An amplification-free electrochemical detection of exosomal miRNA-21 in serum samples

The Analyst ◽  
2018 ◽  
Vol 143 (7) ◽  
pp. 1662-1669 ◽  
Author(s):  
Kseniia Boriachek ◽  
Muhammad Umer ◽  
Md Nazmul Islam ◽  
Vinod Gopalan ◽  
Alfred K. Lam ◽  
...  
Keyword(s):  
Electrochemical Detection ◽  
Biological Samples ◽  
Serum Samples ◽  
Exosomal Mirnas ◽  
Exosomal Mirna

A sensitive and amplification-free assay for the electrochemical detection of exosomal miRNAs in complex biological samples.

scholarly journals DNA tetrahedron-mediated immune-sandwich assay for rapid and sensitive detection of PSA through a microfluidic electrochemical detection system

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Dezhi Feng ◽  
Jing Su ◽  
Yi Xu ◽  
Guifang He ◽  
Chenguang Wang ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Reaction Time ◽  
Electrochemical Detection ◽  
Detection System ◽  
Detection Performance ◽  
Pdms Layer ◽  
Microfluidic Chips ◽  
Serum Samples ◽  
Gold Nanoflowers ◽  
Dna Tetrahedron

AbstractProstate-specific antigen (PSA) is the most widely used biomarker for the early diagnosis of prostate cancer. Existing methods for PSA detection are burdened with some limitations and require improvement. Herein, we developed a novel microfluidic–electrochemical (μFEC) detection system for PSA detection. First, we constructed an electrochemical biosensor based on screen-printed electrodes (SPEs) with modification of gold nanoflowers (Au NFs) and DNA tetrahedron structural probes (TSPs), which showed great detection performance. Second, we fabricated microfluidic chips by DNA TSP-Au NF-modified SPEs and a PDMS layer with designed dense meandering microchannels. Finally, the μFEC detection system was achieved based on microfluidic chips integrated with the liquid automatic conveying unit and electrochemical detection platform. The μFEC system we developed acquired great detection performance for PSA detection in PBS solution. For PSA assays in spiked serum samples of the μFEC system, we obtained a linear dynamic range of 1–100 ng/mL with a limit of detection of 0.2 ng/mL and a total reaction time <25 min. Real serum samples of prostate cancer patients presented a strong correlation between the “gold-standard” chemiluminescence assays and the μFEC system. In terms of operation procedure, cost, and reaction time, our method was superior to the current methods for PSA detection and shows great potential for practical clinical application in the future.

scholarly journals Regulation of Serum Exosomal MicroRNAs in Mice Infected with Orientia tsutsugamushi

2020 ◽  
Vol 9 (1) ◽  
pp. 80
Author(s):  
Le Jiang ◽  
Tatyana Belinskaya ◽  
Zhiwen Zhang ◽  
Teik-Chye Chan ◽  
Wei-Mei Ching ◽  
...  
Keyword(s):  
Post Infection ◽  
Viral Infections ◽  
Scrub Typhus ◽  
Late Phase ◽  
Many Body ◽  
Host Immune Responses ◽  
Lethal Infection ◽  
Exosomal Mirnas ◽  
Exosomal Mirna ◽  
Intercellular Communications

Exosomes are small extracellular vesicles that carry proteins, lipids, and nucleic acids. They are circulated in many body fluids and play an important role in intercellular communications. MicroRNAs (miRNAs), as major components of exosomes, are often regulated in many diseases including bacterial and viral infections. Functionally, exosome-carried miRNAs interact with various immune cells and affect their behavior. Little is known whether exosomal miRNAs are regulated during scrub typhus, a potentially lethal infection caused by intracellular bacteria, Orientiatsutsugamushi. In the present study, we utilized a scrub typhus mouse model and collected serum at various time points post infection. A custom quantitative PCR array covering 92 murine miRNAs was used to profile serum exosomal miRNAs. A total of 12 miRNAs were found to be significantly up- or down-regulated at least at one time point post infection when compared to uninfected animals. Further analysis identified multiple miRNAs in the let-7 family that were consistently down-regulated at early and late phase of infection. Functionally, serum exosomes isolated from infected mice displayed strong proinflammatory effect when incubated with bone marrow-derived macrophages. Our data revealed dynamic regulations of serum exosomal miRNA during scrub typhus infection, which could significantly influence host immune responses and disease outcome.

