Project description:Serum of LCMV infected mice. Data was generated on a Thermo Q Exactive and C18 RP UHPLC. Positive polarity acquisition on LC-MS/MS.
Project description:Update of dataset of serum of LCMV infected mice. No Phree Kit used in extraction protocol. Data was generated on a Thermo Q Exactive and C18 RP UHPLC. Positive polarity acquisition on LC-MS/MS.
Project description:We used data independent acquisition (DIA) mass spectrometry (MS) to profile ~800 proteinsfrom 122 serum samples Dengue or Zika Trinidadian patients. Two time points were collectedper patient. The DIA MS data were matched against a spectral library generated from high pH/low pH separated pooled serum samples.
Project description:Wildtype mice and Autotaxin-overexpressing transgenic mice, both with and without spontaneous tumor formation, were maintained in a colony. Serum samples from mice in the aforementioned three groups were collected and a random subset of mice were selected for microarray analysis. RNA was extracted from serum and subsequently cDNA was generated and run on two 384-well plates probing for 373 viable targets, including microRNAs, control probes and interplate calibrators. Raw Ct values obtained from qPCR analysis were analyzed using GenEx and graphed using GraphPad Prism.
Project description:This study examined microRNA expression of cells maintained in media prepared with replete serum (EVR) or with serum depleted of extracellular vesicles (EVs) by either of two methods: standard overnight ultracentrifugation (UC-EVD) or a proprietary method used by Thermo Fisher (TF-EVD).
Project description:We carried out a prospective, longitudinal, single-center, observational cohort study of patients with confirmed acute methanol poisoning that were treated in hospitals during a mass methanol poisoning outbreak in the Czech Republic in 2012. Venous blood for proteomic analysis was obtained from 24 patients with confirmed acute methanol poisoning upon admission to the hospital (group M (“Methanol”)) with heparin administration for hemodialysis and ethanol or fomepizole administration as the antidote to block ADH. In the follow-up group of survivors of methanol poisoning (group S (“Survivors”)), venous blood samples for proteomic analysis were obtained from 46 patients during the examination, which took place 4 years after discharge from the hospital. For the control group not exposed to methanol, 24 healthy subjects were recruited (group C, “Controls”). Blood samples were spun, the serum was separated, and the samples were frozen to −80 °C until the analyses. Blood serum samples were depleted of most abundant serum proteins using Agilent MARS 14 column, samples fractionated and fractions containing proteins of interest precipitated. Samples were analyzed using LC-MS/MS Thermo Orbitrap Fusion (UHPLC-ESI-Q-OT-qIT) and identified proteins with differential expression.
Project description:A miRNA microarray was performed from HCV infected patient serum samples of bothe genotype 1b and genotype 3a, which are prevalent in India, with the aim of identifying a set of miRNAs which are uniquely differentially expressed during HCV infection. miR-320c, miR-483-5p, miR-134 and miR-198 were found to be upregulated in the patient samples as compared to the controls and are currenty being validated.
Project description:Background: Multiple sclerosis (MS) is a demyelinating autoimmune disease of the central nervous system and the leading cause of lasting neurological disabilities in young adults. Increasing evidence suggests that early treatment prevents the development of disability. However, there have been no reliable serum markers to assist the early diagnosis. In addition, interferon (IFN)-β, which is the major treatment for MS, is not always effective. Therefore, the development of simple serological test to help the early diagnosis and predict responsiveness to IFN-β is of clinical importance. On the other hand, a transmembrane-type semaphorin, Sema4A, has been implicated in experimental autoimmune encephalomyelitis (EAE) by regulating helper T (Th) cell differentiation. Thus, we aimed to identify the implications of Sema4A in diagnosis and pathogenesis of MS. Methods: We assayed serum Sema4A in 59 patients with relapsing-remitting MS (RRMS), 22 patients with clinically isolated syndrome (CIS) and 126 patients with other neurological diseases (OND) by developing a sandwich ELISA. To identify a source of soluble Sema4A and characteristics of MS patients with high levels of Sema4A, we analyzed peripheral blood mononuclear cells (PBMCs) from MS patients and healthy controls by flow cytometry (FACS) and gene chip analysis. The effect of Sema4A was examined in vitro and in vivo using an EAE model. Findings: Sema4A was significantly increased in sera of patients with MS and CIS compared to controls. Sema4A expression was increased on the surface of DCs in MS patients and shed from these cells in a metalloproteinase-dependent manner, affecting the Th17skewing. In addition, patients with high Sema4A levels exhibited more severe disabilities, and IFN-β treatment was not beneficial to those patients. Interpretation: Measuring Sema4A is a practical laboratory test to help diagnose MS and to predict responsiveness to IFN-β therapy. Microarray analysis showed that metalloproteinases such as ADAM 10 and MMPs 1, 3, 9, 12 and 25, which are reported to be involved in the pathogenesis of MS, were increased in PBMCs from MS patients with high Sema4A levels compared to those with low Sema4A and healthy controls. However, ADAM 17 and MMP 2 and 7 did not correlate with Sema4A levels. Taken together, these findings strongly suggest that Sema4A, abundantly expressed on monocytes and DCs in MS patients, is enzymatically cleaved and shed in a portion of the patients, which contributes to the high soluble Sema4A detected by the ELISA. Multiple sclerosis patients were classified according to Serum Sema4A levels. RNA from peripheral blood from 4 healthy controls, 3 MS patients with high serum Sema4A levels and 3 MS patients with low serum Sema4A levels was extracted and hybridized on Affymetrix microarrays. We sought to see whether there is corrlated genes with serum Sema4A levels or not, especially for genes for varions proteases.
Project description:Serum proteomes of healthy and Leishmania-infected dogs were analyzed by DIA-MS approach using non-depleted serum and 60 min gradient time in order to study host response to infection.
Project description:Purpose: Idenfication of microRNA biomarkers of Chronic wasting disease in serum from infected elk Methods: Illumina next generation was used to profile abundance of serum miRNA in elk naturally infected with chronic wasting disease and Hamsters experimentally infected with the 263K scrapie prion strains Results: A signature of 21 miRNAs with diagnostic potential was found to be altered in abundance in serum from CWD infected elk. Of these, 6 were similarily altered in the serum from the 263K infected hamsters Conclusion: These altered miRNA signatures may serve as the basis for non-invasive diagnostic assays for chronic wasting disease and may shed light on the pathogenesis of prion infection