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1.
Prostanoids comprising prostaglandins (PGs) and thromboxanes (TXs) have been shown to play physiological and pathological roles in zebrafish. However, the molecular basis of zebrafish prostanoid receptors has not been established. Here, we demonstrate that there exist at least five ‘contractile’ (Ca2+-mobilizing) and one ‘inhibitory’ (Gi-coupled) prostanoid receptors in zebrafish; five ‘contractile’ receptors consisting of two PGE2 receptors (EP1a and EP1b), two PGF receptors (FP1 and FP2), and one TXA2 receptor TP, and one ‘inhibitory’ receptor, the PGE2 receptor EP3. [3H]PGE2 specifically bound to the membranes of cells expressing zebrafish EP1a, EP1b and EP3 with a Kd of 4.8, 1.8 and 13.6 nM, respectively, and [3H]PGF specifically bound to the membranes of cells expressing zebrafish FP1 and FP2, with a Kd of 6.5 and 1.6 nM, respectively. U-46619, a stable agonist for human and mouse TP receptors, significantly increased the specific binding of [35S]GTPγS to membranes expressing the zebrafish TP receptor. Upon agonist stimulation, all six receptors showed an increase in intracellular Ca2+ levels, although the increase was very weak in EP1b, and pertussis toxin abolished only the EP3-mediated response. Zebrafish EP3 receptor also suppressed forskolin-induced cAMP formation in a pertussis toxin-sensitive manner. In association with the low structural conservation with mammalian receptors, most agonists and antagonists specific for mammalian EP1, EP3 and TP failed to work on each corresponding zebrafish receptor. This work provides further insights into the diverse prostanoid actions mediated by their receptors in zebrafish.  相似文献   

2.
Ubiquitin-activating enzyme E1 (UBE1) catalyzes the first step in the ubiquitination reaction, which targets a protein for degradation via a proteasome pathway. UBE1 plays an important role in metabolic processes. In this study, full-length cDNA and DNA sequences of UBE1 gene, designated CrUBE1, were obtained from ‘Wuzishatangju’ (self-incompatible, SI) and ‘Shatangju’ (self-compatible, SC) mandarins. 5 amino acids and 8 bases were different in cDNA and DNA sequences of CrUBE1 between ‘Wuzishatangju’ and ‘Shatangju’, respectively. Southern blot analysis showed that there existed only one copy of the CrUBE1 gene in genome of ‘Wuzishatangju’ and ‘Shatangju’. The temporal and spatial expression characteristics of the CrUBE1 gene were investigated using semi-quantitative RT-PCR (SqPCR) and quantitative real-time PCR (qPCR). The expression level of the CrUBE1 gene in anthers of ‘Shatangju’ was approximately 10-fold higher than in anthers of ‘Wuzishatangju’. The highest expression level of CrUBE1 was detected in pistils at 7 days after self-pollination of ‘Wuzishatangju’, which was approximately 5-fold higher than at 0 h. To obtain CrUBE1 protein, the full-length cDNA of CrUBE1 genes from ‘Wuzishatangju’ and ‘Shatangju’ were successfully expressed in Pichia pastoris. Pollen germination frequency of ‘Wuzishatangju’ was significantly inhibited with increasing of CrUBE1 protein concentrations from ‘Wuzishatangju’.  相似文献   

