The solution structure of the disulphide-linked homodimer of the human trefoil protein TFF1

AbstractThe trefoil factor family protein, TFF1, forms a homodimer, via a disulphide linkage, that has greater activity in wound healing assays than the monomer. Having previously determined a high-resolution solution structure of a monomeric analogue of TFF1, we now investigate the structure of the homodimer formed by the native sequence. The two putative receptor/ligand recognition domains are found to be well separated, at opposite ends of a flexible linker. This contrasts sharply with the known fixed and compact arrangement of the two trefoil domains of the closely related TFF2, and has significant implications for the mechanism of action and functional specificity of the TFF of proteins

Autoantibodies against the replication protein A complex in systemic lupus erythematosus and other autoimmune diseases

Replication protein A (RPA), a heterotrimer with subunits of molecular masses 70, 32, and 14 kDa, is a single-stranded-DNA-binding factor involved in DNA replication, repair, and recombination. There have been only three reported cases of anti-RPA in systemic lupus erythematosus (SLE) and Sjögren syndrome (SjS). This study sought to clarify the clinical significance of autoantibodies against RPA. Sera from 1,119 patients enrolled during the period 2000 to 2005 were screened by immunoprecipitation (IP) of (35)S-labeled K562 cell extract. Antigen-capture ELISA with anti-RPA32 mAb, immunofluorescent antinuclear antibodies (ANA) and western blot analysis with purified RPA were also performed. Our results show that nine sera immunoprecipitated the RPA70–RPA32–RPA14 complex and all were strongly positive by ELISA (titers 1:62,500 to 1:312,500). No additional sera were positive by ELISA and subsequently confirmed by IP or western blotting. All sera showed fine speckled/homogeneous nuclear staining. Anti-RPA was found in 1.4% (4/276) of SLE and 2.5% (1/40) of SjS sera, but not in rheumatoid arthritis (0/35), systemic sclerosis (0/47), or polymyositis/dermatomyositis (0/43). Eight of nine patients were female and there was no racial predilection. Other positive patients had interstitial lung disease, autoimmune thyroiditis/hepatitis C virus/pernicious anemia, or an unknown diagnosis. Autoantibody specificities found in up to 40% of SLE and other diseases, such as anti-nRNP, anti-Sm, anti-Ro, and anti-La, were unusual in anti-RPA-positive sera. Only one of nine had anti-Ro, and zero of nine had anti-nRNP, anti-Sm, anti-La, or anti-ribosomal P antibodies. In summary, high titers of anti-RPA antibodies were found in nine patients (1.4% of SLE and other diseases). Other autoantibodies found in SLE were rare in this subset, suggesting that patients with anti-RPA may form a unique clinical and immunological subset

Frequent coexistence of anti-topoisomerase I and anti-U1RNP autoantibodies in African American patients associated with mild skin involvement: a retrospective clinical study

Introduction: The presence of anti-topoisomerase I (topo I) antibodies is a classic scleroderma (SSc) marker presumably associated with a unique clinical subset. Here the clinical association of anti-topo I was reevaluated in unselected patients seen in a rheumatology clinic setting.Methods: Sera from the initial visit in a cohort of unselected rheumatology clinic patients (n = 1,966, including 434 systemic lupus erythematosus (SLE), 119 SSc, 85 polymyositis/dermatomyositis (PM/DM)) were screened by radioimmunoprecipitation. Anti-topo I-positive sera were also tested with immunofluorescence and RNA immunoprecipitation.Results: Twenty-five (15 Caucasian, eight African American, two Latin) anti-topo I positive patients were identified, and all except one met the ACR SSc criteria. Coexistence of other SSc autoantibodies was not observed, except for anti-U1RNP in six cases. When anti-topo I alone versus anti-topo I + U1RNP groups were compared, African American (21% vs. 67%), overlap with SLE (0 vs. 50%; P = 0.009) or PM/DM (0 vs. 33%; P = 0.05) or elevated creatine phosphokinase (CPK) (P = 0.07) were more common in the latter group. In comparison of anti-topo I-positive Caucasians versus African Americans, the latter more frequently had anti-U1RNP (13% vs. 50%), mild/no skin changes (14% vs. 63%; P = 0.03) and overlap with SLE (0 vs. 38%; P = 0.03) and PM/DM (0 vs. 25%; P = 0.05).Conclusions: Anti-topo I detected by immunoprecipitation in unselected rheumatology patients is highly specific for SSc. Anti-topo I coexisting with anti-U1RNP in African American patients is associated with a subset of SLE overlapping with SSc and PM/DM but without apparent sclerodermatous changes. \ua9 2011 Satoh et al.; licensee BioMed Central Ltd

Increased expression of FcγRI/CD64 on circulating monocytes parallels ongoing inflammation and nephritis in lupus

