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Academic literature on the topic 'Exosomi Urinari'

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Published: 9 March 2023
Last updated: 10 March 2023

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Journal articles on the topic "Exosomi Urinari"

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Huang, Kun, Sudha Garimella, Alyssa Clay-Gilmour, Lucia Vojtech, Bridget Armstrong, Madison Bessonny, and Alexis Stamatikos. "Comparison of Human Urinary Exosomes Isolated via Ultracentrifugation Alone versus Ultracentrifugation Followed by SEC Column-Purification." Journal of Personalized Medicine 12, no. 3 (February 24, 2022): 340. http://dx.doi.org/10.3390/jpm12030340.

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Chronic kidney disease is a progressive, incurable condition that involves a gradual loss of kidney function. While there are no non-invasive biomarkers available to determine whether individuals are susceptible to developing chronic kidney disease, small RNAs within urinary exosomes have recently emerged as a potential candidate to use for assessing renal function. Ultracentrifugation is the gold standard for urinary exosome isolation. However, extravesicular small RNA contamination can occur when isolating exosomes from biological fluids using ultracentrifugation, which may lead to misidentifying the presence of certain small RNA species in human urinary exosomes. Therefore, we characterized human urinary exosomal preparations isolated by ultracentrifugation alone, or via ultracentrifugation followed by size exclusion chromatography (SEC) column-purification. Using nanoparticle tracking analysis, we identified SEC fractions containing robust amounts of exosome-sized particles, that we further characterized using immunoblotting. When compared to exosomal preparations isolated by ultracentrifugation only, SEC fractionated exosomal preparations showed higher levels of the exosome-positive marker CD81. Moreover, while the exosome-negative marker calnexin was undetectable in SEC fractionated exosomal preparations, we did observe calnexin detection in the exosomal preparations isolated by ultracentrifugation alone, which implies contamination in these preparations. Lastly, we imaged SEC fractionated exosomal preparations using transmission electron microscopy to confirm these preparations contained human urinary exosomes. Our results indicate that combining ultracentrifugation and SEC column-purification exosome isolation strategies is a powerful approach for collecting contaminant-free human urinary exosomes and should be considered when exosomes devoid of contamination are needed for downstream applications.
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Abdeen, Ahmed, Hiroko Sonoda, Ragab El-Shawarby, Saki Takahashi, and Masahiro Ikeda. "Urinary excretion pattern of exosomal aquaporin-2 in rats that received gentamicin." American Journal of Physiology-Renal Physiology 307, no. 11 (December 1, 2014): F1227—F1237. http://dx.doi.org/10.1152/ajprenal.00140.2014.

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Urinary exosomes are nano-sized vesicles secreted into urine from all types of renal epithelial cells and are known to contain possible biomarker proteins for renal diseases. Gentamicin has been reported to decrease the level of renal aquaporin (AQP)2, which is known to be mainly expressed in renal collecting ducts and excreted into the urine via exosomes. In the present study, we investigated whether urinary exosomal AQP2 could serve as a potential biomarker for gentamicin-induced nephrotoxicity, especially collecting duct cell dysfunction. Gentamicin was given to rats intraperitoneally once every day starting on day 0. Gentamicin significantly increased the plasma creatinine concentration from day 5 and beyond. Also, gentamicin induced polyuria and a defective urine concentration mechanism on day 7, suggesting gentamicin-induced collecting duct cell dysfunction. Immunoblot analysis showed that gentamicin significantly increased urinary exosomal AQP2 excretion on day 1 but decreased it on day 7 compared with the control group. Similarly, increased excretion of exosomal tumor susceptibility gene 101 protein, frequently used as an exosome marker protein, was observed on day 1. However, gentamicin did not significantly affect the urinary excretion of exosomal tumor susceptibility gene 101 on day 7. Gentamicin slightly decreased renal AQP2 expression on day 2 and markedly decreased it on day 8. These data strongly suggest that the use of urinary exosomal AQP2 as a biomarker may allow detection of gentamicin-induced collecting duct cell dysfunction. Furthermore, urinary exosomal AQP2 might also be useful for the early detection of gentamicin-induced renal injury in addition to collecting duct injury.
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Cheruvanky, Anita, Hua Zhou, Trairak Pisitkun, Jeffrey B. Kopp, Mark A. Knepper, Peter S. T. Yuen, and Robert A. Star. "Rapid isolation of urinary exosomal biomarkers using a nanomembrane ultrafiltration concentrator." American Journal of Physiology-Renal Physiology 292, no. 5 (May 2007): F1657—F1661. http://dx.doi.org/10.1152/ajprenal.00434.2006.

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Urinary exosomes are excreted from all nephron segments and may serve as biomarkers for classifying renal diseases. Isolation of urinary exosomes by the established ultracentrifugation method has some limitations for use in a clinical laboratory. We sought a rapid and simple way to obtain urinary exosomes. We used a commercially available nanomembrane concentrator to enrich exosomes from urine by centrifugation at 3,000 g for 10–30 min. Urinary exosomal markers tumor susceptibility gene 101, aquaporin-2, neuron-specific enolase, annexin V, angiotensin-converting enzyme, and podocalyxin (PODXL) were recovered from the nanomembrane concentrator and detected by Western blotting, and typical features of urinary vesicles were found by electron microscopy. Exosomal markers were detected in as little as 0.5 ml of urine. By the nanomembrane method, exosomal proteins could be recovered from urine samples frozen at −80°C or refrigerated overnight at 4°C then stored at −80°C. By enriching exosomes we could detect PODXL, a podocyte marker, which decreased by 71% in five male patients with focal segmental glomerulosclerosis and abundant proteinuria. We conclude that 1) use of a nanomembrane concentrator simplifies and accelerates the enrichment of urinary exosomes; and 2) the nanomembrane concentrator can concentrate exosomal proteins from clinical urine samples. This enhanced method may accelerate the translation of urinary exosomal biomarkers from bench to bedside for the diagnosis, classification, and prognostication of renal diseases.
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Huang, Zhibin, Yong Zhang, Jianhua Zhou, and Yu Zhang. "Urinary Exosomal miR-193a Can Be a Potential Biomarker for the Diagnosis of Primary Focal Segmental Glomerulosclerosis in Children." BioMed Research International 2017 (2017): 1–6. http://dx.doi.org/10.1155/2017/7298160.