3.
Syntheses and room-temperature single crystal X-ray structural characterizations are recorded for a variety of silver(I) oxyanion (perchlorate, nitrate and trifluoroacetate (‘tfa’) (increasing basicity)) adducts, AgX, with a number of pyridine (‘py’) bases, L, functionalized in the 2-position with N- or O-donor groups, namely 2-amino-, 2-amino-6-methyl-, 2-aminomethyl-, 2-hydroxy-, 2-methoxy- and 2-acetyl- pyridines, ‘2np’, ‘nmp’, ‘amp’, ‘ohp’, ‘mop’, and ‘acp’. A variety of stoichiometries and associated structural types are defined: [Ag(chelate)2]X, L/X = amp,acp/ClO4, [XAg(chelate)2], L/X = acp/tfa, of 1:2 AgX:L stoichiometry; for 1:1 stoichiometry, although a discrete mononuclear complex [(chelate)Ag(O2NO)] is defined for AgNO3: acp (1:1), all others are polymers, successive silver atoms being linked by N,N′-bridging ligands singly (L/X = 2np/ClO4 (?HAgHTAgTHAgH?), amp/ClO4, NO3 (?HTAgHTAg?) (‘H’ ≡ head, ‘T’ = tail)) or pairwise, ?L2AgX2AgL2Ag? (L/X = 2np/tfa, nmp/NO3). More complex polymeric arrays are found with L/X = ohp/NO3, tfa, where interaction with the metal takes place via the O-donor only, the py functionality being protonated, and in adducts of more complex stoichiometry AgNO3:mop (2:3) and AgNO3:2np (3:4).  相似文献   

4.
In this study, apical meristems from adventitious buds of three lily (Lilium L.) cultivars were successfully cryopreserved by droplet-vitrification. The most effective techniques were as follows. Excised meristems from in vitro plantlets which had been sub-cultured for about 2 months were cold-hardened at 4 °C for 1 week, precultured on MS medium supplemented with 0.3 mol L−1 sucrose for 2 days, osmoprotected in loading solution for 20-40 min at room temperature and then soaked in PVS2 solution for 90-120 min at 0 °C, frozen in microdroplets of vitrification solution placed on aluminium foils, which were immersed rapidly in liquid nitrogen. The meristems were then rapidly rewarmed by dilution solution, transferred to regeneration medium and stored in the dark for two weeks at 20 °C, and then cultured under white fluorescent light at an intensity of 2000 lux, with a 16 h photoperiod at 20 °C. The highest post-thaw survival percentages of three cultivars ‘Siberia’ (Lilium × siberia), Lilium lancifolium Thunb. and ‘Snow Queen’ Lilium × longiflorum were 65.0%, 83.8% and 43.3%, and regeneration percentages were 62.0%, 67.6% and 35.0%, respectively. The study demonstrated that cryopreservation by droplet-vitrification increased survival and regeneration percentages of certain lily cultivars compared with vitrification. Thus to cryopreserve lily meristems, droplet-vitrification method is preferable to the vitrification method.  相似文献   

5.
Dominik Mojzita 《FEBS letters》2010,584(16):3540-3544
l-Xylulose reductase is part of the eukaryotic pathway for l-arabinose catabolism. A previously identified l-xylulose reductase in Hypocrea jecorina turned out to be not the ‘true’ one since it was not upregulated during growth on l-arabinose and the deletion strain showed no reduced l-xylulose reductase activity but instead lost the d-mannitol dehydrogenase activity [17]. In this communication we identified the ‘true’ l-xylulose reductase in Aspergillus niger. The gene, lxrA (JGI177736), is upregulated on l-arabinose and the deletion results in a strain lacking the NADPH-specific l-xylulose reductase activity and having reduced growth on l-arabinose. The purified enzyme had a Km for l-xylulose of 25 mM and a νmax of 650 U/mg.  相似文献   

