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A single exosome detection

Chiara Costanza Volpi, Filippo Pietrantonio, Annunziata Gloghini, Giovanni Fucà, Silvia Giordano, Simona Corso, Giancarlo Pruneri, Maria Antista, Chiara Cremolini, Elena Fasano, Serena Saggio, Simona Faraci, Maria Di Bartolomeo, Filippo de Braud, Massimo Di Nicola, Elda Tagliabue, Serenella Maria Pupa, Lorenzo Castagnoli
The HER2 splice variant characterized by the deletion of exon 16 and denominated as d16HER2, is associated with HER2-positive breast cancer (BC) aggressiveness, stemness, and trastuzumab susceptibility and is considered to be a "flag" of HER2 dependence. However, with the exception of quantitative real-time PCR analysis, easily reproducible assays are still lacking to clinically detect and quantify the d16HER2 expression. Further, no data on d16HER2 expression and its potential role are available in HER2-positive gastrointestinal malignancies...
March 5, 2019: Scientific Reports
Chao Liu, Junxiang Zhao, Fei Tian, Jianqiao Chang, Wei Zhang, Jiashu Sun
Extracellular vesicles (EVs) are heavily implicated in diverse pathological processes. Due to their small size, distinct biogenesis, and heterogeneous marker expression, isolation and detection of single EV subpopulations are difficult. Here we develop a λ-DNA and aptamer mediated approach allowing for simultaneous size-selective separation and surface protein analysis of individual EVs. Using a machine learning algorithm to EV signature based on their size and marker expression, we demonstrate that the isolated microvesicles are more efficient than exosomes and apoptotic bodies in discriminating breast cell lines, and Stage II breast cancer patients with varied immunohistochemical expression of HER2...
February 21, 2019: Journal of the American Chemical Society
Yuanfeng Pang, Chaoguang Wang, LuChun Lu, Chongwen Wang, Zhiwei Sun, Rui Xiao
MicroRNAs have been proved to be the biomarker for early detection of pancreatic cancer and the precisely quantitation of the MicroRNA-10b in the blood samples even can distinguish pancreatic cancer from chronic pancreatitis (CP) and normal controls (NC). In this study, we developed a DSN-assisted dual-SERS biosensor for microRNA-10b in exosome and residual plasma of blood samples detection based on the Fe3 O4 @Ag-DNA-Au@Ag@DTNB (SERS tag) conjugates. In presence of target microRNA, it can hybridized with the complementary DNA probes...
January 25, 2019: Biosensors & Bioelectronics
Dinggeng He, See-Lok Ho, Hei-Nga Chan, Huizhen Wang, Luo Hai, Xiaoxiao He, Kemin Wang, Hung-Wing Li
Tumor exosomes (Exo) are presumed to expedite both the growth and metastasis of tumors by actively participating in nearly all aspects of cancer development. Tumor-derived Exos are thus proposed as a resource for diagnostic biomarkers in bodily fluids. However, most Exo assays require large samples and are time-consuming, complicated, and costly, and thus unsuited for practical applications. Herein, we show an ultrasensitive assay that can directly visualize and quantify tumor Exos in plasma microsamples (1 μL) at the single-vesicle level...
February 19, 2019: Analytical Chemistry
Hongmei Cao, Xin Zhou, Yong Zeng
There is an urgent need of sensitive bioanalytical platforms for sensitive and precise quantification of low-abundance microRNA targets in complex biological samples, including liquid biopsies of tumors. Many of current miRNA biosensing methods require laborious sample pretreatment procedures, including extraction of total RNA, which largely limits their biomedical and clinical applications. Herein we developed an integrated Microfluidic Exponential Rolling Circle Amplification (MERCA) platform for sensitive and specific detection of microRNAs directly in minimally processed samples...
January 15, 2019: Sensors and Actuators. B, Chemical
Rana Raoof, Sebastian Bauer, Hany El Naggar, Niamh M C Connolly, Gary P Brennan, Elizabeth Brindley, Thomas Hill, Hazel McArdle, Elaine Spain, Robert J Forster, Jochen H M Prehn, Hajo Hamer, Norman Delanty, Felix Rosenow, Catherine Mooney, David C Henshall
BACKGROUND: There are no blood-based molecular biomarkers of temporal lobe epilepsy (TLE) to support clinical diagnosis. MicroRNAs are short noncoding RNAs with strong biomarker potential due to their cell-specific expression, mechanistic links to brain excitability, and stable detection in biofluids. Altered levels of circulating microRNAs have been reported in human epilepsy, but most studies collected samples from one clinical site, used a single profiling platform or conducted minimal validation...
