Journal: BioMed Research International

Article Title: MicroRNAs as Salivary Markers for Periodontal Diseases: A New Diagnostic Approach?

doi: 10.1155/2016/1027525

Figure Lengend Snippet: Comparison of methods from current investigations regarding miRNAs in periodontal disease.

Article Snippet: Naqvi et al. 2014 [ ] , Human THP-1-differentiated macrophages , miRNeasy kit (Qiagen) , NanoString nCounter miRNA assay (NanoString Technologies) , Quantitative real-time PCR EvaGreen Master Mix (Biotium) , — , RNU6B , Student's t -test (two-tailed) , miR-29b miR-32 miR-146a miR-891.

Techniques: Comparison, RNA Extraction, Biomarker Discovery, Control, In Vitro, Microarray, Isolation, TaqMan microRNA Assay, SYBR Green Assay, Labeling, Real-time Polymerase Chain Reaction, Virus, Quantitative RT-PCR, In Vivo, Expressing, Mann-Whitney U-Test

Journal: Bioengineered

Article Title: Curcumol enhances cisplatin sensitivity of gastric cancer: involvement of microRNA-7 and the nuclear factor-kappa B/snail family transcriptional repressor 1 axis

doi: 10.1080/21655979.2022.2070975

Figure Lengend Snippet: Curcumol upregulates miR-7 expression in GC cells. a, miRNAs with differential expression in cells after curcumol treatment identified by microarray analysis; b, miR-7 expression in cells after 80 μM curcumol treatment examined by RT-qPCR (* p < 0.05; two-way ANOVA); c, miR-7 expression in tumor and the paired normal tissues evaluated by RT-qPCR (n = 33; * p < 0.05; the unpaired t test); d, correlation between miR-7 expression and the patient’s survival (* p < 0.05; Kaplan-Meier analysis); e, miR-7 expression in MKN45 and HGC27 cells after different doses of curcumol treatment examined by RT-qPCR (* p < 0.05, ** p < 0.01; two-way ANOVA).

Article Snippet: This reaction was performed at 37°C for 30 min. After that, the labeled RNA was hybridized with Human miRNA Expression Microarray V4.0 (Arraystar, Rockville, MD, USA) for 24 h. The gene expression data were obtained using a GeneChip TM Scanner 3000 7 G system (#00-0210, 2008, Thermo Fisher Scientific) and analyzed by the R Language Program (Version 3.6.3, R).

Techniques: Expressing, Quantitative Proteomics, Microarray, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Hsa-miR-31 Governs T-Cell Homeostasis in HIV Protection via IFN-γ-Stat1-T-Bet Axis

doi: 10.3389/fimmu.2021.771279

Figure Lengend Snippet: miR-31 is correlated with disease progression during both acute and chronic HIV-1 infection. (A) Unsupervised clustering of the 251 miRNAs. After normalization and filtering of the microarray data, 251 miRNAs were retained for further analysis. Average linkage hierarchical clustering was performed using a centered correlation metric. Twenty-three samples from the FBD study were clustered into 2 groups: the left cluster was a mixture of elite controllers, viremic controllers and progressors; the right was mainly progressors with one exception. (B) Venn diagram showing the numbers of candidate miRNAs filtered with different criteria. The miRNAs in the lower left and right circles were generated by significance analysis of microarrays (SAM) of participants stratified by the CD4+ T cell count (<250 cells/μL vs. >450 cells/μL) and viral load (<2000 copies/mL vs. >10000 copies/mL), respectively. The identified 15 miRNA candidates were marked in red in (A) . (C) Correlation between expression levels of miR-31 and CD4+ T cell counts in HIV-1 infected individuals (FBD, former blood donor cohort). miR-31 expression was quantified by quantitative RT-PCR, and the relationship between relative level of miR-31 and CD4+ T cell count was examined by Spearman correlation (n = 50). Red dots represent patients that eventually reached the defined endpoints. (D) Kaplan-Meier survival curves of FDB patients stratified by median whole blood miR-31 level during the late phase of chronic infection. (E–G) Kaplan-Meier survival curves of another HIV patient cohort (an acute-phase prospective men who have sex with men (MSM) cohort) stratified by plasma miR-31 levels before and after infection. Absolute CD4+ T cell count below 350 cells/μL, initiation of long-term ART, progression to AIDS and death were defined as endpoints of the study. Patients were separated into two groups stratified by the median miR-31 level in plasma collected before infection (E) , during acute infection phase (F) , during early phase of chronic infection (G) .

Article Snippet: PAXgene Blood miRNA Kit (QIAGEN, Hilden, Germany) was used for extraction and purification of total RNA, including miRNA, from whole blood stabilized in PAXgene Blood RNA Tubes (PreAnalytix, BD, UK).

Techniques: Biomarker Discovery, Infection, Microarray, Generated, Cell Counting, Expressing, Quantitative RT-PCR, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: Hsa-miR-31 Governs T-Cell Homeostasis in HIV Protection via IFN-γ-Stat1-T-Bet Axis

doi: 10.3389/fimmu.2021.771279

Figure Lengend Snippet: Loss of miR-31 triggers CD4+ T cell activation. (A) miR-31 levels in different immune cell subtypes. Data were obtained from a miRNA RTqPCR data from Rossi et al ’s work (see the text for reference). (B) Comparison of absolute naïve CD4+ T cell counts in blood of HIV-1 infected individuals (FBD, n = 50) stratified by miR-31 expression. (C, D) Correlation between miR-31 levels and frequencies of CD38+ T cells (C) or HLA-DR+ T cells in blood of HIV infected individuals (FBD, n=44) (E) Gene set enrichment analysis (GSEA) of “naïve” signature in antagomiR-31- versus antagoNC- treated naïve CD4+ T cells. NES, normalized enrichment score. (F) Heatmap of representative genes associated with activation versus naïve state of T cells. Shown is log2 fold changes of gene expression in antagomiR-31-treated naïve CD4+ T cells relative to that in antagoNC-treated cells (n=3). (G) Effects of antagomiR-31 treatment on CD25 expression of naïve CD4+ T cells, assessed by frequency of CD25+ cells and median fluorescent intensity (MFI) of CD25. Average fold changes were 4.45 and 2.25, respectively (n = 8). Blue, antagomir-31 treated group; red, antagoNC-treated group. Representative FACS data for CD25 were shown in the left panel. (H) Schema of the in vitro assay used for examining the role of miR-31 in HIV-1 infection. CD4+ T cells were sorted, followed by transfection with antagomiR-31 or antagoNC. After 48 hours, cells were stimulated with a mix of anti-CD3 and anti-CD28 antibodies and infected with HIV-1 IIIB 5 days later. (I, J) Cells and supernatants were collected on days 5 and 11 post infection and respectively subjected to flow cytometry for determination of P24-expressing CD4+ T cells (I) and ELISA for quantification of released P24 proteins (J) (n = 3).

Article Snippet: PAXgene Blood miRNA Kit (QIAGEN, Hilden, Germany) was used for extraction and purification of total RNA, including miRNA, from whole blood stabilized in PAXgene Blood RNA Tubes (PreAnalytix, BD, UK).

Techniques: Activation Assay, Comparison, Infection, Expressing, Gene Expression, In Vitro, Transfection, Flow Cytometry, Enzyme-linked Immunosorbent Assay