scholarly journals Diagnostic and prognostic potential of eight whole blood microRNAs for equine sarcoid disease

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0261076
Author(s):  
Jeanne Cosandey ◽  
Eman Hamza ◽  
Vinzenz Gerber ◽  
Alessandra Ramseyer ◽  
Tosso Leeb ◽  
...  
Keyword(s):  
Whole Blood ◽  
Biomarker Discovery ◽  
Quantitative Polymerase Chain Reaction ◽  
Multivariable Analysis ◽  
Young Male ◽  
Microrna Expression ◽  
Initial Examination ◽  
Expression Levels ◽  
Diagnosis And Prognosis ◽  
Equine Sarcoid

MicroRNAs have been proposed as biomarkers for equine sarcoids, the most prevalent equine skin tumors globally. This study served to validate the diagnostic and prognostic potential of whole blood microRNAs identified in a previous study for long-term equine sarcoid diagnosis and outcome prediction. Based on findings of a clinical examination at the age of 3 years and a follow-up following a further 5–12 years, 32 Franches-Montagnes and 45 Swiss Warmblood horses were assigned to four groups: horses with regression (n = 19), progression (n = 9), new occurrences of sarcoid lesions (n = 19) and tumor-free control horses (n = 30). The expression levels for eight microRNAs (eca-miR-127, eca-miR-432, eca-miR-24, eca-miR-125a-5p, eca-miR-134, eca-miR-379, eca-miR-381, eca-miR-382) were analyzed through reverse transcription quantitative polymerase chain reaction in whole blood samples collected on initial examination. Associations of sex, breed, diagnosis, and prognosis with microRNA expression levels were examined using multivariable analysis of variance. Sex and breed influenced the expression level of five and two microRNAs, respectively. Eca-miR-127 allowed discrimination between sarcoid-affected and tumor-free horses. No variation in microRNA expression was found when comparing horses with sarcoid regression and progression. Expression levels of eca-miR-125a-5p and eca-miR-432 varied in male horses that developed sarcoids throughout the study period in comparison to male control horses. While none of the investigated miRNAs was validated for predicting the prognosis of sarcoid regression / progression within young horses with this condition, two miRNAs demonstrated potential to predict if young male (though not female) tumor-free horse can develop sarcoids within the following years. Sex- and breed- biased miRNAs exist within the equine species and have an impact on biomarker discovery.

scholarly journals Important considerations for microRNA extraction methods from whole blood and peripheral blood mononuclear cells

F1000Research ◽  
2014 ◽  
Vol 3 ◽  
pp. 183
Author(s):  
Sadaf Atarod ◽  
Hannah Smith ◽  
Anne Dickinson ◽  
Xiao-Nong Wang
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Whole Blood ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Extraction Methods ◽  
Microrna Expression ◽  
Peripheral Blood Mononuclear ◽  
Total Rna ◽  
Expression Levels ◽  
Blood Mononuclear Cells

MicroRNAs are non-coding RNAs that negatively regulate mRNA expression and play significant roles in both health and disease. Differential microRNA expression has been used to aid diagnosis and discriminate disease stages. The accuracy and reliability of microRNA expression measurement is of utmost importance. For diagnostic investigations, microRNA expression in human peripheral blood is commonly detected using total RNA extracted using different methods. To date, no convincing data have been available showing whether microRNA expression levels are comparable when total RNA has been extracted from whole blood or peripheral blood mononuclear cells (PBMCs). The present study examined miR-146a-5p and miR-155-5p expression using total RNA extracted in parallel from whole blood and PBMCs of 14 healthy volunteers. MicroRNA expression levels were significantly different between whole blood and PBMCs. No significant difference was observed in microRNA expression between fresh and cryopreserved PBMCs (p=0.125 for both). Further observations revealed that gender differences did not influence miR-146a-5p or miR-155-5p expression regardless of using whole blood(p = 0.797 and 1.00 respectively) or PBMC (p = 0.190 and 0.898 respectively). Our results demonstrate that microRNA expression could be subjective to the methods used for total RNA extraction which highlights the importance of using uniform extraction methods.