November 2, 2018: EBioMedicine
Eric H Hill, Jingang Li, Linhan Lin, Yaoran Liu, Yuebing Zheng
Lipid vesicles are important biological assemblies, which are critical to biological transport processes, and vesicles prepared in the lab are a workhorse for studies of drug delivery, protein unfolding, biomolecular interactions, compartmentalized chemistry, and stimuli-responsive sensing. The current method of using optical tweezers for holding lipid vesicles in place for single-vesicle studies suffers from limitations such as high optical power, rigorous optics, and small difference in the refractive indices of vesicles and water...
November 6, 2018: Langmuir: the ACS Journal of Surfaces and Colloids
Jacob O Brunkard, Barbara Baker
The cytosolic RNA exosome, a 3'→5' exoribonuclease complex, contributes to mRNA degradation in eukaryotes, limiting the accumulation of poorly-translated, improperly translated, or aberrant mRNA species. Disruption of cytosolic RNA exosome activity allows aberrant RNA species to accumulate, which can then be detected by host antiviral immune systems as a signature of pathogen infection, activating antiviral defenses. SKI7 is a critical component of the cytosolic RNA exosome in yeast, bridging the catalytic exoribonuclease core with the SKI2/SKI3/SKI8 adaptor complex that guides aberrant RNA substrates into the exosome...
2018: Frontiers in Plant Science
Yuting Yang, Guangxia Shen, Hui Wang, Hongxia Li, Ting Zhang, Nongjian Tao, Xianting Ding, Hui Yu
Exosomes play an important role in numerous cellular processes. Fundamental study and practical use of exosomes are significantly constrained by the lack of analytical tools capable of physical and biochemical characterization. In this paper, we present an optical approach capable of imaging single exosomes in a label-free manner, using interferometric plasmonic microscopy. We demonstrate monitoring of the real-time adsorption of exosomes onto a chemically modified Au surface, calculating the image intensity, and determining the size distribution...
October 9, 2018: Proceedings of the National Academy of Sciences of the United States of America
Francesco Martelli, Lisa Macera, Pietro Giorgio Spezia, Chiara Medici, Mauro Pistello, Daniele Guasti, Paolo Romagnoli, Fabrizio Maggi, Simone Giannecchini
BACKGROUND: Torquetenovirus (TTV) belongs to Anelloviridae family, infects nearly all people indefinitely without causing overt disease establishing a fine and successful interaction with the host. Increasing evidence have shown some human viruses exploit extracellular vesicles thereby helping viral persistence in the host. Here, the presence of TTV in extracellular vesicles circulating in human plasma was investigated. METHODS: TTV DNA was quantified in plasma-derived exosomes from 122 samples collected from 97 diseased patients and 25 healthy donors...
September 20, 2018: Virology Journal
Xingyu Lin, Xiao Huang, Yanzhe Zhu, Katharina Urmann, Xing Xie, Michael R Hoffmann
In this work, we introduce an asymmetric membrane as a simple and robust nanofluidic platform for digital detection of single pathogenic bacteria directly in 10 mL of unprocessed environmental water samples. The asymmetric membrane, consisting of uniform micropores on one side and a high density of vertically aligned nanochannels on the other side, was prepared within 1 min by a facile method. The single membrane covers all the processing steps from sample concentration, purification, and partition to final digital loop-mediated isothermal amplification (LAMP)...
October 23, 2018: ACS Nano
Natalie Luhtala, Tony Hunter
Exosomes, extracellular nanovesicles that carry nucleic acids, lipids, and proteins, have been the subject of several studies to assess their ability to transfer functional cargoes to cells. We recently characterized extracellular nanovesicles released from glioblastoma cells that carry active Ras in complex with proteins regulating exosome biogenesis. Here, we investigated whether a functional transfer of Ras from exosomes to other cells can initiate intercellular signaling. We observed that treatment of serum-starved, cultured glioblastoma cells with exogenous glioblastoma exosomes caused a significant increase in cellular viability over time...
2018: PloS One
Deepa Raghu, Joseph A Christodoulides, Marc Christophersen, Jinny L Liu, George P Anderson, Michael Robitaille, Jeff M Byers, Marc P Raphael
Exosomes are secreted nanovesicles which incorporate proteins and nucleic acids, thereby enabling multifunctional pathways for intercellular communication. There is an increasing appreciation of the critical role they play in fundamental processes such as development, wound healing and disease progression, yet because of their heterogeneous molecular content and low concentrations in vivo, their detection and characterization remains a challenge. In this work we combine nano- and microfabrication techniques for the creation of nanosensing arrays tailored toward single exosome detection...
2018: PloS One
Mostak Ahmed, Laura G Carrascosa, Alain Wuethrich, Paul Mainwaring, Matt Trau
It is a well-known phenomenon that cancer cells release key biological information such as DNA, RNA or proteins into body fluids (e.g., blood, urine or saliva). The analysis of these molecules-often encapsulated within nanovesicules called exosomes-is highly attractive, because it could replace current surgical biopsies, which are painful, costly and potentially risky for patients. For example, current strategies in lung cancer diagnosis involve genetic analyses from tumour tissues to detect the presence of underlying DNA mutations, known to alter the phosphorylation status and function of proteins...
