahaningtyas ◽  
Eddy Heraldy ◽  
Rachmadani ◽  
Yuniawan Hidayat ◽  
Indriana Kartini
Keyword(s):  
Electron Microscopy ◽  
Reduction Process ◽  
Particle Morphology ◽  
Zeolite Catalysts ◽  
X Ray Diffraction ◽  
Y Zeolite ◽  
X Ray ◽  
Irregular Particle ◽  
Transmission Electron ◽  
Pure Metals

Abstract The properties of three types of CoMo/USY catalysts with different synthesized methods have been studied. The sequential and co-impregnation methods followed by activation using calcination and reduction process have been conducted. The properties of the catalysts were examined using Fourier-transform-infrared (FTIR) spectroscopy, X-ray diffraction (XRD) with refinement, and surface area analyzer (SAA). The FTIR spectrum study revealed the enhanced intensity of its Bronsted acid site, and the XRD diffractogram pattern verified the composition of pure metals, oxides, and alloys in the catalyst. The SAA demonstrated the mesoporous features of the catalyst. Scanning electron microscopy showed an irregular particle morphology. Additional analysis using the transmission electron microscopy indicated that the metal has successfully impregnated without damaging the USY structure.

scholarly journals A Bacterially-Expressed Recombinant Envelope Protein from Usutu Virus Induces Neutralizing Antibodies in Rabbits

Vaccines ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 157
Author(s):  
Kinga Böszörményi ◽  
Janet Hirsch ◽  
Gwendoline Kiemenyi Kayere ◽  
Zahra Fagrouch ◽  
Nicole Heijmans ◽  
...  
Keyword(s):  
Envelope Protein ◽  
Neutralizing Antibodies ◽  
Size Exclusion ◽  
Vitro Assay ◽  
Usutu Virus ◽  
Transmission Electron ◽  
Exclusion Chromatography ◽  
Efficacious Vaccine ◽  
Human Infections

Background: Recently, an emerging flavivirus, Usutu virus (USUV), has caused an epidemic among birds in Europe, resulting in a massive die-off in Eurasian blackbirds. Currently found only in Europe and Africa, it can be envisioned that Usutu virus will follow the path of other flaviviruses, like West Nile virus and Zika virus, and will spread via its mosquito vectors and bird hosts to other parts of the world. Several cases of human infections by Usutu virus have already been published. Anticipating this spread, development of an efficacious vaccine would be highly desirable. Method: This study describes the production in E. coli, purification, and refolding of a partial USUV envelope protein. Prior to immunization, the protein was characterized using size exclusion chromatography, transmission electron microscopy and dynamic light scattering, showing the limited presence of virus-like structures, indicating that the protein solution is probably a mixture of mono and multimeric envelope proteins. Results: Immunizations of two rabbits with the refolded E-protein fraction, mixed with a strong adjuvant, resulted in the generation of neutralizing antibodies, as evidenced in an in vitro assay. Discussion: The way forward towards a subunit vaccine against Usutu virus infection is discussed.

scholarly journals An optimized and robust PEG precipitation method for detection of SARS-CoV-2 in wastewater

2021 ◽  
Vol 785 ◽  
pp. 147270
Author(s):  
Sylvia A. Sapula ◽  
Jonathan J. Whittall ◽  
Aaron J. Pandopulos ◽  
Cobus Gerber ◽  
Henrietta Venter
Keyword(s):  
Precipitation Method ◽  
Peg Precipitation Method

scholarly journals Electro-Oxidation of Ammonia at Novel Ag2O−PrO2/γ-Al2O3 Catalysts

Coatings ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 257
Author(s):  
Mariam Khan ◽  
Naveed Kausar Janjua ◽  
Safia Khan ◽  
Ibrahim Qazi ◽  
Shafaqat Ali ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Electrochemical Impedance ◽  
Precipitation Method ◽  
Impregnation Method ◽  
X Rays ◽  
Kinetic And Thermodynamic Parameters ◽  
Catalytic Support ◽  
Transmission Electron ◽  
Electro Oxidation ◽  
Oxidation Of Ammonia

