Characterization of epitope specificities of reference antibodies used for the quantification of the birch pollen allergen Bet v 1.

BACKGROUND: Accurate allergen quantification is needed to document the consistency of allergen extracts used for immunotherapy. Herein, we characterize the epitope specificities of two monoclonal antibodies used in an ELISA for the quantification of the major birch pollen allergen Bet v 1, established as a reference by the BSP090 European project. METHODS: The ability of mAbs 5B4 and 6H4 to recognize Bet v 1 isoforms was addressed by immunochromatography. The capacity of each mAb to compete with patients' IgE for binding to Bet v 1 was measured by ELISA inhibition. Epitope mapping was performed by pepscan analysis, site-directed mutagenesis, and hydrogen/deuterium exchange-mass spectrometry. RESULTS: The 5B4 epitope corresponds to a peptide sequence (I56-K68) overlapping with the binding sites of patients' serum IgEs. Mutation of residues P59, E60, and K65 abolishes 5B4 binding to Bet v 1 and reduces the level of IgE recognition. In contrast, 6H4 recognizes a conformational epitope lying opposite to the 5B4 binding site, involving residues located in segments I44-K55 and R70-F79. Substitution of E45 reduces the binding capacity of 6H4, confirming that it is critical for the interaction. Both mAbs interact with >90% of Bet v 1 content present in the birch pollen extract, while displaying a weak cross-reactivity with other allergens of the PR-10 family. CONCLUSIONS: MAbs 5B4 and 6H4 recognize structurally distinct epitopes present in the vast majority of Bet v 1 isoforms. These results support the relevance as a reference method of the Bet v 1-specific quantitative ELISA adopted by the European Pharmacopoeia.

The serologically defined colon cancer antigen-3 interacts with the protein tyrosine phosphatase PTPN13 and is involved in the regulation of cytokinesis.

Cytokinesis is the final step of cell division. Increasing evidence suggests failure of cytokinesis might contribute to the development of cancer. Here, we demonstrate that the serologically defined colon cancer antigen-3 (SDCCAG3) forms a complex with PTPN13, a protein tyrosine phosphatase known to be involved in the regulation of cytokinesis, carcinogenesis and tumor aggressiveness. We show that SDCCAG3 is a novel endosomal protein, primarily localized at the early/recycling endosomal compartment. SDCCAG3 undergoes dynamic localization during cell division with strong accumulation at the midbody during cytokinesis. Overexpression as well as downregulation correlates with the generation of multinucleate cells. Furthermore, we show interaction of SDCCAG3 with the Arf GTPase activating protein GIT1 (G protein-coupled receptor kinase interactor-1). Overexpression of an ArfGAP-negative version of GIT1 also results in an increased number of multinucleate cells suggesting regulation of Arf-mediated vesicular trafficking or signaling via SDCCAG3. Finally, we demonstrate that SDCCAG3 expression levels are elevated in colon cancers. In summary, we have established SDCCAG3 as a novel endosomal protein, which is involved in the regulation of cytokinesis.

Charter for disabled people using hospitals: a completed access audit cycle.

This completed audit cycle assessed access for disabled people to a district general hospital, in relation to standards laid down in the Royal College of Physicians Charter for Disabled People using Hospitals. The project was effective in demonstrating problems and implementing change to overcome them. It was also useful in raising disability awareness in the young investigators, who easily recognised the shortcomings in facilities for disabled people, and is proposed as a possible model for inclusion in medical undergraduate training programmes to raise disability awareness amongst a new generation of doctors.

Cybersemiotics: a transdisciplinary framework for information studies.

This paper summarizes recent attempts by this author to create a transdisciplinary, non-Cartesian and non-reductionistic framework for information studies in natural, social, and technological systems. To confront, in a scientific way, the problems of modern information technology where phenomenological man is dealing with socially constructed texts in algorithmically based digital bit-machines we need a theoretical framework spanning from physics over biology and technological design to phenomenological and social production of signification and meaning. I am working with such pragmatic theories as second order cybernetics (coupled with autopolesis theory), Lakoffs biologically oriented cognitive semantics, Peirce's triadic semiotics, and Wittgenstein's pragmatic language game theory. A coherent synthesis of these theories is what the cybersemiotic framework attempts to accomplish.

A comparison of hip and lumbopelvic inflexibility and low back pain in runners and cyclists.

OBJECTIVE: To study the correlation between the incidence of low back pain in active runners and cyclists to their level of lumbopelvic inflexibility. SETTING: Laboratory setting. SUBJECTS: One hundred eight volunteers were chosen. None had prior surgery to the hip or lumbar spine regions. Along with runners and cyclists, a sedentary control group was chosen. A medical questionnaire focusing on activity levels in sport and the incidence of disability and low back pain was completed. Eight variables were calculated for statistical significance. INTERVENTIONS: None. KNOWN OUTCOME MEASURES: Correlation between all variables were calculated. Additionally, ANOVAs for total group on each dependent variable were conducted, followed by Tukey HSDs where significant. RESULTS: Results indicated significant correlations between hip extension and sit and reach [r (106) = .36, p > .001], year in sport and mileage [r (106) = .39, p > .001], and sex and hip extension [r (106) = .35, p > .001]. CONCLUSION: A correlation between hip and low back inflexibility and the incidence of low back pain and disability in runners and cyclists is not supported by our data. Inconsistency of age control within our subject groups, along with the choice of flexibility testing, may necessitate further investigation on this subject.

Rotator cuff disease: current trends in orthopedic management.

Rotator cuff disease is a clinical entity that is frequently encountered in the typical primary care office. Of particular importance to the chiropractic physician is the close proximity of the shoulder joint to the cervical spine. Due to the relatively high level of morbidity in the population, rotator cuff disease should be looked at carefully on both the physical and advanced diagnostic level. Several factors contributing to rotator cuff pathology include subacromial impingement, the relative microvascular pattern of the cuff mechanism and prior trauma. The natural history of attrition and aging also play a role in rotator cuff disease. Advanced disease is successfully diagnosed through the efficient utilization and proper sequencing of advanced orthopedic imaging studies.

Lyme disease.

A 29-year-old male tennis player was examined for neck pain, limited mobility, and right elbow pain. The patient was treated for cervical facet syndrome by chiropractic manipulative therapy (CMT) for a period of 6 weeks. After this time the patient had a relapse of cervical pain, spinal myofascitis, dermatitis and migratory arthropathy. Several specialists were consulted throughout the case including a rheumatologist, and a tentative diagnosis of psoriatic arthritis and fibromyalgia was made. A follow-up blood evaluation led to a new diagnosis of Lyme disease.