Identification of key pathway and genes expression in the activation of mast cell via calcium flux using bioinformatics analysis
Ontology highlight
ABSTRACT: The purpose of the present study is to identify gene signatures with IgE/antigen and calcium ionophore A23187 on the activation of MMCs. The differentially expressed genes between the two types of samples were identified with the Microarray Analysis. The gene ontology functional and pathway enrichment analyses of differentially-expressed genes were performed using the database for annotation, visualization and integrated discovery software.
Project description:Dysregulation of microRNAs (miRNAs) plays a critical role in the development of intervertebral disc degeneration (IDD). In this study, we present evidence from in vitro and in vivo research to elucidate the mechanism underlying the role of miR-760 in IDD. miRNA microarray and quantitative reverse transcription-polymerase chain reaction were used to determine the miRNA profiles in patients with IDD. Functional analysis was performed to evaluate the role of miR-760 in the pathogenesis of IDD. Luciferase reporter and western blotting assays were used to confirm the miRNA targets. The expression of miR-760 was significantly decreased in degenerative nucleus pulposus (NP) cells and negatively correlated with disc degeneration grade. Functional assays demonstrated that miR-760 delivery significantly increased NP cell proliferation and promoted the expression of collagen II and aggrecan. Moreover, MyD88 was identified as a target gene of miR-760. miR-760 effectively suppressed MyD88 expression by interacting with the 3′-untranslated region, which was abolished by miR-760 binding site mutations. An in vivo experiment using an IDD mouse model showed that the upregulation of miR-760 could effectively suspend IDD. Therefore, miR-760 was found to play an important role in IDD and can be used as a promising therapeutic target for the treatment of patients with IDD.
Project description:miRNA microarray with four controls and eight PTSD patients were included in the study. This study was performed to see if there is any alteration in the miRNA expression profile in the peripheral blood mononuclear cells (PBMCs) of PTSD patients (war veterans in this case). Microarray for the miRNAs was performed by Johns Hopkins Memorial Institute (Deep Sequencing and Microarray Core Facility), Baltimore. Total RNA, including mRNA, miRNA and other small RNA molecules, were isolated from PBMC samples by using total RNA isolation kit (AllPrep DNA/RNA/miRNA Universal Kit) from Qiagen. Next, total RNA samples were used in the analysis of miRNA differential expression by miRNA array hybridization assay using the Affymetrix miRNA-v1 gene chip. Linear fold-changes in miRNA up-regulation or down-regulation were calculated to compare the differences of all the miRNAs expressed between PTSD patients and controls.
Project description:Seed lipid bodies (LB), previously viewed as paradigmatic, possess a restricted proteome and serve carbon and energy storage. Albeit present in minute quantities, LBs are widespread in post-germinative tissues, where they display vigorous movement, mediate transient organellar interactions, and shuttle enzymes, and signaling molecules. Storage as well as motility and signaling are relevant in shoot meristems of woody perennials, which ahead of winter produce buds that like seeds establish dormancy, dehydrate, express OLEOSIN genes and accumulate LBs. Unlike seeds, buds do not completely desiccate and LBs retain potential signaling functions. Here, we mapped bud LB production under dormancy-inducing conditions. We found that over an 8-week period LBs enlarged significantly. Concomitantly, three major OLEOSIN family genes were transiently upregulated at the start of bud formation, while in their wake expression of lipid biosynthesis gene DGAT1a/b and lipase gene SDP1a/b increased. In parallel, LDAPa/b, LDAP2a, and LDAP3a/b were upregulated, especially LDAP1b. All LDAPs were localized to LBs. The conclusion that OLEOSINs were replaced by LDAPs was supported by the finding that PUX10a, PUX10b and CDC498A1, involved in OLESOSIN ubiquitination and degradation, were significantly upregulated. Our proteomic analysis identified 723 putative LB proteins. These included LB-anchored Caleosin and LDIP, peripherally associated LDAP1/3 and OBL1, and numerous proteins that reflect storage, transport, organellar interaction, opportunistic cargo and contaminants. There is a significant overlap with proteins identified earlier in plasmodesmal proteomes. Collectively, the data support a model in which LBs are part of a pre-installed mechanism that serves dormancy release, survival through winter and regrowth.
