Regulation of autoinducer 2 production and luxS expression in a pathogenic Edwardsiella tarda strain

Edwardsiella tarda is a bacterial pathogen that can infect both humans and animals. TX1, an Ed. tarda strain isolated from diseased fish, was found to produce autoinducer 2 (Al-2)-like activity that was growth phase dependent and modulated by growth conditions. The gene coding for the Al-2 synthase was cloned from TX1 and designated luxS(Et). LuxS(Et) was able to complement the Al-2 mutant phenotype of Escherichia coli strain DH5 alpha. Expression Of luxS(Et) correlated with Al-2 activity and was increased by glucose and decreased by elevated temperature. The effect of glucose was shown to be mediated through the cAMP-CRP complex, which repressed luxS(Et) expression. Overexpression of luxS(Et) enhanced Al-2 activity in TX1, whereas disruption of luxS(Et) expression by antisense RNA interference (i) reduced the level of Al-2 activity, (ii) impaired bacterial growth under various conditions, (iii) weakened the expression of genes associated with the type III secretion system and biofilm formation, and (iv) attenuated bacterial virulence. Addition of exogenous Al-2 was able to complement the deficiencies in the expression of TTSS genes and biofilm production but failed to rescue the growth defects. Our results (i) demonstrated that the Al-2 activity in TX1 is controlled at least in part at the level of luxS(Et) expression, which in turn is regulated by growth conditions, and that the temporal expression of luxS(Et) is essential for optimal bacterial infection and survival; and (ii) suggested the existence in Ed. tarda of a LuxS/Al-2-mediated signal transduction pathway that regulates the production of virulence-associated elements.

Regulation of autoinducer 2 production and luxS expression in a pathogenic Edwardsiella tarda strain

Edwardsiella tarda is a bacterial pathogen that can infect both humans and animals. TX1, an Ed. tarda strain isolated from diseased fish, was found to produce autoinducer 2 (Al-2)-like activity that was growth phase dependent and modulated by growth conditions. The gene coding for the Al-2 synthase was cloned from TX1 and designated luxS(Et). LuxS(Et) was able to complement the Al-2 mutant phenotype of Escherichia coli strain DH5 alpha. Expression Of luxS(Et) correlated with Al-2 activity and was increased by glucose and decreased by elevated temperature. The effect of glucose was shown to be mediated through the cAMP-CRP complex, which repressed luxS(Et) expression. Overexpression of luxS(Et) enhanced Al-2 activity in TX1, whereas disruption of luxS(Et) expression by antisense RNA interference (i) reduced the level of Al-2 activity, (ii) impaired bacterial growth under various conditions, (iii) weakened the expression of genes associated with the type III secretion system and biofilm formation, and (iv) attenuated bacterial virulence. Addition of exogenous Al-2 was able to complement the deficiencies in the expression of TTSS genes and biofilm production but failed to rescue the growth defects. Our results (i) demonstrated that the Al-2 activity in TX1 is controlled at least in part at the level of luxS(Et) expression, which in turn is regulated by growth conditions, and that the temporal expression of luxS(Et) is essential for optimal bacterial infection and survival; and (ii) suggested the existence in Ed. tarda of a LuxS/Al-2-mediated signal transduction pathway that regulates the production of virulence-associated elements.

