Project description:In this study, we compared microRNA (miRNA) profiles of salivary exosomes of patients with oral lichen planus with those of healthy controls. Saliva samples from 16 patients with oral lichen planus and 8 healthy controls were divided into 2 sets and were examined by performing miRNA microarray analysis. Examination of 8 oral lichen planus patients and 4 healthy controls. Each patient and control represent pooled RNAs from salivary exosomes of 8 patients and 4 healthy controls, respectively. Please note that each set (i.e. set1 and set2) was analysed independently.
Project description:In this study, we compared microRNA (miRNA) profiles of salivary exosomes of patients with oral lichen planus with those of healthy controls. Saliva samples from 16 patients with oral lichen planus and 8 healthy controls were divided into 2 sets and were examined by performing miRNA microarray analysis. Examination of 8 oral lichen planus patients and 4 healthy controls. Each patient and control represent pooled RNAs from salivary exosomes of 8 patients and 4 healthy controls, respectively. Please note that each set (i.e. set1 and set2) was analysed independently.
Project description:miRNA profiling of human H9-derived neural stem cells (H9-NSCs) comparing control human adult dermal fibroblasts (hDFs), SOX2-transduced human induced neural stem cells (hDF-iNSC (SOX2)), SOX2/HMGA2-transduced human induced neural stem cells (hDF-iNSC (SOX2/HMGA2)). Goal was to determine the global miRNA expression between the groups. H9-NSC vs hDF vs hDF-iNSC(SOX2) vs hDF-iNSC(SOX2/HMGA2)
Project description:Circulating microRNAs have recently emerged as a new class of promising non-invasive cancer biomarkers. The purpose of this study was the identification by a high-throughput approach (miRNA microarray) of circulating miRNAs associated with breast cancer-derived distant metastasis. To achieve this goal, we resorted to archival plasma samples collected from patients in the control arm of a randomized clinical trial on stage I breast cancer. We compared plasma miRNA levels in patients that developed distant metastasis after a radical or conservative surgery and in those long-term disease-free. Microarray results were technically and independently validated by Real Time PCR. Subsequent in vitro, in vivo and in silico analyses were performed to investigate the miRNA biological/clinical significance in relation with breast cancer progression. We demonstrated that high circulating miR-1246 levels were associated to distant metastasis and that high tissue expression of this miRNA was correlated with unfavorable outcome in ER+HER2- breast cancer patients. In addition, miR-1246 expression was also found to be related with stem-like features. 64 samples (32 patients with metastasis and 32 patients with no metastasis) were considered from a total of 208 hybridized samples collected between 1987 and 2004 (including 94 paired samples according to tumor ER status, age at drawing, drawing year, time from surgery, 7 additional samples from 'no evidence of disease' (NED) patients, 10 references, 1 healthy donor and 2 replicated samples)
Project description:Independent studies have reported that circulating miRNAs have the potential as biomarkers; however, no consolidated guidelines for the discovery process are available. We developed a pipeline using innovative applications of existing bioinformatics methods to (1) face the inapplicability of the classical normalization methods, (2) detect global differences of miRNA distributions between the comparison classes and (3) develop a 'robustM-bM-^@M-^Y classifier. 78 samples (26 hemolyzed and 52 non hemolyzed) were 1:2 paired with caliper matching according to disease status, age at drawing and drawing year, starting from a total of 208 hybridized samples collected between 1987 and 2004 (including 94 paired samples according to tumor ER status, age at drawing, drawing year, time from surgery, 7 additional samples from 'no evidence of disease' (NED) patients, 10 references, 1 healthy donor and 2 replicated samples)
Project description:Skin biopsy specimens of skin lesions were profiled for miRNA expression. In this study, we indentified miRNA species that were differentially expressed in the skin lesions of either the lepromatous or tuberculoid forms of leprosy. One miRNA species, hsa-mir-21, found in the lepromatous lesions was capable of downregulating the vitamin D-dependent antimicrobial pathway. Scalpel or punch skin biopsy specimens were obtained after informed consent from patients with tuberculoid leprosy and patients with lepromatous leprosy at the time of diagnosis. Specimens were embedded in OCT medium, snap-frozen in liquid nitrogen and stored at 80°C until sectioning.
Project description:In this study, we examined alterations in microRNA (miRNA) profiles in peripheral blood from male medical students two months and two days before the National Examination for Medical Practitioners. Blood obtained one month after the examination were used as baseline controls. Several miRNAs were significantly elevated during the pre-examination period in association with significant down-regulation of their target mRNAs two days before the examination. Thus, a distinct group of miRNAs in periheral blood may participate in the integrated response to chronic academic stress in healthy young men Total RNA was prepared from venous blood which was taken from young male medical students. 12 samples from 4 subjects at three time points: two months before, two days before, and one month fater the examination were measured on array platform.
