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tissue extract extracellular vesicles

Birke J Benedikter, Freek G Bouwman, Alexandra C A Heinzmann, Tanja Vajen, Edwin C Mariman, Emiel F M Wouters, Paul H M Savelkoul, Rory R Koenen, Gernot G U Rohde, Rene van Oerle, Henri M Spronk, Frank R M Stassen
Airway epithelial cells secrete extracellular vesicles (EVs) under basal conditions and when exposed to cigarette smoke extract (CSE). Getting insights into the composition of these EVs will help unravel their functions in homeostasis and smoking-induced pathology. Here, we characterized the proteomic composition of basal and CSE-induced airway epithelial EVs. BEAS-2B cells were left unexposed or exposed to 1% CSE for 24 h, followed by EV isolation using ultrafiltration and size exclusion chromatography. Isolated EVs were labelled with tandem mass tags and their proteomic composition was determined using nano-LC-MS/MS...
2019: Journal of Extracellular Vesicles
Qinyue Chen, Yiting Chen, Yali Sun, Wenxiu He, Xiaoli Han, Enhao Lu, Xianyi Sha
Breast cancer is a severe threat to the health of women, and the metastasis of tumor cells leads to high mortality in female patients. Evidence shows that leukocytes are recruited by breast tumors through adhesion to inflammatory endothelial cells as well as tumor cells. Moreover, it is known that Pluronic P123 is effective in the reduction of matrix metalloproteinases (MMPs), which play a key role in the degradation of the extracellular matrix (ECM), therefore helping tumor cells to escape from the primary site...
March 8, 2019: Nanoscale
Stephanie N Hurwitz, James M Olcese, David G Meckes
Circulating and interstitial small membrane-bound extracellular vesicles (EVs) represent promising targets for the development of novel diagnostic or prognostic biomarker assays, and likely serve as important players in the progression of a vast spectrum of diseases. Current research is focused on the characterization of vesicles secreted from multiple cell and tissue types in order to better understand the role of EVs in the pathogenesis of conditions including neurodegeneration, inflammation, and cancer. However, globally consistent and reproducible techniques to isolate and purify vesicles remain in progress...
February 7, 2019: Journal of Visualized Experiments: JoVE
YueTong Chen, Liang Liu, Jie Li, Yu Du, Jing Wang, JiangHui Liu
OBJECTIVE: To investigate the relationship between the expression of linc-VLDLR in extracellular vesicles (EVs) and esophageal carcinomas development and drug resistance. METHODS: The expression of linc-VLDLR and ABCG2 mRNA in 60 cases of esophageal carcinoma tissue, para-carcinoma tissue and the normal esophagus tissue were detected using Fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR). Fifty percent inhibiting concentration (IC50) of adriamycin (ADM) to Eca109 cells was detected by MTT assay, after the treatment of different concentrations of adriamycin (ADM) on esophageal squamous cell carcinoma Eca109 cell line for 24 h...
December 31, 2018: Pathology, Research and Practice
Inbar Shlomovitz, Ziv Erlich, Mary Speir, Sefi Zargarian, Noam Baram, Maya Engler, Liat Edry-Botzer, Ariel Munitz, Ben A Croker, Motti Gerlic
Interleukin-33 (IL-33) is a pro-inflammatory cytokine that plays a significant role in inflammatory diseases by activating immune cells to induce type 2 immune responses upon its release. Although IL-33 is known to be released during tissue damage, its exact release mechanism is not yet fully understood. Previously, we have shown that cleaved IL-33 can be detected in the plasma and epithelium of Ripk1-/- neonates, which succumb to systemic inflammation driven by spontaneous receptor-interacting protein kinase-3 (RIPK3)-dependent necroptotic cell death, shortly after birth...