August 21, 2018: Biomaterials Science
Yaokun Xia, Liangliang Wang, Juan Li, Xiangqi Chen, Jianming Lan, An Yan, Yun Lei, Sheng Yang, Huanghao Yang, Jinghua Chen
Recently, sensitive and selective detection of exosomal microRNAs (miRNAs) has been garnering significant attention, because it is related to many complex diseases, including cancer. Herein, we report a ratiometric fluorescent bioprobe based on DNA-labeled carbon dots (DNA-CDs) and 5,7-dinitro-2-sulfo-acridone (DSA) coupling with the target-catalyzing signal amplification for the detection of exosomal miRNA-21. There was high fluorescence resonance energy transfer (FRET) efficiency between carbon dots (CDs) and DSA when the bioprobe was assembled...
August 7, 2018: Analytical Chemistry
Chunchen Liu, Xiaonan Xu, Bo Li, Bo Situ, Weilun Pan, Yu Hu, Taixue An, Shuhuai Yao, Lei Zheng
Exosomes shed by tumor cells have been recognized as promising biomarkers for cancer diagnostics due to their unique composition and functions. Quantification of low concentrations of specific exosomes present in very small volumes of clinical samples may be used for noninvasive cancer diagnosis and prognosis. We developed an immunosorbent assay for digital qualification of target exosomes using droplet microfluidics. The exosomes were immobilized on magnetic microbeads through sandwich ELISA complexes tagged with an enzymatic reporter that produces a fluorescent signal...
July 11, 2018: Nano Letters
Lin Li, Yu Sun, Min Feng, Liang Wang, Jing Liu
Non-small-cell lung cancer (NSCLC) accounts for 85% of all cases of lung cancer. However, the predicted 5-year survival rate of patients with NSCLC is only 15.9%. microRNAs (miRNAs) are single-stranded, noncoding RNA molecules that are easily detectable in blood in a non-invasive manner, with features of stability, reproducibility and consistency in blood. Therefore, miRNAs derived from blood are able to have a significant impact on NSCLC diagnosis, metastasis and targeted therapies. Compared with the clinical protein markers carcinoembryonic antigen, cytokeratin fragment 21-1 and cancer antigen-125, blood-based miRNAs also display a higher diagnostic efficacy in NSCLC...
June 2018: Oncology Letters
Elyahb Allie Kwizera, Ryan O'Connor, Vojtech Vinduska, Melody Williams, Elizabeth R Butch, Scott E Snyder, Xiang Chen, Xiaohua Huang
Exosomes are a potential source of cancer biomarkers. Probing tumor-derived exosomes can offer a potential non-invasive way to diagnose cancer, assess cancer progression, and monitor treatment responses. Novel molecular methods would facilitate exosome analysis and accelerate basic and clinical exosome research. Methods: A standard gold-coated glass microscopy slide was used to develop a miniaturized affinity-based device to capture exosomes in a target-specific manner with the assistance of low-cost 3-D printing technology...
2018: Theranostics
Ramon M Eichenberger, Stephanie Ryan, Linda Jones, Geraldine Buitrago, Ramona Polster, Marcela Montes de Oca, Jennifer Zuvelek, Paul R Giacomin, Lindsay A Dent, Christian R Engwerda, Matthew A Field, Javier Sotillo, Alex Loukas
Gastrointestinal (GI) parasites, hookworms in particular, have evolved to cause minimal harm to their hosts, allowing them to establish chronic infections. This is mediated by creating an immunoregulatory environment. Indeed, hookworms are such potent suppressors of inflammation that they have been used in clinical trials to treat inflammatory bowel diseases (IBD) and celiac disease. Since the recent description of helminths (worms) secreting extracellular vesicles (EVs), exosome-like EVs from different helminths have been characterized and their salient roles in parasite-host interactions have been highlighted...
2018: Frontiers in Immunology
Jie Zhou, Lingchang Meng, Weiran Ye, Qiaolei Wang, Shizhen Geng, Chong Sun
Alzheimer's disease (AD) considered as the third health "killer" has seriously threatened the health of the elderly. However, the modern diagnostic strategies of AD present several disadvantages: the low accuracy and specificity resulting in some false-negative diagnoses, and the poor sensitivity leading to a delayed treatment. In view of this situation, a enzyme-free and target-triggered signal amplification strategy, based on graphene oxide (GO) and entropy-driven strand displacement reaction (ESDR) principle, was proposed...
August 31, 2018: Analytica Chimica Acta
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