Estradiol stimulates growth hormone production in female goldfish

The effects of estradiol (E(2)) on growth hormone (GH) production was investigated in gonad-intact female goldfish. It was first necessary to generate a specific antibody for use in immunocytochemistry, Western, and dot-blot analyses of GH production. To accomplish this, grass carp GH (gcGH) cDNA was cloned by the reverse transcription polymerase chain reaction (RT-PCR) and expressed in Echerichia coli and a specific polyclonal antibody to recombinant gcGH was generated in the rabbit. In Western blot, the anti-gcGH antibody specifically immunoreacted with recombinant gcGH, purified natural common carp GH, and with a single 21.5-kDa GH form from pituitary extracts of grass carp, common carp, goldfish, and zebrafish but not salmon, trout, or tilapia. Intraperitoneal injection of the recombinant gcGH enhanced the growth rates of juvenile common carp demonstrating biological activity of this GH preparation. Electron microscopic studies showed that the anti-gcGH-I antibody specifically reacted with GH localized in the secretory granules of the goldfish somatotroph. Using anti-gcGH-I in a dot-blot assay, it was found that in vivo implantation of solid silastic pellets containing E(2) (100 mu g/g body weight for 5 days) increased pituitary GH content by 150% in female goldfish. In a second, independent study employing a previously characterized anticommon carp GH antibody for radioimmunoassay, it was found that E(2) increased pituitary GH content by 170% and serum GH levels by approximately 350%. The E(2)-induced hypersecretion of GH and increase in pituitary GH levels was not associated with changes in steady-state pituitary GH mRNA levels, suggesting that this sex steroid may enhance GH synthesis at the posttranscriptional or translational level. Previous observations indicate that GH can stimulate ovarian E(2) production. The present results show that E(2) can in turn stimulate GH production, indicating the existence of a novel pituitary GH-ovarian feedback system in goldfish. (C) 1997 Academic Press.

Two-stage photo-biological production of hydrogen by marine green alga Platymonas subcordiformis

A marine green alga, Platymonas subcordiformis, was demonstrated to photobiologically evolve hydrogen (H-2) after the first stage of photosynthesis, when subjected to a two-phase incubation protocol in a second stage of H2 production: anaerobic incubation in the dark followed by the exposure to light illumination. The anaerobic incubation induced hydrogenase activity to catalyse H? evolution in the following phase of light illumination. H,) evolution strongly depended upon the duration of anaerobic incubation, deprivation of sulphur (S) from the medium and the medium pH. An optimal anaerobic incubation period of 32 h gave the maximum H2 evolution in the second phase in the absence of sulphur. Evolution of H,) was greatly enhanced by 13 times when S was deprived from the medium. This result suggests that S plays a critical role in the mediation of H-2 evolution from R subcordiformis. A 14-fold increase in H-2 production was obtained when the medium pH increased from 5 to 8; with a sharp decline at pH above eight. H-2 evolution was enhanced by 30-50% when supplementing the optimal concentrations of 25 mM acetate and 37.5 mM glucose. (C) 2003 Elsevier B.V. All rights reserved.

Production of active oxygen species from Taxus cuspidata induced by far-UV radiation

In order to understand the role of active oxygen species in mediating plant injuries induced by far-UV radiation, seedlings of Taxus cuspidata Sieb. et Zucc. were irradiated by far-UV rays in laboratory for 4 weeks. The production of organic free-radicals in detached needles, and the production of O-2(radical anion) and O-1(2) in isolated chloroplasts were detected weekly by electron spin resonance (ESR) to evaluate their relative importance. The results show that the cumulative effect of far-UV irradiation, is best indicated by the production of organic free radicals in the needles, O-2(radical anion) production in chloroplasts is the next. The enhancement of O-1(2) production in chloroplasts by the cumulative far-UV irradiation seems to be not so important as O-2(radical anion) in mediating injuries induced by, far-UV radiation because of its high background value.

渭北旱塬持续高效农田种植模式研究与示范

根据渭北旱塬自然经济特点 ,提出了该区农业持续发展的关键技术和增产新途径——农田高效优化种植模式 (以双元覆盖小麦为主 )。通过 4a的试验示范与推广 ,结果表明 :该模式可较对照增产小麦6 2 .92 % ;复种黄豆增收 1 5 0 0～ 2 2 5 0 kg/ hm2 ,较对照增产 1 0 0 %。该模式的聚水、保墒、增温和促进有机质矿化、增加土壤养分效应显著 ,可使土壤水、热环境改善 ,充分利用水资源 ,提高用水效率。

Identification and characterization of the AcrR/AcrAB system of a pathogenic Edwardsiella tarda strain