Measuring MicroRNA Expression Levels in Oncology: from Samples to Data Analysis

2013 ◽  
Vol 18 (4) ◽  
pp. 273-287 ◽  
Author(s):  
Loris De Cecco ◽  
Matteo Dugo ◽  
Silvana Canevari ◽  
Maria Grazia Daidone ◽  
Maurizio Callari
Keyword(s):  
Data Analysis ◽  
Microrna Expression ◽  
Expression Levels

Expression Levels of MicroRNA-125b in Serum Exosomes of Patients with Asthma of Different Severity and its Diagnostic Significance

2019 ◽  
Vol 20 (10) ◽  
pp. 781-784 ◽  
Author(s):  
Meizhen Zhao ◽  
Li Juanjuan ◽  
Fan Weijia ◽  
Xie Jing ◽  
Huang Qiuhua ◽  
...  
Keyword(s):  
Correlation Analysis ◽  
Roc Curve ◽  
Quantitative Polymerase Chain Reaction ◽  
Persistent Asthma ◽  
Asthma Severity ◽  
Control Group ◽  
Spearman Correlation ◽  
Diagnostic Efficacy ◽  
Expression Levels ◽  
Serum Exosomes

Background: This study aimed to investigate the expression levels of microRNA (miRNA)-125b in serum exosomes and its diagnostic efficacy for asthma severity. Methods: The study included 80 patients with untreated asthma and 80 healthy volunteers. The patients were divided into 4 groups according to disease severity: 20 with the intermittent state, 20 with the mildly persistent state, 20 with the moderately persistent state, and 20 with the severely persistent state. The expression levels of miRNA-125b in serum exosomes of each group were detected using a quantitative polymerase chain reaction and compared. The Spearman correlation analysis was used to study the correlation between the expression levels of miRNA-125b in serum exosomes and asthma severity. The diagnostic efficacy of the expression levels of miRNA-125b in exosomes for asthma severity was evaluated using the Receiver Operating Characteristic (ROC) curve. Results: The expression levels of miRNA-125b in serum exosomes of patients with intermittent, mildly persistent, moderately persistent, and severely persistent asthma were all higher than those in the healthy control group, with statistically significant differences. The expression levels of miRNA-125b were also statistically significantly different among patients in each group. The Spearman correlation analysis showed a positive correlation of the relative expression of miRNA-125b in serum exosomes with asthma severity. The area under the ROC curve of the diagnostic efficacy of miRNA-125b in serum exosomes for patients with intermittent, mildly, moderately, and severely persistent asthma was 0.7770, 0.8573, 0.9111, and 0.9995, respectively. Conclusion: The expression levels of miRNA-125b in serum exosomes had a high diagnostic efficacy and might serve as a noninvasive diagnostic marker for asthma severity.

scholarly journals Different macrophages equally induce EMT in endometria of adenomyosis and normal

Reproduction ◽  
2017 ◽  
Vol 154 (1) ◽  
pp. 79-92 ◽  
Author(s):  
Min An ◽  
Dong Li ◽  
Ming Yuan ◽  
Qiuju Li ◽  
Lu Zhang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Transforming Growth Factor ◽  
Epithelial Mesenchymal Transition ◽  
Quantitative Polymerase Chain Reaction ◽  
Immunohistochemical Analysis ◽  
Secretory Phase ◽  
Normal Endometrium ◽  
Mesenchymal Transition ◽  
Expression Levels ◽  
Endometrial Cells