Project description:Using a microarray-based method we report on: i) adequate intra- and interarray reproducibility of miRNA profiling; ii) feasibility of using archival plasma samples stored for an extended period of time and available in limited amounts; and iii) good correlation between different batches. A total of 11 archived plasma samples collected between 1988 and 1999 were evaluated in this study. Firstly, RNA extracted from two samples (S1 and S2) were hybridized in triplicate. The same samples were also hybridized in triplicate after increasing the starting material volume from 2 to 2.5. Nine additional plasma samples were processed and RNA was hybridized on two different batches of arrays. Finally, two samples (S3 and S4) were re-hybridized on an array of the same batch of the first hybridization but close to the chip expiration date.
Project description:MicroRNAs (miRNAs) are known to be deregulated in human breast cancer (BC). The purpose of the current study was to investigate the expression of miRNAs in different stages of BC to assess their biological value in BC progression. MiRNA expression was assessed in a series of BC patients (n=7) with distinct stages of tumour progression (Normal, in-situ (DCIS), primary invasive BC and nodal metastases) to evaluate miRNA differential expression. We used an Agilent miRNA microarray based platform which uses miRBase 16 to screen for 1205 Homo sapiens (hsa) and 144 human viral miRNA candidates. To validate the microarray data, the expression of two deregulated miRNAs was measured by TaqMan quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). This study was conducted Formalin Fixed and Paraffin Embedded (FFPE) specimens from 7 female patients. Twenty M-NM-<m thick FFPE sections were cut and mounted on PALM Membrane Slides. Under direct microscopic visualization, tissues of interest were micro-dissected using PALM non-contact Laser catapulsion instrument (P.A.L.M. Microlaser Technologies, Carl Zeiss Ltd). Total RNA, including miRNAs, was extracted with the TRIzol reagent (Invitrogen, supplied by Fisher Scientific UK Ltd) according to the manufacturerM-bM-^@M-^Ys instructions. Total RNA was quantified with Quant-iT RiboGreen RNA Quantitation Kit (Fisher Scientific UK Ltd), which is an accurate method of measuring total RNA with a detection limit of 0.001-1ng/M-BM-5l. RNA purity and RNA integrity number (RIN) were determined on an Agilent 2100 Bioanalyzer and RNA 6000 NanoLabChip Kits (both Agilent Technologies, Santa Clara, USA), and the RIN of all the samples was >2 but <3. This is expected for FFPE breast samples and is acceptable by Agilent Technologies, Santa Clara, USA, for FFPE samples to undergo miRNA microarray analysis. MiRNAs extracted from laser microdissected tissue components were profiled using Agilent microRNA microarray profiling system (Agilent Technologies, Santa Clara, USA). Total RNA samples were spiked using the MicroRNA Spike-In Kit (Agilent Technologies, Santa Clara, USA) to evaluate efficiencies of labelling and hybridisation. Total RNA was treated with Calf Intestinal Alkaline Phosphatase (CIP), and then 100 ng of total RNA was used per sample to initiate a labelling reaction. Ligation master mix for T4 RNA ligase, which includes Cyanine 3-Cytidine biphosphate (Cyanine 3-pCp) (Complete miRNA Labelling and Hyb Kit, Agilent Technologies, Santa Clara, USA) was used to label the dephosphorylated RNA. Cyanine-3-labelled miRNA samples were hybridised to human miRNA microarrays (Release 16.0, 8x60K) (Agilent Technologies, Santa Clara, USA) at 55M-BM-0C for 20 hrs. Microarray slides were washed with increasing stringency (Gene Expression Wash Buffers, Agilent Technologies, Santa Clara), and subsequently dried with acetonitrile (Sigma-Aldrich, St. Louis, USA). Fluorescent signal intensities were detected on an Agilent Microarray Scanner (Agilent Technologies, Santa Clara, USA) using the Scan Control A.8.4.1 Software (Agilent Technologies, Santa Clara, USA) and extracted from the images with the Feature Extraction 10.7.3.1 Software (Agilent Technologies, Santa Clara, USA).
Project description:microRNAs levels were measured in brown adipose tissue of male C57Bl6N mice that were kept at RT or during cold exposure (8°C) for 24 hrs. Several miRNAs including myomirs were identified to be regulated in response to cold. Brown adipose tissue from 2 groups of 8 weeks old male C57Bl6N mice at room temperature (n=3) or after cold exposure n=3) at 8°C for 24 hrs were excised and RNA was extracted using Trizol and the manufactureres recommended procedure. miRNA expression levels were determined using Agilent microRNA expression analysis platform.