6.
Single crystal X-ray structural characterizations are recorded for a number of adducts of MX:dpex (2:3) stoichiometry (MX = simple univalent copper or silver salt; dpex = Ph2E(CH2)xEPh2 (E = P, As)). CuX:dppe (2:3) (X = Cl, Br, I, CN) are binuclear [(dppe-P,P′)CuX(P-dppe-P′)CuX(P,P′-dppe)], all centrosymmetric. AgX:dpex (2:3) (dpex = ‘dpae’ (Ph2As(CH2)2AsPh2), X = Br, F3CCO2 (= ‘tfa’), F3CSO3 (≡ ‘tfs’); dpex = ‘dpape’ (Ph2As(CH2)2PPh2), X = CN, SCN, OClO3) are one-dimensional polymers ?-E′)1AgX(E-dpex-E′)2-AgX(E-dpex-E′)1AgX?, P, As sites scrambled in the latter. AgNO3:dpam (2:3) is also a one-dimensional polymer, ?AgO·NO·OAg(As-dpam-As)AgO·NO·OAg? (‘dpam’ ≡ Ph2As(CH2)2AsPh2). AgX:dpae (2:3) (X = I, CN, ClO4, NO3) and AgX:dpape (2:3) (X = Br, I, NO3) are two-dimensional polymers with large 30-membered macrocyclic rings; similar webs are found for dppx ligands in AgOH:dppb (2:3) and AgNCO, Agtfa:dpph (2:3) with 42- and 54-membered rings. Complexes AgX:dpape (1:3) (X = Cl, Br) are defined as mono-nuclear [XAg(Ph2P(CH2)2AsPh2)3] arrays, the unidentate ligands predominantly P-bound. Synthetic procedures for the adducts are reported, selected compounds being characterized both in solution (1H, 31P NMR, ESI MS) and in the solid state (IR).  相似文献   

7.
Twenty-one adducts of the form AgX:ER3:L (1:1:1) (X = CF3COO (‘tfa’), CH3COO (‘ac’), E = P, As; R = Ph, cy, o-tolyl; L = 2,2′-bipyridyl (‘bpy’)-based ligand) have been synthesized and characterized by analytical, spectroscopic (IR, far-IR, 1H, 19F and 31P NMR) and single crystal X-ray diffraction studies. The resulting complexes are predominantly of the form [(R3E)AgL]+X, with a trigonal EAgN2 coordination environment, the planarity of which may be perturbed by the approach of anion or solvent. The carboxylate anions have been found to be uni-, or semi-bidentate, or also completely ionic, as in the complexes [Ag(PPh3)(bpy)(H2O)](tfa) and [Ag(PPh3)(dpk · H2O)](tfa) (‘dpk · H2O’ = bis(2-pyridyl)ketone (hydrated)). The complexes Agac:PPh3:dpa (1:1:1) and Agac:P(o-tol)3:dpa:MeCN (1:1:1:1) are dinuclear, with bridging unidentate acetate and terminal unidentate dpa (‘dpa’ = bis(2-pyridyl)amine).  相似文献   

8.
The EfeM protein is a component of the putative EfeUOBM iron-transporter of Pseudomonas syringae pathovar syringae and is thought to act as a periplasmic, ferrous-iron binding protein. It contains a signal peptide of 34 amino acid residues and a C-terminal ‘Peptidase_M75’ domain of 251 residues. The C-terminal domain contains a highly conserved ‘HXXE’ motif thought to act as part of a divalent cation-binding site. In this work, the gene (efeM or ‘Psyr_3370’) encoding EfeM was cloned and over-expressed in Escherichia coli, and the mature protein was purified from the periplasm. Mass spectrometry confirmed the identity of the protein (MW 27,772 Da). Circular dichroism spectroscopy of EfeM indicated a mainly α-helical structure, consistent with bioinformatic predictions. Purified EfeM was crystallised by hanging-drop vapor diffusion to give needle-shaped crystals that diffracted to a resolution of 1.6 Å. This is the first molecular study of a peptidase M75 domain with a presumed iron transport role.  相似文献   

9.
Ten novel adducts of the form AgClO4:PR3:L (1:1:1) (R = Ph, cy, o-tolyl; L = 2,2′-bipyridyl (‘bpy’), 2,2′-biquinoline (‘bq’), bis(2-pyridyl)amine (‘dpa’), bis(2-picolyl)amine (‘dpca’)) have been synthesized and characterized by analytical, spectroscopic (IR, far-IR, 1H and 31P NMR) and single crystal X-ray diffraction studies. The solid state molecular structures show that the complexes predominantly take the form [(R3P)AgL]+X, with a trigonal PAgN2 coordination environment, where the approach of the anion or the solvent may perturb the planarity of the silver environment. The ClO4 anion shows uni- or semi-bidentate coordination, except in the complexes AgClO4:PR3:dpca (1:1:1) (R = Ph and o-tolyl), where the anion remains uncoordinated and the dpca donor is a three-coordinate pincer-like ligand.  相似文献   

