The architecture of variant surface glycoprotein gene expression sites in Trypanosoma brucei

Trypanosoma brucei evades the immune system by switching between Variant Surface Glycoprotein (VSG) genes. The active VSG gene is transcribed in one of approximately 20 telomeric expression sites (ESs). It has been postulated that ES polymorphism plays a role in host adaptation. To gain more insight into ES architecture, we have determined the complete sequence of Bacterial Artificial Chromosomes (BACs) containing DNA from three ESs and their flanking regions. There was variation in the order and number of ES-associated genes (ESAGs). ESAGs 6 and 7, encoding transferrin receptor subunits, are the only ESAGs with functional copies in every ES that has been sequenced until now. A BAC clone containing the VO2 ES sequences comprised approximately half of a 330 kb 'intermediate' chromosome. The extensive similarity between this intermediate chromosome and the left telomere of T. brucei 927 chromosome I, suggests that this previously uncharacterised intermediate size class of chromosomes could have arisen from breakage of megabase chromosomes. Unexpected conservation of sequences, including pseudogenes, indicates that the multiple ESs could have arisen through a relatively recent amplification of a single ES.     

EEG versus MEG localization accuracy: Theory and experiment

Summary We first review the theoretical and computer modelling studies concerning localization accuracy of EEG and MEG, both separately and together; the source is here a dipole. The results show that, of the three causes of localization errors, noise and head modelling errors have about the same effect on EEG and MEG localization accuracies, while the results for measurement placement errors are inconclusive. Thus, these results to date show no significant superiority of MEG over EEG localization accuracy. Secondly, we review the experimental findings, where there are again localization accuracy studies of EEG and MEG both separately and together. The most significant EEG-only study was due to dipoles implanted in the heads of patients, and produced an average localization error of 20 mm. Various MEG-only studies gave an average error of 2–3 mm in saline spheres and 4–8 mm in saline-filled skulls. In the one study where EEG and MEG localization were directly compared in the same actual head, again using dipoles implanted in patients, the average EEG and MEG errors of localization were 10 and 8 mm respectively. The MEG error was later confirmed by a similar (but MEG-only) experiment in another study, using a more elaborate MEG system. In summary, both theory and experiment suggests that the MEG offers no significant advantage over the EEG in the task of localizing a dipole source. The main use of the MEG, therefore, should be based on the proven feature that the MEG signal from a radial source is highly suppressed, allowing it to complement the EEG in selecting between competing source configurations. A secondary useful feature is that it handles source modelling errors differently than does the EEG, allowing it to help clarify non-dipolar extended sources.This work was supported by grants RO1NS26433, RO1NS19558 and RO1NS22703 from the National Institutes of Health.     

Development and loss of toluene diisocyanate reactivity: immunologic,pharmacologic, and provocative challenge studies

Toluene diisocyanate (TDI) sensitivity accompanied by nonspecific bronchial hyperresponsiveness occurs in approximately 5% of occupationally exposed workers. We report the case of a 32-yr-old worker followed longitudinally after removal from isocyanate exposure. TDI reactivity was lost 11 mo after removal from exposure and nonspecific bronchial hyperresponsiveness resolved after 17 mo. Bronchial reactivity to radishes (Raphanus sativus), which developed concurrently with TDI reactivity, was lost 2 yr later. Immunopharmacologic results show that the worker's initial decreased ability of lymphocytes to produce cyclic AMP returned to near normal after 2 yr. IgE antibodies to a human serum albumin tolyl monoisocyanate conjugate were still present at this time.     

Diffusion tensor imaging of normal and injured developing human brain - a technical review

The application of diffusion tensor imaging (DTI) to the evaluation of developing brain remains an area of active investigation. This review focuses on the changes in DTI parameters which accompany both brain maturation and injury. The two primary pieces of information available from DTI studies-water apparent diffusion coefficient and diffusion anisotropy measures-change dramatically during development, reflecting underlying changes in tissue water content and cytoarchitecture. DTI parameters also change in response to brain injury. In this context, not only does DTI offer the possibility of detecting injury earlier than conventional imaging methods, but also appears more sensitive to disruption of white matter than any other imaging method. DTI offers unique insight into brain injury and maturation, and does so in a fashion that can be readily applied in a clinical setting.     

