The Spontaneously Adhesive Leukocyte Function-associated Antigen-1 (LFA-1) Integrin in Effector T Cells Mediates Rapid Actin- and Calmodulin-dependent Adhesion Strengthening to Ligand under Shear Flow

Integrins in effector T cells are highly expressed and important for trafficking of these cells and for their effector functions. However, how integrins are regulated in effector T cells remains poorly characterized. Here, we have investigated effector T cell leukocyte function-associated antigen-1 (LFA-1) regulation in primary murine effector T cells. These cells have high LFA-1 integrin expression and display high spontaneous binding to intercellular adhesion molecule-1 (ICAM-1) ligand under static conditions. In addition, these cells are able to migrate spontaneously on ICAM-1. Atomic force microscopy measurements showed that the force required for unbinding of integrin-ligand interactions increases over time (0.5–20-s contact time). The maximum unbinding force for this interaction was ∼140 piconewtons at 0.5-s contact time, increasing to 580 piconewtons at 20-s contact time. Also, the total work required to disrupt the interaction increased over the 20-s contact time, indicating LFA-1-mediated adhesion strengthening in primary effector T cells over a very quick time frame. Effector T cells adhered spontaneously to ICAM-1 under conditions of shear flow, in the absence of chemokine stimulation, and this binding was independent of protein kinase B/Akt and protein kinase C kinase activity, but dependent on calcium/calmodulin signaling and an intact actin cytoskeleton. These results indicate that effector T cell integrins are highly expressed and spontaneously adhesive in the absence of inside-out integrin signaling but that LFA-1-mediated firm adhesion under conditions of shear flow requires downstream integrin signaling, which is dependent on calcium/calmodulin and the actin cytoskeleton.     

How do morphotypes and chemotypes relate to genotypes? The colonial ascidian Cystodytes (Polycitoridae)

Intraspecies variability is widespread in marine invertebrates. Size, colour, texture, general shape and secondary chemistry can differ quite drastically from one individual to another. Cystodytes dellechiajei (Polycitoridae) is a cosmopolitan colonial ascidian with several morphotypes, most of which differ in colour and spicular composition. New molecular tools enable us to assess the taxonomic status of these morphotypes. To determine whether variation observed in Mediterranean Cystodytes has a genetic basis, we sequenced 45 specimens from eight locations of the western Mediterranean and one from Mayotte (Indian Ocean), and obtained a 617 bp fragment of the mitochondrial gene COI. Fifteen different colour morphs were recorded and four kinds of spicules were found: disk-shaped, sphere-shaped, star-shaped and discoidal, thick spicules with a toothed margin. Zooid morphology was remarkably uniform in the whole sample set. Different tree construction methods (distance-based, parsimony-based, and maximum-likelihood-based) yielded consistent results, and recognized six major clades, which had no correspondence with spicule shape and were only partially consistent with colour morphs. Results are discussed in the light of previous knowledge of the chemistry of blue, green, brown and purple colour morphs. In spite of the different colour patterns and spicular variability we concluded, on the basis of chemical and genetic data, that the morphological traits analysed were not consistent enough to be used to differentiate between Cystodytes species. We point out the importance of genetics and chemistry in assessing the taxonomic status of species with variable morphology.     

Lipid Peroxidation in Hyperlipidaemic Patients. A Study of Plasma using an HPLC-Based Thiobarbituric Acid Test

The question of “increased lipid peroxidation” in plasma from hyperlipidaemic patients was investigated using an improved HPLC-based assay for thiobarbituric acid-reactive material. Levels of TBARS in healthy human controls were at or close to zero, provided that butylated hydroxytoluene was added to the sample with the TBA reagents. Levels of plasma TBARS in hyperlipidaemic patients were elevated, although the absolute levels were much lower than those reported previously in the literature.     

Self-assembling Fmoc dipeptide hydrogel for <Emphasis Type="Italic">in situ</Emphasis> 3D cell culturing

Background  

Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derived hydrogel for use as a 3D cell culture scaffold at the microscale.     

Cyclin E overexpression impairs progression through mitosis by inhibiting APC(Cdh1)

Overexpression of cyclin E, an activator of cyclin-dependent kinase 2, has been linked to human cancer. In cell culture models, the forced expression of cyclin E leads to aneuploidy and polyploidy, which is consistent with a direct role of cyclin E overexpression in tumorigenesis. In this study, we show that the overexpression of cyclin E has a direct effect on progression through the latter stages of mitotic prometaphase before the complete alignment of chromosomes at the metaphase plate. In some cases, such cells fail to divide chromosomes, resulting in polyploidy. In others, cells proceed to anaphase without the complete alignment of chromosomes. These phenotypes can be explained by an ability of overexpressed cyclin E to inhibit residual anaphase-promoting complex (APC(Cdh1)) activity that persists as cells progress up to and through the early stages of mitosis, resulting in the abnormal accumulation of APC(Cdh1) substrates as cells enter mitosis. We further show that the accumulation of securin and cyclin B1 can account for the cyclin E-mediated mitotic phenotype.