Cyclic AMP increases COX-2 expression via mitogen-activated kinase in human myometrial cells

Cyclic AMP (cAMP) is the archetypal smooth muscle relaxant, mediating the effects of many hormones and drugs. However, recently PGI(2) , acting via cAMP/PKA, was found to increase contraction-associated protein expression in myometrial cells and to promote oxytocin-driven myometrial contractility. Cyclo-oxygenase-2 (COX-2) is the rate-limiting enzyme in prostaglandin synthesis, which is critical to the onset and progression of human labour. We have investigated the impact of cAMP on myometrial COX-2 expression, synthesis and activity. Three cAMP agonists (8-bromo-cAMP, forskolin and rolipram) increased COX-2 mRNA expression and further studies confirmed that this was associated with COX-2 protein synthesis and activity (increased PGE(2) and PGI(2) in culture supernatant) in primary cultures of human myometrial cells. These effects were neither reproduced by specific agonists nor inhibited by specific inhibitors of known cAMP-effectors (PKA, EPAC and AMPK). We then used shRNA to knockdown the same effectors and another recently described cAMP-effector PDZ-GEF(1-2) , without changing the response to cAMP. We found that MAPK activation mediated the cAMP effects on COX-2 expression and that PGE(2) acts through EP-2 to activate MAPK and increase COX-2. These data provide further evidence in support of a dual role for cAMP in the regulation of myometrial function.     

鲤科鱼蓝黑鲮微卫星位点的种间扩增(英文)

本研究使用105对微卫星引物对7种鲤科鱼类进行跨越种间PCR扩增,共得到14个多态性微卫星位点.其中9个扩增效果较好的位点用于分析来自帕吉勒提河(Bhagirathi, n=20)和戈达瓦里河(Godavari, n=25)的蓝黑鲮(Labeo calbasu)样品的遗传多样性.结果显示,前者在每个位点的平均等位基因数为7.33,而后者为8 1,期望杂合度介于0.795(Bhagirathi)和0.801(Godavari)之间;4个位点MFW11* (Godavari)、R1*(Godavari)、R3* (Bhagirathi) 和 Lr38*(Bhagirathi和Godavari)都表现出明显的杂合子缺失和哈迪温伯格平衡偏离;而任意两位点间都未观测到连锁不平衡现象;位点R3*极可能存在无效等位基因.上述结果表明这些多态性微卫星位点作为共显性标记在蓝黑鲮群体遗传学研究中有着较好的应用前景.     

Structural characteristics of the plasmid-encoded toxin from enteroaggregative Escherichia coli

Intoxication by the plasmid-encoded toxin (Pet) of enteroaggregative Escherichia coli requires toxin translocation from the endoplasmic reticulum (ER) to the cytosol. This event involves the quality control system of ER-associated degradation (ERAD), but the molecular details of the process are poorly characterized. For many structurally distinct AB-type toxins, ERAD-mediated translocation is triggered by the spontaneous unfolding of a thermally unstable A chain. Here we show that Pet, a non-AB toxin, engages ERAD by a different mechanism that does not involve thermal unfolding. Circular dichroism and fluorescence spectroscopy measurements demonstrated that Pet maintains most of its secondary and tertiary structural features at 37 degrees C, with significant thermal unfolding only occurring at temperatures >or=50 degrees C. Fluorescence quenching experiments detected the partial solvent exposure of Pet aromatic amino acid residues at 37 degrees C, and a cell-based assay suggested that these changes could activate an ERAD-related event known as the unfolded protein response. We also found that HEp-2 cells were resistant to Pet intoxication when incubated with glycerol, a protein stabilizer. Altogether, our data are consistent with a model in which ERAD activity is triggered by a subtle structural destabilization of Pet and the exposure of Pet hydrophobic residues at physiological temperature. This was further supported by computer modeling analysis, which identified a surface-exposed hydrophobic loop among other accessible nonpolar residues in Pet. From our data it appears that Pet can promote its ERAD-mediated translocation into the cytosol by a distinct mechanism involving partial exposure of hydrophobic residues rather than the substantial unfolding observed for certain AB toxins.     

The inhibitory action of phospholamban involves stabilization of alpha-helices within the Ca-ATPase.

