Simple and efficient methods for isolation and activity measurement of the recombinant hirudin variant 3 from Bacillus subtilis

A simple purification approach of the recombinant hirudin variant 3 from the Bacillus subtilis was established, by which the hirudin could be purified to the purity of 95% through one-step chromatography with the total recovery rate of 83.9%. A modified Markwardt thrombin titration method for measuring hirudin activity was also set up. Briefly, a series of concentrations of thrombin was prepared and titrated to hirudin sample, respectively and the anti-thrombin activity-range of hirudin was narrowed down by several thrombin solutions at high or low concentration and the optimum group of thrombin concentrations was determined for titration of the hirudin sample. In this modified method, the hirudin activity was determined more accurately, concisely and promptly than the classic Markwardt method.Key words: Hirudin, thrombin titration method, chromatography, purification

The influence of community factors on local entrepreneurs' support for tourism

This study probes the influence of community attachment on local entrepreneurs’ perceptions of tourism’s local impacts. Six latent constructs were derived from social exchange theory and community attachment theory; 11 hypotheses were tested, using structural equation modelling, with data from 297 Taiwanese night market entrepreneurs. Significant theoretical contributions to understanding relationships between entrepreneurs and community were found: effects of community satisfaction on support for tourism were significant and fully mediated by perceived benefits. Community factors and tourist contact frequency were important in entrepreneurs’ decisions on further tourism development. Community factors showed low but significant relationships with the perceived costs of tourism, A case is made for sustainable tourism governance measures, including partnership creation, destination management systems, and visitor experience planning

The effect of calcium on auxin depletion-induced tomato (Lycopersicon esculentum Mill.) pedicel abscission

Indole-3-acetic acid (IAA) and calcium are the most important factors that instigate plant organ abscission. This study aimed to elucidate the mechanisms that underlie the effects of IAA and calcium on delayed abscission in tomato. The results showed a clear trend towards reduced abscission rates with increased concentrations of IAA, and the applications on pedicel proximal or distal side also resulted in a different abscission. IAA combined with calcium significantly improved inhibition in contrast to IAA only, while IAA combined with magnesium exhibited little increased inhibition. 1-NNaphthylphthalamic acid (NPA), a polar auxin transport inhibitor, accelerated the abscission. IAA transported basipetally through an assay with 4 mm long pedicel sections indicated that the average transport intensity of [3H]-IAA applied to the distal pedicel end was 65 Bq h–1 and the average velocity was 5.29 mm h-1. When the proximal side was incubated in [3H]-IAA, its average transport intensity reduced to 19.53 Bq h–1 and the average velocity was only 1.92 mm h-1. Calcium treatment enhanced IAA transportation, as shown by significantly enhancing the transport intensity, but it had no effect on velocity.Keywords: Indole-3-acetic acid (IAA), calcium, abscission, tomat

Inhibition of the tyrosine phosphatase SHP-2 suppresses angiogenesis in vitro and in vivo

Endothelial cell survival is indispensable to maintain endothelial integrity and initiate new vessel formation. We investigated the role of SHP-2 in endothelial cell survival and angiogenesis in vitro as well as in vivo. SHP-2 function in cultured human umbilical vein and human dermal microvascular endothelial cells was inhibited by either silencing the protein expression with antisense-oligodesoxynucleotides or treatment with a pharmacological inhibitor (PtpI IV). SHP-2 inhibition impaired capillary-like structure formation (p < 0.01; n = 8) in vitro as well as new vessel growth ex vivo (p < 0.05; n = 10) and in vivo in the chicken chorioallantoic membrane (p < 0.01, n = 4). Additionally, SHP-2 knock-down abrogated fibroblast growth factor 2 (FGF-2)-dependent endothelial proliferation measured by MTT reduction ( p ! 0.01; n = 12). The inhibitory effect of SHP-2 knock-down on vessel growth was mediated by increased endothelial apoptosis ( annexin V staining, p ! 0.05, n = 9), which was associated with reduced FGF-2-induced phosphorylation of phosphatidylinositol 3-kinase (PI3-K), Akt and extracellular regulated kinase 1/2 (ERK1/2) and involved diminished ERK1/2 phosphorylation after PI3-K inhibition (n=3). These results suggest that SHP-2 regulates endothelial cell survival through PI3-K-Akt and mitogen-activated protein kinase pathways thereby strongly affecting new vessel formation. Thus, SHP-2 exhibits a pivotal role in angiogenesis and may represent an interesting target for therapeutic approaches controlling vessel growth. Copyright (C) 2007 S. Karger AG, Basel

Transfer Functions for Protein Signal Transduction: Application to a Model of Striatal Neural Plasticity