Endometrial cells and microenvironment are two important factors in the pathogenesis of adenomyosis. Our previous study demonstrated that macrophages can induce eutopic epithelial cells of adenomyosis to suffer from epithelial–mesenchymal transition (EMT). The aim of this study is to detect whether macrophages interacting with epithelial cells equally induce the EMT process in normal and eutopic endometria of healthy and adenomyotic patients; and whether macrophages parallelly polarize to M2. We investigated the expression levels of epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), cytokeratin7 (CK7), vimentin, transforming growth factor-β1 (TGFB1), SMAD3 and pSMAD3 using immunohistochemistry and western blot, and then estimated the genetic levels of CD163, IL10 and MMP12 using real-time quantitative polymerase chain reaction (RT-PCR) in macrophages. Eutopic and normal endometrial tissues were obtained from 20 patients with adenomyosis and 11 control patients without adenomyosis, respectively. The immunohistochemical analysis shows distinct EMT in eutopic endometria in secretory phase; the expression levels of TGFB1, SMAD3 and pSMAD3 that indicate signal pathway of EMT were also higher in secretory phase. Macrophages can induce EMT process in primary endometrial epithelial cells derived from normal and eutopic endometria. After co-culturing, THP-1-derived macrophages polarized to M2. Compared with the eutopic endometrium group, further polarization to M2 was observed in the normal endometrium group. These results indicate that adenomyosis may be promoted by the pathologic EMT of epithelial cells, which is induced by macrophages that incapably polarize to M2.

Expression of Cannabinoid Receptors in Myometrium and its Correlation With Dysmenorrhea in Adenomyosis

2019 ◽  
Vol 26 (12) ◽  
pp. 1618-1625 ◽  
Author(s):  
Xue Shen ◽  
Hua Duan ◽  
Sha Wang ◽  
Wei Hong ◽  
Yu-Yan Wang ◽  
...  
Keyword(s):  
Messenger Rna ◽  
Cannabinoid Receptors ◽  
Pain Perception ◽  
Cannabinoid Receptor ◽  
Quantitative Polymerase Chain Reaction ◽  
Premenopausal Women ◽  
Type I ◽  
Junctional Zone ◽  
Tissue Samples ◽  
Expression Levels

The myometrium, especially the junctional zone (JZ), is now well documented to have a role in the pathogenesis of adenomyosis. Cannabinoid receptors have been shown to participate in the establishment of endometriosis and its pain perception. However, its relation to adenomyosis has not been identified yet. The aim of this study was to investigate the expression of cannabinoid receptor type I (CB1) and type II (CB2) in myometrium of uteri with and without adenomyosis and determine the correlation between their levels and clinical parameters of adenomyosis. We collected tissue samples of JZ and the outer myometrium from 45 premenopausal women with adenomyosis and 34 women without adenomyosis. CB1 and CB2 messenger RNA (mRNA) and protein expression levels were evaluated by the use of Western blotting and real-time quantitative polymerase chain reaction from all samples. Clinical information on the severity of dysmenorrhea and other data were collected. We found both CB1 and CB2 mRNA and protein levels in women with adenomyosis were significantly higher than those of controls, and CB1 expression levels in JZ were positively correlated with the severity of dysmenorrhea. These data suggest that cannabinoid receptor CB1 may be involved in the pathogenesis of dysmenorrhea in adenomyosis and may be a potential therapeutic target.

Migrating Progenitor Cells Derived From Injured Cartilage Surface Respond to Damage-Associated Molecular Patterns

Cartilage ◽  
2021 ◽  
pp. 194760352110495
Author(s):  
Lei Ding ◽  
Cheng Zhou ◽  
Hongjun Zheng ◽  
Quanming Wang ◽  
Haiyan Song ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Progenitor Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Critical Role ◽  
Cartilage Degeneration ◽  
Expression Levels ◽  
Chondrogenic Progenitor Cells ◽  
The Difference ◽  
Molecular Patterns ◽  
Damage Associated Molecular Patterns

