10 research outputs found
Circulating miRNAs, isomiRs and small RNA clusters in human plasma and breast milk
Circulating small RNAs, including miRNAs but also isomiRs and
other RNA species, have the potential to be used as non-invasive
biomarkers for communicable and non-communicable diseases. This
study aims to characterize and compare small RNA profiles in
human biofluids. For this purpose, RNA was extracted from plasma
and breast milk samples from 15 healthy postpartum mothers.
Small RNA libraries were prepared with the NEBNext(R) small RNA
library preparation kit and sequenced in an Illumina HiSeq2000
platform. miRNAs, isomiRs and clusters of small RNAs were
annotated using seqBuster/seqCluster framework in 5 plasma and
10 milk samples that passed the initial quality control. The RNA
yield was 81 ng/mL [standard deviation (SD): 41] and 3985 ng/mL
(SD: 3767) for plasma and breast milk, respectively. Mean number
of good quality reads was 4.04 million (M) (40.01% of the reads)
in plasma and 12.5M (89.6%) in breast milk. One thousand one
hundred eighty two miRNAs, 12,084 isomiRs and 1,053 small RNA
clusters that included piwi-interfering RNAs (piRNAs), tRNAs,
small nucleolar RNAs (snoRNA) and small nuclear RNAs (snRNAs)
were detected. Samples grouped by biofluid, with 308 miRNAs,
1,790 isomiRs and 778 small RNA clusters differentially
detected. In summary, plasma and milk showed a different small
RNA profile. In both, miRNAs, piRNAs, tRNAs, snRNAs, and snoRNAs
were identified, confirming the presence of non-miRNA species in
plasma, and describing them for the first time in milk
Quality control and mapped reads to miRNAs [mean and (SD)], by biofluid.
<p>Quality control and mapped reads to miRNAs [mean and (SD)], by biofluid.</p
Main characteristics of study participants.
<p>Main characteristics of study participants.</p
Number of clusters and abundance in milk and plasma.
<p>Number of clusters and abundance in milk and plasma.</p
Relative abundance of miRNA and small RNA clusters.
<p>(A, C) Plasma, (B, D) milk. The x-axis represents miRNAs (A, B) or small RNA clusters (C, D) ordered according to their expression level and the y-axis represents the abundance as percentage of reads (%) (average of all samples). Error bar shows SD.</p
Number and relative abundance of different types of isomiRs, as combinations of different editions, in milk and plasma.
<p>Number and relative abundance of different types of isomiRs, as combinations of different editions, in milk and plasma.</p
Differential levels of miRNAs by biofluid—top 10 ordered by p value.
<p>Differential levels of miRNAs by biofluid—top 10 ordered by p value.</p
Differential levels of small RNA clusters by biofluid—top 10 ordered by p value.
<p>Differential levels of small RNA clusters by biofluid—top 10 ordered by p value.</p
Circulating miRNAs, isomiRs and small RNA clusters in human plasma and breast milk
Circulating small RNAs, including miRNAs but also isomiRs and other RNA species, have the potential to be used as non-invasive biomarkers for communicable and non-communicable diseases. This study aims to characterize and compare small RNA profiles in human biofluids. For this purpose, RNA was extracted from plasma and breast milk samples from 15 healthy postpartum mothers. Small RNA libraries were prepared with the NEBNext® small RNA library preparation kit and sequenced in an Illumina HiSeq2000 platform. miRNAs, isomiRs and clusters of small RNAs were annotated using seqBuster/seqCluster framework in 5 plasma and 10 milk samples that passed the initial quality control. The RNA yield was 81 ng/mL [standard deviation (SD): 41] and 3985 ng/mL (SD: 3767) for plasma and breast milk, respectively. Mean number of good quality reads was 4.04 million (M) (40.01% of the reads) in plasma and 12.5M (89.6%) in breast milk. One thousand one hundred eighty two miRNAs, 12,084 isomiRs and 1,053 small RNA clusters that included piwi-interfering RNAs (piRNAs), tRNAs, small nucleolar RNAs (snoRNA) and small nuclear RNAs (snRNAs) were detected. Samples grouped by biofluid, with 308 miRNAs, 1,790 isomiRs and 778 small RNA clusters differentially detected. In summary, plasma and milk showed a different small RNA profile. In both, miRNAs, piRNAs, tRNAs, snRNAs, and snoRNAs were identified, confirming the presence of non-miRNA species in plasma, and describing them for the first time in milk
Circulating miRNAs, isomiRs and small RNA clusters in human plasma and breast milk
Circulating small RNAs, including miRNAs but also isomiRs and other RNA species, have the potential to be used as non-invasive biomarkers for communicable and non-communicable diseases. This study aims to characterize and compare small RNA profiles in human biofluids. For this purpose, RNA was extracted from plasma and breast milk samples from 15 healthy postpartum mothers. Small RNA libraries were prepared with the NEBNext® small RNA library preparation kit and sequenced in an Illumina HiSeq2000 platform. miRNAs, isomiRs and clusters of small RNAs were annotated using seqBuster/seqCluster framework in 5 plasma and 10 milk samples that passed the initial quality control. The RNA yield was 81 ng/mL [standard deviation (SD): 41] and 3985 ng/mL (SD: 3767) for plasma and breast milk, respectively. Mean number of good quality reads was 4.04 million (M) (40.01% of the reads) in plasma and 12.5M (89.6%) in breast milk. One thousand one hundred eighty two miRNAs, 12,084 isomiRs and 1,053 small RNA clusters that included piwi-interfering RNAs (piRNAs), tRNAs, small nucleolar RNAs (snoRNA) and small nuclear RNAs (snRNAs) were detected. Samples grouped by biofluid, with 308 miRNAs, 1,790 isomiRs and 778 small RNA clusters differentially detected. In summary, plasma and milk showed a different small RNA profile. In both, miRNAs, piRNAs, tRNAs, snRNAs, and snoRNAs were identified, confirming the presence of non-miRNA species in plasma, and describing them for the first time in milk