INTRODUCTION: The high-affinity receptor for IgG Fcγ/CD64 is critical for the development of lupus nephritis (LN). Cross-linking Fc receptor on recruited monocytes by IgG-containing immune complexes is a key step in immune-complex-mediated nephritis in systemic lupus erythematosus (SLE). The goal of this study was to determine whether expression of Fc receptor (FcγR) I on circulating monocytes is associated with systemic inflammation and renal disease in SLE patients. METHODS: We studied 205 SLE patients (132 with LN and 73 without LN) along with 74 healthy control individuals. Surface expression of CD14 (monocytes), FcγRI/CD64, FcγRII/CD32, and FcγRIII/CD16 was evaluated by flow cytometry. Monocyte function was assessed by determining the migratory capacity and the ability to produce CCL2 (monocyte chemotractic protein 1). High-sensitivity C-reactive protein, C3 and C4 were measured by nephelometry. RESULTS: There was little difference in the expression of FcγRIII/CD16 or FcγRIII/CD32 on circulating monocytes between patients with SLE and control individuals. In contrast, FcγRI/CD64 expression was significantly higher in SLE patients and even higher in patients with LN. FcγRI/CD64 expression was positively associated with serum creatinine and indicators of systemic inflammation. Monocytes from patients with high FcγRI/CD64 expression also exhibited increased chemotaxis and capacity to produce monocyte chemotractic protein 1. CONCLUSIONS: Increased FcγRI/CD64 expression on circulating monocytes parallels systemic inflammation and renal disease in SLE patients. We propose that circulating monocytes activated by immune complexes and/or proinflammatory mediators upregulate surface expression of FcγRI/CD64 in SLE. The enhanced chemotactic and inflammatory potential of the activated monocytes may participate in a vicious cycle of immune cell recruitment and renal injury in SLE

Governing and accelerating transformative entrepreneurship: exploring the potential for small business innovation on urban sustainability transitions

The alluring yet nebulous concept of transformative change is increasingly gaining traction in conversations about pathways to more sustainable futures. As such, new conceptual tools are needed to illuminate variety of actors, interests, and capacities at play in potentially radical experiments. This paper draws upon multi-level governance theory, sustainability transitions scholarship, and sustainability entrepreneurship literature, to interrogate the transformative potential of small and medium-sized enterprises (SMEs). We (1) identify characteristics of SMEs that might make them relatively more able to produce radical innovations, (2) explore dimensions of the broader socio-political context that influence the likelihood of this potential to be translated into action in urban spaces, and (3) discuss implications of these dynamics for transformative sustainability governance

Governing and accelerating transformative entrepreneurship: exploring the potential for small business innovation on urban sustainability transitions

The alluring yet nebulous concept of transformative change is increasingly gaining traction in conversations about pathways to more sustainable futures. As such, new conceptual tools are needed to illuminate variety of actors, interests, and capacities at play in potentially radical experiments. This paper draws upon multi-level governance theory, sustainability transitions scholarship, and sustainability entrepreneurship literature, to interrogate the transformative potential of small and medium-sized enterprises (SMEs). We (1) identify characteristics of SMEs that might make them relatively more able to produce radical innovations, (2) explore dimensions of the broader socio-political context that influence the likelihood of this potential to be translated into action in urban spaces, and (3) discuss implications of these dynamics for transformative sustainability governance

Insights to scaling remote plasma sources sustained in NF3 mixtures

Remote plasma sources (RPSs) are being developed for low damage materials processing during semiconductor fabrication. Plasmas sustained in NF3 are often used as a source of F atoms. NF3 containing gas mixtures such as NF3/O2 and NF3/H2 provide additional opportunities to produce and control desirable reactive species such as F and NO. In this paper, results from computational investigations of RPS sustained in capacitively coupled plasmas are discussed using zero-dimensional global and two-dimensional reactor scale models. A comprehensive reaction mechanism for plasmas sustained in Ar/NF3/O2 was developed using electron impact cross sections for NF2 and NF calculated by ab initio molecular R-matrix methods. For validation of the reaction mechanism, results from the simulations were compared with optical emission spectroscopy measurements of radical densities. Dissociative attachment and dissociative excitation of NFx are the major sources of F radicals. The exothermicity from these Franck–Condon dissociative processes is the dominant gas heating mechanism, producing gas temperatures in excess of 1500 K. The large fractional dissociation of the feedstock gases enables a larger variety of end-products. Reactions between NFx and O atom containing species lead to the formation of NO and N2O through endothermic reactions facilitated by the gas heating, followed by the formation of NO2 and FNO from exothermic reactions. The downstream composition in the flowing afterglow is an ion–ion plasma maintained by, in oxygen containing mixtures, [F−] ≈ [NO+] since NO has the lowest ionization potential and F has the highest electron affinity among the major neutral species

Genetic Programming for Reverse Engineering

a particular emphasis on the growing importance of recent developments in genetic programming and genetic improvement for reverse engineering. This includes work on SBSE for remodularisation, refactoring, regression testing, syntax-preserving slicing and dependence analysis, concept assignment and feature location, bug fixing, and code migration. We also explore the possibilities for new directions in research using GP and GI for partial evaluation, amorphous slicing, and product lines, with a particular focus on code transplantation. This paper accompanies the keynote given by Mark Harman at the 20 th Working Conference on Reverse Engineering (WCRE 2013). I