Objective: To delineate the response of migrating chondrogenic progenitor cells (CPCs) that arose from the surface of mechanically injured articular cartilage to proinflammatory damage-associated-molecular-patterns (DAMPs). Design: Bovine CPCs and non-CPC chondrocytes isolated from either impacted or scratched articular cartilage were studied. Those 2 types of cells were treated with mitochondrial DAMPs (MTDs; 10 nM fMLF and 10 µg/mL CpG DNA), or 10 nM HMGB1, or 10 ng/mL IL-1b for 24 hours. At the end of experiments, conditioned media and cell lysates were collected for analysis of expression levels of matrix metalloproteinases (MMPs), chemokines, and cytokines that are associated with cartilage degeneration with Western blotting and quantitative polymerase chain reaction. The difference of expression levels was compared by Welch’s t-test. Results: Our data indicated that HMGB1 and MTDs remarkably upregulated pro-MMP-13 expression in CPCs. Compared with non-CPCs, CPCs expressed significantly more baseline mRNAs of MMP-13, CXCL12, and IL-6. MTDs greatly increased the expression of MMP-13 and IL-6 in CPCs by over 100-fold ( P < 0.001). MTDs also significantly increased IL-8 expression in CPCs to a similar extent ( P < 0.001). However, when IL-1b was present, CPCs expressed less MMP-3 and active MMP-13 proteins as well as less CCL2 and IL-6 than did non-CPCs. Conclusions: We concluded that CPCs were more sensitive than non-CPCs in response to DAMPs, especially MTDs. The proinflammatory nature of CPCs implied their critical role in the early phase of posttraumatic osteoarthritis development.

scholarly journals Abnormal MicroRNA Expression in Ts65Dn Hippocampus and Whole Blood: Contributions to Down Syndrome Phenotypes

2011 ◽  
Vol 33 (5) ◽  
pp. 451-467 ◽  
Author(s):  
Jennifer Keck-Wherley ◽  
Deepak Grover ◽  
Sharmistha Bhattacharyya ◽  
Xiufen Xu ◽  
Derek Holman ◽  
...  
Keyword(s):  
Down Syndrome ◽  
Whole Blood ◽  
Microrna Expression

scholarly journals Prognostic value of microRNA expression levels in pancreatic adenocarcinoma: a review of the literature

Oncotarget ◽  
2017 ◽  
Vol 8 (42) ◽  
pp. 73345-73361 ◽  
Author(s):  
Patrick Wald ◽  
X. Shawn Liu ◽  
Cory Pettit ◽  
Mary Dillhoff ◽  
Andrei Manilchuk ◽  
...  
Keyword(s):  
Pancreatic Adenocarcinoma ◽  
Prognostic Value ◽  
Microrna Expression ◽  
Review Of The Literature ◽  
Expression Levels

scholarly journals Whole Blood Holding Time Prior to Plasma Processing Alters microRNA Expression Profile

2022 ◽  
Vol 12 ◽  
Author(s):  
Sung Hye Kim ◽  
David A. MacIntyre ◽  
Lynne Sykes ◽  
Maria Arianoglou ◽  
Phillip R. Bennett ◽  
...  
Keyword(s):  
Mirna Expression ◽  
Whole Blood ◽  
Biomarker Discovery ◽  
Expression Profiles ◽  
Holding Time ◽  
Circulating Mirnas ◽  
Sample Collection ◽  
Aberrant Expression ◽  
Discovery Research ◽  
Mirna Expression Profiles

