The cytokine transforming growth factor-beta (TGF-beta) was initially purified from human platelets, a rich source of this protein. In addition to platelets, TGF-beta1 is also found in other blood fractions, including plasma and the circulating leukocytes. However, more than 15 years after the initial isolation of TGF-beta1, there remains no consensus on how much TGF-beta1 is present in normal human plasma. Here we review the difficulties associated with measuring TGF-beta concentrations in complex biological fluids, and discuss the current state of knowledge on the distribution of TGF-beta isoforms in various blood fractions as well as the nature of the TGF-beta-containing protein complexes.

Animals↗

PubMedDate not supplied

Degenerate perturbations of protein structure as the mechanism of anaesthetic action.

The interaction of the n-alkanols with lipid bilayers and excitable membranes shows that there is no simple correlation between conduction block and any of the perturbations of bilayer structure currently proposed as unitary mechanisms of local anaesthetic action. We propose instead that the n-alkanols act by direct interaction with target proteins to cause perturbations which depend directly on the precise structure of the alcohol.

Alcohols↗

PubMedDate not supplied

Mitogens increase phosphorylation of phosphoinositides in thymocytes.

In many cell systems the interaction of ligands with their receptors causes rapid breakdown and resynthesis of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2). Recent work has focused on the role of the degradation products of PtdIns(4,5)P2 as intermediates in the activation of cell function and growth: inositol trisphosphate (InsP3) can release Ca2+ from intracellular stores and diacylglycerol is thought to activate protein kinase C. This enzyme is also activated by phorbol esters (for example, 12-O-tetradecanoyl phorbol 13-acetate, TPA) and this is assumed to account for the pleiotropic effects of TPA on cell function and growth. Mouse thymocytes are not mitogenically stimulated by TPA alone, but it is a potent co-mitogen in combination with either concanavalin A (Con A) or A23187 (A. N. Corps and J.C.M., unpublished observations). Here we show that mitogenic concentrations of TPA, A23187 and Con A each cause an increase in the net phosphorylation of phosphatidylinositol (PtdIns) to PtdIns(4,5)P2 in mouse thymocytes. This is consistent with simulation by the mitogens of the same phosphoinositide phosphorylations in intact cells as recently demonstrated for the isolated products of the src and ros viral oncogenes in a cell-free system.

Animals↗

PubMedDate not supplied

A common sequence of calcium and pH signals in the mitogenic stimulation of eukaryotic cells.

When normal quiescent (G0) cells are stimulated by mitogens to enter the cell cycle, the metabolic derepression which occurs is similar in a variety of cells. The mechanisms initiating these responses and their relationship to subsequent progression through G1 to DNA synthesis in S phase, however, are generally undefined. The clearest evidence has been obtained in sea urchin eggs, where fertilization by sperm causes a rapid, transient increase in the concentration of free cytoplasmic Ca2+ [(Ca]i), followed by a sustained increase in cytoplasmic pH (pHi). It has been demonstrated clearly that these ionic responses are obligatory for progression to DNA synthesis by the normal pathway after fertilization, although the Ca2+ signal can be bypassed by parthenogenetic agents which elevate directly pHi (for example, NH+4 ions). These observations raise the questions of whether other eukaryotic cells show the same sequence of ionic responses when stimulated by mitogens and whether such signals are an obligatory component of their mitogenic pathways. We show here that a common sequence of [Ca]i and pHi responses occurs in both quiescent mouse thymocytes and Swiss 3T3 fibroblasts stimulated by appropriate mitogens. Furthermore, 'opportunistic' mitogens (those that do not act on the cells in vivo, such as concanavalin A (Con A), the Ca2+ ionophore A23187 and 12-o-tetradecanoyl phorbol 13-acetate CTPA] that are mitogenic for both mouse thymocytes and 3T3 fibroblast, each produce characteristic ionic responses that are the same in both types of cell.

Animals↗

PubMedDate not supplied

A common phenotype associated with atherogenesis in diverse mouse models of vascular lipid lesions.

The introduction of a range of different genetic modifications in mice results in altered lipoprotein metabolism and the development of vascular lipid lesions. At present, however, it is unclear to what extent the molecular events underlying lipid lesion formation are similar in these different mouse models of atherosclerosis. The aim of this study was to compare the protein expression pattern of lipid lesions from seven different mouse lines with varying susceptibility to vascular lipid lesion development, to determine to what extent lesions induced by different genetic interventions have a similar composition. The proteins we have measured, using quantitative immunofluorescence, are proteins whose expression is known to be modulated during atherogenesis in humans, including plasminogen activator inhibitor (PAI)-1, transforming growth factor (TGF)-beta 1, osteopontin and the macrophage marker CD11b. In all the mice lines we have investigated, PAI-1 was elevated wherever lesions developed. Active TGF-beta was depressed in the vessel wall of mice which developed lipid lesions, particularly in the intima. In contrast, TGF-beta 1 antigen (active plus latent TGF-beta 1) was increased at lesion sites. Accumulation of osteopontin and, with the marked exception of apolipoprotein(a) transgenic mice, tissue macrophages occurred at sites of lipid deposition in the vessel wall. Each lesion, irrespective of its size and the mouse strain in which it developed, had similar amounts of PAI-1, active TGF-beta and osteopontin per unit area of lesion. These data are consistent with a common phenotype accompanying atherogenesis, irrespective of the genetic basis of susceptibility.

Animals↗