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Background. Glomerular upregulation of miR-193a has been detected in primary focal segmental glomerulosclerosis (FSGS) but not in other glomerular diseases. We aimed to isolate exosomes from urine of children with primary FSGS and to assess the diagnostic potential of urinary exosomal miR-193a for primary FSGS. Methods. The first morning urine samples were collected from children with primary FSGS (n=8) and minimal change disease (MCD, n=5). Isolated urinary exosomes were confirmed by electron microscopy and Western blotting. Urinary exosomal microRNA was extracted, and the expression levels of exosomal miR-193a were quantified by real-time PCR. The diagnosis value of urinary exosomal miR-193a levels for primary FSGS was evaluated by ROC analysis. Results. The isolated vesicles were qualitatively compatible with exosomes. The levels of urinary exosomal miR-193a were significantly higher in children with primary FSGS than those in children with MCD. Moreover, the area under the ROC for the diagnosis of primary FSGS using urinary exosomal miR-193a was 0.85. Conclusions. A significant increase in the levels of urinary exosomal miR-193a in primary FSGS patients compared to those in MCD ones was observed. This study suggests that urinary exosomal miR-193a may be a new noninvasive biomarker for the diagnosis of primary FSGS.
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Garcia-Vives, Eloi, Cristina Solé, Teresa Moliné, Marta Vidal, Irene Agraz, Josep Ordi-Ros, and Josefina Cortés-Hernández. "The Urinary Exosomal miRNA Expression Profile is Predictive of Clinical Response in Lupus Nephritis." International Journal of Molecular Sciences 21, no. 4 (February 18, 2020): 1372. http://dx.doi.org/10.3390/ijms21041372.

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Data on exosomal-derived urinary miRNAs have identified several miRNAs associated with disease activity and fibrosis formation, but studies on prognosis are lacking. We conducted a qPCR array screening on urinary exosomes from 14 patients with biopsy-proven proliferative lupus glomerulonephritis with a renal outcome of clinical response (n = 7) and non-response (n = 7) following therapy. Validation studies were performed by qRT-PCR in a new lupus nephritis (LN) cohort (responders = 22 and non-responders = 21). Responder patients expressed significantly increased levels of miR-31, miR-107, and miR-135b-5p in urine and renal tissue compared to non-responders. MiR-135b exhibited the best predictive value to discriminate responder patients (area under the curve = 0.783). In vitro studies showed exosome-derived miR-31, miR-107, and miR-135b-5p expression to be mainly produced by tubular renal cells stimulated with inflammatory cytokines (e.g IL1, TNFα, IFNα and IL6). Uptake of urinary exosomes from responders by mesangial cells was superior compared to that from non-responders (90% vs. 50%, p < 0.0001). HIF1A was identified as a potential common target, and low protein levels were found in non-responder renal biopsies. HIF1A inhibition reduced mesangial proliferation and IL-8, CCL2, CCL3, and CXCL1 mesangial cell production and IL-6/VCAM-1 in endothelial cells. Urinary exosomal miR-135b-5p, miR-107, and miR-31 are promising novel markers for clinical outcomes, regulating LN renal recovery by HIF1A inhibition.
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Yu, Yanting, Zhiyun Ren, Anni Xie, Yutao Jia, Ying Xue, Ping Wang, Daxi Ji, and Xiaoyan Wang. "Assessment of Urinary Exosomal NHE3 as a Biomarker of Acute Kidney Injury." Diagnostics 12, no. 11 (October 30, 2022): 2634. http://dx.doi.org/10.3390/diagnostics12112634.

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The diagnosis of acute kidney injury (AKI) traditionally depends on the serum creatinine (Scr) and urine output, which lack sufficient sensitivity and specificity. Using urinary exosomes as a biomarker has unique advantages. To assess whether urinary exosomal Na+/H+ exchanger isoform 3 (NHE3) protein could serve as a biomarker of AKI, we constructed four AKI rat models: cisplatin (7.5 mg/kg) injected intraperitoneally (IP), furosemide (20 mg/kg, IP) with a low-NaCl (0.03%) diet, a low-NaCl (0.03%) diet with candesartan (1 mg/kg, IP) and bilateral ischemia and reperfusion (I/R) injury for 40 min. Additionally, we assessed six sepsis-associated AKI patients and six healthy volunteers. Urinary exosomes were extracted by ultracentrifugation, and the NHE3 protein abundance was tested by immunoblotting for all the AKI rats and human subjects. The isolated cup-shaped particles with an average diameter of 70 nm and enrichment in CD63 were identified as exosomes. NHE3 abundance was six times higher in exosomes than in the whole urine. In cisplatin-induced AKI rats, urinary exosomal NHE3 was increased on day 2, one day earlier than the increases in Scr and blood urea nitrogen (BUN). In additional rats, urinary exosomal NHE3 decreased along with the decline in Scr after EPO pretreatment. In volume-depletion AKI induced by furosemide injection with a low-NaCl diet, the urinary exosomal NHE3 expression was higher than that in the control. Under a low-NaCl diet with candesartan-related AKI, the urinary exosomal NHE3 was elevated on day 5, earlier than Scr. In I/R-injury AKI, the urinary exosomal NHE3 was also raised compared with that in the control. In humans, the urinary exosomal NHE3 level was also elevated in sepsis-associated AKI patients in comparison with that in the healthy volunteers. The urinary exosomal NHE3 was increased in multiple AKI; it may be used as a diagnostic biomarker of AKI.
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Cao, Yuhan, Yuanhui Shi, Yuwei Wang, Yanlang Yang, Wenjun Guo, Cuifeng Zhang, Wenjun Pei, and Cong Fu. "Exosomal hsa_circ_0008925 from Urine Is Related to Chronic Renal Fibrosis." Disease Markers 2022 (February 18, 2022): 1–10. http://dx.doi.org/10.1155/2022/1899282.

