Similar Datasets
Project description:To better characterise adipose cells from a child with ROHHAD syndrome, we performed next-generation sequencing (NGS) and bioinformatic analyses on these cells compared to tissue from an age- and sex-matched control. The two most up-/down-regulated genes were validated using QPCR.
2023-07-19 | GSE208445 | GEO
Project description:Most proteogenomic approaches for mapping single amino acid polymorphisms (SAPs) require construction of a sample-specific database containing protein variants predicted from the next-generation sequencing (NGS) data. We present a new strategy for direct SAP detection without relying on NGS data. Among the 348 putative SAP peptides identified in an industrial yeast strain, 85.6% of SAP sites were validated by genomic sequencing.
2018-04-18 | PXD003101 | Pride
Project description:Cushing’s syndrome (CS) is a rare disease with high morbidity and mortality. Diagnosis and subtyping are complex and challenging. Circulating microRNAs were described to be useful as minimally invasive diagnostic markers. Our aim was to determine and compare the circulating microRNA expression profiles of patients with CS and controls. We included three groups of patients of the German Cushing’s registry: A.) patients with florid adrenal dependent CS scheduled for adrenalectomy (CPA); B.) patients with florid pituitary dependent CS scheduled for surgery (CD); and C.) patients in whom CS had been ruled out (controls). Next-generation sequencing was performed in the 30 CS serum samples before and after curative surgery, respectively, and in 10 baseline samples of controls. No significant differential expression was observed between all the CS samples and controls by NGS as well as by QPCR. The sequencing of the preoperative samples revealed a significant differential expression of miR-182-5p (p=0.02) between CD and controls. The differential expression was validated by QPCR in the discovery cohort and in an independent validation cohort. MiR-96-5p, miR-146b-5p, miR-183-5p, miR-185-5p, miR-616-5p and miR-629-5p were found to be significantly upregulated in CPA samples in comparison to CD group of the preoperative group in NGS. However, similar upregulation pattern could not be observed by QPCR in the discovery and validation cohorts. Additionally, miR-96-5p and miR-185-5p were found to be modulated by dexamethasone in controls. Thus, our study reports miR-182-5p as a possible biomarker for CD, which has to be validated in a prospective cohort.
2020-08-25 | GSE156693 | GEO
Project description:Next-generation RNA sequencing (NGS) was used for the combinatorial analysis of mRNA-miRNA gene expression profiles in sacral chordoma and nucleus pulposus samples, and to predict miRNA-mRNA regulatory networks with altered activity in chordoma. We identified a gene set including key regulators of the Hippo pathway, which is targeted by differently expressed miRNAs, and validated their altered expression by RT-qPCR. These newly identified miRNA/RNA interactions are predicted to have a role in the self-renewal process of chordoma stem cells, which might sustain the high rate of recurrence for this tumor.
2022-07-28 | GSE209695 | GEO
Project description:Interventions: An observational study at the Dutch Screening for Breast Cancer will be performed in 66 postmenopausal women without breast cancer. By acquiring insight into the intestinal microbiota composition of postmenopausal women without breast cancer, a control group will be set up for already existing research lines in microbiota research in breast cancer patients at MUMC+. Fecal samples and questionnaires will be collected. The intestinal microbiota composition and absolute abundance of the fecal samples will be analyzed by with 16S rRNA Next Generation Sequencing (NGS) with subsequent qPCR to convert relative abundance to absolute abundance.
Primary outcome(s): The primary endpoints include the microbiota composition.
Study Design: N/A , unknown, Other
| 21684 | ecrin-mdr-crc
Project description:Evaluting HLA loss of heterozygosity, a key immune escape mechanism, using NGS and immunopeptidomics.
2021-11-04 | PXD022323 | Pride
Project description:Background: Extramedullary disease (EM) in multiple myeloma (MM) is defined by escape of malignant plasma cells out of the bone marrow. The mechanisms of extramedullary spread are not well understood. MicroRNAs (miRNA) were proven to be involved in the pathogenesis of MM; nevertheless, there is a lack of knowledge about their role in EM. Methods: Using NGS analysis, we identified ten differentially expressed miRNA in plasma cells of MM versus EM patients. We performed qPCR in validation phase, using TaqMan Advanced MiRNA Assays. Results: In this study, we identified 10 miRNAs differentially expressed between MM and EM patiens.
2022-12-14 | GSE129918 | GEO
Project description:Purpose: In testis the effects of exposure to mixtures of Endocrine disruptors compounds (EDCs) upon expression of miRNAs were not addressed. Objective: To identify the expression profiles of the 'miRNome' in mice testis chronic exposed to a defined mixture of five EDCs. Methods: Pregnant mice from 0.5 post-coital day were exposed in the drinking water to a mixture containing 0.3 mg/Kg-bw/day of each phthalate (DEHP, DBP, BBP), plus 0.05 mg/Kg-bw/day of each alkylphenol (NP, OP) until adulthood of male mouse (60 days old). We characterized the 'miRNome' by next generation sequence (NGS). Results: In mouse testis exposed to EDCs mixture we detected by NGS 2 up-regulated and 8 down-regulated miRNAs along to 36 isomiRs differentially expressed; these results were validated by RT-qPCR. and functional analysis showed deregulation of testicular hormonal status, spermatogenesis disruption and germ cells apoptosis. Conclusions: Here we provide the first association between deregulation of miRNAs, isomiRs, with histopathological and hormonal alterations in adult mice testis exposed to mixture of EDCs.
2017-07-31 | GSE84695 | GEO
Project description:Characterization of microorganisms and quantification of histamine present in Whitemouth croaker (Micropogonias furnieri) by NGS, qPCR and HPLC-DAD
| PRJEB33351 | ENA
Project description:Libraries of single-stranded oligodeoxynucleotides (ssODNs) can be enriched for sequences that specifically bind molecules on naïve complex biological samples like cells or tissues. Depending on the enrichment strategy, the ssODNs can identify molecules specifically associated with a defined biological condition, for example a pathological phenotype, and thus are potentially useful for biomarker discovery. We performed ADAPT, a variant of SELEX, on exosomes secreted by VCaP prostate cancer cells. A library of ~1011 ssODNs was enriched for those that bind to VCaP exosomes and discriminate them from exosomes derived from LNCaP prostate cancer cells. Next-generation sequencing (NGS) identified the best discriminating ssODNs, nine of which were resynthesized and their discriminatory ability confirmed by qPCR. Affinity purification with one of the sequences (Sequence 7) combined with LC-MS/MS identified its molecular target complex, whereof most proteins are part of or associated with the multiprotein ESCRT complex participating in exosome biogenesis. Within this complex, YBX1 was identified as the directly bound target protein. ADAPT thus is able to differentiate exosomes from cancer cell subtypes from the same lineage. The composition of ESCRT complexes in exosomes from VCaP versus LNCaP cells might constitute a discriminatory element between these prostate cancer subtypes.
2020-01-13 | PXD011496 | Pride