CoFe2O4 supported g-C3N4 nanocomposite for sensitive electrochemical detection of dopamine

2021 ◽  
Author(s):  
Benadict Joseph Xavier ◽  
Umesh N ◽  
Sea-Fue Wang ◽  
Antolin Jesilaa Jesu Amalraj
Keyword(s):  
Electrochemical Detection ◽  
Electrode Material ◽  
Biological Samples ◽  
Selective Electrode ◽  
High Demand

The quantification of the neurotransmitter dopamine in human biological samples has gained more importance. A good antifouling, and highly selective electrode material is still in high demand. So, to resolve...

scholarly journals Plasma Exosomal miRNA Levels after Radiotherapy Are Associated with Early Progression and Metastasis of Cervical Cancer: A Pilot Study

2021 ◽  
Vol 10 (10) ◽  
pp. 2110
Author(s):  
Oyeon Cho ◽  
Do-Wan Kim ◽  
Jae-Youn Cheong
Keyword(s):  
Cervical Cancer ◽  
Pilot Study ◽  
Biological Functions ◽  
Patients With Cancer ◽  
Initial Stage ◽  
Exosomal Mirnas ◽  
Early Progression ◽  
Before And After ◽  
Microenvironmental Factors ◽  
Exosomal Mirna

Plasma exosomal miRNAs are key regulators of cell-cell interactions associated with several biological functions in patients with cancer. This pilot study aimed to investigate the log2 fold change (log2FC) of the expression of exosomal miRNAs and related mRNAs in the blood of patients with cervical cancer to identify prognostic markers better than those currently available. We sequenced plasma exosomal RNA from 56 blood samples collected from 28 patients with cervical cancer, who had been treated with concurrent chemoradiotherapy (CCRT). Changes in the expression of miRNAs and mRNAs before and after CCRT were represented as log2FC. Their biological functions were studied by miRNA-mRNA network analysis, using ingenuity pathway analysis, after the selection of two groups of miRNAs, each associated with early progression (EP) and metastasis, also described as initial stage. Seven patients experienced EP, three of whom died within four months after progression. Reduced levels of miR-1228-5p, miR-33a-5p, miR-3200-3p, and miR-6815-5p and increased levels of miR-146a-3p in patients with EP revealed unresolved inflammation, with accompanying increased expression of PCK1 and decreased expression of FCGR1A. Increased levels of miR-605-5p, miR-6791-5p, miR-6780a-5p, and miR-6826-5p and decreased levels of miR-16-1-3p (or 15a-3p) were associated with the degree of metastasis and led to the systemic activation of myeloid, endothelial, and epithelial cells, as well as neurons, phagocytes, and platelets. Log2FCs in the expression of miRNAs and mRNAs from plasma exosomes after CCRT are associated with EP and metastasis, reflecting unresolved inflammation and systemic microenvironmental factors, respectively. However, this study, supported by preliminary data insufficient to reach clear conclusions, should be verified in larger prospective cohorts.

scholarly journals Exosomes derived from miR-126-3p-overexpressing synovial fibroblasts suppress chondrocyte inflammation and cartilage degradation in a rat model of osteoarthritis

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Yan Zhou ◽  
Jianghua Ming ◽  
Yaming Li ◽  
Bochun Li ◽  
Ming Deng ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Apoptotic Cell ◽  
Synovial Fibroblasts ◽  
Cartilage Degeneration ◽  
Cartilage Degradation ◽  
Exosomal Mirnas ◽  
Exosomal Mirna ◽  
And Control ◽  
And Cartilage

