Immunological detection of Bacteroides fragilis in clinical-samples

A monospecific polyclonal antiserum, prepared against Bacteroides fragilis common polysaccharide antigen purified by polyacrylamide gel immunoblot detected B. fragilis, B. thetaiotaomicron, B. ovatus and Prevotella melaninogenica in pus samples from various anatomical sites by immunofluorescence microscopy of the pus. With standard clinical laboratory culture methods, 36% of 147 samples were positive for one or more of the above bacteria. Of these, B. fragilis accounted for 33%. By immunofluorescent labelling of pus with the common antigen antiserum the detection of these bacteria in the samples increased to 50%. All nine of the blood cultures in which B. fragilis was detected by culture contained bacteria positive for the common antigen. Immunofluorescent labelling of pus samples with a selection of monoclonal antibodies specific for surface polysaccharides which are known to be antigenically variable in culture in vitro and in an animal model of infection showed that these polysaccharides are also variable in natural infection. The results indicate that the common polysaccharide antigen, in contrast to the variable surface polysaccharides, is a suitable target for the immunodetection of B, fragilis in clinical samples from a range of anatomical sites.

Immunological variants of the aerobactin-cloacin DF13 outer membrane protein receptor IutA among enteric bacteria

Mouse monoclonal antibodies (MAbs) were generated against a 76-kDa IutA receptor of pathogenic avian Escherichia coli 15972. Six of the eight IutA-specific MAbs isolated (AB1 to AB6) were shown to be directed toward membrane-exposed conformational epitopes, although they did not interfere with the uptake of ferric aerobactin and cloacin DF13 as assessed by competition experiments with purified ligands. The two remaining IutA MAbs (AB9 and AB10) recognized linear epitopes buried in the IutA molecule. The panel of IutA MAbs was used to characterize IutA variants occurring in strains of E. coli, Klebsiella pneumoniae, Enterobacter spp., and Shigella spp., resulting in the identification of four immunological groups of IutAs. MAb AB9 defined an epitope conserved in all IutA variants. In addition, the panel of IutA MAbs served to identify the presence of IutA in wild-type bacteria grown in the presence of diphenylamine to reduce the expression of O-specific polysaccharide.

Immunological homologues of the Arabidopsis thaliana beta 1 tubulin are polyglutamylated in Nicotiana tabacum

Peptide-specific antibody AABI, raised to the C-terminal 13 amino acids of Arabidopsis thaliana beta 1 tubulin, identifies a single electrophoretically separable beta-tubulin on 2-D-gel Western blots of total protein extracts from A. thaliana seedlings. We show that AABI crossreacts with two of the eight polyglutamylated beta-tubulin isoforms present in purified Nicotiana tabacum tubulin fractionated by high-resolution isoelectric focussing. Immunolocalisation studies using AAB1 revealed that the two N. tabacum polyglutamylated beta 1-tubulin isoforms are utilised in all four plant microtubule arrays (the interphase cortical array, the preprophase band, the spindle and the phragmoplast) indicating that there is no apparent subcellular sorting of these isotypes.

Inflammatory and immunological biomarkers are not related to survival in adults with Cystic Fibrosis

Background
Chronic Pseudomonas aeruginosa pulmonary infection is associated with a decline in lung function and reduced survival in people with Cystic Fibrosis (CF). Damaging inflammatory and immunological mediators released in the lungs can be used as markers of chronic infection, inflammation and lung tissue damage.

Methods
Clinical samples were collected from CF patients and healthy controls. Serum IgG and IgA anti-Pseudomonas antibodies, sputum IL-8 and TNFα, plasma IL-6 and urine TNFr1 were measured by ELISA. Sputum neutrophil elastase (NE), cathepsin S and cathepsin B were measured by spectrophotometric and fluorogenic assays. The relationship between IgG and IgA, inflammatory mediators and long-term survival was determined.

Results
IgG and IL-6 positively correlated with mortality. However, multivariate analysis demonstrated that after adjusting for FEV1, IgG was not independently related to mortality. A relationship was observed between IgG and IL-6, TNFα, TNFr1 and between IgA and IL8, cathepsin S and cathepsin B.

Conclusions
These data indicate that biomarkers of inflammation are not independent predictors of survival in people with CF.