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At present, there is no noninvasive biomarker of renal fibrosis. The potential diagnostic value of urinary exosome-derived circRNAs from glomerular disease patients for renal fibrosis is still uncertain. Here, we first detected the expression of hsa_circ_0008925 in TGF-β1-cultured HK-2 cell-derived exosomes. Secondly, we collected urine samples from 95 biopsy-proven glomerular disease patients and 34 healthy controls. The expression of hsa_circ_0008925 was analyzed, and the correlation with renal function and pathological changes was calculated. The receiver operating characteristic (ROC) curve for the diagnosis of renal fibrosis was performed. The results showed that in exosomes derived from TGF-β1-cultured HK-2 cells, the expression of hsa_circ_0008925 was increased compared with normal cultured. Further, the expression level of hsa_circ_0008925 was increased in urinary exosomes from renal fibrosis patients and correlated with serum creatinine, blood urea nitrogen (BUN), estimated glomerular filtration rate, and cystatin C. The level of hsa_circ_0008925 was furthermore correlated with the score of tubulointerstitial fibrosis (TIF) and the score of glomerular sclerosis. The ROC curve showed that hsa_circ_0008925 can diagnose renal fibrosis at a cut-off value of 0.093 with a sensitivity of 52.2% and specificity of 96.4%. In summary, we indicated that urinary exosomal hsa_circ_0008925 could be acted as a noninvasive biomarker for renal fibrosis in glomerular diseases patients.
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Chen, Chen, Anquan Shang, Zujun Sun, Yuting Gao, Jingjuan Huang, Yili Ping, Wenjing Chang, et al. "Urinary Exosomal Long Noncoding RNA TERC as a Noninvasive Diagnostic and Prognostic Biomarker for Bladder Urothelial Carcinoma." Journal of Immunology Research 2022 (January 25, 2022): 1–9. http://dx.doi.org/10.1155/2022/9038808.

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Purpose. Bladder cancer is one of the most common urological malignancies worldwide, and approximately 90% of bladder cancer cases are histologically typed as bladder urothelial carcinoma (BLCA). Exosomes are 30 to 200 nm extracellular vesicles that transport microRNAs, long noncoding RNAs (lncRNAs), mRNAs, circular RNAs, and proteins across tissues and through circulation. Urinary exosomes may contain genetic information from tumor cells. Herein, we explored the clinical significance of urinary exosomal lncRNA telomerase RNA component (TERC) levels to provide an urgently needed diagnostic and prognostic biomarker for BLCA. Materials and Methods. In this study, we used RNA-sequencing of samples from four BLCA patients and three healthy controls to identify that TERC was differentially expressed in urinary exosomes. We then used quantitative PCR in different types of clinical samples to validate the biomarker and analyzed results using receiver operating characteristic curves. Results. We found that TERC was significantly upregulated in urinary exosomes from BLCA patients compared with those from healthy controls ( P < 0.0001 ). Urinary exosomal TERC showed higher sensitivity (78.65%) and accuracy (77.78%) than existing indicators including nuclear matrix protein-22 and urine cytometry. Using the cut-off value 4.302, the area under the curve for urinary exosomal TERC was 0.836 (95% confidence interval: 0.768–0.891, P < 0.0001 ). Furthermore, this noninvasive assay could distinguish low-grade and high-grade tumors ( P = 0.0153 ). Conclusions. TERC is enriched in urinary exosomes from BLCA patients. Urinary exosomal TERC could become a diagnostic and prognostic biomarker for BLCA that allows clinicians to realize noninvasive detection of BLCA.
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Li, Guorong, Nora Mallouk, Pascale Flandrin, Arnauld Garcin, Claude Lambert, Sid Ali Berremila, Hocine Habchi, and Nicolas Mottet. "Presence of Urinary Exosomes for Liquid Biopsy of Clear Cell Renal Cell Carcinoma: Protocol for a Pilot Feasibility Study." JMIR Research Protocols 10, no. 7 (July 20, 2021): e24423. http://dx.doi.org/10.2196/24423.

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Background Approximately 70%-80% of kidney cancers are clear cell renal cell carcinomas (CCRCCs). Patient management is based on imaging (abdominal ultrasound and computerized tomography), surgical excision of the tumor, and pathological analysis. A tissue biopsy is therefore necessary to confirm the diagnosis and avoid unnecessary nephrectomy. For metastatic cancers, a tissue biopsy is essential for establishing the targeted therapy. This biopsy of tumor material is invasive and painful. Other techniques such as liquid biopsy would help reduce the need for tissue biopsy. The development of a simple biological test for diagnosis is essential. CA9 is a powerful marker for the diagnosis of CCRCC. Exosomes have become a major source of liquid biopsy because they carry tumor proteins, RNA, and lipids. Urine is the most convenient biological liquid for exosome sampling. Objective The aim of this study (PEP-C study) is mainly to determine whether it is possible to detect urinary exosomal CA9 for the molecular diagnosis of CCRCC. Methods This study will include 60 patients with CCRCC and 40 noncancer patients. Exosomes will be isolated from urine samples and exosomal CA9 will be detected by transmission electron microscopy, flow cytometry, and reverse transcription-quantitative polymerase chain reaction. Results This study is currently underway with funding support from the CHU Saint-Etienne of France. Conclusions We expect to demonstrate that urinary tumor exosomes could be a novel liquid biopsy to diagnose CCRCC and to guide clinicians in treatment decision-making. Trial Registration ClinicalTrials.gov NCT04053855; https://clinicaltrials.gov/ct2/show/NCT04053855 International Registered Report Identifier (IRRID) DERR1-10.2196/24423
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Riffo-Campos, Angela L., Javier Perez-Hernandez, Olga Martinez-Arroyo, Ana Ortega, Ana Flores-Chova, Josep Redon, and Raquel Cortes. "Biofluid Specificity of Long Non-Coding RNA Profile in Hypertension: Relevance of Exosomal Fraction." International Journal of Molecular Sciences 23, no. 9 (May 6, 2022): 5199. http://dx.doi.org/10.3390/ijms23095199.

