Immuno-assays

Immuno-assays are helpful serological techniques for the laboratory diagnosis of autoimmune blistering diseases (AIBD) and for monitoring disease activity of individual patients. The three main immuno-assays are immunoblotting, immunoprecipitation and ELISA. All three make use of the ability of the autoimmune IgG to bind to the target antigen. Immunoblotting is used to visualize the apparent molecular mass of the antigen thereby enabling its identification. The same goes for immunoprecipitation that, although being a more laborious technique than immunoblotting, has the advantage that it recognizes more epitopes on the autoantigen than immunoblotting. Whereas immunoblotting and immunoprecipitation are qualitative assays ELISA is a rapid and easy quantitative technique. ELISA therefore enables monitoring of the antibody titer during the disease course. Two recently developed techniques, keratinocyte binding assay and keratinocyte footprint assay are the most sensitive assays for pemphigus and anti-laminin-332 mucous membrane pemphigoid.</p

Systems biology in animal sciences

Systems biology is a rapidly expanding field of research and is applied in a number of biological disciplines. In animal sciences, omics approaches are increasingly used, yielding vast amounts of data, but systems biology approaches to extract understanding from these data of biological processes and animal traits are not yet frequently used. This paper aims to explain what systems biology is and which areas of animal sciences could benefit from systems biology approaches. Systems biology aims to understand whole biological systems working as a unit, rather than investigating their individual components. Therefore, systems biology can be considered a holistic approach, as opposed to reductionism. The recently developed ‘omics’ technologies enable biological sciences to characterize the molecular components of life with ever increasing speed, yielding vast amounts of data. However, biological functions do not follow from the simple addition of the properties of system components, but rather arise from the dynamic interactions of these components. Systems biology combines statistics, bioinformatics and mathematical modeling to integrate and analyze large amounts of data in order to extract a better understanding of the biology from these huge data sets and to predict the behavior of biological systems. A ‘system’ approach and mathematical modeling in biological sciences are not new in itself, as they were used in biochemistry, physiology and genetics long before the name systems biology was coined. However, the present combination of mass biological data and of computational and modeling tools is unprecedented and truly represents a major paradigm shift in biology. Significant advances have been made using systems biology approaches, especially in the field of bacterial and eukaryotic cells and in human medicine. Similarly, progress is being made with ‘system approaches’ in animal sciences, providing exciting opportunities to predict and modulate animal traits

LHC and B physics probes of neutrinoless double beta decay in supersymmetry without R-parity

In the event of an observation of neutrinoless double beta decay, a relevant question would be: what lepton number violating physics is responsible for the decay? The exchange of Majorana neutrinos and/or supersymmetric particles may contribute. We point out that measurements of supersymmetric signals at the LHC, including single slepton production, could be used to help bound some supersymmetric processes contributing to neutrinoless double beta decay. LHC information about the supersymmetric spectrum could be combined with Bd-Bd bar mixing data in order to bound a competing neutrinoless double beta decay process involving sbottom exchange

Direct immunofluorescence microscopy

Direct immunofluorescence plays an important role in the diagnosis of autoimmune blistering diseases. The purpose of direct immunofluorescence microscopy is to detect in vivo antibodies in patient's skin or mucosa. Direct immunofluorescence of pemphigus shows depositions of immunoglobulins and/or complement on the epithelial cell surface of keratinocytes, whereas pemphigoid shows linear deposition of immunoglobulins along the epidermal basement membrane zone. This linear deposition can be separated in an n-serrated pattern and a u-serrated pattern. An n-serrated pattern is seen in blistering diseases with binding above the lamina densa with antibodies against hemidesmosomal components, e.g. bullous pemphigoid, while a u-serrated pattern points to a sublamina densa binding diseases caused by autoantibodies against type VII collagen, e.g. epidermolysis bullosa acquisita. Finally, dermatitis herpetiformis shows a granular IgA deposition along the epidermal basement membrane zone.</p