December 21, 2018: FEBS Journal
Jong Sik Lee, Jae Young Hur, In Ae Kim, Hee Joung Kim, Chang Min Choi, Jae Chol Lee, Wan Seop Kim, Kye Young Lee
BACKGROUND: EGFR genotyping in pulmonary adenocarcinoma patients who develop pleural effusions is mostly performed using cytology or cell block slides with low sensitivity. Liquid biopsy using the supernatant of pleural effusions may be more effective because they contain many components released by cancer cells. Extracellular vesicles (EVs) are known to carry oncogenic double-stranded DNA that is considered a notable biomarker. Here, we investigate the efficiency of liquid biopsy using cell-free DNA (cfDNA) and extracellular vesicle-derived DNA (EV-derived DNA) from the supernatant of pleural effusions for EGFR genotyping in patients with pulmonary adenocarcinoma...
December 10, 2018: BMC Cancer
Kentaro Jingushi, Motohide Uemura, Kosuke Nakano, Yujiro Hayashi, Cong Wang, Yu Ishizuya, Yoshiyuki Yamamoto, Takuji Hayashi, Toshiro Kinouchi, Kyosuke Matsuzaki, Taigo Kato, Atsunari Kawashima, Takeshi Ujike, Akira Nagahara, Kazutoshi Fujita, Koji Ueda, Kazutake Tsujikawa, Norio Nonomura
Renal cell carcinoma (RCC) is the most common type of kidney cancer in adults, responsible for approximately 90‑95% of cases. We previously reported a novel method that enables direct extraction of extracellular vesicles (EVs) from surgically resected viable tissues, yielding what we term tissue‑exudative extracellular vesicles (Te‑EVs). Quantitative LC/MS analysis identified 3,871 proteins in Te‑EVs, among which leukocyte‑associated immunoglobulin‑like receptor 1 (LAIR1) was highly enriched in tumor Te‑EVs...
November 20, 2018: Oncology Reports
A Safwat, D Sabry, A Ragiae, E Amer, R H Mahmoud, R M Shamardan
This study aimed to evaluate the effect of mesenchymal stem cells (MSCs)-derived exosomes in retina regeneration of experimentally induced diabetes mellitus (DM) in a rabbit model. Exosomes are extracellular vesicles that contain many microRNAs (micRNAs), mRNAs, and proteins from their cells of origin. DM was induced by intravenous (IV) injection of streptozotocin in rabbits. MSCs were isolated from adipose tissue of rabbits. Exosomes were extracted from MSCs by ultracentrifugation. Exosomes were injected by different routes (IV, subconjunctival (SC), and intraocular (IO))...
January 2018: Journal of Circulating Biomarkers
Lisa N van der Vorm, Joost E I G Brouwers, Ceráya Mondria, Bas de Laat, Philip G de Groot, Jasper A Remijn
Background: Upon tooth extraction, extravascular tissue factor (TF) initiates coagulation to arrest bleeding. Additionally, saliva is in constant contact with the wound and contains extracellular vesicle-derived procoagulant TF. Since the duration of postextraction bleeding is highly variable between patients, we hypothesized this may be caused by variation in saliva-derived TF-induced clotting activity. Objectives: We aimed to assess the variability of saliva-induced thrombin generation (TG) in healthy individuals...
October 2018: Research and Practice in Thrombosis and Haemostasis
Sippy Kaur, Ahmed G Abu-Shahba, Riku O Paananen, Heidi Hongisto, Hanna Hiidenmaa, Heli Skottman, Riitta Seppänen-Kaijansinkko, Bettina Mannerström
Extracellular vesicles (EVs) are reported to be involved in stem cell maintenance, self-renewal, and differentiation. Due to their bioactive cargoes influencing cell fate and function, interest in EVs in regenerative medicine has rapidly increased. EV-derived small non-coding RNA mimic the functions of the parent stem cells, regulating the maintenance and differentiation of stem cells, controlling the intercellular regulation of gene expression, and eventually affecting the cell fate. In this study, we used RNA sequencing to provide a comprehensive overview of the expression profiles of small non-coding transcripts carried by the EVs derived from human adipose tissue stromal/stem cells (AT-MSCs) and human pluripotent stem cells (hPSCs), both human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSC)...