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Non-coding RNA (ncRNA)-mediated targeting of various genes regulates the molecular mechanisms of the pathogenesis of hypertension (HTN). However, very few circulating long ncRNAs (lncRNAs) have been reported to be altered in essential HTN. The aim of our study was to identify a lncRNA profile in plasma and plasma exosomes associated with urinary albumin excretion in HTN by next-generation sequencing and to assess biological functions enriched in response to albuminuria using GO and KEGG analysis. Plasma exosomes showed higher diversity and fold change of lncRNAs than plasma, and low transcript overlapping was found between the two biofluids. Enrichment analysis identified different biological pathways regulated in plasma or exosome fraction, which were implicated in fatty acid metabolism, extracellular matrix, and mechanisms of sorting ncRNAs into exosomes, while plasma pathways were implicated in genome reorganization, interference with RNA polymerase, and as scaffolds for assembling transcriptional regulators. Our study found a biofluid specific lncRNA profile associated with albuminuria, with higher diversity in exosomal fraction, which identifies several potential targets that may be utilized to study mechanisms of albuminuria and cardiovascular damage.
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Dissertations / Theses on the topic "Exosomi Urinari"

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CORBETTA, SAMUELE. "Proteomica degli exosomi urinari per la ricerca di biomarcatori nella nefropatia diabetica e nelle tubulopatie ereditarie." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2015. http://hdl.handle.net/10281/76004.

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Le urine costituiscono il fluido biologico di elezione nella ricerca di biomarcatori per le patologie renali in quanto possono essere raccolte in modo semplice e non invasivo; una strategia per la semplificazione del proteoma urinario è rappresentata dall’isolamento degli exosomi urinari (UE), nanovescicole di membrana (30-100 nm) rilasciate dalle cellule epiteliali nello spazio urinario. In questo lavoro abbiamo focalizzato l’attenzione sulla nefropatia diabetica (DN), una comune complicazione del diabete ed una delle cause più frequenti di insufficienza renale terminale (ESDR), e sulle tubulopatie ereditarie (SLTs), un gruppo eterogeneo di malattie genetiche rare dell’età pediatrica caratterizzate da difetti in proteine coinvolte nel riassorbimento di sodio-cloruro a livello dell’ansa di Henle e/o dei tubuli renali distali, come i cotrasportatori NCC NKCC2, alterati rispettivamente nella sindrome di Gitelman (GS) e nella sindrome di Bartter1 (B1). L’obiettivo che ci siamo proposti è stato quello di studiare il proteoma degli UE in: 1) un modello animale di DN, i ratti ZDF (Zucker Diabetic Fatty), e corrispondenti controlli, per l’identificazione di potenziali biomarcatori diagnostici/prognostici e 2) pazienti affetti da SLTs e controlli sani al fine di proporre un approccio diagnostico complementare/alternativo all’analisi genetica e fornire un punto di partenza per la ricerca di biomarcatori. Per quanto riguarda lo studio della DN, sono state raccolte le urine delle 24 ore da 7 ratti ZDF e controlli a differenti età per monitorare l’evoluzione della DN, gli UE sono stati isolati mediante ultracentrifugazione, seguita da caratterizzazione biochimica. Dopo l’allestimento di un pool rappresentativo di UE di ratti ZDF e controlli a 20 settimane, ne è stato analizzato il proteoma tramite LC-ESI-MS/MS portando all’identificazione ed alla quantificazione label-free di 286 proteine. Il contenuto differenziale di alcune di queste proteine è stato confermato tramite immunoblotting; il contenuto della Major-Urinary-Protein-1 è risultato significativamente più alto, quello della proteina Xaa-Pro-Dipeptidase più basso e quello della Neprilisina invariato, rispettivamente, negli UE di ratti ZDF rispetto ai controlli. Per quanto riguarda le SLTs, la casistica era formata da 32 pazienti SLTs già studiati dal punto di vista genetico e biochimico-clinico, da 4 pazienti SLTs non classificabili e da 22 controlli sani. Dopo la raccolta delle seconde urine del mattino, sono stati isolati e caratterizzati gli UE. I risultati hanno mostrato che il segnale relativo al cotrasportatore NCC risulta significativamente ridotto o assente negli UE dei pazienti GS rispetto ai controlli, e allo stesso modo il cotrasportatore NKCC2 per i pazienti B1. Le differenze nei livelli di queste due proteine negli UE ci consentono il riconoscimento dei pazienti GS e B1 rispetto ai controlli e, in combinazione con i dati biochimico-clinici, rispetto agli altri pazienti Bartter. E’ stato quindi proposto e validato statisticamente un approccio diagnostico che può risultare utile in casi di SLTs a diagnosi incerta. Per quanto riguarda la proteina NCC è stata inoltre effettuata una correlazione tra la quantità del trasportatore presente negli UE e la gravità della mutazione corrispondente. Per l’identificazione di biomarcatori delle altre due forme di SLTs studiate, la sindrome di Bartter2 (B2) e di Bartter3 (B3), abbiamo allestito un pool di UE, abbiamo separato le proteine mediante elettroforesi bidimensionale selezionando alcuni spot differenziali da analizzare tramite LC-ESI-MS/MS; il contenuto differenziale negli UE di alcune proteine identificate verrà validato mediante IB. In conclusione possiamo affermare che la composizione proteica degli UE risulta alterata in maniera riproducibile in presenza di nefropatia diabetica e di tubulopatie ereditarie; le differenze evidenziate possono costituire un punto di partenza per l’identificazione di biomarcatori diagnostici/prognostici e per il chiarimento dei meccanismi patogenetici di tali malattie.
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Hiemstra, Thomas Francois. "Investigating the biology of urinary exosomes." Thesis, University of Cambridge, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.610116.

