Inhibition of PI3K prevents the proliferation and differentiation of human lung fibroblasts into myofibroblasts: the role of class I P110 isoforms

Idiopathic pulmonary fibrosis (IPF) is a progressive fibroproliferative disease characterized by an accumulation of fibroblasts and myofibroblasts in the alveolar wall. Even though the pathogenesis of this fatal disorder remains unclear, transforming growth factor-β (TGF-β)-induced differentiation and proliferation of myofibroblasts is recognized as a primary event. The molecular pathways involved in TGF-β signalling are generally Smad-dependent yet Smad-independent pathways, including phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt), have been recently proposed. In this research we established ex-vivo cultures of human lung fibroblasts and we investigated the role of the PI3K/Akt pathway in two critical stages of the fibrotic process induced by TGF-β: fibroblast proliferation and differentiation into myofibroblasts. Here we show that the pan-inhibitor of PI3Ks LY294002 is able to abrogate the TGF-β-induced increase in cell proliferation, in α- smooth muscle actin expression and in collagen production besides inhibiting Akt phosphorylation, thus demonstrating the centrality of the PI3K/Akt pathway in lung fibroblast proliferation and differentiation. Moreover, for the first time we show that PI3K p110δ and p110γ are functionally expressed in human lung fibroblasts, in addition to the ubiquitously expressed p110α and β. Finally, results obtained with both selective inhibitors and gene knocking-down experiments demonstrate a major role of p110γ and p110α in both TGF-β-induced fibroblast proliferation and differentiation. This finding suggests that specific PI3K isoforms can be pharmacological targets in IPF.     

Mycorrhizal responses in wheat: shading decreases growth but does not lower the contribution of the fungal phosphate uptake pathway

Effects have been investigated of reduced C supply (induced by shade) on arbuscular mycorrhizal (AM) colonisation, mycorrhizal growth responses (MGRs) and on AM-mediated and direct uptake of phosphate (Pi) (using ³²P) in wheat, a plant that does not usually respond positively to AM colonisation. Shading markedly reduced growth and shoot/root dry weight ratios of both AM and non-mycorrhizal wheat, indicating decreased photosynthetic C supply. However, shading had very little effect on percent root length colonised by Rhizophagus irregularis or Gigaspora margarita or on MGRs, which remained slightly positive or zero, regardless of shade; there were no growth depressions under shade. By 6 weeks, when the contributions of the AM pathway were measured with ³²P supplied in small hyphal compartments, R. irregularis had supplied 23 to 28 % of shoot P with no significant effect of shading. Data show that reduced C availability did not reduce the contribution of the AM pathway to plant P, so the fungi were not acting physiologically as parasites. These results support our previous hypothesis that lack of positive MGR is not necessarily the outcome of excessive C use by the fungi or failure to deliver P via the AM pathway.     

The O-polysaccharide of Pseudomonas syringae pv. mori NCPPB 1656 is a beta-(1-->2)-linked homopolymer of L-rhamnose

The O-polysaccharide from the lipopolysaccharide of the phytopathogenic bacterium Pseudomonas syringae pv. mori NCPPB 1656 was studied by sugar analysis along with 1H and 13C NMR spectroscopy and found to be a new beta-(1-->2)-linked homopolymer of L-rhamnose.     

A spectroscopy approach for the study of the interactions of bioactive vanadium species with bovine serum albumin

The interest in biological functions (benefits or toxics effects) of vanadium species has grown enormously in recent years. In this work, different spectroscopic methods were applied to study the effects of the interaction of vanadyl and vanadate species with bovine serum albumin (BSA), considered as the most abundant plasma protein. UV-Vis, Fourier transform infrared (FT-IR), and FT-Raman spectroscopies were used to investigate changes in secondary and tertiary structures of BSA induced by the binding of oxovanadium(IV) and vanadate(V) species (VO(2+) and VO3(-), respectively). Correlations between the metal ion binding mode, protein conformational transitions, and structural variations were established.     

α-Synuclein and Its A30P Mutant Affect Actin Cytoskeletal Structure and Dynamics

The function of α-synuclein, a soluble protein abundant in the brain and concentrated at presynaptic terminals, is still undefined. Yet, α-synuclein overexpression and the expression of its A30P mutant are associated with familial Parkinson''s disease. Working in cell-free conditions, in two cell lines as well as in primary neurons we demonstrate that α-synuclein and its A30P mutant have different effects on actin polymerization. Wild-type α-synuclein binds actin, slows down its polymerization and accelerates its depolymerization, probably by monomer sequestration; A30P mutant α-synuclein increases the rate of actin polymerization and disrupts the cytoskeleton during reassembly of actin filaments. Consequently, in cells expressing mutant α-synuclein, cytoskeleton-dependent processes, such as cell migration, are inhibited, while exo- and endocytic traffic is altered. In hippocampal neurons from mice carrying a deletion of the α-synuclein gene, electroporation of wild-type α-synuclein increases actin instability during remodeling, with growth of lamellipodia-like structures and apparent cell enlargement, whereas A30P α-synuclein induces discrete actin-rich foci during cytoskeleton reassembly. In conclusion, α-synuclein appears to play a major role in actin cytoskeletal dynamics and various aspects of microfilament function. Actin cytoskeletal disruption induced by the A30P mutant might alter various cellular processes and thereby play a role in the pathogenesis of neurodegeneration.     

Time‐resolved spectral measurements of delayed luminescence from a single soybean seed: effects of thermal damage and correlation with germination performance

Delayed luminescence from a single dry soybean seed was investigated in both spectral and time domains, under different excitation wavelengths. Emission spectra were collected, under 337 nm laser excitation, from native and artificially deteriorated seeds and the time‐dependence of different spectral components was analyzed in detail. The single seed viability was evaluated through observation of germination properties after imbibition and compared with different parameters related to the luminescence kinetics. The significant correlation found between single seed delayed luminescence parameters and germination capability strongly validates the connection of this phenomenon with the functional state of the system and suggests the development of a non‐invasive technique for seed quality determination. Copyright © 2009 John Wiley & Sons, Ltd.     

Are behavioral responses to eyespots in sticklebacks influenced by the visual environment? An experimental examination

Eyespots are taxonomically widespread color patterns consisting of large concentric rings that are commonly assumed to protect prey by influencing the behaviors of predators. Although there is ample experimental evidence supporting an anti‐predator function of eyespots in terrestrial animals, whether eyespots have a similar deterring function in aquatic animals remains unclear. Furthermore, studies in terrestrial systems suggest that the protective function of eyespots depends on ambient light conditions where predators encounter them, but this effect has never been tested in aquatic environments. Here, we examine how eyespots influence behavioral responses in an aquatic environment under different visual environments, using laboratory‐reared three‐spined sticklebacks (Gasterosteus aculeatus) as model predators. Specifically, we experimentally examined behavioral responses of sticklebacks toward artificial prey patterns (control vs. eyespots) under two different light environment treatments (low vs. high). We found that eyespots did not postpone attacks from sticklebacks. However, sticklebacks approaching eyespots stopped more frequently than sticklebacks approaching prey items with a control pattern. Sticklebacks were (marginally) slower to attack prey in the low‐light treatment, but the light level did not influence stickleback behavioral responses toward eyespots. We conclude that eyespots can modulate some behaviors of an aquatic predator, albeit with a different functional role from that previously demonstrated in terrestrial species.