scholarly journals Extracellular Vesicles Analysis in the COVID-19 Era: Insights on Serum Inactivation Protocols towards Downstream Isolation and Analysis

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 544
Author(s):  
Roberto Frigerio ◽  
Angelo Musicò ◽  
Marco Brucale ◽  
Andrea Ridolfi ◽  
Silvia Galbiati ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Analytical Techniques ◽  
Heat Inactivation ◽  
Force Microscopy ◽  
Atomic Force ◽  
Droplet Pcr ◽  
Detergent Treatment ◽  
Routine Procedures ◽  
The Impact ◽  
Mirna Content

Since the outbreak of the COVID-19 crisis, the handling of biological samples from confirmed or suspected SARS-CoV-2-positive individuals demanded the use of inactivation protocols to ensure laboratory operators’ safety. While not standardized, these practices can be roughly divided into two categories, namely heat inactivation and solvent-detergent treatments. These routine procedures should also apply to samples intended for Extracellular Vesicles (EVs) analysis. Assessing the impact of virus-inactivating pre-treatments is therefore of pivotal importance, given the well-known variability introduced by different pre-analytical steps on downstream EVs isolation and analysis. Arguably, shared guidelines on inactivation protocols tailored to best address EVs-specific requirements will be needed among the analytical community, yet deep investigations in this direction have not yet been reported. We here provide insights into SARS-CoV-2 inactivation practices to be adopted prior to serum EVs analysis by comparing solvent/detergent treatment vs. heat inactivation. Our analysis entails the evaluation of EVs recovery and purity along with biochemical, biophysical and biomolecular profiling by means of a set of complementary analytical techniques: Nanoparticle Tracking Analysis, Western Blotting, Atomic Force Microscopy, miRNA content (digital droplet PCR) and tetraspanin assessment by microarrays. Our data suggest an increase in ultracentrifugation (UC) recovery following heat treatment; however, it is accompanied by a marked enrichment in EVs-associated contaminants. On the other hand, solvent/detergent treatment is promising for small EVs (<150 nm range), yet a depletion of larger vesicular entities was detected. This work represents a first step towards the identification of optimal serum inactivation protocols targeted to EVs analysis.

scholarly journals Extracellular Vesicles Analysis in the COVID-19 Era: Insights on Serum Inactivation Protocols Towards Downstream Isolation and Analysis

2020 ◽  
Author(s):  
Roberto Frigerio ◽  
Angelo Musicò ◽  
Marco Brucale ◽  
Andrea Ridolfi ◽  
Silvia Galbiati ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Heat Inactivation ◽  
Force Microscopy ◽  
Atomic Force ◽  
Droplet Pcr ◽  
Detergent Treatment ◽  
Relevant Topic ◽  
Routine Procedures ◽  
The Impact ◽  
Mirna Content

AbstractSince the outbreak of COVID-19 crisis, the handling of biological samples from confirmed or suspected SARS-CoV-2 positive individuals demanded the use of inactivation protocols to ensure laboratory operators safety. While not standardized, these practices can be roughly divided in two categories, namely heat inactivation and solvent-detergent treatments. As such, these routine procedures should also apply to samples intended for Extracellular Vesicles (EVs) analysis. Assessing the impact of virus inactivating pre-treatments is therefore of pivotal importance, given the well-known variability introduced by different pre-analytical steps on downstream EVs isolation and analysis. Arguably, shared guidelines on inactivation protocols tailored to best address EVs-specific requirements will be needed among the EVs community, yet deep investigations in this direction haven’t been reported so far.In the attempt of sparking interest on this highly relevant topic, we here provide preliminary insights on SARS-CoV-2 inactivation practices to be adopted prior serum EVs analysis by comparing solvent/detergent treatment vs. heat inactivation. Our analysis entailed the evaluation of EVs recovery and purity along with biochemical, biophysical and biomolecular profiling by means of Nanoparticle Tracking Analysis, Western Blotting, Atomic Force Microscopy, miRNA content (digital droplet PCR) and tetraspanin assessment by microarrays. Our data suggest an increase in ultracentrifugation (UC) recovery following heat-treatment, however accompanied by a marked enrichment in EVs-associated contaminants. On the contrary, solvent/detergent treatment is promising for small EVs (< 150 nm range), yet a depletion of larger vesicular entities was detected. This work represents a first step towards the identification of optimal serum inactivation protocols targeted to EVs analysis.

Study of NSCLC cell migration promoted by NSCLC-Derived Extracellular Vesicle using atomic force microscopy

2021 ◽  
Author(s):  
Shuwei Wang ◽  
Jiajia Wang ◽  
Tuoyu Ju ◽  
Kaige Qu ◽  
Fan Yang ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Cell Migration ◽  
Cancer Cells ◽  
Extracellular Vesicles ◽  
Extracellular Vesicle ◽  
Cancer Microenvironment ◽  
Biological Processes ◽  
Force Microscopy ◽  
Atomic Force ◽  
Cellular Components

Extracellular Vesicles (EVs) secreted by cancer cells have a key role in the cancer microenvironment and progression. Previous studies have mainly focused on molecular functions, cellular components and biological processes...