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Garcia, Vives Eloi. "Exosomas urinarios: identificación de biomarcadores de respuesta clínica en la nefritis lúpica." Doctoral thesis, Universitat Autònoma de Barcelona, 2019. http://hdl.handle.net/10803/669399.

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Antecedentes Los biomarcadores actuales en la nefritis lúpica (NL) no son lo suficientemente sensibles o específicos para predecir la evolución renal. Datos recientes abalan el uso de los exosomas urinarios como fuente de biomarcadores de origen renal, con especial relevancia en su contenido de miRNAs. Algunos estudios han identificado varios miRNAs asociados con la actividad de la enfermedad y la formación de fibrosis, pero no se han realizado estudios sobre biomarcadores de pronóstico. Métodos Para identificar biomarcadores pronóstico, realizamos una micromatriz de miRNA en exosomas urinarios de pacientes con NL activa, analizando los resultados en función de la respuesta al tratamiento presentada por los pacientes (7 respondedores y 7 no respondedores). Los estudios de validación se realizaron mediante técnica de RT-qPCR en una nueva cohorte de LN (respondedores = 21 y no respondedores = 22). Entre los miRNAs de interés se realizó un estudio comparativo con los niveles séricos así como con una cohorte de pacientes sanos y otra de nefropatías no lúpicas. Posteriormente se realizó hibridación in situ de tejido renal y estudios in vitro para identificar el origen, diana celular y el mecanismo de acción de estos exosomas a nivel renal. Para cada miRNA expresado diferencialmente, los genes diana potenciales fueron predichos a través del estudio de conjuntos de datos de miRNA. Resultados Los pacientes que respondieron a la pauta de tratamiento estándar expresaron niveles significativamente mayores de miR-31-5p, miR-107 y miR-135b-5p en la orina y en tejido renal en comparación con los no respondedores. El miR-135b-5p mostró el mejor valor predictivo para discriminar a los pacientes que respondieron (AUC = 0.783 [intervalo de confianza del 95%, 0.640 - 0.926], al valor de corte > 0.0884 con una sensibilidad del 77.8% y especificidad del 71.4%). La expresión de los miRNAs se localizó principalmente en la estructura tubular, demostrándose, además, una mayor formación de exosomas enriquecidos con miR-31-5p, miR- 107 y miR-135b-5p a nivel de esta estirpe en comparación con la de células endoteliales glomerulares o mesangiales (p < 0,0001). En cuanto a la diana de estos exosomas, su observó una velocidad de internalización de los derivados de orina de pacientes respondedores superior a los de no respondedores tanto a nivel de las células endoteliales glomerulares como mesangiales (p = 0,001 y 0,0002), siendo además superior la capacidad de internalización a nivel de las células mesangiales englobadas (90% frente a 50%, p < 0,0001). El miR-135b-5p demostró capacidad en la inhibición de la proliferación de las células mesangiales (p < 0.01). Además, el análisis de las distintas dianas biológicas pudo demostrar de forma global un papel de estos miRNAs en la modulación de la respuesta inflamatoria, con reducción de los niveles de citoquinas, quimioquinas y moléculas de adhsesión, un cierto efecto en la modulación sobre la angiogénesis, así como una marcada capacidad antifibrosante. El análisis de la vía identificó nueve dianas comunes relevantes para la recuperación renal, una de ellas, HIF-1α, común para los 3 miRNA de interés. Los 3 miRNAs demostraron un efecto modulador sobre la vía del HIF- 1α a través de su inhibición. Conclusiones Los niveles de miR-135b-5p, miR-31-5p y miR-107 son capaces de predecir la respuesta al tratamiento en el momento del brote y evolutivamente en los pacientes afectos de NL, de forma específica. El origen de estos parecería situarse a nivel de las células tubulares mientras que las mesangiales y endoteliales glomerulares se presentarían como las principales efectoras de sus acciones biológicas. HIF-1α fue identificado como diana común de estos 3 miRNAs, ocasionando la presencia de estos miRNAs una supresión a nivel de la expresión de esta vía, considerando clave en la patogénesis de la NL.
Background Current biomarkers in Lupus nephritis (LN) are not sensitive or specific enough to predict renal outcome. Recent data support the use of urinary exosomes as a source of biomarkers of renal origin, with special relevance in their miRNA content. Some studies have identified several miRNAs associated with disease activity and fibrosis formation, but no studies on prognostic biomarkers have been conducted. Methods To identify prognostic biomarkers, we performed a miRNA microarray on urinary exosomes from patients with active LN, analyzing their results according to the response to standard therapy (7 responders and 7 non-responders). Validation studies were performed by RT-qPCR technique in a new LN cohort (responders=21 and non-responders=22). Among the miRNA of interest, a comparative study was conducted with serum levels in the same cohort as well as with a healthy control group and non-lupus nephropathy cohort. Subsequently, in situ renal tissue hybridization and in vitro studies were performed to identify the origin and cell target of these exosomes at renal level and to understand their mechanism of action. For each differentially expressed miRNA, potential target genes were predicted through miRNA-target datasets. Results Responder patients expressed significantly increased levels of miR-31-5p, miR-107 and miR- 135b-5p in urine and renal tissue compared to non-responders. MiR-135b-5p exhibited the best predictive value to discriminate responder patients (AUC= 0.783 [95% confidence interval, 0.640 - 0.926], cut-off > 0.0884 with 77.8% sensitivity and 71.4% specificity). MiRNAs expression was mainly located in the tubular structure, demonstrating, in addition, greater formation on exosomes enriched with miR-31-5p, miR-107 and miR-135b-5p present in this lineage compared to endothelial glomerular and mesangial cells (p < 0.0001). Regarding the target of these exosomes, a faster rate of internalization of responders' urinary-derived exosomes was observed in both endothelial glomerular and mesangial cells (p = 0,001 y 0,0002), and there was greater capacity of internalization on mesangial cells (90% vs. 50%, p < 0.0001). The miR-135b-5p demonstrated ability to inhibit the proliferation of mesangial cells (p < 0.01). In addition, the analysis of the different biological targets demonstrate globally a role of these miRNAs in the modulation of the inflammatory response, with reduction of cytokines, chemokines and adhesion molecules levels, a modulatory effect on angiogenesis, as well as a marked antifibrosing capacity. The pathway analysis identified nine common targets relevant to renal recovery, one of them, HIF-1α, common for the 3 miRNA of interest. The 3 miRNAs demonstrated a modulating effect on the HIF-1α pathway through its inhibition. Conclusions Levels of miR-135b-5p, miR-31-5p and miR-107 are able to predict the response to treatment at the time of the renal flare and during follow-up in patients with NL, specifically. The origin of the exosomes enriched with these miRNAs appears to be at the level of tubular cells while the mesangial and glomerular endothelial cells appear as the main effectors of their biological actions. HIF-1α was identified as the common target of these 3 miRNAs, causing the presence of these miRNAs suppression of the expression levels of this pathway, considered key in the pathogenesis of NL.
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Burballa, Tàrrega Carla 1988. "Discovery of putative prognostic and therapeutic miRNA in uEVs of Dent's Disease 1 patients and characterisation of cellular models of the disease." Doctoral thesis, TDX (Tesis Doctorals en Xarxa), 2021. http://hdl.handle.net/10803/671807.