Using Graphs to Visualise and Detect Anomalies in Access Control System Transactions

Physical Access Control Systems are commonly used to secure doors in buildings such as airports, hospitals, government buildings and offices. These systems are designed primarily to provide an authentication mechanism, but they also log each door access as a transaction in a database. Unsupervised learning techniques can be used to detect inconsistencies or anomalies in the mobility data, such as a cloned or forged Access Badge, or unusual behaviour by staff members. In this paper, we present an overview of our method of inferring directed graphs to represent a physical building network and the flows of mobility within it. We demonstrate how the graphs can be used for Visual Data Exploration, and outline how to apply algorithms based on Information Theory to the graph data in order to detect inconsistent or abnormal behaviour.

European Recommendations for Primary Prevention of Congenital Anomalies: A Joined Effort of EUROCAT and EUROPLAN Projects to Facilitate Inclusion of This Topic in the National Rare Disease Plans

Congenital anomalies (CA) are the paradigm example of rare diseases liable to primary prevention actions due to the multifactorial etiology of many of them, involving a number of environmental factors together with genetic predispositions. Yet despite the preventive potential, lack of attention to an integrated preventive strategy has led to the prevalence of CA remaining relatively stable in recent decades. The 2 European projects, EUROCAT and EUROPLAN, have joined efforts to provide the first science-based and comprehensive set of recommendations for the primary prevention of CA in the European Union. The resulting EUROCAT-EUROPLAN 'Recommendations on Policies to Be Considered for the Primary Prevention of Congenital Anomalies in National Plans and Strategies on Rare Diseases' were issued in 2012 and endorsed by EUCERD (European Union Committee of Experts on Rare Diseases) in 2013. The recommendations exploit interdisciplinary expertise encompassing drugs, diet, lifestyles, maternal health status, and the environment. The recommendations include evidence-based actions aimed at reducing risk factors and at increasing protective factors and behaviors at both individual and population level. Moreover, consideration is given to topics specifically related to CA (e.g. folate status, teratogens) as well as of broad public health impact (e.g. obesity, smoking) which call for specific attention to their relevance in the pre- and periconceptional period. The recommendations, reported entirely in this paper, are a comprehensive tool to implement primary prevention into national policies on rare diseases in Europe.

Immunological determination of the pharmaceutical diclofenac in environmental and biological samples

A highly sensitive and specific competitive ELISA on 96-microwell plates was developed for the analysis of the nonsteroidal anti-inflammatory drug diclofenac. Within the water cycle in Europe, this is one of the most frequently detected pharmaceutically active compounds. The LOD at a signal-tonoise ratio (S/N) of 3, and the IC ₅₀, were found to be 6 ng/L and 60 ng/L respectively in tap water. In a comparative study using ELISA and GC-MS, diclofenac levels in wastewater from 21 sewage treatment plants were determined and a good correlation between these methods was found (ELISA vs. GCMS: r = 0.70, slope = 0,90, intercept = 0.37, n = 24). An average degradation rate of -25% can be calculated. Labscale-experiments on the elimination of diclofenac in continuous pilot sewage plants revealed a removal rate of only 5% over a period of 13 weeks. In a further study, the ELISA was applied to a number of extracts of various animal tissues from a range of species, and again a very good relationship between ELISA and LC-ESI/MS data sets was obtained (r = 0.90, p<0.0001; n = 117). The ELISA has proven to be a simple, rapid, reliable and affordable alternative to otherwise costly and advanced techniques for the detection of diclofenac in matrix diverse water samples and tissue extracts after only relatively simple sample preparation. © 2007 American Chemical Society.

Detecting localized homogeneous anomalies over spatio-temporal data

The last decade has witnessed an unprecedented growth in availability of data having spatio-temporal characteristics. Given the scale and richness of such data, finding spatio-temporal patterns that demonstrate significantly different behavior from their neighbors could be of interest for various application scenarios such as – weather modeling, analyzing spread of disease outbreaks, monitoring traffic congestions, and so on. In this paper, we propose an automated approach of exploring and discovering such anomalous patterns irrespective of the underlying domain from which the data is recovered. Our approach differs significantly from traditional methods of spatial outlier detection, and employs two phases – i) discovering homogeneous regions, and ii) evaluating these regions as anomalies based on their statistical difference from a generalized neighborhood. We evaluate the quality of our approach and distinguish it from existing techniques via an extensive experimental evaluation.