Top-down Mass Spectrometry Analysis of Human Serum Autoantibody Antigen-Binding Fragments.

Detecting autoimmune diseases at an early stage is crucial for effective treatment and disease management to slow disease progression and prevent irreversible organ damage. In many autoimmune diseases, disease-specific autoantibodies are produced by B cells in response to soluble autoantigens due to defects in B cell tolerance mechanisms. Autoantibodies accrue early in disease development, and several are so disease-specific they serve as classification criteria.

The prevention, screening and treatment of congenital heart block from neonatal lupus: a survey of provider practices.

Objective: To survey an international sample of providers to determine their current practices for the prevention, screening, and treatment of congenital heart block (CHB) due to maternal Ro/SSA antibodies.

Methods: A survey was designed by the organizing committee of the 9th International Conference of Reproduction, Pregnancy and Rheumatic Diseases. It was sent to attendants of the conference and authors of recent publications or abstracts at ACR 2012, 2013 or 2014 on rheumatic diseases and pregnancy.

Stained Gels Can Be Stored for Several Months in Nonsealed Polyethylene Bags.

It is customary to dry gels (SDS-PAGE, native gels, or two-dimensional gels) after staining for record keeping purposes. This is typically carried out with gel dryers or by drying between two cellophane sheets held together by acrylic frames. Here, we report a simple method to store a variety of stained gels without any storage buffer within flexible nonsealed polyethylene bags.

Paper Adsorbents Remove Coomassie Blue from Gel Destain and Used Gel Stain in an Environment-Friendly Manner.

Coomassie Brilliant Blue (CBB), used to stain protein gels, is known to be toxic. Therefore, laboratories do not discard used CBB into the sink owing to the possibility of it contaminating drinking water supplies. We tested the ability of various paper adsorbents to adsorb CBB released from gels during destaining.

Destaining Coomassie Brilliant Blue-Stained Sodium Dodecyl Sulfate-Polyacrylamide Protein Gels Using a Household Detergent.

We report here a one-step method using the household detergent Vim Ultra to destain sodium dodecyl sulfate-polyacrylamide protein gels stained with Coomassie Brilliant Blue. This method, originally described by Pal and group, uses a 5% suspension of the detergent to destain gels efficiently. This method is cheap and user-friendly compared to the commonly used methanol-acetic acid-water containing destaining solvent.

Heat/Pressure Treatment with Detergents Significantly Increases Curcumin Solubility and Stability: Its Use as an Environment-Friendly Protein Gel Stain.

Curcumin, the principal curcuminoid in the food spice turmeric, is insoluble in water at room temperature. We have previously solubilized curcumin in water with the application of heat (100 °C) and found that this solubilized curcumin could be used as a gel protein stain. However, heat solubilization in water solubilized only a small fraction of the curcuminoid (1.

Curcumin/Turmeric as an Environment-Friendly Protein Gel Stain.

Curcumin, the main curcuminoid in food spice turmeric, is insoluble in water at room temperature. We showed that curcumin can be solubilized in water with the application of heat (100 °C). Here we demonstrate that heat-solubilized curcumin can serve as a nontoxic and environment-friendly fluorescent/colorimetric reversible protein stain.

Application of Heat to Quickly Stain and Destain Proteins Stained with Coomassie Blue.

Proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis have been visualized reliably by staining with Coomassie Brilliant Blue. In this chapter, we show that it is possible to drastically reduce protein staining and destaining time, while simultaneously increasing detection sensitivity, with the application of heat. It took 5 min to stain proteins at 55, 62.

Peptidoglycan induces disseminated intravascular coagulation in baboons through activation of both coagulation pathways.

Anthrax infections exhibit progressive coagulopathies that may contribute to the sepsis pathophysiology observed in fulminant disease. The hemostatic imbalance is recapitulated in primate models of late-stage disease but is uncommon in toxemic models, suggesting contribution of other bacterial pathogen-associated molecular patterns (PAMPs). Peptidoglycan (PGN) is a bacterial PAMP that engages cellular components at the cross talk between innate immunity and hemostasis.

Unique clinical characteristics, autoantibodies and medication use in Native American patients with systemic lupus erythematosus.

Objective: Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with varied morbidity and mortality. We assessed clinical presentations, autoantibody specificities and therapeutic interventions in Native American (NA) patients with SLE.

Methods: Patients with SLE meeting 1997 American College of Rheumatology classification criteria (n=3148) were enrolled between 1992 and 2010 in the multiethnic, cross-sectional Lupus Family Registry and Repository.

The impact of menopause on functional status in women with rheumatoid arthritis.

Objective: The aim of this study was to investigate the association of menopause with functional status outcomes in women with RA.

Methods: Participants were women in a US-wide observational cohort who developed RA before menopause. The HAQ measured functional status.

APache Is an AP2-Interacting Protein Involved in Synaptic Vesicle Trafficking and Neuronal Development.

Synaptic transmission is critically dependent on synaptic vesicle (SV) recycling. Although the precise mechanisms of SV retrieval are still debated, it is widely accepted that a fundamental role is played by clathrin-mediated endocytosis, a form of endocytosis that capitalizes on the clathrin/adaptor protein complex 2 (AP2) coat and several accessory factors. Here, we show that the previously uncharacterized protein KIAA1107, predicted by bioinformatics analysis to be involved in the SV cycle, is an AP2-interacting clathrin-endocytosis protein (APache).

Amelioration of ongoing experimental autoimmune encephalomyelitis with fluoxetine.