We present a novel formulation for biochemical reaction networks in the context of signal transduction. The model consists of input-output transfer functions, which are derived from differential equations, using stable equilibria. We select a set of 'source' species, which receive input signals. Signals are transmitted to all other species in the system (the 'target' species) with a specific delay and transmission strength. The delay is computed as the maximal reaction time until a stable equilibrium for the target species is reached, in the context of all other reactions in the system. The transmission strength is the concentration change of the target species. The computed input-output transfer functions can be stored in a matrix, fitted with parameters, and recalled to build discrete dynamical models. By separating reaction time and concentration we can greatly simplify the model, circumventing typical problems of complex dynamical systems. The transfer function transformation can be applied to mass-action kinetic models of signal transduction. The paper shows that this approach yields significant insight, while remaining an executable dynamical model for signal transduction. In particular we can deconstruct the complex system into local transfer functions between individual species. As an example, we examine modularity and signal integration using a published model of striatal neural plasticity. The modules that emerge correspond to a known biological distinction between calcium-dependent and cAMP-dependent pathways. We also found that overall interconnectedness depends on the magnitude of input, with high connectivity at low input and less connectivity at moderate to high input. This general result, which directly follows from the properties of individual transfer functions, contradicts notions of ubiquitous complexity by showing input-dependent signal transmission inactivation.Comment: 13 pages, 5 tables, 15 figure

Thermodynamic Properties of Supported and Embedded Metallic Nanocrystals: Gold on/in SiO2

We report on the calculations of the cohesive energy, melting temperature and vacancy formation energy for Au nanocrystals with different size supported on and embedded in SiO2. The calculations are performed crossing our previous data on the surface free energy of the supported and embedded nanocrystals with the theoretical surface-area-difference model developed by W. H. Qi for the description of the size-dependent thermodynamics properties of low-dimensional solid-state systems. Such calculations are employed as a function of the nanocrystals size and surface energy. For nanocrystals supported on SiO2, as results of the calculations, we obtain, for a fixed nanocrystal size, an almost constant cohesive energy, melting temperature and vacancy formation energy as a function of their surface energy; instead, for those embedded in SiO2, they decreases when the nanocrystal surface free energy increases. Furthermore, the cohesive energy, melting temperature and vacancy formation energy increase when the nanocrystal size increases: for the nanocrystals on SiO2, they tend to the values of the bulk Au; for the nanocrystals in SiO2 in correspondence to sufficiently small values of their surface energy, they are greater than the bulk values. In the case of the melting temperature, this phenomenon corresponds to the experimentally well-known superheating process

A peptide derived from TIMP-3 inhibits multiple angiogenic growth factor receptors and tumour growth and inflammatory arthritis in mice

The binding of vascular endothelial growth factor (VEGF) to VEGF receptor-2 (VEGFR-2) on the surface of vascular endothelial cells stimulates many steps in the angiogenic pathway. Inhibition of this interaction is proving of value in moderating the neovascularization accompanying age-related macular degeneration and in the treatment of cancer. Tissue inhibitor of metalloproteinases-3 (TIMP-3) has been shown to be a natural VEGFR-2 specific antagonist—an activity that is independent of its ability to inhibit metalloproteinases. In this investigation we localize this activity to the C-terminal domain of the TIMP-3 molecule and characterize a short peptide, corresponding to part of this domain, that not only inhibits all three VEGF-family receptors, but also fibroblast growth factor and platelet-derived growth factor receptors. This multiple-receptor inhibition may explain why the peptide was also seen to be a powerful inhibitor of tumour growth and also a partial inhibitor of arthritic joint inflammation in vivo

Allelopathic effects of Ulva pertusa, Corallina pilulifera and Sargassum thunbergii on the growth of the dinoflagellates Heterosigma akashiwo and Alexandrium tamarense

The allelopathic effects of fresh tissue, dry powder and aqueous extracts of three macroalgae, Ulva pertusa, Corallina pilulifera and Sargassum thunbergii, on the growth of the dinoflagellates Heterosigma akashiwo and Alexandrium tamarense were evaluated using coexistence culture systems in which concentrations of the three macroalga were varied. The results of the coexistence assay showed that the growth of the two microalgae was strongly inhibited by using fresh tissue, dry powder and aqueous extracts of the three macroalga; the allelochemicals were lethal to H. akashiwo at relatively higher concentrations of the three macroalga. The macroalgae showing the most allelopathic effect on H. akashiwo and A. tamarense using fresh tissue were U. pertusa and S. thunbergii, using dry powder were S. thunbergii and U. pertusa, and using aqueous extracts were U. pertusa and C. pilulifera. We also examined the potential allelopathic effect on the two microalgae of culture filtrate of the three macroalga; culture medium filtrate initially exhibited no inhibitory effects when first added but inhibitory effects became apparent under semi-continuous addition, which suggested that continuous release of small quantities of rapidly degradable allelochemicals from the fresh macroalgal tissue were essential to effectively inhibit the growth of the two microalgae

W18O49 Nanowires as Ultraviolet Photodetector

Photodetectors in a configuration of field effect transistor were fabricated based on individual W18O49 nanowires. Evaluation of electrical transport behavior indicates that the W18O49 nanowires are n-type semiconductors. The photodetectors show high sensitivity, stability and reversibility to ultraviolet (UV) light. A high photoconductive gain of 104 was obtained, and the photoconductivity is up to 60 nS upon exposure to 312 nm UV light with an intensity of 1.6 mW/cm2. Absorption of oxygen on the surface of W18O49 nanowires has a significant influence on the dark conductivity, and the ambient gas can remarkably change the conductivity of W18O49 nanowire. The results imply that W18O49 nanowires will be promising candidates for fabricating UV photodetectors