AbstractMicroRNAs (miRNAs) encapsulated within exosomes can serve as essential regulators of intercellular communication and represent promising biomarkers of several aging-associated disorders. However, the relationship between exosomal miRNAs and osteoarthritis (OA)-related chondrocytes and synovial fibroblasts (SFCs) remain to be clarified. Herein, we profiled synovial fluid-derived exosomal miRNAs and explored the effects of exosomal miRNAs derived from SFCs on chondrocyte inflammation, proliferation, and survival, and further assessed their impact on cartilage degeneration in a surgically-induced rat OA model. We identified 19 miRNAs within synovial fluid-derived exosomes that were differentially expressed when comparing OA and control patients. We then employed a microarray-based approach to confirm that exosomal miRNA-126-3p expression was significantly reduced in OA patient-derived synovial fluid exosomes. At a functional level, miRNA-126-3p mimic treatment was sufficient to promote rat chondrocyte migration and proliferation while also suppressing apoptosis and IL-1β, IL-6, and TNF-α expression. SFC-miRNA-126-3p-Exos were able to suppress apoptotic cell death and associated inflammation in chondrocytes. Our in vivo results revealed that rat SFC-derived exosomal miRNA-126-3p was sufficient to suppress the formation of osteophytes, prevent cartilage degeneration, and exert anti-apoptotic and anti-inflammatory effects on articular cartilage. Overall, our findings indicate that SFC exosome‐delivered miRNA-126-3p can constrain chondrocyte inflammation and cartilage degeneration. As such, SFC-miRNA-126-3p-Exos may be of therapeutic value for the treatment of patients suffering from OA.

scholarly journals A facile graphene oxide based sensor for electrochemical detection of prostate anti-cancer (anti-testosterone) drug flutamide in biological samples

RSC Advances ◽  
2017 ◽  
Vol 7 (41) ◽  
pp. 25702-25709 ◽  
Author(s):  
R. Karthik ◽  
Mani Govindasamy ◽  
Shen-Ming Chen ◽  
Tse-Wei Chen ◽  
J. Vinoth kumar ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Electrochemical Sensor ◽  
Electrochemical Detection ◽  
Glassy Carbon ◽  
Biological Samples ◽  
Cancer Drug ◽  
Carbon Electrode ◽  
Modified Glassy Carbon Electrode ◽  
Anti Cancer

An electrochemical sensor based on graphene oxide modified glassy carbon electrode for the determination of anti-cancer drug flutamide.

scholarly journals Milk exosome-derived miRNAs from water buffalo are implicated in immune response and metabolism process

2020 ◽  
Author(s):  
Zujing Chen ◽  
Yueqin Xie ◽  
Junyi Luo ◽  
Ting Chen ◽  
Qianyun Xi ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Buffalo Milk ◽  
Cow Milk ◽  
Lactation Period ◽  
Bioactive Substances ◽  
Physiological Importance ◽  
Exosomal Mirnas ◽  
Potential Applications ◽  
Multiple Species ◽  
Exosomal Mirna

Abstract Background: Buffalo milk is rich in various nutritional components and bioactive substances that provide more essential health benefits to human body. Recently, exosome identified in the breast milk has been reported as a neotype nutrient and can mediate intercellular communication with exosomal miRNAs. In the present study, we therefore hypothesized that exosome-derived miRNAs from buffalo milk would play the potential physiological importance of consumption of buffalo milk.Results: We isolated exosomes from buffalo and cow milk samples that were obtained at mid-lactation period, and the exosomal miRNA profiles were then generated using miRNA-seq. In addition, miRNAomes of pig, human and panda milk exosomes were downloaded from GEO database. Finally, a total of 27 milk exosomal miRNA profiles that included 4 buffalo, 4 cow, 8 pig, 4 human and 7 panda were analyzed using the miRDeep2 program. A total of 558 unique miRNA candidates existed across all species, and the top 10 highly expressed miRNA were evolutionarily conserved across multiple species. Functional analysis revealed that these milk enriched miRNAs targeted 400 putative sites to modulate disease resistance, immune responsiveness and basic metabolism events. In addition, a total of 32 miRNAs in buffalo milk were significantly up-regulated compared with non-buffalo milks, while 16 were significantly down-regulated. Of interest, functional analysis showed that up-regulated miRNA were mainly related to host metabolism processes, while the predicted functions of down-regulated miRNAs were enriched in immune response.Conclusion: In this study, we explored the exosomal miRNAome differences between milks of different animals, expanding the theoretical basis for potential applications of the miRNA-containing vesicles.

