The importance of shoot morphological plasticity on plant coexistence: a pot experiment.

Plant morphological plasticity affects species coexistence by enhancing local coexistence. Here, we test the importance of plasticity to light availability for species coexistence. We hypothesise that high average plasticity in a species assemblage promotes coexistence and tested for the effect of differential plasticity on the competitive success of neighbouring species. Sixteen herbaceous species with known morphological plasticity were grown pairwise in 95 combinations in 285 pots. We calculated mean plasticity and difference of plasticity for each pair of species in a pot using previously estimated degree of plasticity in leaf number, leaf length, leaf area and SLA. We then related these to biomass-based evenness of abundance in a pot and to competitive success of the 16 species. Unexpectedly, average plasticity did not affect biomass production between coexisting species. Instead, large differences in plasticity among two competitive neighbours predicted low diversity (high degree of dominance) in an assemblage. Higher than neighbour plasticity generally predicted competitive superiority in an assemblage. The opposite was true for plasticity of SLA, where species with low plasticity tended to dominate. Unlike earlier field studies, our results show that phenotypic plasticity in various plant traits pose opposite effects to interspecific competition. Subsequently, these effects possibly affect species composition and richness through which plasticity has significant consequences for plant communities and, therefore, should be accounted for in relevant studies in plant ecology.

N-Cadherin modified lipid bilayers promote neural network formation and circuitry.

Neural adhesion, maturation, and the correct wiring of the brain to establish each neuron's intended connectivity are controlled by complex interactions of bioactive molecules such as ligands, growth factors, or enzymes. The correct pairing of adjacent neurons is thought to be highly regulated by ligand-mediated cell-cell adhesion proteins, which are known to induce signaling activities. We developed a new platform consisting of supported lipid bilayers incorporated with Fc-chimera synaptic proteins like ephrinA5 or N-cadherin. We extensively characterized their function employing a quartz crystal microbalance with dissipation (QCM-D), calcium imaging, and immunofluorescence analysis. Our biomimetic platform has been shown to promote neural cell adhesion and to improve neural maturation at day in vitro 7 (DIV7) as indicated by an elevated expression of synaptophysin.

Smac mimetic sensitizes glioblastoma cells to Temozolomide-induced apoptosis in a RIP1- and NF-κB-dependent manner.

Inhibitor of apoptosis (IAP) proteins are expressed at high levels in many cancers and therefore represent attractive targets for therapeutic intervention. Here, we report for the first time that the second mitochondria-derived activator of caspases (Smac) mimetic BV6 sensitizes glioblastoma cells toward Temozolomide (TMZ), the first-line chemotherapeutic agent in the treatment of glioblastoma. BV6 and TMZ synergistically reduce cell viability and trigger apoptosis in glioblastoma cells (combination index <0.4-0.8), which is accompanied by increased loss of mitochondrial-membrane potential, cytochrome c release, caspase activation and caspase-dependent apoptosis. Analysis of the molecular mechanisms reveals that BV6 causes rapid degradation of cIAP1, leading to stabilization of NF-κB-inducing kinase and NF-κB activation. BV6-stimulated NF-κB activation is critically required for sensitization toward TMZ, as inhibition of NF-κB by overexpression of the mutant IκBα super-repressor profoundly reduces loss of mitochondrial membrane potential, cytochrome c release, caspase activation and apoptosis. Of note, BV6-mediated sensitization to TMZ is not associated with increased tumor necrosis factor alpha (TNFα) production. Also, TNFα, CD95 or TRAIL-blocking antibodies or knockdown of TNFR1 have no or little effect on combination treatment-induced apoptosis. Interestingly, BV6 and TMZ cooperate to trigger the formation of a RIP1 (receptor activating protein 1)/caspase-8/FADD complex. Knockdown of RIP1 by small interfering RNA significantly reduces BV6- and TMZ-induced caspase-8 activation and apoptosis, showing that RIP1 is necessary for apoptosis induction. By demonstrating that BV6 primes glioblastoma cells for TMZ in a NF-κB- and RIP1-dependent manner, these findings build the rationale for further (pre)clinical development of Smac mimetics in combination with TMZ.

cIAP1 and TAK1 protect cells from TNF-induced necrosis by preventing RIP1/RIP3-dependent reactive oxygen species production.

