The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation

Carboxyfluorescein succinimidyl ester (CFSE) is an effective and popular means to monitor lymphocyte division1-3. CFSE covalently labels long-lived intracellular molecules with the fluorescent dye, carboxyfluorescein. Thus, when a CFSE-labeled cell divides, its progeny are endowed with half the number of carboxyfluorescein-tagged molecules and thus each cell division can be assessed by measuring the corresponding decrease in cell fluorescence via Flow cytometry. The capacity of CFSE to label lymphocyte populations with a high fluorescent intensity of exceptionally low variance, coupled with its low cell toxicity, make it an ideal dye to measure cell division. Since it is a fluorescein-based dye it is also compatible with a broad range of other fluorochromes making it applicable to multi-color flow cytometry. This article describes the procedures typically used for labeling mouse lymphocytes for the purpose of monitoring up to 8 cell divisions. These labeled cells can be used both for in vitro and in vivo studies

Bystander B cells rapidly acquire antigen receptors from activated B cells by membrane transfer: a novel mechanism for enhancing specific antigen presentation

The B cell antigen receptor (BCR) efficiently facilitates the capture and processing of a specific antigen for presentation on MHC class II molecules to antigen specific CD4+ T cells (1). Despite this, the majority of B cells are only thought to play a limited role in CD4+ T cell activation since BCRs are clonotypically expressed. Here we show, however, that activated B cells can, both in vitro and in vivo, rapidly donate their BCR to bystander B cells, a process that is mediated by direct membrane transfer between adjacent B cells and is amplified by the interaction of the BCR with specific antigen. This results in a dramatic expansion in the number of antigen-binding B cells in vivo, with the transferred BCR endowing recipient B cells with the ability to present specific antigen to antigen-specific CD4+ T cells

The Use of CFSE-like Dyes for Measuring Lymphocyte Proliferation : Experimental Considerations and Biological Variables

The measurement of CFSE dilution by flow cytometry is a powerful experimental tool to measure lymphocyte proliferation. CFSE fluorescence precisely halves after each cell division in a highly predictable manner and is thus highly amenable to mathematical modelling. However, there are several biological and experimental conditions that can affect the quality of the proliferation data generated, which may be important to consider when modelling dye dilution data sets. Here we overview several of these variables including the type of fluorescent dye used to monitor cell division, dye labelling methodology, lymphocyte subset differences, in vitro versus in vivo experimental assays, cell autofluorescence, and dye transfer between cells.This work was supported by a Project Grant to BQ and CP and a Program Grant to CP from the National Health and Medical Research Council (NHMRC) of Australia

Anisotropic diffusion in continuum relaxation of stepped crystal surfaces

We study the continuum limit in 2+1 dimensions of nanoscale anisotropic diffusion processes on crystal surfaces relaxing to become flat below roughening. Our main result is a continuum law for the surface flux in terms of a new continuum-scale tensor mobility. The starting point is the Burton, Cabrera and Frank (BCF) theory, which offers a discrete scheme for atomic steps whose motion drives surface evolution. Our derivation is based on the separation of local space variables into fast and slow. The model includes: (i) anisotropic diffusion of adsorbed atoms (adatoms) on terraces separating steps; (ii) diffusion of atoms along step edges; and (iii) attachment-detachment of atoms at step edges. We derive a parabolic fourth-order, fully nonlinear partial differential equation (PDE) for the continuum surface height profile. An ingredient of this PDE is the surface mobility for the adatom flux, which is a nontrivial extension of the tensor mobility for isotropic terrace diffusion derived previously by Margetis and Kohn. Approximate, separable solutions of the PDE are discussed.Comment: 14 pages, 1 figur

Modelling and forecasting the UK tourism growth cycle in Algarve

Over the past three decades, Portugal has developed a strong economic dependence on tourism, which has several implications for the country's overall economic development. Tourism is an activity that is interrelated strongly with the economic system since Portugal as a whole and specific regions in particular rely on the performance of tourism for their economic activity. Moreover, because economic cycles affect tourism development, it is highly vulnerable to economic fluctuations. Most tourists who visit Portugal are from the European Union, especially Western Europe. Statistics are based on the number of overnight stays in hotel accommodation and other similar establishments. In 2005, the main source markets were the UK (30.7%), Germany (16.5%), Spain (11.5%), the Netherlands (6.8%), France (4.7%), Ireland (3.6%) and Italy (3.1%). These values show that the UK has the greatest share of visitors to Algarve. The purpose of this paper is to propose a modelling approach that best fits the tourism flow pattern in order to support forecasting. The paper contributes to our understanding of the relationship between economic cycles and tourism flows to Portugal (Algarve) and explores the potential of applying the diffusion index model proposed by Stock and Watson (1999, 2002) for tourism demand forecasting

Asymptotics for Panel Models with Common Shocks

This article develops a novel asymptotic theory for panel models with common shocks. We assume that contemporaneous correlation can be generated by both the presence of common regressors among units and weak spatial dependence among the error terms. Several characteristics of the panel are considered: cross-sectional and time-series dimensions can either be fixed or large; factors can either be observable or unobservable; the factor model can describe either a cointegration relationship or a spurious regression, and we also consider the stationary case. We derive the rate of convergence and the limit distributions for the ordinary least square (OLS) estimates of the model parameters under all the aforementioned cases

N-[(E)-4-Chloro­benzyl­idene]-2,4-dimethyl­aniline

The title mol­ecule, C15H14ClN, exists in a trans configuration with respect to the C=N bond [1.2813 (16) Å]. The dihedral angle between the benzene rings is 52.91 (6)°. The crystal structure is stabilized by weak inter­molecular C—H⋯π inter­actions

2-[(E)-(2,4,6-Trichloro­phen­yl)imino­meth­yl]phenol

The title mol­ecule, C13H8Cl3NO, exists in a trans configuration with respect to the C=N bond [1.278 (2) Å]. The benzene rings form a dihedral angle of 24.64 (11)°. The mol­ecular structure is stabilized by an intra­molecular O—H⋯N hydrogen bond, which generates an S(6) ring motif. In the crystal, π–π stacking inter­actions [centroid–centroid distances = 3.6893 (14) Å] are observed

2-[(E)-(2,4-Dimethyl­phen­yl)imino­meth­yl]phenol

The asymmetric unit of the title compound, C15H15NO, contains two independent mol­ecules, both of which exist in trans configurations with respect to the C=N bonds [1.278 (2) and 1.279 (2) Å]. In each mol­ecule, intra­molecular O—H⋯N hydrogen bonds generate S(6) ring motifs. In one mol­ecule, the benzene rings form a dihedral angle of 13.38 (9)°, while in the other mol­ecule the dihedral angle is 30.60 (10)°. In the crystal, the two independent mol­ecules are linked via weak inter­molecular C—H⋯O hydrogen bonds