Structuring of Silicon Wafer Surfaces on the Sub-100 nm Scale by Hydrogen Plasma Treatments

2003 ◽  
Vol 788 ◽  
Author(s):  
R. Job ◽  
Y. Ma ◽  
A. G. Ulyashin
Keyword(s):  
Hydrogen Plasma ◽  
Structural Defects ◽  
Process Conditions ◽  
Force Microscopy ◽  
Czochralski Silicon ◽  
Atomic Force ◽  
Nano Structures ◽  
Plasma Treatments ◽  
The Impact ◽  
Wafer Surfaces

ABSTRACTHydrogen plasma treatments applied on standard Czochralski silicon (Cz Si) wafers cause a structuring of the surface regions on the sub-100 nm scale, i.e. a thin ‘nano-structured’ Si layer is created up to a depth of ∼ 150 nm. The formation of the ‘nano-structures’ and their evolution in dependence on the process conditions was studied. The impact of post-hydrogenation annealing on the morphology of the structural defects was studied up to 1200 °C. The H-plasma treated and annealed samples were analyzed at surface and sub-surface regions by scanning electron microscopy (SEM), atomic force microscopy (AFM), and μ-Raman spectroscopy.

scholarly journals Small Extracellular Vesicles Isolated from Serum May Serve as Signal-Enhancers for the Monitoring of CNS Tumors

2020 ◽  
Vol 21 (15) ◽  
pp. 5359 ◽  
Author(s):  
Gabriella Dobra ◽  
Matyas Bukva ◽  
Zoltan Szabo ◽  
Bella Bruszel ◽  
Maria Harmati ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Serum Proteins ◽  
Lumbar Disc ◽  
Principal Component ◽  
Cns Tumors ◽  
Serum Samples ◽  
Force Microscopy ◽  
Isolation And Characterization ◽  
Atomic Force ◽  
Patient Groups

Liquid biopsy-based methods to test biomarkers (e.g., serum proteins and extracellular vesicles) may help to monitor brain tumors. In this proteomics-based study, we aimed to identify a characteristic protein fingerprint associated with central nervous system (CNS) tumors. Overall, 96 human serum samples were obtained from four patient groups, namely glioblastoma multiforme (GBM), non-small-cell lung cancer brain metastasis (BM), meningioma (M) and lumbar disc hernia patients (CTRL). After the isolation and characterization of small extracellular vesicles (sEVs) by nanoparticle tracking analysis (NTA) and atomic force microscopy (AFM), liquid chromatography -mass spectrometry (LC-MS) was performed on two different sample types (whole serum and serum sEVs). Statistical analyses (ratio, Cohen’s d, receiver operating characteristic; ROC) were carried out to compare patient groups. To recognize differences between the two sample types, pairwise comparisons (Welch’s test) and ingenuity pathway analysis (IPA) were performed. According to our knowledge, this is the first study that compares the proteome of whole serum and serum-derived sEVs. From the 311 proteins identified, 10 whole serum proteins and 17 sEV proteins showed the highest intergroup differences. Sixty-five proteins were significantly enriched in sEV samples, while 129 proteins were significantly depleted compared to whole serum. Based on principal component analysis (PCA) analyses, sEVs are more suitable to discriminate between the patient groups. Our results support that sEVs have greater potential to monitor CNS tumors, than whole serum.

scholarly journals Synthesis of Metarhizium anisopliae–Chitosan Nanoparticles and Their Pathogenicity against Plutella xylostella (Linnaeus)

2021 ◽  
Vol 10 (1) ◽  
pp. 1
Author(s):  
Jianhui Wu ◽  
Cailian Du ◽  
Jieming Zhang ◽  
Bo Yang ◽  
Andrew G. S. Cuthbertson ◽  
...  
Keyword(s):  
Plutella Xylostella ◽  
Metarhizium Anisopliae ◽  
Chitosan Nanoparticles ◽  
Analytical Techniques ◽  
X Ray ◽  
Synthesis Of Nanoparticles ◽  
Force Microscopy ◽  
Atomic Force ◽  
Afm Analysis

Nanotechnology is increasingly being used in areas of pesticide production and pest management. This study reports the isolation and virulence of a new Metarhizium anisopliae isolate SM036, along with the synthesis and characterization of M. anisopliae–chitosan nanoparticles followed by studies on the efficacy of nanoparticles against Plutella xylostella. The newly identified strain proved pathogenic to P. xylostella under laboratory conditions. The characterization of M. anisopliae–chitosan nanoparticles through different analytical techniques showed the successful synthesis of nanoparticles. SEM and HRTEM images confirmed the synthesis of spherical-shaped nanoparticles; X-ray diffractogram showed strong peaks between 2θ values of 16–30°; and atomic force microscopy (AFM) analysis revealed a particle size of 75.83 nm for M. anisopliae–chitosan nanoparticles, respectively. The bioassay studies demonstrated that different concentrations of M. anisopliae–chitosan nanoparticles were highly effective against second instar P. xylostella under laboratory and semi-field conditions. These findings suggest that M. anisopliae–chitosan nanoparticles can potentially be used in biorational P. xylostella management programs.