scholarly journals Differential Expression of Serum Exosomal miRNAs in Breast Cancer Patients and Healthy Controls

2021 ◽  
Author(s):  
Rahim Asgari ◽  
Jafar Rezaie
Keyword(s):  
Breast Cancer ◽  
Cancer Patients ◽  
Circulatory System ◽  
Health Concern ◽  
Healthy Controls ◽  
Flow Cytometry Analysis ◽  
Breast Cancer Patients ◽  
Serum Samples ◽  
Exosomal Mirnas ◽  
And Control

Purpose: Breast cancer has become as a serious public health concern worldwide. Breast cancer cells release exosomes into the circulatory system, which are easily accessible for further analysis like cancer diagnosis. In this study, we aimed to investigate expression of circulating exosomal miRNAs (miRs) in the serum of individuals with breast cancer and healthy controls. Methods: Exosomes were collected from serum samples using a commercial kit and characterized by scanning electron microscopy (SEM) and flow cytometry analysis. Expression of miRs such as miR-21, miR-155, miR-182, miR-373, and miR-126 were evaluated by real-time PCR. Results: The result showed that the expression level of exosomal miR-21, miR-155, miR-182, and miR-373 in the serum of breast cancer patients was higher than of those controls (P<0.05). However, expression of miR-126 did not change between breast cancer and control individuals (P>0.05). Conclusion: Our results showed a different miRs expression pattern between breast cancer and healthy samples, supposing potential biomarkers for breast cancer. Further studies focusing on these miRs are required to confirm our findings.

PSV-4 Determination of Deoxynivalenol (DON) Content in Biological Samples as an Indicator of DON Intake in Grower-finisher Pigs

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 206-207
Author(s):  
Michael O Wellington ◽  
Michael A Bosompem ◽  
Veronika Nagl ◽  
Daniel A Columbus
Keyword(s):  
Mass Spectrometry ◽  
Biological Samples ◽  
High Performance ◽  
Tandem Mass ◽  
Serum Samples ◽  
Blood Samples ◽  
Swine Diets ◽  
Direct Quantification ◽  
Serum And Urine

Abstract Due to difficulties in obtaining consistent and/or reliable measures of deoxynivalenol (DON) in complete swine diets, we investigated whether measuring DON in biological samples could be used as an indicator of DON ingestion in pigs. In this study, graded levels of DON (1, 3, or 5 ppm) were fed to grower-finisher pigs for a period of 77-d. On d 35 and 77 of the study, urine samples were quantitatively collected over a 24-h period and blood samples were collected between 3 – 4 h after the morning meal on each of those days for serum DON analysis. For direct quantification of DON in urine, high-performance liquid chromatography with tandem mass spectrometry was performed. For serum samples, indirect quantification of DON was performed via enzymatic hydrolysis. We observed that DON content in urine increased linearly as intake of DON increased (Fig.1A; P &lt; 0.05). Analysis of DON in serum follow a similar trend, where serum DON content was increased as DON intake increased (Fig.1B; P &lt; 0.05). An average of 30% of DON ingested was recovered as DON in urine over a 24-h period. In summary, there was a linear relationship between DON intake and DON content in both urine and blood serum, therefore, analyzing DON concentration in serum and urine could be used as a tool to estimate for DON exposure in pigs under controlled conditions.

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