Project description:We compared transcriptional differences between Periostin siRNA treated and GFP-siRNA treated in OP9 cells using Affymetrix mouse 430_2 array.
Project description:Human embryonic stem cells (hESCs) exhibit a skewed X chromosome inactivation (XCI) have recently been reported. Whether there is any copy number variation (CNV), loss of heterozygousity (LOH) and single nucleotide variant (SNV) in those epigenetically distinct cells and whether these abnormalities have any correlation to skewed XCI in hESCs is currently unknown.
Project description:This experiment focused on the miRNA expressions within the tissues specific niches of mice afflicted with melanoma, compared to littermates without malignancy. BRafCA/ PtenloxP/ Tyr::CreERT2 or genetically matched littermates without Cre recombinase were treated with 2µl 4-hydroxytamoxifen (5mM), every other day, for on days 0, 2, and 4, on the shaved right flank of the abdomen of the animal. Mice slowly grow melanoma at the treated site, and some metastases distal to treatment location (eg. Ear or opposite side of abdomen). Endpoint criteria was designated as a Samples were taken when tumors reached 1.5mm diameter. Total RNA was isolated from tumors or tamoxifen treated healthy skin, skin adjacent to the treatment site, or the ear ( representing a metastatic site). Total RNA was prepared with FLAG-TAG kit and miRNA microarray 4.0 was performed.
Project description:A rare chromosome 20 with additional G-positive band was found during prenatal diagnosis. To identified the rare chromosome 20, standard cytogenetic and molecular cytogenetic analysis of the patients was performed. Array comparative genomic hybridization (aCGH) were performed to exclude genomic imbalance.
Project description:PTCs cause a multitude of human diseases and there are no established therapeutic options for their therapeutic management. Herein, we report the high-throughput cloning and identification, characterization and functional analysis of anticodon-edited tRNA which display efficacious PTC reversion in eukaryotic cells and mouse skeletal muscle. Notably, our screen identifies ACE-tRNA, in total, with the potential to repair a vast majority of known human disease-causing PTC, but this therapeutic will require overcoming tissue and delivery specific challenges. However, the engineered tRNA, once delivered, faithfully encode their cognate amino acid, thus abrogating spurious effects on downstream protein stability, folding, and trafficking, and consequently negating the need for tandem therapies involving protein folding or trafficking agents. When transfected as cDNA, ACE-tRNAs rescued multiple full-length proteins via PTC suppression; a NLuc luciferase reporter, a model protein HDH, and two disease nonsense mutations in CFTR.
Project description:Our understanding of how human skin cells differ according to body site and tumour formation is limited. To address this we have created a multi-scale spatial atlas of healthy skin and basal cell carcinoma (BCC), incorporating in vivo optical coherence tomography, single cell RNA sequencing, spatial global transcriptional profiling and in situ sequencing. Computational spatial deconvolution and projection revealed the localisation of distinct cell populations to specific tissue contexts. Although cell populations were conserved between healthy anatomical sites and in BCC, mesenchymal cell populations including fibroblasts and pericytes retain signatures of developmental origin. Spatial profiling and in silico lineage tracing support a hair follicle origin for BCC and demonstrate that cancer-associated fibroblasts are an expansion of a POSTN+ subpopulation associated with hair follicles in healthy skin. RGS5+ pericytes are also expanded in BCC suggesting a role in vascular remodelling during cancer neovascularization. Our findings suggest that the identity of mesenchymal cell populations is regulated by signals emanating from adjacent structures and that these signals are repurposed to promote the expansion of skin cancer stroma. The resource we have created is publicly available in an interactive format for the research community.