scholarly journals Isolation and characterization of extracellular vesicles in Candida albicans

2020 ◽  
pp. 99-108
Author(s):  
Virginia Pérez-Doñate ◽  
Facundo Pérez-Giménez ◽  
Lucas Del Castillo Agudo ◽  
Juan Alberto Castillo-Garit ◽  
Mar Soria-Merino ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Fungal Infections ◽  
Protein Composition ◽  
Yeast Cultures ◽  
Isolation And Characterization ◽  
Fungal Organisms ◽  
Systemic Infections ◽  
Molecular Components

Background: The occurrence of systemic infections due to C. albicans has increased especially in critically ill patients. In fungal infections, secretory mechanisms are key events for disease establishment. Recent findings demonstrate that fungal organisms release many molecular components to the extracellular space in extracellular vesicles.Aims: We develop a method to obtain exosomes from yeast cultures of the Candida albicans.Methods: Yeast strains used in this work were C. albicans SC5314, C. parapsilosis (ATCC 22019) and C. krusei (ATCC 6258). Yeasts were grown at 37.º in liquid YPD medium. The cell cultures were centrifuged and the supernatant filtered through sterile nitrocellulose. Filtrates were concentrated and centrifuged using an ultracentrifuge. The sediment was analyzed by electron microscopy of transmission.Results: The transmission of electron microscopy and nanoparticle tracking analysis confirmed the presence of extracellular vesicles (exosomes) of sizes between 100 and 200 nm and the absence of cellular contaminants. This was ratified by the characterization of proteins performed through the western blot technique, where the absence of cell contamination in the preparations was assessed.Conclusions: The method proves to be highly effective due to the homogeneity and purity of the obtained microvesicles. The protocol developed in this paper proves to be effective for obtaining exosomes of other Candida species, which will allow future studies to determine its protein composition and the role that these vesicles can play.

scholarly journals 109 Isolation and characterization of myometrium and decidual cell-derived extracellular vesicles

2021 ◽  
Vol 224 (2) ◽  
pp. S75-S76
Author(s):  
Megan Shepherd ◽  
Enkhtuya Radnaa ◽  
Rheanna Urrabaz-Garza ◽  
Talar Kechichian ◽  
Ourlad Alzeus G. Tantengco ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Decidual Cell ◽  
Isolation And Characterization

scholarly journals Emerging techniques in the isolation and characterization of extracellular vesicles and their roles in cancer diagnostics and prognostics

The Analyst ◽  
2016 ◽  
Vol 141 (2) ◽  
pp. 371-381 ◽  
Author(s):  
Vijaya Sunkara ◽  
Hyun-Kyung Woo ◽  
Yoon-Kyoung Cho
Keyword(s):  
Extracellular Vesicles ◽  
Cancer Diagnostics ◽  
Clinical Implementation ◽  
Characterization Methods ◽  
Cancer Management ◽  
Isolation And Characterization

We present an overview of current isolation, detection, and characterization methods of extracellular vesicles and their applications and limitations as a potential emerging biomarker in cancer management and their clinical implementation.

Isolation and Characterization of Extracellular Vesicles in Stem Cell-Related Studies

2017 ◽  
pp. 205-223
Author(s):  
Zezhou Zhao ◽  
Dillon C. Muth ◽  
Vasiliki Mahairaki ◽  
Linzhao Cheng ◽  
Kenneth W. Witwer
Keyword(s):  
Stem Cell ◽  
Extracellular Vesicles ◽  
Isolation And Characterization

scholarly journals Isolation and characterization of platelet-derived extracellular vesicles

2014 ◽  
Vol 3 (1) ◽  
pp. 24692 ◽  
Author(s):  
Maria T. Aatonen ◽  
Tiina Öhman ◽  
Tuula A. Nyman ◽  
Saara Laitinen ◽  
Mikaela Grönholm ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Isolation And Characterization

scholarly journals Isolation and Characterization of Cellulose Nanofibers fromGigantochloa scortechiniias a Reinforcement Material

