scholarly journals Salting-Out Approach Is Worthy of Comparison with Ultracentrifugation for Extracellular Vesicle Isolation from Tumor and Healthy Models

Biomolecules ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1857
Author(s):  
Simona Serratì ◽  
Antonio Palazzo ◽  
Annamaria Lapenna ◽  
Helena Mateos ◽  
Antonia Mallardi ◽  
...  
Keyword(s):  
Culture Media ◽  
Human Cancer ◽  
Wide Spectrum ◽  
Good Alternative ◽  
Extracellular Vesicle ◽  
Performance Ratio ◽  
Salting Out ◽  
Good Efficiency ◽  
Protein Depletion ◽  
Isolation Techniques

The role of extracellular vesicles (EVs) has been completely re-evaluated in the recent decades, and EVs are currently considered to be among the main players in intercellular communication. Beyond their functional aspects, there is strong interest in the development of faster and less expensive isolation protocols that are as reliable for post-isolation characterisations as already-established methods. Therefore, the identification of easy and accessible EV isolation techniques with a low price/performance ratio is of paramount importance. We isolated EVs from a wide spectrum of samples of biological and clinical interest by choosing two isolation techniques, based on their wide use and affordability: ultracentrifugation and salting-out. We collected EVs from human cancer and healthy cell culture media, yeast, bacteria and Drosophila culture media and human fluids (plasma, urine and saliva). The size distribution and concentration of EVs were measured by nanoparticle tracking analysis and dynamic light scattering, and protein depletion was measured by a colorimetric nanoplasmonic assay. Finally, the EVs were characterised by flow cytometry. Our results showed that the salting-out method had a good efficiency in EV separation and was more efficient in protein depletion than ultracentrifugation. Thus, salting-out may represent a good alternative to ultracentrifugation.

Supporting Distance Users of Mobile Learning Technology

2011 ◽  
pp. 248-261
Author(s):  
Yong Liu ◽  
Hongxiu Li
Keyword(s):  
Mobile Learning ◽  
Social Transformation ◽  
Wide Spectrum ◽  
Good Alternative ◽  
Traditional Education ◽  
Learning Technology ◽  
Distance Learner ◽  
Mobile Learning Technology ◽  
Demographic Shifts ◽  
Rapid Deployment

With the rapid deployment of mobile devices, mobile learning emerges as a promising approach giving rise to a wide spectrum of new education possibilities. It serves as an effective conduit to deliver education to civilians of all social-economic levels, in particular the learners previously unreachable from traditional education systems, such as problem teenagers, social employees, and ageing people. Hence, unlike traditional education approaches, it is considered to be a good alternative to deal with the challenges posed by demographic shifts and social transformation. The purpose of this chapter is to: (i) identify the theoretical and technological underpinnings for delivering mobile learning to the distance learner, and (ii) discuss the possible learner communities that can benefit from mobile learning technology, with regard to their unique learning requirements and features.

scholarly journals Methodological Approaches to Study Extracellular Vesicle miRNAs in Epstein–Barr Virus-Associated Cancers

2018 ◽  
Vol 19 (9) ◽  
pp. 2810 ◽  
Author(s):  
Li Sun ◽  
David Meckes
Keyword(s):  
Epstein Barr Virus ◽  
Biomarker Discovery ◽  
Immune Escape ◽  
Human Cancer ◽  
Biological Fluids ◽  
Extracellular Vesicle ◽  
Cellular Mirnas ◽  
Barr Virus ◽  
Diagnosis And Prognosis ◽  
Epstein Barr

Epstein Barr-virus (EBV) was the first virus identified to be associated with human cancer in 1964 and is found ubiquitously throughout the world’s population. It is now established that EBV contributes to the development and progression of multiple human cancers of both lymphoid and epithelial cell origins. EBV encoded miRNAs play an important role in tumor proliferation, angiogenesis, immune escape, tissue invasion, and metastasis. Recently, EBV miRNAs have been found to be released from infected cancer cells in extracellular vesicles (EVs) and regulate gene expression in neighboring uninfected cells present in the tumor microenvironment and possibly at distal sites. As EVs are abundant in many biological fluids, the viral and cellular miRNAs present within EBV-modified EVs may serve as noninvasion markers for cancer diagnosis and prognosis. In this review, we discuss recent advances in EV isolation and miRNA detection, and provide a complete workflow for EV purification from plasma and deep-sequencing for biomarker discovery.

