scholarly journals Extracellular Vesicles-Based Biomarkers Represent a Promising Liquid Biopsy in Endometrial Cancer

Cancers ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 2000 ◽  
Author(s):  
Carolina Herrero ◽  
Alexandre de la Fuente ◽  
Carlos Casas-Arozamena ◽  
Victor Sebastian ◽  
Martin Prieto ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Extracellular Vesicles ◽  
Biological Fluids ◽  
Low Cost ◽  
New Technology ◽  
Response To Therapy ◽  
Clinical Implementation ◽  
Clinical Value ◽  
Specific Content ◽  
Technical Performance

Tumor-derived extracellular vesicles (EVs) are secreted in large amounts into biological fluids of cancer patients. The analysis of EVs cargoes has been associated with patient´s outcome and response to therapy. However, current technologies for EVs isolation are tedious and low cost-efficient for routine clinical implementation. To explore the clinical value of circulating EVs analysis we attempted a proof-of-concept in endometrial cancer (EC) with ExoGAG, an easy to use and highly efficient new technology to enrich EVs. Technical performance was first evaluated using EVs secreted by Hec1A cells. Then, the clinical value of this strategy was questioned by analyzing the levels of two well-known tissue biomarkers in EC, L1 cell adhesion molecule (L1CAM) and Annexin A2 (ANXA2), in EVs purified from plasma in a cohort of 41 EC patients and 20 healthy controls. The results demonstrated the specific content of ANXA2 in the purified EVs fraction, with an accurate sensitivity and specificity for EC diagnosis. Importantly, high ANXA2 levels in circulating EVs were associated with high risk of recurrence and non-endometrioid histology suggesting a potential value as a prognostic biomarker in EC. These results also confirmed ExoGAG technology as a robust technique for the clinical implementation of circulating EVs analyses.

scholarly journals The Role of Extracellular Vesicles as Modulators of the Tumor Microenvironment, Metastasis and Drug Resistance in Colorectal Cancer

Cancers ◽  
2019 ◽  
Vol 11 (6) ◽  
pp. 746 ◽  
Author(s):  
Kodappully S. Siveen ◽  
Afsheen Raza ◽  
Eiman I. Ahmed ◽  
Abdul Q. Khan ◽  
Kirti S. Prabhu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Drug Resistance ◽  
Tumor Microenvironment ◽  
Extracellular Vesicles ◽  
Significant Role ◽  
Biological Fluids ◽  
Therapeutic Targets ◽  
Response To Therapy ◽  
High Morbidity

Colorectal cancer (CRC) is one of the most common cancers worldwide, with high morbidity and mortality rates. A number of factors including modulation of the tumor microenvironment, high metastatic capability, and resistance to treatment have been associated with CRC disease progression. Recent studies have documented that tumor-derived extracellular vesicles (EVs) play a significant role in intercellular communication in CRC via transfer of cargo lipids, proteins, DNA and RNAs to the recipient tumor cells. This transfer influences a number of immune-related pathways leading to activation/differentiation/expression of immune cells and modulation of the tumor microenvironment that plays a significant role in CRC progression, metastasis, and drug resistance. Furthermore, tumor-derived EVs are secreted in large amounts in biological fluids of CRC patients and as such the expression analysis of EV cargoes have been associated with prognosis or response to therapy and may be a source of therapeutic targets. This review aims to provide a comprehensive insight into the role of EVs in the modulation of the tumor microenvironment and its effects on CRC progression, metastasis, and drug resistance. On the other hand, the potential role of CRC derived EVs as a source of biomarkers of response and therapeutic targets will be discussed in detail to understand the dynamic role of EVs in CRC diagnosis, treatment, and management.

scholarly journals Isolation of Extracellular Vesicles from Biological Fluids via the Aggregation–Precipitation Approach for Downstream miRNAs Detection

Diagnostics ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 384
Author(s):  
Maria Y. Konoshenko ◽  
Evgeniy A. Lekchnov ◽  
Olga E. Bryzgunova ◽  
Elena Kiseleva ◽  
Inna A. Pyshnaya ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Biological Fluids ◽  
Low Cost ◽  
Precipitation Method ◽  
Medical Applications ◽  
Non Invasive ◽  
Clinical Laboratories ◽  
Healthy Donors ◽  
Invasive Diagnostics ◽  
Productive Method

