Project description:BACKGROUND:The biologic attributes of the endocrine pancreas and the comparative endocrinology of islet amyloid polypeptide (IAPP) of fish are not well described in the literature. This study describes the endocrine pancreasof one teleostean fish. Ten captive Atlantic wolffish (Anarhichas lupus)from the Montreal Biodome were submitted for necropsy and their pancreata were collected. RESULTS:Grossly, all the fish pancreata examined contained 1-3 nodules of variable diameter (1-8 mm). Microscopically, the nodules were uniform, highly cellular, and composed of polygonal to elongated cells. Immunofluorescence for pancreatic hormones was performed. The nodules were immunoreactive for insulin most prominent centrally, but with IAPP and glucagon only in the periphery of the nodules. Exocrine pancreas was positive for chromogranin A. Not previously recognized in fish, IAPP immunoreactivity occurred in α, glucagon-containing, cells and did not co-localize with insulin in β cells. The islet tissues were devoid of amyloid deposits. IAPP DNA sequencing was performed to compare the sequence among teleost fish and the potency to form amyloid fibrils. In silico analysis of the amino acid sequences 19-34 revealed that it was not amyloidogenic. CONCLUSIONS:Amyloidosis of pancreatic islets would not be expected as a spontaneous disease in the Atlantic wolffish. Our study underlines that this teleost fish is a potential candidate for pancreatic xenograft research.
Project description:Raw Data for: Lipidome changes indicate oxidative stress, inflammation, and specific loss of phosphatidylserine inflammatory protection in patients with lupus
Abstract:
Systemic lupus erythematosus (SLE, lupus) is a chronic autoimmune disease which has a complex etiology and suffers from both high false positive rates and false negative rates in diagnosis and classification. Substantial lipid changes have been observed previously in lupus patients, and hence we carried out the most comprehensive lipidomics study in lupus to date using LipidMatch Flow. In this study, we investigated various sub-categories of lupus including lupus nephritis, active versus non-active lupus, as well as comparisons to non-lupus controls. A total of 1,105 unique lipids spanning 36 lipid classes (or sub-classes) were annotated in blood plasma samples; of these, 111 lipids changed significantly between controls and active lupus. We determined for the first-time specific oxidized lipid markers, with oxidized triacylglycerols being the most significantly increased lipid sub-class in active lupus as compared to controls. Other indicators of oxidative stress included decreased lipids containing ether linkages and/or polyunsaturated fatty acids. Increased ceramide (d18:1/16:0) and decreased 20-22 carbons containing species (especially 20:4) indicated an inflammatory response in patients with active lupus. Furthermore, we determined significant downregulation of phosphatidylserines, which are known inflammation suppressors, in patients with lupus and a decrease in Coenzyme Q9 and Q10 with supplementation of lupus patients with these compounds shown to be protective. Several unique lipids with unknown biology are also shown to significantly change in active lupus. Many of these trends were also observed in non-active lupus, suggesting lipidomics related changes may occur early in disease development. In conclusion, this comprehensive lipidomics study expands our knowledge of the lipid alterations associated with lupus, providing insights into disease pathogenesis and depleted lipids which could serve as therapeutic targets.
Project description:Membranous lupus nephritis is a frequent cause of nephrotic syndrome in patients with systemic lupus erythematosus. Unlike phospholipase A2 receptor or thrombospondin type 1 domain containing 7A-associated membranous nephropathy, where known antibodies can be detected within sera by indirect immunofluorescence and/or enzyme-linked immunosorbent assay, it is not possible to monitor disease activity in membranous lupus nephritis where the target autoantigens are mostly unknown. Determination of the target autoantigen has diagnostic significance, informs prognosis, and allows for non-invasive monitoring of disease activity in serum. We utilized mass spectrometry for antigen discovery of laser capture microdissected glomeruli from formalin-fixed paraffin embedded tissue and tissue IgG immunoprecipitation studies from frozen kidney biopsy tissue. We identified neural cell adhesion molecule 1 (NCAM1) to be a target antigen in membranous lupus nephritis and within rare cases of primary membranous nephropathy. The prevalence of NCAM1-associated membranous neuropathy was 5.7% of cases of membranous lupus nephritis. NCAM1 co-localizes with IgG within glomerular immune deposits. Additionally, serum from NCAM1 patients showed reactivity to NCAM1 recombinant protein. The presence of anti-NCAM1 antibodies in sera could allow for non-invasive monitoring of the disease. We propose that NCAM1 is a target autoantigen in a subset of patients with membranous lupus nephritis. Future studies are needed to determine whether anti-NCAM1 antibody levels correlate with disease activity or response to therapy.
Project description:Genome-wide alternative splice analysis of RNA from lupus and its severe form lupus nephritis We aimed to explore the genome-wide peripheral blood transcriptome of lupus (SLE) and its severe form lupus nephritis (LN) cases compared to healthy subjects (HC) using high density Affymetrix Human Exon1.0.ST arrays. Analysis revealed 15 splice variants that are differentially expressed between SLE/HC and 99 variants between LN/HC (pâ¤0.05,SI>orâ¤0.5,Benjamin Hochberg-False discovery rate correction). Comparison between LN/SLE revealed 7 variants that are differentially expressed with pâ¤0.05,SI>0.5,Benjamin Hochberg-FDR correction. Pathway analysis of differentially spliced genes revealed 11 significant pathways in SLE and 12 in LN (p<0.05). Analysis of peripheral blood transcriptome revealed signature causative genes that are alternatively spliced, signifying their clinical relevance in the pathophysiology of disease. The extent of differential splicing was found to be higher in LN than in SLE, signifying the need for further in-depth research in the same domain. Present study is the first to reveal the significance of alternative variants in susceptibility to SLE and LN. We analyzed blood from 11 female subjects (5 lupus, 3 lupus nephritis and 3 healthy control) using the Affymetrix Human Exon 1.0 ST platform. Array data was processed by Alt Analyze and Genespring software. No techinical replicates were performed. One of the outiler sample (HC2) was excluded from further analysis.
Project description:MicroRNAs (miRNA) have emerged as an important new class of modulators of gene expression. In this sudy we investigated miRNA that are differentially expressed in lupus nephritis. Microarray technology was used to investigate differentially expressed miRNA in PBMCs and EBV-transformed cell lines obtained from lupus nephritis patients and controls. TaqMan-based stem-loop real-time PCR was used for validation. Microarray analysis of miRNA expressed in African Americans (AA) derived lupus nephritis samples revealed 29 differentially expressed miRNA, of 850 tested. Microarray analysis of miRNA expressed in European American (EA) derived lupus nephritis samples revealed 50 differentially expressed miRNA, of 850 tested.