scholarly journals Bladder Cancer in the Genomic Era

2019 ◽  
Vol 143 (6) ◽  
pp. 695-704 ◽  
Author(s):  
Charles C. Guo ◽  
Bogdan Czerniak
Keyword(s):  
Bladder Cancer ◽  
Molecular Mechanisms ◽  
Complex Disease ◽  
Human Cancer ◽  
Wide Spectrum ◽  
Clinicopathologic Features ◽  
Pathologic Features ◽  
Genomic Characterization

Context.— Bladder cancer is a heterogeneous disease that exhibits a wide spectrum of clinical and pathologic features. The classification of bladder cancer has been traditionally based on morphologic assessment with the aid of immunohistochemistry. However, recent genomic studies have revealed that distinct alterations of DNA and RNA in bladder cancer may underlie its diverse clinicopathologic features, leading to a novel molecular classification of this common human cancer. Objective.— To update recent developments in genomic characterization of bladder cancer, which may shed insights on the molecular mechanisms underlying the origin of bladder cancer, dual-track oncogenic pathways, intrinsic molecular subtyping, and development of histologic variants. Data Sources.— Peer-reviewed literature retrieved from PubMed search and authors' own research. Conclusions.— Bladder cancer is likely to arise from different uroprogenitor cells through papillary/luminal and nonpapillary/basal tracks. The intrinsic molecular subtypes of bladder cancer referred to as luminal and basal exhibit distinct expression signatures, clinicopathologic features, and sensitivities to standard chemotherapy. Genomic characterization of bladder cancer provides new insights to understanding the biological nature of this complex disease, which may lead to more effective treatment.

scholarly journals Biochemical Evaluation and Green Synthesis of Nano Silver Using Peroxidase fromEuphorbia(Euphorbia amygdaloides) and Its Antibacterial Activity

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Semra Cicek ◽  
Azize Alaylı Gungor ◽  
Ahmet Adiguzel ◽  
Hayrunnisa Nadaroglu
Keyword(s):  
Silver Nanoparticles ◽  
Antibacterial Activity ◽  
Green Synthesis ◽  
Yersinia Pseudotuberculosis ◽  
Wide Spectrum ◽  
Good Alternative ◽  
Uv Spectrum ◽  
Simple Method ◽  
Time Period ◽  
Peroxidase Enzyme

Silver nanoparticles are used an increased attention for various biomedical and medical applications. In this study, green synthesis of silver nanoparticles was made with simple method by using peroxidase enzyme partially purified fromEuphorbia(Euphorbia amygdaloides) plant. Optimum pH, temperature and time period were determined to obtain silver nanoparticles using the peroxidase enzyme. The result shows that higher silver nanoparticle was synthesized for 4 hours and at 20°C and pH 8. Also, optimal concentration of metal ions was found as 0.5 mM. The synthesized silver nanoparticles were characterized by UV spectrum, scanning electron microscope (SEM) and X-ray diffraction. Antibacterial activity of silver nanoparticles was measured against some microorganisms such asSerratia marcescens, Yersinia pseudotuberculosis, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Listeria monocytogenes, and Escherichia coli. Synthesized silver nanoparticles have wide spectrum antibacterial activity in low concentration and may be a good alternative therapeutic approach in medicine and pharmaceutical fields in future.

scholarly journals LN-6: a monoclonal antibody to vimentin expressed in non-hematopoietic mesenchymal cells and derived tumors and reactive in B5-fixed, paraffin-embedded tissues.