Cross-linking of cell surface ganglioside GM1 induces the selective apoptosis of mature CD+ T lymphocytes

Gangliosides are glycosphingolipids found ubiquitously on thesurface of mammalian cells. They contain a ceramide tail thatis inserted into the membrane and exposed carbohydrate and sialicacid moleties. The non-toxic B subunit oligomer (EtxB) of Escherichiacoli heat-labile enterotoxin (Etx) is a potent immunogen invivo and has profound modulatory effects on EtxB-primed lymphocytesin vitro, properties which are dependent on its ability to bindto GM1 ganglioside receptors. Here, it is shown that cross-linkingGM1 by EtxB causes a differential effect on mature CD4⁺ andCD8⁺ T cells from lymph node cultures proliferating in responseto an unrelated antigen, ovalbumin. Addition of EtxB to suchcultures led to the complete depletion of CD8⁺ T cells comparedwith enhanced activation of CD4⁺ T cells [as measured by expressionof CD25 (IL-2R)]. By contrast, addition of a mutant EtxB, EtxB(G33D),which does not bind to GM1, failed to trigger CD8⁺ T cell depletion.When EtxB was added to isolated non-immune CD8⁺ lymphocytesrapid (12–18 h) alterations in nuclear morphology andthe appearance of sub-G₀/G₁ levels of DNA were induced; propertieswhich are characteristic of cells undergoing apoptosis. EtxB(G33D)failed to trigger apoptosis, indicating that the induction ofthe apoptotic signal was dependent on the binding of GM1. Thesefindings provide an insight into the potent immunogenicity andimmunomodulatory properties of E. coli enterotoxins as wellas heralding a novel method for the selective induction of apoptosisin mature CD8⁺ T lymphocytes.     

Development of an inhaled endotoxin challenge protocol for characterizing evoked cell surface phenotype and genomic responses of airway cells in allergic individuals.

BACKGROUND: Environmental exposure to endotoxin is a known cause of exacerbation of asthma. Inhaled endotoxin protocols have been used to evaluate airway cell surface phenotypes associated with antigen presentation and innate immunity in healthy volunteers, but not in allergic volunteers. OBJECTIVES: To establish the safety of challenge with low-dose endotoxin (10,000 endotoxin units) (lipopolysaccharide [LPS]) inhalation in allergic individuals, to measure airway cell surface phenotypes associated with antigen presentation and innate immunity in induced sputum (IS) after LPS challenge, and to conduct gene expression profiling in IS cells to determine which host genetic networks are modified by LPS inhalation. METHODS: Induced sputum was obtained before and 6 hours after LPS inhalation in 10 allergic volunteers (8 with asthma and 2 with rhinitis). Flow cytometry was used to examine cell surface phenotypes on IS cells. Genomic expression was analyzed on a subset of IS samples (n = 10) using microarray and ingenuity pathway analysis. RESULTS: A total of 10,000 endotoxin units of LPS induced significant up-regulation of membrane CD14, CD11b, CD16, HLA-DR, CD86, and Fcepsilon receptor 1 on sputum phagocytes and increased expression of genes that influence antigen-presenting surface molecules (HLA-DR, chemokine ligand 2 or monocyte chemoattractant protein 1, v-rel reticuloendotheliosis viral oncogene homolog, prostaglandin-endoperoxide synthase 2 or cyclooxygenase 2, and transforming growth factor beta), immune activation (CD14, interleukin 1beta, and regulated upon activation, normal T cell expressed and secreted), and inflammation (intracellular adhesion molecule 1 and inhibitory kappaBalpha). Gene profiles for nuclear factor kappaB, interleukin 1, and tumor necrosis factor pathways were also significantly affected. CONCLUSIONS: Low-dose inhaled endotoxin challenge is safe in allergic individuals with mild to moderate disease. It enhances airway cell surface phenotypes and expression of genes associated with antigen presentation, innate immunity, and inflammation. Microarray with ingenuity pathway analysis can be successfully applied to sputum cells to characterize genetic responses to inhaled exacerbants.     

Variant Creutzfeldt–Jakob disease: a summary of current scientific knowledge in relation to public health

THE PRION DISEASES POSE UNIQUE SCIENTIFIC, medical, veterinary and regulatory challenges. Here, we summarize current information bearing on the natural history, pathobiology and epidemiology of these disorders and public policy responses to the potential threats to public health posed, particularly, by bovine spongiform encephalopathy and variant Creutzfeldt–Jakob disease (vCJD). Six years after the first case reports of vCJD, there is still no clear indication of the magnitude of the primary epidemic, or of the likelihood of lateral transmission of this untreatable disease by iatrogenic means, particularly by blood and blood products. However, the unsettling nature of the available evidence warrants prudence regarding public health policy and regulation, as well as a forward-looking approach to research.