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Chaturbhuj K. Saurabh ◽  
Asniza Mustapha ◽  
M. Mohd. Masri ◽  
A. F. Owolabi ◽  
M. I. Syakir ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cellulose Nanofibers ◽  
X Ray Diffraction ◽  
X Ray ◽  
Isolation And Characterization ◽  
Transmission Electron ◽  
Reinforcement Material ◽  
Bamboo Fibers ◽  
Ft Ir

Cellulose nanofibers (CNF) were isolated fromGigantochloa scortechiniibamboo fibers using sulphuric acid hydrolysis. This method was compared with pulping and bleaching process for bamboo fiber. Scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and thermogravimetric analysis were used to determine the properties of CNF. Structural analysis by FT-IR showed that lignin and hemicelluloses were effectively removed from pulp, bleached fibers, and CNF. It was found that CNF exhibited uniform and smooth morphological structures, with fiber diameter ranges from 5 to 10 nm. The percentage of crystallinity was significantly increased from raw fibers to cellulose nanofibers, microfibrillated, along with significant improvement in thermal stability. Further, obtained CNF were used as reinforcement material in epoxy based nanocomposites where tensile strength, flexural strength, and modulus of nanocomposites improved with the addition of CNF loading concentration ranges from 0 to 0.7%.

Isolation and characterization of the peptidoglycans from selected Gram-positive and Gram-negative periodontal pathogens

1985 ◽  
Vol 31 (2) ◽  
pp. 154-160 ◽  
Author(s):  
M. R. Barnard ◽  
S. C. Holt
Keyword(s):  
Electron Microscopy ◽  
Diaminopimelic Acid ◽  
Muramic Acid ◽  
Periodontal Pathogens ◽  
Gram Positive ◽  
Gram Negative ◽  
Isolation And Characterization ◽  
Bacteroides Gingivalis ◽  
Scanning Electron

The peptidoglycans from several Gram-negative and Gram-positive periodontal pathogens were isolated, purified, and characterized both morphologically and chemically. In addition, the effects of the mureolytic enzymes, lysozyme, M-1 N-acetyl-muramidase, and the AM-3 endopeptidase, on the peptidoglycans were examined. These enzymes were found to be highly effective in the degradation of the purified peptidoglycans; however, a Bacteroides capillus peptidoglycan–protein complex exhibited a greater resistance to these enzymes. Morphologically, the peptidoglycans consisted of large saccular sheets which, when viewed by scanning electron microscopy, contained numerous holes and tears. Chemically, the peptidoglycans consisted of muramic acid, glucosamine, alanine, glutamic acid, and meso-diaminopimelic acid (DAP). One Bacteroides species, Bacteroides gingivalis strain W, contained glycine and LL-DAP, suggestive of an indirectly cross-linked A3γ peptidoglycan.

scholarly journals Genome sequence and characterization of five bacteriophages infecting Streptomyces coelicolor and Streptomyces venezuelae: Alderaan, Coruscant, Dagobah, Endor1 and Endor2

2020 ◽  
Author(s):  
Aël Hardy ◽  
Vikas Sharma ◽  
Larissa Kever ◽  
Julia Frunzke
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Streptomyces Coelicolor ◽  
Morphological Differentiation ◽  
Viral Diversity ◽  
Isolation And Characterization ◽  
Phage Evolution ◽  
Transmission Electron ◽  
Family Based

AbstractStreptomyces are well-known antibiotic producers, and are also characterized by a complex morphological differentiation. Streptomyces, like all bacteria, are confronted with the constant threat of phage predation, which in turn shapes bacterial evolution. However, despite significant sequencing efforts recently, relatively few phages infecting Streptomyces have been characterized compared to other genera. Here, we present the isolation and characterization of five novel Streptomyces phages. All five phages belong to the Siphoviridae family, based on their morphology as determined by transmission electron microscopy. Genome sequencing revealed that four of them were temperate phages, while one had a lytic lifestyle. Moreover, one of the newly sequenced phages shows very little homology to already described phages, highlighting the still largely untapped viral diversity. Altogether, this study expands the number of characterized phages of Streptomyces and sheds light on phage evolution and phage-host dynamics in Streptomyces.