MicroRNAs (miRNAs) can exhibit aberrant expression under different physiological and pathological conditions. Therefore, differentially expressed circulating miRNAs have been a focus of biomarker discovery research. However, the use of circulating miRNAs comes with challenges which may hinder the reliability for their clinical application. These include varied sample collection protocols, storage times/conditions, sample processing and analysis methods. This study focused on examining the effect of whole blood holding time on the stability of plasma miRNA expression profiles. Whole blood samples were collected from healthy pregnant women and were held at 4°C for 30 min, 2 h, 6 h or 24 h prior to processing for plasma isolation. Plasma RNA was extracted and the expression of 179 miRNAs were analyzed. Unsupervised principal component analysis demonstrated that whole blood holding time was a major source of variation in miRNA expression profiles with 53 of 179 miRNAs showing significant changes in expression. Levels of specific miRNAs previously reported to be associated with pregnancy-associated complications such as hsa-miR-150-5p, hsa-miR-191-5p, and hsa-miR-29a-3p, as well as commonly used endogenous miRNA controls, hsa-miR-16-5p, hsa-miR-25-3p, and hsa-miR-223-3p were significantly altered with increase in blood holding time. Current protocols for plasma-based miRNA profiling for diagnostics describe major differences in whole blood holding periods ranging from immediately after collection to 26 h after. Our results demonstrate holding time can have dramatic effects on analytical reliability and reproducibility. This highlights the importance of standardization of blood holding time prior to processing for plasma in order to minimize introduction of non-biological variance in miRNA profiles.

scholarly journals Oral Dextran Sulfate Sodium Administration Induces Peripheral Spondyloarthritis Features in SKG Mice Accompanied by Intestinal Bacterial Translocation and Systemic Th1 and Th17 Cell Activation.

2021 ◽  
Author(s):  
Yuya Tabuchi ◽  
Masao Katsushima ◽  
Yuri Nishida ◽  
Mirei Shirakashi ◽  
Hideaki Tsuji ◽  
...  
Keyword(s):  
Bacterial Translocation ◽  
Whole Blood ◽  
Th17 Cell ◽  
Cell Activation ◽  
Dextran Sulfate ◽  
Dextran Sulfate Sodium ◽  
Quantitative Polymerase Chain Reaction ◽  
Bacterial Species ◽  
Bacterial Dna ◽  
Anti Fungal

Abstract Background: Spondyloarthritis (SpA) is an autoimmune and autoinflammatory musculoskeletal disease characterised by systemic enthesitis. Recent research has focused on subclinical inflammatory bowel disease (IBD) in SpA pathogenesis. SKG mice, harbouring the Zap70 W163C mutation, increase autoreactive Th17 cells intrinsically, and show SpA features, including enteritis, after peritoneal injection of β-1,3- glucan under SPF conditions. In a conventional environment, they exhibit spontaneous arthritis with fungal factors. This study aimed to clarify whether oral dextran sulfate sodium (DSS) administration, utilised in IBD model mice, can provoke SpA features in SKG mice under SPF conditions, focusing on the relationship between gut microorganisms and SpA pathogenesis.Methods: SKG and BALB/c mice were administered oral DSS, and their body weights, arthritis, and enthesitis scores were recorded. In another cohorts, antibiotics (meropenem and vancomycin) or an anti-fungal agent (amphotericin B) were administered orally before DSS administration. The splenic Th1 and Th17 cell populations were examined before and after DSS administration using flow cytometry. Furthermore, the amount of circulating bacterial DNA in whole blood was measured by absolute quantitative polymerase chain reaction (qPCR), and the number and characteristics of bacterial species corresponding to these circulating DNA were analised by next-generation sequencing (NGS).Results: Ankle enthesitis as a peripheral SpA feature was elicited in half of DSS-administered SKG mice, and none of the BALB/c mice. Pre-administration of antibiotics suppressed enthesitis, whilst an anti-fungal agent could not. Th1 and Th17 cell levels in the spleen increased after DSS administration, and this was suppressed by pre-administration of antibiotics. SKG mice have a larger amount of bacterial DNA in whole blood than BALB/c mice before and one day after the initiation of DSS administration. The number of bacterial species in whole blood increased after DSS administration in SKG and BALB/c mice. Some genera and species significantly specific to the DSS-treated SKG mice group were also detected. Conclusion: Oral DSS administration alone elicited peripheral enthesitis in SKG mice with bacterial translocation accompanied by increased splenic Th1 and Th17 cell levels. Pre-administration of antibiotics ameliorated these DSS-induced SpA features. These findings suggest that intestinal bacterial leakage plays a pivotal role in SpA pathogenesis.

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