In patients with multiple sclerosis, the selective serotonin reuptake inhibitor, fluoxetine, resulted in less acute disease activity. We tested the immune modulating effects of fluoxetine in a mouse model of multiple sclerosis, i.e.

Mutations in KEOPS-complex genes cause nephrotic syndrome with primary microcephaly.

Galloway-Mowat syndrome (GAMOS) is an autosomal-recessive disease characterized by the combination of early-onset nephrotic syndrome (SRNS) and microcephaly with brain anomalies. Here we identified recessive mutations in OSGEP, TP53RK, TPRKB, and LAGE3, genes encoding the four subunits of the KEOPS complex, in 37 individuals from 32 families with GAMOS. CRISPR-Cas9 knockout in zebrafish and mice recapitulated the human phenotype of primary microcephaly and resulted in early lethality.

Use of SLICC criteria in a large, diverse lupus registry enables SLE classification of a subset of ACR-designated subjects with incomplete lupus.

Objective: SLE is traditionally classified using the American College of Rheumatology (ACR) criteria. The Systemic Lupus International Collaborating Clinics (SLICC) recently validated an alternative system. This study examined large cohorts of subjects with SLE and incomplete lupus erythematosus (ILE) to compare the impact of ACR and SLICC criteria.

Systemic lupus erythematosus biomarkers: the challenging quest.

SLE, a multisystem heterogeneous disease, is characterized by production of antibodies to cellular components, with activation of both the innate and the adaptive immune system. Decades of investigation of blood biomarkers has resulted in incremental improvements in the understanding of SLE. Owing to the heterogeneity of immune dysregulation, no single biomarker has emerged as a surrogate for disease activity or prediction of disease.

Unique Inflammatory Mediators and Specific IgE Levels Distinguish Local from Systemic Reactions after Anthrax Vaccine Adsorbed Vaccination.

Although the U.S. National Academy of Sciences concluded that anthrax vaccine adsorbed (AVA) has an adverse event (AE) profile similar to those of other adult vaccines, 30 to 70% of queried AVA vaccinees report AEs.

GenomeRunner web server: regulatory similarity and differences define the functional impact of SNP sets.

Motivation: The growing amount of regulatory data from the ENCODE, Roadmap Epigenomics and other consortia provides a wealth of opportunities to investigate the functional impact of single nucleotide polymorphisms (SNPs). Yet, given the large number of regulatory datasets, researchers are posed with a challenge of how to efficiently utilize them to interpret the functional impact of SNP sets.

Results: We developed the GenomeRunner web server to automate systematic statistical analysis of SNP sets within a regulatory context.

Altered type II interferon precedes autoantibody accrual and elevated type I interferon activity prior to systemic lupus erythematosus classification.

Objectives: The relationship of immune dysregulation and autoantibody production that may contribute to systemic lupus erythematosus (SLE) pathogenesis is unknown. This study evaluates the individual and combined contributions of autoantibodies, type I interferon (IFN-α) activity, and IFN-associated soluble mediators to disease development leading to SLE.

Methods: Serial serum specimens from 55 individuals collected prior to SLE classification (average timespan=4.

Cytokine profiles show heterogeneity of interferon-β response in multiple sclerosis patients.

Objective: To evaluate serum cytokine profiles for their utility to determine the heterogeneous responses to interferon (IFN)-β treatment in patients with multiple sclerosis (MS).

Methods: Patients with relapsing-remitting MS (RRMS) or clinically isolated syndrome receiving de novo IFN-β treatment were included in this prospective, observational study. Number of relapses and changes in disability were assessed 2 years prior to and 2 years after initiation of treatment.

Antibody Reactivity of B Cells in Lupus Patients with Increased Disease Activity and ARID3a Expression.

Earlier studies showed that the DNA-binding protein, Bright/ARID3a bound to a subset of human and mouse immunoglobulin heavy chain promoters where it enhanced expression. Indeed, mice with transgenic expression of ARID3a in all B lymphocytes have expanded MZ B cells and produce anti-nuclear antibodies (ANAs). Consistent with our findings in mice, we observed that human systemic lupus erythematosus (SLE) patients had expanded numbers of peripheral blood ARID3a B cells that were associated with increased disease activity (p = 0.

A highlight from the LUPUS 2014 meeting: eight great ideas.

This review describes eight 'great ideas' regarding bench-to-bedside considerations in systemic lupus erythematosus (SLE) presented at the second international LUPUS meeting in Quebec, September 2014. The topics included: correcting the impaired clearance of apoptotic fragments; optimisation of clinical trial design: the PERFECT (Pre Evaluation Reducing Frighteningly Elevated Coverable Targets) study; lipidomics and metabolomics in SLE; importance of the inflammasome; identification and treatment of asymptomatic autoimmunity: prevention of SLE; combining low doses of hydroxychloroquine and quinacrine for long-term maintenance therapy of SLE; reducing emergency room visits and the critical relevance of the autoantigen.

IL-18 acts in synergy with IL-7 to promote ex vivo expansion of T lymphoid progenitor cells.

Although IL-18 has not previously been shown to promote T lymphopoiesis, results obtained via a novel data mining algorithm (global microarray meta-analysis) led us to explore a predicted role for this cytokine in T cell development. IL-18 is a member of the IL-1 cytokine family that has been extensively characterized as a mediator of inflammatory immune responses. To assess a potential role for IL-18 in T cell development, we sort-purified mouse bone marrow-derived common lymphoid progenitor cells, early thymic progenitors (ETPs), and double-negative 2 thymocytes and cultured these populations on OP9-Delta-like 4 stromal layers in the presence or absence of IL-18 and/or IL-7.