1989 ◽  
Vol 37 (9) ◽  
pp. 1363-1370 ◽  
Author(s):  
E Stathopoulos ◽  
G S Naeve ◽  
C R Taylor ◽  
A L Epstein
Keyword(s):  
Monoclonal Antibody ◽  
Human Cancer ◽  
Wide Spectrum ◽  
Mesenchymal Cells ◽  
Lymphoid Cells ◽  
Normal Cells ◽  
Immunohistochemical Diagnosis ◽  
Pathological Tissues

We generated a monoclonal antibody (MAb), designated LN-6, directed against human vimentin, which retains its immunoreactivity in B5-fixed, paraffin-embedded tissues. Like other anti-vimentin MAb, LN-6 was found to be reactive with a wide spectrum of human sarcomas and normal cells of mesenchymal derivation. However, unlike other similar reagents, LN-6 was unreactive with normal and malignant human lymphoid cells and therefore displays a more restricted immunoreactivity. Because of its ability to stain routinely processed pathological tissues and its marked reactivity with human sarcomas, LN-6 is a unique reagent for the immunohistochemical diagnosis of human cancer.

scholarly journals Serum extracellular vesicle depletion processes affect release and infectivity of HIV-1 in culture

2016 ◽  
Author(s):  
Zhaohao Liao ◽  
Dillon C. Muth ◽  
Erez Eitan ◽  
Meghan Travers ◽  
Elin Lehrmann ◽  
...  
Keyword(s):  
Culture Media ◽  
Virus Production ◽  
Cell Types ◽  
Extracellular Vesicle ◽  
Substantial Effect ◽  
Primary Cells ◽  
Antiviral Responses ◽  
Health And Disease ◽  
Infected Cells ◽  
Hiv 1

ABSTRACTExtracellular vesicles (EVs, including exosomes and microvesicles) are involved in intercellular communication in health and disease and affect processes including immune and antiviral responses. Ultracentrifuged serum is depleted of EVs and, when used in culture media, reduces growth and viability of some cell types. In this study, we examined the effects of serum EV depletion processes on HIV-1 replication in primary cells and cell lines, including two HIV-1 latency models. Increased HIV-1 production was observed in certain EV-depleted conditions, along with cell morphology changes and decreased cell viability. Add-back of ultracentrifuge pellets rescued baseline HIV-1 production. Primary cells appeared to be less sensitive to EV depletion. ACH-2 and U1 latency models produced more HIV-1 under EV-depleted conditions, while virus produced under processed serum conditions was more infectious. Finally, changes in cellular metabolism and gene expression were associated with EV-depleted culture. In conclusion, the EV environment of HIV-1 infected cells has a substantial effect on virus production and infectivity. EV-dependence of cell cultures should be examined carefully along with other experimental variables. However, EVs may not be the only particles depleted by ultracentrifugation or other processes. Effects of EVs may be accompanied by or confused with those of closely associated or physically similar particles.

scholarly journals Brevinin-1GHd: a novel Hylarana guentheri skin secretion-derived Brevinin-1 type peptide with antimicrobial and anticancer therapeutic potential

2020 ◽  
Vol 40 (5) ◽  
Author(s):  
Yangyang Jiang ◽  
Yue Wu ◽  
Tao Wang ◽  
Xiaoling Chen ◽  
Mei Zhou ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Human Cancer ◽  
Wide Spectrum ◽  
Antimicrobial Properties ◽  
Skin Secretion ◽  
Anticancer Activities ◽  
Peptide Analog ◽  
Wide Range ◽  
Low Cytotoxicity ◽  
Short Time

Abstract Host-defense antimicrobial peptides (AMPs) from amphibians are usually considered as one of the most promising next-generation antibiotics because of their excellent antimicrobial properties and low cytotoxicity. In the present study, one novel Brevinin-1 type peptide, Brevinin-1GHd, was isolated and characterized from the skin secretion of the frog, Hylarana guentheri. Brevinin-1GHd was found to possess a wide range of antimicrobial activity through penetrating the bacterial membrane within a short time while showing low hemolysis at bactericidal concentrations, even against the resistant strains. It also inhibited and eradicated biofilms that are thought to be closely related to the rise in resistance. Meanwhile, Brevinin-1GHd exhibited wide-spectrum anti-proliferation activity toward human cancer lines. Taken together, these results indicate that Brevinin-1GHd with its excellent antimicrobial and anticancer activities is a promising candidate for a novel antibiotic agent, and study of its structure–activity relationships also provided a rational template for further research and peptide analog design.

scholarly journals Extracellular Vesicle Integrins Distinguish Unique Cancers

2019 ◽  
Author(s):  
Stephanie N. Hurwitz ◽  
David G. Meckes Jr.
Keyword(s):  
Human Cancer ◽  
Tumor Stage ◽  
Extracellular Vesicle ◽  
Preliminary Evidence ◽  
Integrin Receptors ◽  
Primary Tumors ◽  
Human Cancer Cell Lines ◽  
Tumor Growth And Metastasis ◽  
Metastatic Sites ◽  
Cell Expression