Extracellular vesicles (EVs) have high potential as sources of biomarkers for non-invasive diagnostics. Thus, a simple and productive method of EV isolation is demanded for certain scientific and medical applications of EVs. Here we aim to develop a simple and effective method of EV isolation from different biofluids, suitable for both scientific, and clinical analyses of miRNAs transported by EVs. The proposed aggregation–precipitation method is based on the aggregation of EVs using dextran blue and the subsequent precipitation of EVs using 1.5% polyethylene glycol solutions. The developed method allows the effective isolation of EVs from plasma and urine. As shown using TEM, dynamic light scattering, and miRNA analyses, this method is not inferior to ultracentrifugation-based EV isolation in terms of its efficacy, lack of inhibitors for polymerase reactions and applicable for both healthy donors and cancer patients. This method is fast, simple, does not need complicated equipment, can be adapted for different biofluids, and has a low cost. The aggregation–precipitation method of EV isolation accessible and suitable for both research and clinical laboratories. This method has the potential to increase the diagnostic and prognostic utilization of EVs and miRNA-based diagnostics of urogenital pathologies.

Separation of Extracellular Vesicles by DNA-Directed Immunocapturing Followed by Enzymatic Release

2020 ◽  
Author(s):  
Dario Brambilla ◽  
Laura Sola ◽  
Elisa Chiodi ◽  
Natasa Zarovni ◽  
Diogo Fortunato ◽  
...  
Keyword(s):  
Cell Culture ◽  
Extracellular Vesicles ◽  
Culture Media ◽  
Biological Fluids ◽  
Imaging Techniques ◽  
Cell Communication ◽  
Disease Diagnosis ◽  
Cell Culture Supernatant ◽  
Transmission Electron ◽  
Downstream Analysis

Extracellular vesicles (EVs) have attracted great interest among researchers due to their role in cell-cell communication, disease diagnosis, and drug delivery. In spite of their potential in the medical field, there is no consensus on the best method for separating microvesicles from cell culture supernatant and complex biological fluids. Obtaining a good recovery yield and preserving physical characteristics is critical for the diagnostic and therapeutic use of EVs. The separation is made complex by the fact that blood and cell culture media, contain a large number of nanoparticles in the same size range. Methods that exploit immunoaffinity capture provide high purity samples and overcome the issues of currently used separation methods. However, the release of captured nanovesicles requires harsh conditions that hinder their use in certain types of downstream analysis. Herein, a novel capture and release approach for small extracellular vesicles (sEVs), based on DNAdirected immobilization of antiCD63 antibody is presented. The flexible DNAlinker increases the capture efficiency and allows releasing of EVs by exploiting the endonucleasic activity of DNAse I. This separation protocol works under mild conditions, enabling the release of intact vesicles that can be successfully analyzed by imaging techniques. In this article sEVs recovered from plasma were characterized by established techniques for EVs analysis including nanoparticle tracking and transmission electron microscopy.<br>

Microextraction and Chromatographic Analysis of Budesonide Epimers in Exhaled Breath Condensate

2020 ◽  
Vol 16 (8) ◽  
pp. 1032-1040
Author(s):  
Laleh Samini ◽  
Maryam Khoubnasabjafari ◽  
Mohamad M. Alimorad ◽  
Vahid Jouyban-Gharamaleki ◽  
Hak-Kim Chan ◽  
...  
Keyword(s):  
Biological Fluids ◽  
Exhaled Breath Condensate ◽  
Low Cost ◽  
Exhaled Breath ◽  
Chromatography Method ◽  
Extraction Solvent ◽  
Drug Concentrations ◽  
Dispersive Liquid Liquid Microextraction ◽  
Breath Condensate

Background: Analysis of drug concentrations in biological fluids is required in clinical sciences for various purposes. Among other biological samples, exhaled breath condensate (EBC) is a potential sample for follow up of drug concentrations. Methods: A dispersive liquid-liquid microextraction (DLLME) procedure followed by a validated liquid chromatography method was employed for the determination of budesonide (BDS) in EBC samples collected using a homemade setup. EBC is a non-invasive biological sample with possible applications for monitoring drug concentrations. The proposed analytical method is validated according to the FDA guidelines using EBC-spiked samples. Its applicability is tested on EBC samples collected from healthy volunteers receiving a single puff of BDS. Results: The best DLLME conditions involved the use of methanol (1 mL) as a disperser solvent, chloroform (200 μL) as an extraction solvent, and centrifugation rate of 3500 rpm for 5 minutes. The method was validated over a concentration range of 21-210 μg·L-1 in EBC. Inter- and intra-day precisions were less than 10% where the acceptable levels are less than 20%. The validated method was successfully applied for the determination of BDS in EBC samples. Conclusion: The findings of this study indicate that the developed method can be used for the extraction and quantification of BDS in EBC samples using a low cost method.