October 19, 2018: Scientific Reports
Wanessa Scopel, Fernando Luis Cônsoli
Symbionts are widely distributed in eukaryotes, and potentially affect the physiology, ecology and evolution of their host. Most insects harbour free-living bacteria in their haemocoel and gut lumen, intracellular-living bacteria in a range of tissues or bacteria in host-derived specialized cells. Stinkbugs, as do many arthropods, harbour extracellular bacteria in the gut that may affect the fitness of their host. This study identified the culturable symbionts associated with the ovaries, spermatheca, seminal vesicle and posterior midgut region (V4) of males and females of Euschistus heros (F...
July 17, 2018: Antonie Van Leeuwenhoek
Jaganmohan R Jangamreddy, Michel K C Haagdorens, M Mirazul Islam, Philip Lewis, Ayan Samanta, Per Fagerholm, Aneta Liszka, Monika K Ljunggren, Oleksiy Buznyk, Emilio I Alarcon, Nadia Zakaria, Keith M Meek, May Griffith
Short collagen-like peptides (CLPs) are being proposed as alternatives to full-length collagen for use in tissue engineering, on their own as soft hydrogels, or conjugated to synthetic polymer for mechanical strength. However, despite intended clinical use, little is known about their safety and efficacy, mechanism of action or degree of similarity to the full-length counterparts they mimic. Here, we show the functional equivalence of a CLP conjugated to polyethylene glycol (CLP-PEG) to full-length recombinant human collagen in vitro and in promoting stable regeneration of corneal tissue and nerves in a pre-clinical mini-pig model...
March 15, 2018: Acta Biomaterialia
Kentaro Jingushi, Motohide Uemura, Naomi Ohnishi, Wataru Nakata, Kazutoshi Fujita, Takuya Naito, Risa Fujii, Naomi Saichi, Norio Nonomura, Kazutake Tsujikawa, Koji Ueda
Cancer-associated extracellular vesicles (EVs) are intimately involved in establishment of tumor microenvironment and occurrence of metastasis. However, previous studies have mainly relied on experiments with cultured cell lines or mouse models, making it difficult to gain a full understanding of EV functions in human body. Hence, we extracted EVs directly from surgically resected viable clear cell renal cell carcinoma (ccRCC) tissues and adjacent normal renal tissues (n = 20). Quantitative LC/MS analysis identified 3,871 tissue-exudative EV (Te-EV) proteins, among which azurocidin (AZU1) was highly enriched in tumor Te-EVs (p = 2...
February 1, 2018: International Journal of Cancer. Journal International du Cancer
Jennifer Ngolab, Ivy Trinh, Edward Rockenstein, Michael Mante, Jazmin Florio, Margarita Trejo, Deborah Masliah, Anthony Adame, Eliezer Masliah, Robert A Rissman
Proteins implicated in neurodegenerative conditions such as Alzheimer's disease (AD) and Dementia with Lewy Bodies (DLB) have been identified in bodily fluids encased in extracellular vesicles called exosomes. Whether exosomes found in DLB patients can transmit pathology is not clear. In this study, exosomes were successfully harvested through ultracentrifugation from brain tissue from DLB and AD patients as well as non-diseased brain tissue. Exosomes extracted from brains diagnosed with either AD or DLB contained aggregate-prone proteins...
June 9, 2017: Acta Neuropathologica Communications
Javier M Figueroa, Johan Skog, Johnny Akers, Hongying Li, Ricardo Komotar, Randy Jensen, Florian Ringel, Isaac Yang, Steven Kalkanis, Reid Thompson, Lori LoGuidice, Emily Berghoff, Andrew Parsa, Linda Liau, William Curry, Daniel Cahill, Chetan Bettegowda, Frederick F Lang, E Antonio Chiocca, John Henson, Ryan Kim, Xandra Breakefield, Clark Chen, Karen Messer, Fred Hochberg, Bob S Carter
Background: RNAs within extracellular vesicles (EVs) have potential as diagnostic biomarkers for patients with cancer and are identified in a variety of biofluids. Glioblastomas (GBMs) release EVs containing RNA into cerebrospinal fluid (CSF). Here we describe a multi-institutional study of RNA extracted from CSF-derived EVs of GBM patients to detect the presence of tumor-associated amplifications and mutations in epidermal growth factor receptor (EGFR). Methods: CSF and matching tumor tissue were obtained from patients undergoing resection of GBMs...