10.
Competitive abilities of Lagarosiphon major (Ridley) Moss (invasive in Belgium) and native Ceratophyllum demersum L. were assessed experimentally in relation to sediment dredging. We mimicked these conditions by taking undisturbed sediment (‘before dredging’ treatment) and by using restored sediment where the uppermost nutrient rich top layer was removed (‘after dredging’ treatment). Both the species were allowed to grow for seven weeks in monocultures and mixed cultures at different planting densities. Overall, invasive L. major performed better than native C. demersum independent of the characteristics of the growth environment. L. major achieved a higher relative growth rate (RGR) in both treatments based on total length (0.17-0.21 week−1) and weight (0.10-0.19 week−1) compared to C. demersum (length: 0.04-0.07 week−1; weight: 0.03-0.17 week−1). The better performance of L. major was due to a high plasticity under stressful conditions of low free CO2 and high pH. Intraspecific competition and niche partitioning were observed between the two species indicating that species coexistence is favoured instead of competitive exclusion. L. major performed better in the ‘after dredging’ treatment. Consequently, we deduce that sediment dredging will not lead to a decline of the invasive L. major.  相似文献   

11.
12.
The aqueous solution behaviour of the equilibrium related cis-[PdCl2(PTA)2] and [PdCl(PTA)3]Cl complexes has been investigated in the presence of acid and iodide ions. Several of the resulting species were identified and a reaction scheme accounting for identified complexes is proposed. The crystal structures of trans-[PdI2(PTA-H)2][PdI3(PTA)]2 · 2H2O (1) (PTA-H+ = protonated form of PTA) and trans-[PdI2(PTA)2] (2) are reported. The geometry around the Pd(II) metal centre in 1 (for both the cation and anion) and 2 is distorted square planar. The PTA ligands occupy a trans orientation in the cation of 1 and in complex 2. Compound 1 represents a rare example of a Pd(II) system wherein the cation:anion pair, in a 1:2 ratio, are both coordination complexes. It is the first d8 Ni-triad square planar complex containing only one PTA ligand and only the second platinum group metal complex. For the cation in 1, the bond distances and angles are Pd(1)-P(1) = 2.2864(16) Å, Pd(1)-I(1) = 2.6216(7) Å, P(1)-Pd(1)-P(1)′ = 180.00(7)° and P(1)-Pd(1)-I(1) = 87.62(4)°, while in the anion the bond distances are Pd(2)-P(2) = 2.2377(15) Å, Pd(2)-I(4) = 2.5961(13) Å, Pd(2)-I(2) = 2.6328(13) Å, Pd(2)-I(3) = 2.6513(8) Å, while the angles are P(2)-Pd(2)-I(4) = 90.00(5)°, P(2)-Pd(2)-I(2) = 89.69(5)°, I(4)-Pd(2)-I(2) = 179.57(2)°, P(2)-Pd(2)-I(3) = 175.19(4)°, I(4)-Pd(2)-I(3) = 90.29(4)° and I(2)-Pd(2)-I(3) = 90.05(4)°. Bond distances and angles of the coordination polyhedron in 2 are Pd-P = 2.327(3) Å, Pd-I = 2.5916(10) Å, P-Pd-I = 89.13(7)° and P-Pd-P = 180.00(13)°. The average effective- and Tolman cone angles for the two ligands, calculated from the crystallographic data, are 115° and 117° for PTA and PTA-H, respectively.  相似文献   