The proteomic profile of extracellular vesicles (EVs) has been of increasing interest, particularly in understanding cancer growth, drug resistance, and metastatic behavior. Emerging data suggests that cancer-derived EVs may carry an array of oncogenic cargo, including certain integrin proteins that may, in turn, promote cell detachment, migration, and selection of future metastatic sites. We previously reported a large comparison of secreted vesicle protein cargo across sixty diverse human cancer cell lines. Here, we analyze the distinct integrin profiles of these cancer EVs. We further demonstrate the enrichment of integrin receptors in breast cancer EVs compared to vesicles secreted from benign breast epithelial cells. Total EV integrin levels, including the quantity of integrins α2, αv, β4, and β5 correlate with breast tumor stage. In particular, integrin α2 also largely reflects progenitor cell expression, highlighting the utility of this integrin protein as a potential circulating biomarker of primary tumors. This study provides preliminary evidence of the value of vesicle-associated integrin proteins in cancer diagnosis and prediction of tumor stage. Differential expression of integrins across cancer cells, and selective packaging of integrins into EVs may contribute to further understanding the development and progression of tumor growth and metastasis across a variety of cancer types.

scholarly journals CRISPR screens in physiologic medium reveal conditionally essential genes in human cells

2020 ◽  
Author(s):  
Nicholas J. Rossiter ◽  
Kimberly S. Huggler ◽  
Charles H. Adelmann ◽  
Heather R. Keys ◽  
Ross W. Soens ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Culture Media ◽  
Human Cancer ◽  
Cancer Cell Lines ◽  
Medium Composition ◽  
Human Cells ◽  
Genetic Screens ◽  
Gene Essentiality ◽  
Human Cancer Cell Lines

SUMMARYForward genetic screens across hundreds of diverse cancer cell lines have started to define the genetic dependencies of proliferating human cells and how these vary by genotype and lineage. Most screens, however, have been carried out in culture media that poorly resemble metabolite availability in human blood. To explore how medium composition influences gene essentiality, we performed CRISPR-based screens of human cancer cell lines cultured in traditional versus human plasma-like medium (HPLM). Sets of medium-dependent fitness genes span several cellular processes and can vary with both natural cell-intrinsic diversity and the specific combination of basal and serum components that comprise typical culture media. Notably, we traced the causes for each of three conditional growth phenotypes to the availability of metabolites uniquely defined in HPLM versus traditional media. Our findings reveal the profound impact of medium composition on gene essentiality in human cells, and also suggest general strategies for using genetic screens in HPLM to uncover new cancer vulnerabilities and gene-nutrient interactions.

Synthesis and Characterization of L-Lysin Coated Iron Oxide Nanoparticles as Appropriate Choices for Cell Immobilization and Magnetic Separation

2019 ◽  
Vol 9 (4) ◽  
pp. 462-466 ◽  
Author(s):  
Mohammad Javad Raee ◽  
Alireza Ebrahiminezhad ◽  
Mohammad Bagher Ghoshoon ◽  
Ahmad Gholami ◽  
Younes Ghasemi
Keyword(s):  
Iron Oxide ◽  
Magnetic Separation ◽  
Iron Oxide Nanoparticles ◽  
Cell Separation ◽  
Culture Media ◽  
Cell Immobilization ◽  
Good Alternative ◽  
Oxide Nanoparticles ◽  
Bacterial Cells ◽  
Separation Processes

Introduction:Cell separation is one of the important steps of purification in downstream processes. Some separation techniques such as centrifugation and filtration are expensive and would affect cell viability. Magnetic separation can be a good alternative for laboratory and industrial cell separation processes.Methods:For this purpose, L-lysine coated Iron Oxide Nanoparticles (IONs) were synthesized and used for magnetic separation of Escherichia coli as the most applied microbial cell in biotechnological processes.Results:IONs have successfully decorated the bacterial cells and cells were completely separated by applying an external magnetic field.Conclusion:This study showed that coating of E. coli cells with IONs could help to isolate cells from culture media without using expensive instruments.

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