Edwardsielia tarda is one of the leading marine pathogens that can infect a wide range of cultured marine species. In this study, the acrR-acrAB cluster was cloned from TX1, a pathogenic E. tarda strain isolated from diseased fish. AcrR and AcrAB were found to be involved in resistance against acriflavine and methyl viologen, which positively regulate the expression of acrAB. AcrR negatively regulates its own expression and the expression of the acrAB operon, most likely by interacting with a 24-bp operator site that overlaps the putative promoter of acrA (PacrA). The repressive effect of AcrR on PacrA could be relieved by acriflavine, methyl viologen, and ethidium bromide, the presence of each of which enhanced transcription from PacrA. Interruption of the regulated expression of acrR by introducing into TX1 a plasmid that overexpresses acrR affected growth under stress conditions, AI-2 production, and bacterial virulence. In addition, mutational analyses identified a constitutively active AcrR mutant (named N215), which exhibits full repressor activity but is impaired in its ability to interact with the inducer. Overexpression of N215 produced the same kind of but moderately stronger effect on TX1 compared to that produced by overexpression of the wild-type acrR.

Regulation of the Edwardsiella tarda Hemolysin Gene and luxS by EthR

Edwardsiella tarda is a pathogen with a broad host range that includes human and animals. The E. tarda hemolysin (Eth) system, which comprises EthA and EthB, is a noted virulence element that is widely distributed in pathogenic isolates of E. tarda. Previous study has shown that the expression of ethB is regulated by iron, which suggests the possibility that the ferric uptake regulator (Fur) is involved in the regulation of ethB. The work presented in this report supports the previous findings and demonstrates that ethB expression was decreased under conditions when the E. tarda Fur (Fur(Et)) was overproduced, and enhanced when Fur(Et) was inactivated. We also identified a second ethB regulator, EthR, which is a transcription regulator of the GntR family. EthR represses ethB expression by direct interaction with the ethB promoter region. In addition to ethB, EthR also modulates, but positively, luxS expression and AI-2 production by binding to the luxS promoter region. The expression of ethR itself is subject to negative autoregulation; interference with this regulation by overexpressing ethR during the process of infection caused (i) drastic changes in ethB and luxS expressions, (ii) vitiation in the tissue dissemination and survival ability of the bacterium, and (iii) significant attenuation of the overall bacterial virulence. These results not only provide new insights into the regulation mechanisms of the Eth hemolysin and LuxS/AI-2 quorum sensing systems but also highlight the importance of these systems in bacterial virulence.

Macrozoobenthos in 2 shallow mesotrophic Chinese lakes with contrasting sources of primary production

Comparative studies on macrozoobenthos were done in 2 shallow mesotrophic lakes in the middle basins of the Yangtze River, China: Lake Biandantang where macrophytes were abundant, and Lake Houhu where macrophytes were scarce Samples were taken monthly at 4 stations in each lake from April 1997 to March 1999, and a total of 67 and 31 tara of macrozoobenthos were recorded in Lake Biandantang and Lake Houhu, respectively. Both annual mean density and biomass of macrozoobenthos were higher in Lake Biandantang than in Lake Houhu: 780 vs 532 indivials/m(2) and 37.1 vs 25.9 g wet mass/m(2), respectively. Abundance of functional feeding groups followed the order: scraper > collector > predator > shredder in Lake Biandantang, and collector > predator > scraper > shredder in Lake Houhu. Only 1 density peak occurred from winter to early spring in Lake Houhu; however, in Lake Biandantang, there were 2 peaks, the winter peak and spring peak. K-dominance curves and Shannon-Wiener, Simpson, and Margelef indices indicated that macrozoobenthos were more diverse in Lake Biandantang than in Lake Houhu Our study suggests that, in shallow lakes, submerged macrophytes are essential for the maintenance of biodiversity of macrozoobenthos mainly because the macrophytes increase habit heterogeneity and availability of suitable food, and may also decrease predation by fish on the macrozoobenthos.