Three members of the IAP family (X-linked inhibitor of apoptosis (XIAP), cellular inhibitor of apoptosis proteins-1/-2 (cIAP1 and cIAP2)) are potent suppressors of apoptosis. Recent studies have shown that cIAP1 and cIAP2, unlike XIAP, are not direct caspase inhibitors, but block apoptosis by functioning as E3 ligases for effector caspases and receptor-interacting protein 1 (RIP1). cIAP-mediated polyubiquitination of RIP1 allows it to bind to the pro-survival kinase transforming growth factor-ß-activated kinase 1 (TAK1) which prevents it from activating caspase-8-dependent death, a process reverted by the de-ubiquitinase CYLD. RIP1 is also a regulator of necrosis, a caspase-independent type of cell death. Here, we show that cells depleted of the IAPs by treatment with the IAP antagonist BV6 are greatly sensitized to tumor necrosis factor (TNF)-induced necrosis, but not to necrotic death induced by anti-Fas, poly(I:C) oxidative stress. Specific targeting of the IAPs by RNAi revealed that repression of cIAP1 is responsible for the sensitization. Similarly, lowering TAK1 levels or inhibiting its kinase activity sensitized cells to TNF-induced necrosis, whereas repressing CYLD had the opposite effect. We show that this sensitization to death is accompanied by enhanced RIP1 kinase activity, increased recruitment of RIP1 to Fas-associated via death domain and RIP3 (which allows necrosome formation), and elevated RIP1 kinase-dependent accumulation of reactive oxygen species (ROS). In conclusion, our data indicate that cIAP1 and TAK1 protect cells from TNF-induced necrosis by preventing RIP1/RIP3-dependent ROS production.

Structure-function analysis of a phage display-derived peptide that binds to insulin-like growth factor binding protein 1.

Highly structured, peptide antagonists of the interaction between insulin-like growth factor 1 (IGF-I) and IGF binding protein 1 (IGFBP-1) have recently been discovered by phage display of naïve peptide libraries [Lowman, H. B., et al. (1998) Biochemistry 37, 8870--8878]. We now report a detailed analysis of the features of this turn-helix peptide motif that are necessary for IGFBP-1 binding and structural integrity. Further rounds of phage randomization indicate the importance of residues contributing to a hydrophobic patch on one face of the helix. Alanine-scanning substitutions confirm that the hydrophobic residues are necessary for binding. However, structural analysis by NMR spectroscopy indicates that some of these analogues are less well folded. Structured, high-affinity analogues that lack the disulfide bond were prepared by introducing a covalent constraint between side chains at positions i and i + 7 or i + 8 within the helix. Analogues based on this scaffold demonstrate that a helical conformation is present in the bound state, and that hydrophobic side chains in this helix, and residues immediately preceding it, interact with IGFBP-1. By comparison of alanine scanning data for IGF-I and the turn-helix peptide, we propose a model for common surface features of these molecules that recognize IGFBP-1.

Barium ions reduce contraction kinetics in rat tracheal smooth muscle.

In isolated rat tracheal smooth muscle, contraction was induced by electrical field stimulation (= EL; frequency = 30 Hz; pulse duration = 0.17 ms), 10 mumol/l acetylcholine (= ACh), potassium depolarization (137 mmol/l K), or by 10 mmol/l barium (= BA). Contraction kinetics were studied by analyzing tension recovery after cessation of a 1.8 s length vibration (100 Hz sinusoidal; amplitude = 6% of the muscle length). The contraction speed was high during EL as can be seen from the short time constant of post-vibration tension recovery (tau = 5.90 +/- 0.14 s) found 30 s after onset of stimulation. The time constants of tension recovery during long-term (50 min) activation averaged 12.88 +/- 0.32 s (K) and 13.24 +/- 0.17 s (ACh) when the vibration was stopped 8-45 min after onset of activation. As both of these stimuli act mainly via cholinergic receptors, similar down-regulated contraction kinetics occur under steady-state conditions of tonic contraction. However, during barium activation steady-state conditions need a 45 min agonist incubation, and the time constant of post-vibration tension recovery was extended to 34.05 +/- 2.21 s. Thus, calcium may be replaced by barium in the force generation process but it produces slower cycling of cross-bridges.