An Ag2O(x)−PrO2(y)/γ-Al2O3 electrocatalyst series (X:Y is for Ag:Pr from 0 to 10) was synthesized, to use synthesized samples in electrochemical applications, a step in fuel cells advancements. Ag2O(x)−PrO2(y)/γ-Al2O3/Glassy-Carbon was investigated for electrochemical oxidation of ammonia in alkaline medium and proved to be highly effective, having high potential utility, as compared to commonly used Pt-based electrocatalysts. In this study, gamma alumina as catalytic support was synthesized via precipitation method, and stoichiometric wt/wt.% compositions of Ag2O−PrO2 were loaded on γ-Al2O3 by co-impregnation method. The desired phase of γ-Al2O3 and supported nanocatalysts was obtained after heat treatment at 800 and 600 °C, respectively. The successful loadings of Ag2O−PrO2 nanocatalysts on surface of γ-Al2O3 was determined by X-rays diffraction (XRD), Fourier-transform Infrared Spectroscopy (FTIR), and energy dispersive analysis (EDX). The nano-sized domain of the sample powders sustained with particle sizes was calculated via XRD and scanning electron microscopy (SEM). The surface morphology and elemental compositions were examined by SEM, transmission electron microscopy (TEM) and EDX. The conductive and electron-transferring nature was investigated by cyclic voltammetry and electrochemical impedance (EIS). Cyclic voltammetric profiles were observed, and respective kinetic and thermodynamic parameters were calculated, which showed that these synthesized materials are potential catalysts for ammonia electro-oxidation. Ag2O(6)−PrO2(4)/γ-Al2O3 proved to be the most proficient catalyst among all the members of the series, having greater diffusion coefficient, heterogeneous rate constant and lesser Gibbs free energy for this system. The catalytic activity of these electrocatalysts is revealed from electrochemical studies which reflected their potentiality as electrode material in direct ammonia fuel cell technology for energy production.

scholarly journals Phycocyanin content and nutritional profile of Arthrospira platensis from Mexico: efficient extraction process and stability evaluation of phycocyanin

BMC Chemistry ◽  
2021 ◽  
Vol 15 (1) ◽  
Author(s):  
Sanghamitra Khandual ◽  
Edgar Omar Lopez Sanchez ◽  
Hugo Espinosa Andrews ◽  
Jose Daniel Padilla de la Rosa
Keyword(s):  
Storage Temperature ◽  
Extraction Process ◽  
Arthrospira Platensis ◽  
Extraction Methods ◽  
Precipitation Method ◽  
Partial Purification ◽  
Natural Food ◽  
Solvent Ratio ◽  
C Storage ◽  
Dry Biomass

AbstractPhycocyanin is a blue natural food colorant with multiple health benefits. Here we propose an efficient phycocyanin extraction method from Arthrospira platensis from Mexico. Three extraction methods were applied to optimize the extraction process, using water and buffer as solvents, with three pH values at two agitation times. The highest phycocyanin, 54.65 mg/g, was extracted from dry biomass with water as a solvent using an ultrasonication bar. The optimum condition of extraction was determined to be 1:50 biomass/solvent ratio for dry biomass, with the freeze/thaw method for 20 min repeated twice, and then agitated at 120 rpm for 24 h. The phycocyanin content was 48.88 mg/g biomass, with a purity of 0.47. For scalable phycocyanin productivity, the sonication method is recommended as there is no statistical difference. The phycocyanin stability was best at − 20 °C storage temperature at pH 7 for 35 days. Partial purification with ammonium sulfate was found to be suitable as a fractional precipitation method, first at 0–20% and then 20–65%, to get purity nearly 1. Total protein was found to be 55.52%, and total amino acids after phycocyanin extraction was 33%. The maximum phycocyanin yield using water as a solvent was the most interesting result regardless of the method used for extraction.

Effectiveness of Calcium Deficiency in Nanosized Hydroxyapatite for Removal of Fe(II), Cu(II), Ni(II) and Cr(VI) Ions from Aqueous Solutions

2019 ◽  
Vol 56 ◽  
pp. 17-27
Author(s):  
Van Dat Doan ◽  
Van Thuan Le ◽  
Hoang Sinh Le ◽  
Dinh Hien Ta ◽  
Hoai Thuong Nguyen
Keyword(s):  
Electron Microscopy ◽  
Ion Exchange ◽  
Aqueous Solutions ◽  
Calcium Deficiency ◽  
Exchange Mechanism ◽  
Precipitation Method ◽  
X Ray ◽  
Transmission Electron ◽  
Bet Method ◽  
Langmuir Isotherm Model

In this work, nanosized calcium deficient hydroxyapatite (nCDHA) was synthesized by the precipitation method, and then utilized as an adsorbent for removal of Fe (II), Cu (II), Ni (II) and Cr (VI) ions from aqueous solutions after characterizing it by various techniques as scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDX) and BET method. A possible structure of synthesized nCDHA was proposed. The adsorption study indicated that the adsorption equilibrium is well fitted with Langmuir isotherm model with the maximum adsorption capacities followed the order of Fe (II) > Cu (II) > Ni (II) > Cr (VI) with the values of 137.23, 128.02, 83.19 and 2.92 mg/g, respectively. The ion-exchange mechanism was dominant for the adsorption of metal ions onto nCDHA at initial metal concentrations lower than 0.01 mol/L. Along with the ion-exchange mechanism, there was an additional precipitation occurred on the surface of nCDHA in the case of Fe (II) and Cu (II) at initial concentrations higher than 0.01 mol/L.

Sign in / Sign up

Export Citation Format

Share Document