Defect production in silicon carbide irradiated with Ne and Xe ions with energy of 2.3 MeV/amu

In the present work specimens of mono-crystalline silicon carbide (4H polytype) were irradiated to three successively increasing ion fluences ranging from 7.2 x 10(14) to 6.0 x 10(16) ions/cm(2) (corresponding to the peak displacement damage of 1, 4 and 13 dpa) with Ne and Xe ions respectively with the energy of 2.3 MeV/amu. The irradiated specimens were subsequently annealed at temperatures of 1173 and 1273 K. Defect structure was investigated with transmission electron microscopy (TEM) using a cross-sectional specimen preparation technique. The typical microstructures of the annealed specimens irradiated with Ne or Xe ions to high fluences are characterized by small gas bubbles in high concentration in the peak damage region and black dots and dislocation loops (located in the basal plane) in a shallower and broader depth region. Larger dislocation loops were observed in the Xe-ion irradiated specimen than in the Ne-ion irradiated specimen at the same peak damage level. The enhanced formation of dislocation loops in the case of Xe-ion irradiation is understandable by assuming stronger inclination of heavier inert-gas atoms to occupy substitute site in the peak damage region.

Phenomenological analysis of the double-pion production in nucleon-nucleon collisions up to 2.2 GeV

With an effective Lagrangian approach, we analyze several NN -> NN pi pi channels by including various resonances with mass up to 1.72 GeV. For the channels with the pion pair of isospin zero, we confirm the dominance of N*(1440) -> N sigma in the near-threshold region. At higher energies and for channels with the final pion pair of isospin one, we find large contributions from N*(1440) -> Delta pi, double-Delta, Delta(1600) -> N*(1440)pi, Delta(1600) -> Delta pi and Delta(1620) -> Delta pi. There are also sizable contributions from Delta -> Delta pi, Delta -> N pi, N -> Delta pi, and nucleon pole at energies close to the threshold. We give a good reproduction to the total cross sections up to beam energies of 2.2 GeV except for the pp -> pp pi(0)pi(0) channel at energies around 1.1 GeV and our results agree with the existing data of differential cross sections of pp -> pp pi(+)p pi(-), pp -> nn pi(+)pi(+), and pp -> pp pi(0)pi(0) which are measured at CELSIUS and COSY.

Identified particle production, azimuthal anisotropy, and interferometry measurements in Au plus Au collisions at root s(NN)=9.2 GeV

We present the first measurements of identified hadron production, azimuthal anisotropy, and pion interferometry from Au + Au collisions below the nominal injection energy at the BNL Relativistic Heavy-Ion Collider (RHIC) facility. The data were collected using the large acceptance solenoidal tracker at RHIC (STAR) detector at root s(NN) = 9.2 GeV from a test run of the collider in the year 2008. Midrapidity results on multiplicity density dN/dy in rapidity y, average transverse momentum < p(T)>, particle ratios, elliptic flow, and Hanbury-Brown-Twiss (HBT) radii are consistent with the corresponding results at similar root s(NN) from fixed-target experiments. Directed flow measurements are presented for both midrapidity and forward-rapidity regions. Furthermore the collision centrality dependence of identified particle dN/dy, < p(T)>, and particle ratios are discussed. These results also demonstrate that the capabilities of the STAR detector, although optimized for root s(NN) = 200 GeV, are suitable for the proposed QCD critical-point search and exploration of the QCD phase diagram at RHIC.

Reducing H2S production by O-2 feedback control during large-scale sewage sludge composting

Hydrogen sulfide (H2S) production patterns and the influence of oxygen (O-2) concentration were studied based on a well operated composting plant. A real-time, online multi-gas detection system was applied to monitor the concentrations of H2S and O-2 in the pile during composting. The results indicate that H2S was mainly produced during the early stage of composting, especially during the first 40 h. Lack of available O-2 was the main reason for H2S production. Maintaining the O-2 concentration higher than 14% in the pile could reduce H2S production. This study suggests that shortening the interval between aeration or aerating continuously to maintain a high O-2 concentration in the pile was an effective strategy for restraining H2S production in sewage sludge composting. (C) 2010 Elsevier Ltd. All rights reserved.