scholarly journals Platelet-Derived Extracellular Vesicles for Regenerative Medicine

2021 ◽  
Author(s):  
Miquel Antich-Rosselló ◽  
Maria Antònia Forteza-Genestra ◽  
Marta Monjo ◽  
Joana Maria Ramis
Keyword(s):  
Wound Healing ◽  
Regenerative Medicine ◽  
Extracellular Vesicles ◽  
Cellular Differentiation ◽  
Neural Regeneration ◽  
Isolation And Characterization ◽  
Biomedical Field ◽  
New Treatments ◽  
Different Sources

Extracellular vesicles (EVs) present a great potential for the development of new treatments in the biomedical field. To be used as therapeutics, many different sources have been used for EVs obtention, while only few studies have addressed the use of platelet derived EVs (pEVs). In fact, pEVs have been shown to intervene in different healing responses, thus some studies have evaluated their regenerative capability in wound healing or hemorrhagic shock. Even more, pEVs have proven to induce cellular differentiation, enhancing musculoskeletal or neural regeneration. However, the obtention and characterization of pEVs is widely heterogeneous and differs from the recommendations of the International Society for Extracellular Vesicles. Therefore, in this review, we aim to present the main advances in the therapeutical use of pEVs in the regenerative medicine field while highlighting the isolation and characterization steps followed. The main goal of this review is to portray the studies performed in order to enhance the translation of the pEVs research into feasible therapeutical applications.

scholarly journals Orally Administered 5-aminolevulinic Acid for Isolation and Characterization of Circulating Tumor-Derived Extracellular Vesicles in Glioblastoma Patients

Cancers ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3297
Author(s):  
Sybren L. N. Maas ◽  
Thomas S. van Solinge ◽  
Rosalie Schnoor ◽  
Anudeep Yekula ◽  
Joeky T. Senders ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Aminolevulinic Acid ◽  
Culture Media ◽  
Protoporphyrin Ix ◽  
Liquid Biopsies ◽  
Guided Surgery ◽  
Tumor Visualization ◽  
Isolation And Characterization ◽  
Droplet Pcr

Background: In glioblastoma (GB), tissue is required for accurate diagnosis and subtyping. Tissue can be obtained through resection or (stereotactic) biopsy, but these invasive procedures provide risks for patients. Extracellular vesicles (EVs) are small, cell-derived vesicles that contain miRNAs, proteins, and lipids, and possible candidates for liquid biopsies. GB-derived EVs can be found in the blood of patients, but it is difficult to distinguish them from circulating non-tumor EVs. 5-aminolevulinic acid (5-ALA) is orally administered to GB patients to facilitate tumor visualization and maximal resection, as it is metabolized to fluorescent protoporphyrin IX (PpIX) that accumulates in glioma cells. In this study, we assessed whether PpIX accumulates in GB-derived EVs and whether these EVs could be isolated and characterized to enable a liquid biopsy in GB. Methods: EVs were isolated from the conditioned media of U87 cells treated with 5-ALA by differential ultracentrifugation. Blood samples were collected and processed from healthy controls and patients undergoing 5-ALA guided surgery for GB. High-resolution flow cytometry (hFC) enabled detection and sorting of PpIX-positive EVs, which were subsequently analyzed by digital droplet PCR (ddPCR). Results: PpIX-positive EVs could be detected in conditioned cell culture media as well as in patient samples after administration of 5-ALA. By using hFC, we could sort the PpIX-positive EVs for further analysis with ddPCR, which indicated the presence of EVs and GB-associated miRNAs. Conclusion: GB-derived EVs can be isolated from the plasma of GB patients by using 5-ALA induced fluorescence. Although many challenges remain, our findings show new possibilities for the development of blood-based liquid biopsies in GB patients.

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