13.
Single-crystal X-ray structural characterizations of MX:dpam (1:1) (‘dpam’ = Ph2AsCH2AsPh2) are reported for MX = AgCl, Br; CuI, CN/Cl (all isomorphous) and AgI, AgSCN, CuSCN arrays, all being of the novel form [(μ-X){M(μ-X)(As-dpam-As′)2M′}], essentially the familiar M(E-dpem-E′)2M′ binuclear array with both ‘bridging’ and (linking) ‘terminal’ (pseudo-)halides involved in the polymer. A different arrangement of bridging and linking entities is found with AgX:dpae (1:1)2(∞|∞), X = Br, NCO, ‘dpae’ = Ph2As(CH2)2AsPh2, now comprising [M(μ-X)2(As-dpae-As)M] kernels linked by As-dpae-As′, while in the thiocyanate analogue units are linked by the dpae ligands into a two-dimensional web. Synthetic procedures for all adducts have been reported. All compounds have been characterized both in solution (1H, 13C, 31P NMR, ESI MS) and in the solid state (IR).  相似文献   

14.
The first total synthesis of caminoside B, a novel marine antimicrobial glycolipid isolated from the marine sponge Caminus sphaeroconia, was developed. This marine small molecule inhibitor (IC50 = 20 μM) targeting type III secretory pathway of bacterial pathogenesis was assembled in good yield via a ‘2+2+1’ strategy based on stereocontrolled construction of the four glycosidic linkages.  相似文献   

15.
The binuclear mixed valence copper(I/II) compound [CuI(CN)3CuII(tn)2] (1) (tn = propane-1,3-diamine) and its acetonitrile adduct [CuI(CN)3CuII(tn)2] · 2MeCN (2) have been synthesized. Complex 1 crystallizes triclinic, space group , a = 8.117(2) Å, b = 8.389(2) Å, c = 11.920(2) Å, α = 108.728(3)°, β = 100.024(3)°, γ = 104.888(4)°, Z = 2, and compound 2 monoclinic, space group P21/m, a = 8.752(2) Å, b = 13.243(3) Å, c = 9.549(2) Å, β = 114.678(4)°, Z = 2. In both crystal structures, the binuclear [CuI(CN)3CuII(tn)2] complex with slightly different bonding geometries is formed. One of the three nitrogen atoms of a CuI(CN)3 moiety is coordinated to Cu(II) at the apex of a square-pyramid with two chelating ligands tn on its base. The shortest intramolecular CuII?CuII distance in 1 is 5.640(7) Å. The EPR behaviour of 1 has been investigated at room temperature and at 77 K. The magnetic properties were measured in the temperature range 1.8-300 K.  相似文献   

16.
The effects of temperature, water level and burial depth on seed germination of two submerged species, Myriophyllum spicatum and Potamogeton malaianus, were investigated under controlled laboratory conditions. There was no significant difference in final germination of M. spicatum among water level treatments, but P. malaianus germinations at 1 cm and 12 cm water levels were better than at 0 cm water level at temperatures of 20 °C and 30 °C. Little to no germination was observed for either species at the temperature of 10 °C. At 15 °C, however, germination increased significantly to 66.3-70.6% for M. spicatum and to 29.4-48.1% for P. malaianus under all three water level treatments. Increased temperature from 15 °C to 30 °C had no significant effect on the final germination of M. spicatum except at the 1 cm water level, but enhanced significantly the germination of P. malaianus. Analysis of the mean time to germination revealed that M. spicatum was a faster germinator relative to P. malaianus. The two species’ germination differed markedly in response to burial depth. Germination percentage of M. spicatum was 71.3% at 0 cm burial depth, but decreased to 5.0% and to 2.5% at depths of 1 cm and 2 cm, respectively; whereas germination percentages of P. malaianus were 40.0%, 23.8%, 12.5%, 7.5% and 1.3% at depths of 0 cm, 1 cm, 2 cm, 3 cm and 5 cm, respectively. We concluded that the two species respond differently to germination strategies. The findings provided further insight into how germination strategy contributes to the seed bank formation and species invasion.  相似文献   

