Intracellular Cytokine Staining and Flow Cytometry: Considerations for Application in Clinical Trials of Novel Tuberculosis Vaccines.

Intracellular cytokine staining combined with flow cytometry is one of a number of assays designed to assess T-cell immune responses. It has the specific advantage of enabling the simultaneous assessment of multiple phenotypic, differentiation and functional parameters pertaining to responding T-cells, most notably, the expression of multiple effector cytokines. These attributes make the technique particularly suitable for the assessment of T-cell immune responses induced by novel tuberculosis vaccines in clinical trials. However, depending upon the particular nature of a given vaccine and trial setting, there are approaches that may be taken at different stages of the assay that are more suitable than other alternatives. In this paper, the Tuberculosis Vaccine Initiative (TBVI) TB Biomarker Working group reports on efforts to assess the conditions that will determine when particular assay approaches should be employed. We have found that choices relating to the use of fresh whole blood or peripheral blood mononuclear cells (PBMC) and frozen PBMC; use of serum-containing or serum-free medium; length of stimulation period and use of co-stimulatory antibodies can all affect the sensitivity of intracellular cytokine assays. In the case of sample material, frozen PBMC, despite some loss of sensitivity, may be more advantageous for batch analysis. We also recommend that for multi-site studies, common antibody panels, gating strategies and analysis approaches should be employed for better comparability.

Didactical reflections about some proofs of the pythagorean proposition

This theoretical paper presents some dimensions considered in the literature to analyze proof in the teaching and learning of mathematics. In order to show how different types of proofs can be used with students of different levels, we use these dimensions to analyze four proofs of the Pythagorean proposition.

Basic components in the scienctific didactical training of the secondary school mathematics teachers

Secondary mathematics teacher training in Spain is currently the subject of a heated revision debate. The speed of social, cultural, scientific and economic changes have left a hundred years old teacher training model well behind. However, academical inertia and professional interests are impeding a real new training of the mathematics teacher as an autonomous mathematical educator. Teachers of Didactic of Mathematics and the Spanish Associations of mathematics teachers have recently been discussing the issue. Their conclusions are included here.

HyperJoseph: the hypertextual organization. Epistemological considerations

HYPERJOSEPH combines hypertext, information retrieval, literary studies, Biblical scholarship, and linguistics. Dialectically, this paper contrasts hypertextual form (the extant tool) and AI-captured content (a desideratum), in the HYPERJOSEPH project. The discussion is more general and oriented to epistemology.

Ontological considerations of time, meta-predicates and temporal propositions

A natural approach to representing and reasoning about temporal propositions (i.e., statements with time-dependent truth-values) is to associate them with time elements. In the literature, there are three choices regarding the primitive for the ontology of time: (1) instantaneous points, (2) durative intervals and (3) both points and intervals. Problems may arise when one conflates different views of temporal structure and questions whether some certain types of temporal propositions can be validly and meaningfully associated with different time elements. In this paper, we shall summarize an ontological glossary with respect to time elements, and diversify a wider range of meta-predicates for ascribing temporal propositions to time elements. Based on these, we shall also devise a versatile categorization of temporal propositions, which can subsume those representative categories proposed in the literature, including that of Vendler, of McDermott, of Allen, of Shoham, of Galton and of Terenziani and Torasso. It is demonstrated that the new categorization of propositions, together with the proposed range of meta-predicates, provides the expressive power for modeling some typical temporal terms/phenomena, such as starting-instant, stopping-instant, dividing-instant, instigation, termination and intermingling etc.

Elemental analysis of soil samples for forensic purposes by inductively coupled plasma spectrometry - precision considerations

Major and trace elemental composition provides a powerful basis for forensic comparison of soils, sediments and rocks. However, it is important that the potential 'errors' associated with the procedures are fully understood and quantified, and that standard protocols are applied for sample preparation and analysis. This paper describes such a standard procedure and reports results both for instrumental measurement precision (repeatability) and overall 'method' precision (reproducibility). Results obtained both for certified reference materials and example soils show that the instrumental measurement precision (defined by the coefficient of variation, CV) for most elements is better than 2-3%. When different solutions were prepared from the same sample powder, and from different sub-sample powders prepared from the same parent sample, the CV increased to c. 5-6% for many elements. The largest variation was found in results for certified reference materials generated from 23 instrument runs over an 18 month period (mean CV=c. 11%). Some elements were more variable than others. W was found to be the most variable and the elements V, Cr, Co, Cu, Ni and Pb also showed higher than average variability. SiO2, CaO, Al2O3 and Fe2O3, Rb, Sr, La, Ce, Nd and Sm generally showed lower than average variability, and therefore provided the most reliable basis for inter-sample comparison. It is recommended that, whenever possible, samples relating to the same investigation should be analysed in the same sample run, or at least sequential runs.

Some Practical Considerations In The Measurement Of Pollution Effects On Bivalve Mollusks And Some Possible Ecological Consequences

A consideration of some physiological (rates of oxygen consumption, the scope for growth) and cellular (the cytochemical latency of a lysosomal enzyme) processes in bivalve molluscs suggests that animal size and seasonal changes related to the gametogenic cycle are important sources of natural variability. Correcting for size using regression techniques, and limiting measurements to one part of the gametogenic cycle, reduces observed natural variability considerably. Differences between populations are then still apparent, but the results of laboratory experiments with hydrocarbons from crude oil suggest that it should be possible to detect sub-lethal effects due to pollution (the ‘signal’) in the presence of the remaining natural variability (the ‘noise’). Statistical considerations, taken together with results from current studies on Mytilus edulis and Scobicularia plana, indicate that sample sizes of 10–15 individuals should suffice for the detection of possible pollution effects. The physiological effects to be expected in the presence of sub-lethal levels of polluting hydrocarbons are on a scaie that can cause significant ecological damage to a population through a reduction in fecundity and the residual reproductive value of the individuals.