October 19, 2017: Neuro-oncology
Valerie Jeanneret, Fang Wu, Paola Merino, Enrique Torre, Ariel Diaz, Lihong Cheng, Manuel Yepes
Tissue-type plasminogen activator (tPA) is a serine proteinase released by the presynaptic terminal of cerebral cortical neurons following membrane depolarization (Echeverry et al., 2010). Recent studies indicate that the release of tPA triggers the synaptic vesicle cycle and promotes the exocytosis (Wu et al., 2015) and endocytic retrieval (Yepes et al., 2016) of glutamate-containing synaptic vesicles. Here we used electron microscopy, proteomics, quantitative phosphoproteomics, biochemical analyses with extracts of the postsynaptic density (PSD), and an animal model of cerebral ischemia with mice overexpressing neuronal tPA to study whether the presynaptic release of tPA also has an effect on the postsynaptic terminal...
2016: Frontiers in Molecular Neuroscience
Shanli Zhu, Sai Wang, Yu Lin, Pengyue Jiang, Xiaobin Cui, Xinye Wang, Yuanbin Zhang, Weiqing Pan
BACKGROUND: Schistosoma japonicum is a parasitic flatworm that causes human schistosomiasis. Secreted extracellular vesicles (EVs) play a key role in pathogen-host interfaces. Previous studies have shown that S. japonicum adult worms can release microRNA (miRNA)-containing EVs, which can transfer their cargo to mammalian cells and regulate gene expression in recipient cells. Tissue-trapped eggs are generally considered the major contributor to the severe pathology of schistosomiasis; however, the interactions between the host and parasite in this critical stage remain largely unknown...
November 8, 2016: Parasites & Vectors
Samantha Sheller, John Papaconstantinou, Rheanna Urrabaz-Garza, Lauren Richardson, George Saade, Carlos Salomon, Ramkumar Menon
At term, the signals of fetal maturity and feto-placental tissue aging prompt uterine readiness for delivery by transitioning quiescent myometrium to an active stage. It is still unclear how the signals reach the distant myometrium. Exosomes are a specific type of extracellular vesicle (EVs) that transport molecular signals between cells, and are released from a wide range of cells, including the maternal and fetal cells. In this study, we hypothesize that i) exosomes act as carriers of signals in utero-placental compartments and ii) exosomes reflect the physiologic status of the origin cells...
2016: PloS One
M A Crookenden, C G Walker, H Peiris, Y Koh, A Heiser, J J Loor, K M Moyes, A Murray, V S R Dukkipati, J K Kay, S Meier, J R Roche, M D Mitchell
Biomarkers that identify prepathological disease could enhance preventive management, improve animal health and productivity, and reduce costs. Circulating extracellular vesicles, particularly exosomes, are considered to be long-distance, intercellular communication systems in human medicine. Exosomes provide tissue-specific messages of functional state and can alter the cellular activity of recipient tissues through their protein and microRNA content. We hypothesized that exosomes circulating in the blood of cows during early lactation would contain proteins representative of the metabolic state of important tissues, such as liver, which play integral roles in regulating the physiology of cows postpartum...
September 2016: Journal of Dairy Science
Xavier Gallart-Palau, Aida Serra, Siu Kwan Sze
BACKGROUND: Extracellular vesicles (EVs) act as key mediators of intercellular communication and are secreted and taken up by all cell types in the central nervous system (CNS). While detailed study of EV-based signaling is likely to significantly advance our understanding of human neurobiology, the technical challenges of isolating EVs from CNS tissues have limited their characterization using 'omics' technologies. We therefore developed a new Protein Organic Solvent Precipitation (PROSPR) method that can efficiently isolate the EV repertoire from human biological samples...
May 23, 2016: Molecular Neurodegeneration
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