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Dent disease 1 (DD1) is a rare renal tubulopathy caused by CLCN5 mutations and characterized by low molecular weight proteinuria, variable hypercalciuria, nephrocalcinosis and/or nephrolithiasis and progression to kidney failure. The underlying mechanisms linking ClC-5 loss-of-function and endocytosis impairment in the renal proximal tubule (and other DD1 phenotypes) remain unknown. In this thesis we have followed three approaches to identify altered pathways by ClC-5 mutations: (1) conduct a European survey to analyse the prevalence and DD1 clinical features, (2) study miRNA expression profiles from DD1 patients’ urinary exosome-like vesicles (uEVs) to get insight into DD1 pathophysiological mechanisms and (3) characterisation of a DD1 cell model. The European survey showed that DD1 has a variable presentation. Our study of uEVs miRNA identified new pathophysiological pathways, which may lead to identify putative diagnostic and prognostic biomarkers. Finally, our cell model with different mutations provides a valuable prototype for additional investigation of impaired pathways.
La malaltia de Dent 1 (DD1) és una tubulopatia renal rara causada per mutacions en el gen CLCN5 i caracteritzada per proteinuria de baix pes molecular, hipercalciuria, nefrocalcinosi i/o litiasis renals així com progrés a insuficiència renal. Els mecanismes que causen la pèrdua de funció de ClC-5 i el defecte en l’endocitosi en el túbul proximal (entre d’altres fenotips de DD1) no es coneixen. En aquesta Tesi hem desenvolupat tres aproximacions per identificar vies alterades per mutacions en ClC-5. (1) hem fet una enquesta europea per analitzar la prevalença i les característiques clíniques de DD1, (2) hem estudiat l’expressió de miRNA en vesícules exosome-like urinàries (uEVs) per entendre els mecanismes fisiopatològics de la malaltia i (3) hem caracteritzat un model cel·lular de DD1. L’enquesta europea mostrà que DD1 té una presentació variable. El nostre estudi de miRNA en uEVs va permetre identificar nous mecanismes fisiopatològics que poden ser potencials biomarcadors diagnòstics i pronòstics de DD1. Finalment, el nostre model cel·lular amb diferents mutacions provà representar un prototip vàlid per investigacions addicionals del mecanismes desregulats.
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Lai, Yue Fan, and 賴岳汎. "Searching Potential Protein Biomarkers of Bladder Cancer from Urinary Exosomes." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/58768233351293356294.

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碩士
長庚大學
生物醫學研究所
99
Bladder cancer mortality increases annually in Taiwan. The main detection tool for bladder cancer is cystoscopy. However, this method is an invasive and costly procedure. Urine is stored in bladder for hours and can be collected non-invasively. Therefore, urine can be a good material for discovery of bladder cancer biomarkers. Multivesicular body derived exosomes present in saliva, plasma, and urine. Previous studies pointed out that exosomes correlate with diseases, and can be a material for biomarker searching. In this work, we compared the proteomic profiles of urinary exosomes between controls and bladder cancer patients using isotopic dimethyl labelling and two-dimensional LC/MS/MS. The differentially-expressed exosomal proteins will be potential non-invasive biomarkers for detection of bladder cancer. Exosomes extracted from 9 hernia and 9 bladder cancer patients were pooled as a control and a bladder cancer samples, respectively. After digestion of exosomal proteins, peptides of hernia and bladder cancer samples were labelled with light and heavy dimethylation reagents, respectively. The labelled peptides were analyzed by on-line SCX/RP LC/MS/MS. The quantification of proteins were performed by MaxQuant software. We further used the dimethylation platform to compare the exosome proteins expressions of 9 hernia and 9 bladder cancer individuals without pooling samples. We have indentified total 3873 exosomal proteins in two comparisons, and 107 proteins show higher different concentration levels between bladder cancer and hernia patients. 29 of 107 proteins in urinary exosomes could directly quantified by multiple-reaction-monitoring (MRM)-MS. MRM-MS validated 29 proteins expression levels between 5 hernia and 5 bladder cancer patients’ exosomes, and most results were consistent with that we have seen in discovery phase. Then we validated more clinical sample. We quantified 29 proteins expression levels in 12 hernia, 28 bladder cancer, 5 hematuria and 3 urinary tract infection patients’ exosomes. 22 of 29 proteins had significant differentially expressed level between hernia and bladder cancer patient, and higher expressed level of 3 proteins in bladder cancer patients were not due to hematuria or urinary tract infection. In this work, we implemented two comparisons to searching differentially-expressed exosomal proteins, and we validated these proteins by MRM-MS. The validation results are consistent with prediction. In the future, we will increase the sample number to raise the reliability of these potential bladder cancer biomarkers.
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Lin, Shih-Yi, and 林詩怡. "Investigate the proteomics of urinary exosomes for biomarkers of urothelial carcinoma." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/59sj6j.