Clinical Value of Computer-Assisted Taylor Three-dimensional External Fixation in the Treatment of Tibiofibular Fracture (Preprint)

2020 ◽  
Author(s):  
Hongfeng Sheng ◽  
Weixing Xu ◽  
Bin Xu ◽  
Hongpu Song ◽  
Di Lu ◽  
...  
Keyword(s):  
Retrospective Study ◽  
Internal Fixation ◽  
External Fixation ◽  
External Fixator ◽  
Theoretical Basis ◽  
New Technology ◽  
Three Dimensional ◽  
Computer Assisted ◽  
Clinical Value

UNSTRUCTURED The retrospective study of Taylor's three-dimensional external fixator for the treatment of tibiofibular fractures provides a theoretical basis for the application of this technology. The paper collected 28 patients with tibiofibular fractures from the Department of Orthopaedics in our hospital from March 2015 to June 2018. After the treatment, the follow-up evaluation of Taylor's three-dimensional external fixator for the treatment of tibiofibular fractures and concurrency the incidence of the disease, as well as the efficacy and occurrence of the internal fixation of the treatment of tibial fractures in our hospital. The results showed that Taylor's three-dimensional external fixator was superior to orthopaedics in the treatment of tibiofibular fractures in terms of efficacy and complications. To this end, the thesis research can be concluded as follows: Taylor three-dimensional external fixation in the treatment of tibiofibular fractures is more effective, and the incidence of occurrence is low, is a new technology for the treatment of tibiofibular fractures, it is worthy of clinical promotion.

Azo (Hydrazone) pigments: general principles

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Robert Christie
Keyword(s):  
High Performance ◽  
Metal Salt ◽  
Low Cost ◽  
Cost Effective ◽  
Precipitation Process ◽  
Ambient Temperatures ◽  
Technical Performance ◽  
Chemical Structures ◽  
General Chemical ◽  
Physical Form

Abstract This paper presents an overview of the general chemical principles underlying the structures, synthesis and technical performance of azo pigments, the dominant chemical class of industrial organic pigments in the yellow, orange, and red shade areas, both numerically and in terms of tonnage manufactured. A description of the most significant historical features in this group of pigments is provided, starting from the discovery of the chemistry on which azo colorants are based by Griess in the mid-nineteenth century, through the commercial introduction of the most important classical azo pigments in the early twentieth century, including products known as the Hansa Yellows, β-naphthol reds, including metal salt pigments, and the diarylide yellows and oranges, to the development in the 1950s and 1960s of two classes of azo pigments that exhibit high performance, disazo condensation pigments and benzimidazolone-based azo pigments. A feature that complicates the description of the chemical structures of azo pigments is that they exist in the solid state as the ketohydrazone rather than the hydroxyazo form, in which they have been traditionally been illustrated. Numerous structural studies conducted over the years on an extensive range of azo pigments have demonstrated this feature. In this text, they are referred to throughout as azo (hydrazone) pigments. Since a common synthetic procedure is used in the manufacture of virtually all azo (hydrazone) pigments, this is discussed in some detail, including practical aspects. The procedure brings together two organic components as the fundamental starting materials, a diazo component and a coupling component. An important reason for the dominance of azo (hydrazone) pigments is that they are highly cost-effective. The syntheses generally involve low cost, commodity organic starting materials and are carried out in water as the reaction solvent, which offers obvious economic and environmental advantages. The versatility of the approach means that an immense number of products may be prepared, so that they have been adapted structurally to meet the requirements of many applications. On an industrial scale, the processes are straightforward, making use of simple, multi-purpose chemical plant. Azo pigments may be produced in virtually quantitative yields and the processes are carried out at or below ambient temperatures, thus presenting low energy requirements. Finally, provided that careful control of the reaction conditions is maintained, azo pigments may be prepared directly by an aqueous precipitation process that can optimise physical form, with control of particle size distribution, crystalline structure, and surface character. The applications of azo pigments are outlined, with more detail reserved for subsequent papers on individual products.

