Quantitative Proteomics by iTRAQ-PRM based Reveals the New Characterization for Gout.

BackgroundGout is a common and complex form of immunoreactive arthritis based on hyperuricemia, while the symptoms would turn to remission or even got worse. So, it is hard to early identify whether an asymptomatic hyperuricemia (AHU) patient will be susceptible to get acute gout attack and it is also hard to predict the process of gout remission to flare. Here, we report that the plasma proteins profile can distinguish among acute gout (AG), remission of gout (RG), AHU patients, and healthy controls.MethodsWe established an isobaric tags for relative and absolute quantification (iTRAQ) and parallel reaction monitoring (PRM) based method to measure the plasma proteins for AG group (n=8), RG group(n=7), AHU group(n=7) and healthy controls(n=8).Resultseleven differentially expressed proteins such as Histone H2A, Histone H2B, Thrombospondin-1 (THBS1), Myeloperoxidase (MPO), Complement C2, Complement component C8 beta chain (C8B), Alpha-1-acid glycoprotein 1 (ORM1), Inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4), Carbonic anhydrase 1 (CA1), Serum albumin (ALB) and Multimerin-1 (MMRN1) were identified. Histone H2A, Histone H2B and THBS1 might be the strongest influential regulator to maintain the balance and stability of the gout process. The complement and coagulation cascades is one of the main functional pathways in the mechanism of gout process.ConclusionsHistone H2A, Histone H2B and THBS1 may be novel biomarkers in discriminating gout attack from AHU or RG, providing new theoretical insights for the prognosis, treatment, and management of gout process.Trial registrationThis study is not a clinical trial.

VALIDATION OF SIGNIFICANT PROTEIN MARKERS FROM THE PLASMA OF SEVERE DENGUE INFECTED ADULT PATIENTS USING SELECTED REACTION MONITORING ASSAY

ABSTRACT Background :Infection with dengue virus (DENV) causes a spectrum of clinical manifestations ranging from mild dengue fever (DF) to the potentially lethal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Dengue is endemic to the tropical and sub-tropical regions of the world, which are home to over half the population of the world as well as being popular tourist destinations Aim :To study about information of the pathogenesis and clinical profile of dengue virus and Validation of significant protein markers of dengue infected adult patients using selected reaction monitoring assays. Material &Methods :This study was conducted in Department of Medicine, Sardar Patel Medical College, Bikaner from July 2017 to December 2018. Total 26 cases with features of dengue illness along with positive dengue serology in this duration were admitted in hospital. This was a hospital based study. Results : Proteomic analysis in severe dengue patients (DHF/DSS) shows down regulation of three significant proteins CD-44 antigen, complement component C8 beta chain and leucine-rich alpha-2-glycoprotein as compared to healthy controls. According to dengue agglutination test, 100% cases had IgM positive while 69.2% cases had NS1 positive. No case had IgG positive. Mean age in DHF group was 30.4812.58 and in DSS group mean age was 25.0012.39 years and this difference was found statistically insignificant (p>0.05). Most common clinical manifestation was fever, abdominal pain and myalgia (100%) while rash was present in 96.2%, vomiting and headache were present in 92.3% cases each and least common clinical manifestation was retro-orbital pain (88.5%).Gum bleeding, epistaxis and petechiae were found in most cases.According to sensorium 96.2% cases were found normal and 3.8% cases were altered sensorium.According to outcome, 20 cases were cured successfully while 6 cases discharge on request. Patients were on telephonically follow up for one week, there was no mortality. Conclusion : In present study revealed that validating 26 serum samples used for optimization by using selected reaction monitoring. Three significant proteins, CD44 Antigen, Complement component C8 beta chain, Leucine-rich alpha-2-glycoprotein are downregulated in severe dengue (DHF/DSS) patients as compared to healthy controls. These are predictive biomarkers of severe dengue fever (DHF/DSS).

Proteomic screening of plasma identifies potential noninvasive biomarkers associated with significant/advanced fibrosis in patients with nonalcoholic fatty liver disease

Noninvasive biomarkers are clinically useful for evaluating liver fibrosis stage in patients with nonalcoholic fatty liver disease (NAFLD). The aim of the present study was to compare plasma proteins in patients with early nonalcoholic steatohepatitis (NASH) (F0-F1) versus NASH with significant/advanced fibrosis (F2–F4) to determine whether candidate proteins could be used as potential noninvasive biomarkers. Nineteen biopsy-proven NAFLD patients including ten early NASH patients and nine NASH patients with significant/advanced fibrosis were enrolled in the present study. High-resolution proteomics screening of plasma was performed with the SCIEX TripleTOF 5600 System. Proteins were quantified using two different software platforms, Progenesis Qi and Scaffold Q+, respectively. Progenesis Qi analysis resulted in the discovery of 277 proteins compared with 235 proteins in Scaffold Q+. Five consensus proteins (i.e. Complement component C7; α-2-macroglobulin; Complement component C8 γ chain; Fibulin-1; α-1-antichymotrypsin) were identified. Complement component C7 was three-fold higher in the NASH group with significant/advanced fibrosis (F2–F4) compared with the early NASH (F0-F1) group (q-value = 3.6E-6). Complement component C7 and Fibulin-1 are positively correlated with liver stiffness (P=0.000, P=0.002, respectively); whereas, Complement component C8 γ chain is negatively correlated (P=0.009). High levels of Complement C7 are associated with NASH with significant/advanced fibrosis and Complement C7 is a perfect classifier of patients included in this pilot study. Further studies will be needed in a larger validation cohort to confirm the utility of complement proteins as biomarkers or mechanistic determinants of NASH with significant/advanced fibrosis.

Differentially expressed proteins in the blood serum of piglets in response to a diet supplemented with inulin

In the present study we introduced a two-dimensional electrophoresis and matrix-assisted laser desorption/ionisation time of flight mass spectrometry-based proteomic workflow to identify proteins that show altered expression as a result of the addition of 2% of water extract of inulin-type fructans to the diet of growing piglets. This analysis allowed us to detect an average of 240 spots per gel with a mass range from 10 to 250 kDa and a pH ranging from 3 to 10. Twenty protein spots were found to show statistically significant differences in their expression. Of these, 7 protein spots were up-regulated, whereas 13 showed down-regulation in response to the experimental diet. In total, 13 spots were identified, representing 8 distinct gene products. The experimental diet caused a significant change in proteins directly or indirectly involved in hemostasis and the innate immune response. Increased levels of fibrinogen along with decreased plasminogen expression may indicate that a fructan-rich diet favours the deposits of fibrin and promotes blood clotting. We also found increased expression of vitronectin and the alpha subunit of the complement component C8 which may protect the host organism against excessive cytolitic activity of the activated complement. The piglets from the experimental group had slightly increased values of IgG and IgA, whereas the IgM level tended to be decreased. The fructan-rich diet did not have any influence on plasma total cholesterol, HDL and LDL cholesterol and triglyceride levels.