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博士
中國醫藥大學
臨床醫學研究所博士班
105
Purpose: Studies have focused on establishing noninvasive, rapid methods for discovering urothelial carcinoma (UC) biomarkers. The urinary exosome proteome is believed to directly reflect the proteome of UC, providing a suitable investigation resource. Increasingly applied matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) spectrometry facilitates reliable clinical diagnosis of bacteria. MALDI-TOF spectrometry, a rapid analytical platform, has not been used for urinary exosome analysis. Therefore, we used it for determining UC biomarkers. Experimental Design: From 2012 to 2015, we enrolled 129 consecutive patients with UC and 62 participants without UC. Exosomes from their urine were isolated, and the protein extracts of these exosomes were analyzed through MALDI-TOF spectrometry. Moreover, immunohistochemical (IHC) analysis of another 122 UC and 26 non-UC tissues was conducted to verify the discovered biomarkers. Results: Two peaks at m/z 5593 (fragmented peptide of alpha-1-antitrypsin; sensitivity, 50.4%; specificity, 96.9%) and m/z 5947 (fragmented peptide of histone H2B1K sensitivity, 62.0%; specificity, 92.3%) were identified as UC diagnosis exosome biomarkers. UC patients with detectable histone H2B1K showed 2.29- and 3.11-fold increased risks of recurrence and progression, respectively, compared with those with nondetectable histone H2B1K. Verification results of IHC staining revealed significantly higher expression of alpha 1-antitrypsin (p = 0.038) and H2B1K (p = 0.005) in UC tissues than in normal tissues. The expression of alpha 1-antitrypsin and H2B1K in UC tissues was significantly correlated with UC grades (p < 0.05). Conclusion: Urinary exosome proteins alpha 1-antitrypsin and histone H2B1K, which are identified through MALDI-TOF analysis, could facilitate rapid diagnosis and prognosis of UC.
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Miao, Yuxuan. "Mechanisms of Bacterial Expulsion as a Cell Autonomous Defense Strategy In the Bladder Epithelium." Diss., 2015. http://hdl.handle.net/10161/9862.

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Due to its close proximity to the gastrointestinal tract, the human urinary tract is

subjected to constant barrage by gut-­associated bacteria. However, for the most part, this tract has resisted infection by various microbes. The impregnability of the urinary tract to microbial colonization is attributable to the ability of the bladder to promptly sense and mount robust responses to microbial challenge. A powerful mechanism for the elimination of invading bacteria was recently described in bladder epithelial cells, involving non-­lytic ejection of intracellular bacteria back into the extracellular milieu. In spite of the effectiveness of this defense strategy, much of the underlying mechanisms surrounding how this powerful cellular defense activity detects intracellular UPEC and shuttles them from their intracellular location to the plasma membrane of BECs to be exported remains largely a mystery.

Here, we describe uropathogenic E.coli (UPEC) expelled from infected bladder

epithelium cells (BECs) within membrane-­bound vesicles as a distinct cellular defense

response. Examination of the intracellular UPEC revealed that intracellular bacteria were

initially processed via autophagy, the conventional degradative pathway, then delivered

into multivesicular bodies (MVBs) and encapsulated in nascent intraluminal vesicle membrane. We further show the bacterial expulsion is triggered when intracellular UPEC follow the natural degradative trafficking pathway and reach lysosomes and attempt to neutralize its pH to avoid degradation. This pathogen-­mediated activity is detected by mucolipin TRP channel 3 (TRPML3), a transient receptor potential cation channel localized on lysosomes, which spontaneously initiates lysosome exocytosis resulting in expulsion of exosome-­encased bacteria. These studies reveal a cellular default system for lysosome homeostasis and also, how it is coopted by the autonomous defense program to clear recalcitrant pathogens.


Dissertation
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Book chapters on the topic "Exosomi Urinari"

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Salvi, Samanta, Erika Bandini, and Francesco Fabbri. "Urinary Exosomes in Prostate Cancer." In Urinary Biomarkers, 115–20. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1354-2_10.

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Alvarez-Llamas, Gloria, and Irene Zubiri. "Proteome of Human Urinary Exosomes in Diabetic Nephropathy." In Biomarkers in Kidney Disease, 347–67. Dordrecht: Springer Netherlands, 2016. http://dx.doi.org/10.1007/978-94-007-7699-9_22.

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Alvarez-Llamas, Gloria, and Irene Zubiri. "Proteome of Human Urinary Exosomes in Diabetic Nephropathy." In Biomarkers in Kidney Disease, 1–21. Dordrecht: Springer Netherlands, 2015. http://dx.doi.org/10.1007/978-94-007-7743-9_22-1.

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Abdeen, Ahmed, Hiroko Sonoda, Ayae Tanaka, and Masahiro Ikeda. "Urinary Exosomes as a Possible Source of Kidney Disease Biomarkers." In Role of Exosomes in Biological Communication Systems, 221–44. Singapore: Springer Singapore, 2020. http://dx.doi.org/10.1007/978-981-15-6599-1_10.

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Pitto, Marina, Samuele Corbetta, and Francesca Raimondo. "Preparation of Urinary Exosomes: Methodological Issues for Clinical Proteomics." In Methods in Molecular Biology, 43–53. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-1872-0_3.