We have used attenuated total reflection Fourier transform infrared (ATR-FTIR) and circular dichroism (CD) spectroscopies to identify secondary and dynamic structural changes within the Ca-ATPase that result from the functional inhibition of transport activity by phospholamban (PLB). Isotopically labeled [(13)C]PLB was expressed and purified from Escherichia coli and was functionally reconstituted with unlabeled Ca-ATPase, permitting the resolution of the amide I and II absorbance bands of the Ca-ATPase from those of [(13)C]PLB. Upon co-reconstitution of the Ca-ATPase with PLB, spectral shifts are observed in both the CD spectra and the amide I and II bands associated with the Ca-ATPase, which are indicative of increased alpha-helical stability. Corresponding changes in the kinetics of H/D exchange occur upon association with PLB, indicating that 100 +/- 20 residues in the Ca-ATPase that normally undergo rapid amide H/D exchange become exchange resistant. There are no corresponding large changes in the secondary structure of PLB. The affinity of the structural interaction between PLB and the Ca-ATPase is virtually identical to that associated with functional inhibition (K(d) = 140 +/- 30 microM), confirming that the inhibitory regulation of the Ca-ATPase by PLB involves the stabilization of alpha-helices within the Ca-ATPase.     

Xylanase-induced reduction of chlorine dioxide consumption during elemental chlorine-free bleaching of different pulp types

The potential of crude xylanase from Thermomyces lanuginosus and Xylanase P (a commercial xylanase) was evaluated in bleaching of various paper pulp types. Xylanases released chromophores and reducing sugars and decreased kappa number of pulps. Chlorine-bleached, alkali-extracted bagasse and post-oxygen kraft pulps, pretreated with enzymes, gained over 5 brightness points over controls. Biobleaching of soda-aq pulp with Xylanase P produced chlorine dioxide savings of up to 30% or 4.5 kg chlorine dioxide t^–1 pulp.     

Structure and Function of Oak Forests in Central Himalaya. II. Nutrient Dynamics

This paper elucidates nutrient dynamics in oak forests previouslyinvestigated for dry matter dynamics. The nutrient concentrationsin different life forms were of the order: herb > shrub >tree, whereas the standing state of nutrients were of the order:tree > shrub > herb. Soil, litter and vegetation, respectively,accounted for 32·4–98·0 %; 0·3–3·5%, and 10·2–66·6 % of the total nutrientsin the system. Considerable reductions (8·5–41·7%)in concentrations of nutrients in leaves occurred during senescence.The uptake of nutrients by vegetation, and also by differentcomponents with and without adjustment for internal recycling,has been calculated separately. Annual transfer of litter (above+ below ground) to the soil by vegetation was 115·9–187N, 7·5–15·6 P, 122·7–195·1Ca, 36·1–48·8 K and 2·88–5·16Na kg ha^–1 yr^–1. Turnover rate and turnover timefor different nutrients ranged between 0·66–0·84yr^–1 and 1·19–1·56 yr^–1, respectively.Compartment models for nutrient dynamics have been developedto represent the distribution of nutrient contents and net annualfluxes within the system. Quercus leucotrichophora forest, Q.floribunda forest, Q. lanuginosa forest, Nutrient concentration, standing state, uptake, internal cycling, turnover     

Standing State and Cycling of Nitrogen in Soil-vegetation Components of Prairie Ecosystems

In the present paper, distribution of nitrogen in differentplant compartments and in the top 20 cm soil from three grasslandsites for a 3-year period at the time of peak standing crophas been studied. The sites represent short-grass, mixed-grass,and tall-grass prairies located, respectively, in Colorado,South Dakota, and Oklahoma. Partitioning, uptake, transfers,and release of nitrogen in the above-ground and below-groundcompartments were evaluated with respect to the abiotic factors.A positive linear relationship was found with the total nitrogencontent in shoots (g m^–2) and the annual precipitationas well as with the annual actual evapotranspiration. On theother hand, a negative linear relationship was found with thenitrogen per gram dry weight in shoots, and the annual precipitationand evapotranspiration. Inter-seasonal and inter-site variationsin the nitrogen content (g m^–2) in different plant compartmentsindicated significant differences. The short-grass and mixed-grassprairies indicated greater nitrogen content in the above-groundand below-ground plant tissues as compared to the tall-grassprairie. However the total nitrogen content in shoots from thetall-grass prairie was greater than that of short-grass or mixed-grassprairies which was mainly due to greater shoot biomass in thetall-grass prairie. The major portion of nitrogen (over 90 percent) in the system was retained in the soil while a fractionof nitrogen (4–8 per cent) resided in the biological material.The distribution of nitrogen in different plant compartments,the uptake, transfer, and release of nitrogen with respect toabiotic factors have been discussed in detail.