scholarly journals Upregulation of Serum lncRNA DLEU1 Predicts Progression of Premalignant Endometrial Lesion and Unfavorable Clinical Outcome of Endometrial Cancer

2020 ◽  
Vol 19 ◽  
pp. 153303382096558
Author(s):  
Lixia Shan ◽  
Tao Zhao ◽  
Yu Wang
Keyword(s):  
Endometrial Cancer ◽  
Endometrial Hyperplasia ◽  
Cox Regression ◽  
Quantitative Polymerase Chain Reaction ◽  
Critical Role ◽  
Disease Free Survival ◽  
Lymphocytic Leukemia ◽  
Healthy Controls ◽  
Clinical Value ◽  
Cox Regression Analysis

Objective: Long non-coding RNAs (lncRNAs) play a critical role in tumorigenesis. Upregulation of lncRNA deleted in lymphocytic leukemia 1 (DLEU1) has been reported in endometrial cancer (EC) tissues. This prospective study aimed to determine the potential clinical significance of serum lncRNA DLEU1 in EC. Methods: The serum lncRNA DLEU1 level was detected in EC patients, patients with endometrial hyperplasia and healthy controls by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Then its clinical value in EC was further evaluated. Results: Our results demonstrated that serum lncRNA DLEU1 levels were significantly increased in patients with EC, and serum lncRNA DLEU1 showed good performance for discriminating EC patients from patients with endometrial hyperplasia and healthy controls. In addition, EC patients with advanced clinicopathological features had higher circulating lncRNA DLEU1 level than those with favorable clinical characteristics. Moreover, EC patients in the high serum lncRNA DLEU1 group suffered worse overall survival and disease-free survival than those in the low serum lncRNA DLEU1 group. Furthermore, multivariate cox regression analysis displayed that the serum lncRNA DLEU1 served as an independent prognostic factor for EC. Conclusions: Collectively, our study suggests that serum lncRNA DLEU1 is a novel and promising biomarker for prognostic estimation of EC.

scholarly journals Compact Miniaturized Bioluminescence Sensor Based on Continuous Air-Segmented Flow for Real-Time Monitoring: Application to Bile Salt Hydrolase (BSH) Activity and ATP Detection in Biological Fluids

Chemosensors ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 122
Author(s):  
Aldo Roda ◽  
Pierpaolo Greco ◽  
Patrizia Simoni ◽  
Valentina Marassi ◽  
Giada Moroni ◽  
...  
Keyword(s):  
Bile Salt ◽  
Light Emission ◽  
Ex Vivo ◽  
Biological Fluids ◽  
Low Cost ◽  
Bile Salt Hydrolase ◽  
Segmented Flow ◽  
Active Substrate ◽  
Flat Spiral ◽  
Bsh Activity

A simple and versatile continuous air-segmented flow sensor using immobilized luciferase was designed as a general miniaturized platform based on sensitive biochemiluminescence detection. The device uses miniaturized microperistaltic pumps to deliver flows and compact sensitive light imaging detectors based on BI-CMOS (smartphone camera) or CCD technology. The low-cost components and power supply make it suitable as out-lab device at point of need to monitor kinetic-related processes or ex vivo dynamic events. A nylon6 flat spiral carrying immobilized luciferase was placed in front of the detector in lensless mode using a fiber optic tapered faceplate. ATP was measured in samples collected by microdialysis from rat brain with detecting levels as low as 0.4 fmoles. The same immobilized luciferase was also used for the evaluation of bile salt hydrolase (BSH) activity in intestinal microbiota. An aminoluciferin was conjugatated with chenodeoxycholic acid forming the amide derivative aLuc-CDCA. The hydrolysis of the aLuc-CDCA probe by BSH releases free uncaged aminoluciferin which is the active substrate for luciferase leading to light emission. This method can detect as low as 0.5 mM of aLuc-CDCA, so it can be used on real faecal human samples to study BSH activity and its modulation by diseases and drugs.

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.

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