Indirect immunofluorescence microscopy

The purpose of indirect immunofluorescence microscopy is to detect circulating antibodies in patient's serum. For this purpose an adequate substrate is necessary to visualize these antibodies. Monkey esophagus is the most widely used substrate for detecting of circulating autoantibodies in patients with autoimmune blistering diseases. In all variants of pemphigus antibodies show an epithelial cell surface pattern, resulting from present autoantibodies against the desmosomal molecules desmoglein 1 and/or 3. This pattern is also called chicken wire or honeycomb pattern. In pemphigoid a linear deposition along the epithelial basement membrane can be observed, caused by autoantibodies against hemidesmosomes or their connecting proteins underneath. Human salt split skin is a valuable substrate in the diagnosis of subepidermal autoimmune blistering diseases. Important antigens in the roof of salt split skin are type XVII collagen (BP180) and BP230, whereas laminin 332, p200 and type IV collagen are situated in the floor of the blister. This implies that bullous pemphigoid, mucous membrane pemphigoid, pemphigoid gestationis, and lichen planus pemphigoides show staining of IgG on the epidermal side of the blister. On the other hand anti-laminin 332 pemphigoid, anti-p200 pemphigoid, epidermolysis bullosa acquisita, and bullous SLE show staining on the dermal side. Other less used, but valuable substrates in some instances, are rat bladder and knock-out skin.</p

S₃ flavor symmetry at the LHC

Discrete symmetries employed to explain neutrino mixing and mass hierarchies are often associated with an enlarged scalar sector which might lead to exotic Higgs decay modes. We explore such a possibility in a scenario with S3 flavor symmetry which requires three scalar SU(2) doublets. The spectrum is fixed by minimizing the scalar potential, and we observe that the symmetry of the model leads to tantalizing Higgs decay models potentially observable at the CERN Large Hadron Collider (LHC)

Non-bullous Lichen Planus Pemphigoides:A Case Report

is missing (Short communication)

Keratinocyte footprint assay discriminates antilaminin-332 pemphigoid from all other forms of pemphigoid diseases

Background Antilaminin-332 mucous membrane pemphigoid is a chronic severe pemphigoid disease characterized by autoantibodies to laminin-332. At present no commercial assay is available to demonstrate antilaminin-332 antibodies, and diagnosis relies on in-house techniques with limited sensitivities. Objectives In order to move, keratinocytes cultured in vitro secrete laminin-332 to attach to the culture dish. In that way, they leave behind a unique footprint trail of laminin-332. We aimed to develop a sensitive and specific laboratory assay to determine antilaminin-332 autoantibodies in patient serum based on binding of patient IgG to these unique footprints. Methods Normal human keratinocytes were grown on glass coverslips and incubated with patient or control serum for 1 h. The binding of IgG was then investigated by immunofluorescence. After validating the test for its ability to identify antilaminin-332 autoantibodies it was converted into a daily available test based on binding of IgG to dried coverslips that can be stored frozen. The staining patterns of sera from patients with antilaminin-332 pemphigoid were then compared with those of sera from patients with other autoimmune bullous diseases and normal human sera. Results IgG of all antilaminin-332 pemphigoid sera (n = 16) bound to laminin-332 footprints, while all normal human controls (n = 55) were negative. From the sera of patients with other diseases (n = 72) four sera tested positive. The footprint assay was also positive for sera that were negative by salt-split skin analysis, demonstrating that it is a very sensitive technique. Conclusions The keratinocyte footprint assay is a fast and specific assay to confirm or rule out the presence of antilaminin-332 autoantibodies. What's already known about this topic? Antilaminin-332 mucous membrane pemphigoid is a severe form of pemphigoid, and patients may have an increased risk of malignancies. The diagnosis of antilaminin-332 mucous membrane pemphigoid is complicated by the lack of specific commercial tests for antilaminin-332 antibodies and can be confirmed only in specialized laboratories. Keratinocytes in culture need laminin-332 for adhesion and migration and therefore deposit it on the bottom of the culture dish. What does this study add? The keratinocyte footprint assay detects antilaminin-332 autoantibodies in patient serum using the native laminin-332 produced by cultured keratinocytes. What is the translational message? The keratinocyte footprint assay is a fast and specific assay to confirm or rule out the presence of antilaminin-332 autoantibodies