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Barigazzi, Elisa, Lucia Santorelli, W. Morello, F. Raimondo, B. Crapella, L. Ghio, C. Tamburello, G. Montini, and M. Pitto. "New Insight into Idiopathic Nephrotic Syndrome: Strategy Based on Urinary Exosomes." In Toxic Chemical and Biological Agents, 217–18. Dordrecht: Springer Netherlands, 2020. http://dx.doi.org/10.1007/978-94-024-2041-8_13.

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Santorelli, Lucia, Elisa Barigazzi, M. Pitto, and F. Raimondo. "Investigation of the N-Glycoproteome in the Urinary Exosomes: Technical Challenges." In Toxic Chemical and Biological Agents, 257–58. Dordrecht: Springer Netherlands, 2020. http://dx.doi.org/10.1007/978-94-024-2041-8_26.

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Conde-Vancells, Javier, and Juan M. Falcon-Perez. "Isolation of Urinary Exosomes from Animal Models to Unravel Noninvasive Disease Biomarkers." In Liver Proteomics, 321–40. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-959-4_21.

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DiStefano, Johanna K., Rupesh Kanchi Ravi, and Mahdieh Khosroheidari. "Urinary Exosomes as Potential Source for Identification of Biomarkers for Kidney Damage: Comparing Methodologies." In Biomarkers in Disease: Methods, Discoveries and Applications, 939–54. Dordrecht: Springer Netherlands, 2015. http://dx.doi.org/10.1007/978-94-007-7696-8_47.

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Alvarez, M. Lucrecia. "Isolation of Urinary Exosomes for RNA Biomarker Discovery Using a Simple, Fast, and Highly Scalable Method." In RNA Mapping, 145–70. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-1062-5_13.

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Conference papers on the topic "Exosomi Urinari"

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Berner, K., M. Hirschfeld, G. Rücker, D. Weiß, A. Ritter, M. Jäger, I. Juhasz-Böss, and T. Erbes. "Urinary exosomal microRNAs as potential non-invasive biomarkers in breast cancer detection." In 40. Jahrestagung der Deutschen Gesellschaft für Senologie e.V. © Georg Thieme Verlag KG, 2020. http://dx.doi.org/10.1055/s-0040-1710603.

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Berner, K., M. Hirschfeld, G. Rücker, D. Weiß, A. Ritter, I. Juhasz-Böss, and T. Erbes. "Urinary exosomal microRNAs as potential non-invasive biomarkers in breast cancer detection." In Kongressabstracts zur Tagung 2020 der Deutschen Gesellschaft für Gynäkologie und Geburtshilfe (DGGG). © 2020. Thieme. All rights reserved., 2020. http://dx.doi.org/10.1055/s-0040-1717838.

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Blondal, Thorarinn, Anne I. Rasmussen, Anni R. Thomsen, Michael Borre, Jacob Fredsøe, Ditte Andreasen, Torben Falck Ørntoft, Karina D. Sørensen, and Peter Mouritzen. "Abstract 1943: A microRNA signature in urinary exosomes for diagnosis of prostate cancer." In Proceedings: AACR 107th Annual Meeting 2016; April 16-20, 2016; New Orleans, LA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.am2016-1943.

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Mouritzen, Peter, Jacob Christian Fredsøe, Thorarinn Blondal, Anne Karin Rasmussen, Michael Borre, Christa Haldrup, Ditte Andreasen, Niels Tolstrup, Torben Falck Ørntoft, and Karina Dalsgaard Sørensen. "Abstract B40: A two-microRNA signature in urinary exosomes for diagnosis of prostate cancer." In Abstracts: AACR Special Conference on Noncoding RNAs and Cancer: Mechanisms to Medicines; December 4-7, 2015; Boston, MA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.nonrna15-b40.

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Baumgart, Sophie, Joana Heinzelmann, Michael Stoeckle, Marie Stampe Ostenfeld, and Kerstin Junker. "Abstract 5182: Characterization of miRNA expression pattern fromin-vitroobtained exosomes of different urinary bladder cancer cell lines." In Proceedings: AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA. American Association for Cancer Research, 2015. http://dx.doi.org/10.1158/1538-7445.am2015-5182.

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Wilson, Brittany, Rebekah Betar, Alexander Martin, Zackaria Niazi, Michael Boyer, Lori Winter, Victor Babich, Francesca Di Sole, and Elitsa Ananieva. "Abstract 2377: microRNA expression profile in urinary exosomes is dependent on non-invasive lymphoma induction in mice." In Proceedings: AACR Annual Meeting 2021; April 10-15, 2021 and May 17-21, 2021; Philadelphia, PA. American Association for Cancer Research, 2021. http://dx.doi.org/10.1158/1538-7445.am2021-2377.

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Solé-Marcé, Cristina, Eloi Garcia-Vives, Irene Agraz, Josep Ordi-Ros, and Josefina Cortés-Hernández. "THU0236 URINARY EXOSOMAL MIR-31, MIR-107 AND MIR-135B-5P FROM TUBULAR RENAL CELLS AS RESPONDER BIOMARKER IN LUPUS NEPHRITIS." In Annual European Congress of Rheumatology, EULAR 2019, Madrid, 12–15 June 2019. BMJ Publishing Group Ltd and European League Against Rheumatism, 2019. http://dx.doi.org/10.1136/annrheumdis-2019-eular.5346.

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Berrondo, Claudia, Jonathan Flax, Aisha Siebert, Victor Kucherov, Alex Rosenberg, Christopher Fucile, and Carla Beckham. "Abstract B31: Bladder cancer patient urinary exosomes and tumors contain long noncoding RNA that may serve as therapeutic targets and biomarkers." In Abstracts: AACR Special Conference on Noncoding RNAs and Cancer: Mechanisms to Medicines; December 4-7, 2015; Boston, MA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.nonrna15-b31.

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