Comparison of Methodologies to Detect Low Levels of Hemolysis in Serum for Accurate Assessment of Serum microRNAs

Shah, Jaynish S.; Soon, Patsy S.; Marsh, Deborah J.

doi:10.1371/journal.pone.0153200

Cited by 158 publications

(149 citation statements)

References 33 publications

Supporting

Mentioning

146

Contrasting

Order By: Relevance

“…Stable reference (miR‐103a‐3p, miR‐191‐5p, miR‐425‐5p, miR‐223‐3p, miR‐423‐3p; Exiqon's biofluid guidelines and refs. and hemolysis (miR‐23a, miR‐451a) miRNA controls were included in the plasma analysis. The relative expression level of miRNAs was calculated using the 2 −ΔΔCt method.…”

Section: Methodsmentioning

confidence: 99%

Circulating Brain‐Enriched MicroRNAs for Detection and Discrimination of Idiopathic and Genetic Parkinson's Disease

et al. 2019

View full text Add to dashboard Cite

Background A minimally invasive test for early detection and monitoring of Parkinson's disease (PD) is a highly unmet need for drug development and planning of patient care. Blood plasma represents an attractive source of biomarkers. MicroRNAs (miRNAs) are conserved noncoding RNA molecules that serve as posttranscriptional regulators of gene expression. As opposed to ubiquitously expressed miRNAs that control house‐keeping processes, brain‐enriched miRNAs regulate diverse aspects of neuron development and function. These include neuron‐subtype specification, axonal growth, dendritic morphogenesis, and spine density. Backed by a large number of studies, we now know that the differential expression of neuron‐enriched miRNAs leads to brain dysfunction. Objectives The aim was to identify subsets of brain‐enriched miRNAs with diagnostic potential for familial and idiopathic PD as well as specify the molecular pathways deregulated in PD. Methods Initially, brain‐enriched miRNAs were selected based on literature review and validation studies in human tissues. Subsequently, real‐time reverse transcription polymerase chain reaction was performed in the plasma of 100 healthy controls and 99 idiopathic and 53 genetic (26 alpha‐synucleinA53T and 27 glucocerebrosidase) patients. Statistical and bioinformatics analyses were carried out to pinpoint the diagnostic biomarkers and deregulated pathways, respectively. Results An explicit molecular fingerprint for each of the 3 PD cohorts was generated. Although the idiopathic PD fingerprint was different from that of genetic PD, the molecular pathways deregulated converged between all PD subtypes. Conclusions The study provides a group of brain‐enriched miRNAs that may be used for the detection and differentiation of PD subtypes. It has also identified the molecular pathways deregulated in PD. © 2019 International Parkinson and Movement Disorder Society

show abstract

Section: Methodsmentioning

confidence: 99%

Circulating Brain‐Enriched MicroRNAs for Detection and Discrimination of Idiopathic and Genetic Parkinson's Disease

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Plasma samples were collected from the jugular vein in EDTA tubes (Becton Dickinson, USA) on Days 0, 8, 16 and 60 after insemination, as described before [23, 25]. The ratio between miR-451 and miR-23a was used as an indicator of haemolysis [26]; most samples had a miR ratio < 5, indicating absence of haemolysis (S1 Fig). All animal procedures were carried out under the UK Home Office Animals (Scientific Procedures) Act 1986, license 60/4604, with approval by the Ethical Review Committee, University of Edinburgh.…”

Section: Methodsmentioning

confidence: 99%

Changes in circulating microRNA levels can be identified as early as day 8 of pregnancy in cattle

Ioannidis

Donadeu

2017

PLoS ONE

View full text Add to dashboard Cite

Poor reproductive performance remains a major issue in the dairy industry, with low conception rates having a significant impact on milk production through extended calving intervals. A major limiting factor is the lack of reliable methods for early pregnancy diagnosis. Identification of animals within a herd that fail to conceive within 3 weeks after insemination would allow early re-insemination and shorten calving intervals. In a previous study, we found an increase in plasma miR-26a levels in Day 16-pregnant relative to non-pregnant heifers, however changes in miRNA levels that early during pregnancy were very small which likely prevented the identification of robust biomarkers. In this study, we extended our analyses to a wider interval during pregnancy (Days 8 to 60, n = 11 heifers) with the rationale that this may facilitate the identification of additional early pregnancy miRNA biomarkers. Using small RNA sequencing we identified a total of 77 miRNAs that were differentially expressed on Day 60 relative to Day 0 of pregnancy. We selected 14 miRNAs for validation by RT-qPCR and confirmed significant differences in the expression of let-7f, let-7c, miR-30c, miR-101, miR-26a, miR-205 and miR-143 between Days 0 and 60. RT-qPCR profiling throughout Days 0, 8, 16 and 60 of pregnancy showed a distinct increase in circulating levels of miR-26a (3.1-fold, P = 0.046) as early as Day 8 of pregnancy. In summary, in contrast to earlier stages of pregnancy (≤ Day 24), marked differences in the levels of multiple miRNAs can be detected in circulation by Day 60 in cattle. Retrospective analyses showed miR-26a levels to be increased in circulation as early as Day 8, sooner than previously reported in any species, suggesting a biological role for this miRNA in the very early events of pregnancy.

show abstract

“…PBS was used as a blank buffer. A free hemoglobin calibration curve was generated by measuring the absorption of various concentrations of pure human hemoglobin dissolved in PBS solution [28] and employed as a reference for experimental samples.…”

Section: Methodsmentioning

confidence: 99%

Trapped Chromatin Fibers Damage Flowing Red Blood Cells

et al. 2018

View full text Add to dashboard Cite

Neutrophils are the most abundant white blood cells in the circulation and serve antimicrobial functions. One of their antimicrobial mechanisms involves the release of neutrophil extracellular traps (NETs), long chromatin fibers decorated with antimicrobial granular proteins that contribute to the elimination of pathogens. However, the release of NETs has also been associated with disease processes. While recent research has focused on biochemical reactions catalyzed by NETs, significantly less is known about the mechanical effect of NETs in circulation. Here, microfluidic devices and biophysical models are employed to study the consequences of the interactions between NETs trapped in channels and red blood cells (RBCs) flowing in blood over the NETs. It has been found that the RBCs can be deformed and ruptured after interactions with NETs, generating RBC fragments. Significant increases in the number of RBC fragments have also been found in the circulation of patients with conditions in which NETs have been demonstrated to be present in circulation, including sepsis and kidney transplant. Further studies will probe the potential utility of RBC fragments in the diagnostic, monitoring, and treatment of diseases associated with the presence of NETs in circulation.

show abstract

Comparison of Methodologies to Detect Low Levels of Hemolysis in Serum for Accurate Assessment of Serum microRNAs

Cited by 158 publications

References 33 publications

Circulating Brain‐Enriched MicroRNAs for Detection and Discrimination of Idiopathic and Genetic Parkinson's Disease

Circulating Brain‐Enriched MicroRNAs for Detection and Discrimination of Idiopathic and Genetic Parkinson's Disease

Changes in circulating microRNA levels can be identified as early as day 8 of pregnancy in cattle

Trapped Chromatin Fibers Damage Flowing Red Blood Cells

Contact Info

Product

Resources

About