scholarly journals Deswelling Induced Morphological Changes in Dual pH and Temperature Responsive Ultra-Low Crosslinked Poly (N-Isopropyl Acrylamide)-Co-Acrylic Acid Microgels

2018 ◽  
Author(s):  
Molla Islam ◽  
Maddie Tumbarello ◽  
Andrew Lyon
Keyword(s):  
Light Scattering ◽  
Acrylic Acid ◽  
Morphological Changes ◽  
Scattering Data ◽  
Ionization State ◽  
Force Microscopy ◽  
Temperature Responsive ◽  
Atomic Force ◽  
Isopropyl Acrylamide ◽  
The Impact

<div>We demonstrated the deswelling induced morphological change in dual pH and Temperature responsive ultra-low crosslinked Poly (N-isopropyl acrylamide)-co-acrylic acid microgels. The responsivity with pH and temperature were studied by light scattering and atomic force microscopy. Light scattering data suggest that at pH 4.5 the microgels undergo multiple transitions associated with collapse of pNIPAm-rich segments and repulsion between the AAc-rich segments. The evolution of punctate structures around the periphery or throughout the whole microgels at pH 4.5 and 6.5 respectively was revealed by AFM, further illustrating the heterogeneous deswelling present in the ionized copolymer microgels.</div><div>The impact of this study and understanding how ionization state of copolymer dictates the overall structural properties of microgels will widen our understanding for their applications in biotechnology</div><div><b><br></b></div>

scholarly journals Effects of suspended micro- and nanoscale particles on zooplankton functional diversity of drainage system reservoirs at an open-pit mine

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Anna Maria Goździejewska ◽  
Monika Gwoździk ◽  
Sławomir Kulesza ◽  
Mirosław Bramowicz ◽  
Jacek Koszałka
Keyword(s):  
Functional Diversity ◽  
Drainage System ◽  
Open Pit ◽  
Nanoscale Particles ◽  
X Ray ◽  
Force Microscopy ◽  
Suspension Parameters ◽  
Atomic Force ◽  
Size Of Particles ◽  
The Impact

Abstract Water from mining drainage is turbid because of suspensions. We tested the hypothesis that the chemical composition as well as shape and size of particles in suspensions of natural origin affect the density and functional diversity of zooplankton. The suspensions were analyzed with atomic force microscopy (AFM), energy dispersive X-ray spectroscopy (EDS), scanning electron microscopy (SEM), and optical microscopy. Elements found in the beidellite clays were also identified in the mineral structure of the particles. As the size of the microparticles decreased, the weight proportions of phosphorus, sulfur, and chlorine increased in the suspensions. These conditions facilitated the biomass growth of large and small microphages and raptorials. As the size of the nanoparticles decreased, the shares of silicon, aluminum, iron, and magnesium increased. These conditions inhibited raptorials the most. Ecosystem functionality was the highest with intermediate suspension parameters, which were at the lower range of the microphase and the upper range of the nanophase. The functional traits of zooplankton demonstrate their potential for use as sensitive indicators of disruptions in aquatic ecosystems that are linked with the presence of suspensions, and they facilitate gaining an understanding of the causes and scales of the impact of suspensions.

scholarly journals Natural and Activated Allophane Catalytic Activity Based on the Microactivity Test in Astm Norm 3907/D3907M-2019

2020 ◽  
Vol 10 (9) ◽  
pp. 3035
Author(s):  
Edward Henry Jiménez Calderón ◽  
Ana Emperatriz Paucar Tipantuña ◽  
Paulina Fernanda Herrera Mullo ◽  
Daniel Alejandro Hidalgo Cháfuel ◽  
Washington Ruiz ◽  
...  
Keyword(s):  
Analytical Techniques ◽  
Bet Surface Area ◽  
Navier Stokes ◽  
Maximum Temperature ◽  
X Ray Diffraction ◽  
X Ray ◽  
Force Microscopy ◽  
Catalytic Activation ◽  
Atomic Force ◽  
Activating Agent

The optimal conditions of the catalytic activation of allophane were evaluated for possible use as a catalyst within a fluidized bed catalytic cracking unit (FCC). The physicochemical properties of natural allophane and activated allophane were studied by using an alkaline activating agent, followed by a hydrothermal treatment. For the characterization, analytical techniques were used: Fourier transform infrared spectroscopy, particle size, (BET) surface area, thermogravimetry (TGA), X-ray diffraction (XRD), chemisorption, X-ray fluorescence (XRF), atomic force microscopy (AFM), and chromatography. The catalytic evaluation was determined by the (MAT) micro activity test equipment constructed according to ASTM D-3907/D3907M-2019. In addition, the Navier–Stokes 3D equations (nonlinear partial derivatives) were studied, which allow studying molecular dynamics contributing substantively to chemical kinetics describing the process of decomposition of crude oil in thermal cracking, determining the maximum temperature at which it retains its properties through the action of heat.

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