17.
Detailed optical and EPR analyses of states induced in dark-adapted PS II membranes by cryogenic illumination permit characterization and quantification of all pigment derived donors and acceptors, as well as optically silent (in the visible, near infrared) species which are EPR active. Near complete turnover formation of QA is seen in all centers, but with variable efficiency, depending on the donor species. In minimally detergent-exposed PS II membranes, negligible (< 5%) oxidation of chlorophyll or carotenoid centers occurs for illumination temperatures 5-20 K. An optically silent electron donor to P680+ is observed with the same decay kinetics as the S1 split signal. Cryogenic donors to P680+ seen are: (i) transient (t1/2 ∼ 150 s) tyrosine related species, including ‘split signals’ (∼ 15% total centers), (ii) reduced cytochrome b559 (∼ 30-50% centers), and (iii) an organic donor, possibly an amino acid side chain, (∼ 30% centers).  相似文献   

18.
We investigated the effect of five day exposure to CO2-acidified sea water treatments (pHNBS = 7.89 [control], 7.44, 7.16 and 6.78, T = 9.5 °C) on the extracellular acid-base balance of the northern sea urchin Strongylocentrotus dröebachiensis. In each case there was an uncompensated respiratory acidosis which increased in intensity with decreasing environmental pH. This was very similar to results for another sea urchin species, Psammechinus miliaris (8 d exposure, T = 15 °C). However, there were some important differences in the response to low seawater pH between the two urchin species S. dröebachiensis and P. miliaris. At the lowest pH tested (6.78) there was a metabolic component to this acidosis recorded (correlated with a significant increase in l-lactate) in S. dröebachiensis but not P. miliaris. The acidosis was accompanied by a very small, but significant increase in coelomic fluid calcium. Also the water used in our study was (controlling for pH) markedly undersaturated with respect to carbonate compared with that used in the Psammechinus study, highlighting the need for the environmental context to be assessed in future comparative studies.  相似文献   

19.
Conidial tolerance to the upper thermal limits of summer is important for fungal biocontrol agents, whose conidia are formulated into mycoinsecticides for field application. To develop an efficient assay system, aerial conidia of eight Metarhizium anisopliae, four M. anisopliae var. anisopliae, and six M. anisopliae var. acridum isolates with different host and geographic origins were wet-stressed for ≤180 min at 48 °C or incubated for 14 d colony growths at 10-35 °C. The survival ratios (relative to unstressed conidia) of each isolate, examined at 15-min intervals, fit a logistic equation (r2 ≥ 0.975), yielding median lethal times (LT50s) of 14.3-150.3 min for the 18 isolates stressed at 48 °C. Seven grasshopper isolates from Africa had a mean LT50 of 110 (73-150) min, but could not grow at 10 or 15 °C. The mean LT50 of five non-grasshopper isolates capable of growing at 10-35 °C was 16 (10-26) min only. Three isolates with typically low (type I), medium (type II), and high (type III) levels of tolerance to 48 °C were further assayed for ≤4-d tolerance of their conidia to the wet stress at 38, 40, 42, or 45 °C. The resultant LT50s decreased to 20, 53 and 167 min at 48 °C from 507, 1612, and 8256 min at 38 °C for types I, II and III, respectively. For the distinguished types, the logarithms of the LT50s were significantly correlated to the temperatures of 38-48 °C with an inverse linearity (r2 ≥ 0.88). The method developed to assay quantitatively fungal thermotolerance would be useful for screening of fungal candidates for improved pest control in summer.  相似文献   

20.
In this study, we present the activities of Al2O3 supported CaO and MgO catalysts in the transesterification of lipid of yellow green microalgae, Nannochloropsis oculata, as a function of methanol amount and the CaO and MgO loadings at 50 °C. We found that pure CaO and MgO were not active and CaO/Al2O3 catalyst among all the mixed oxide catalysts showed the highest activity. Not only the basic site density but also the basic strength is important to achieve the high biodiesel yield. Biodiesel yield over 80 wt.% CaO/Al2O3 catalyst increased to 97.5% from 23% when methanol/lipid molar ratio was 30.  相似文献   

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