Specific sensing between inositol epimers by a bis(boronate)

Bis(boronates) that utilize internal photoinduced electron transfer (PET) quenching mechanisms can specifically signal the binding of chiro-inositol without responding to its epimer, myo-inositol.     

Synthesis of organic osmolytes and salt tolerance mechanisms in Paspalum vaginatum

Synthesis of organic compounds in response to salinity stress and their contribution to organic osmotic adjustment were investigated in seashore paspalum (Paspalum vaginatum Swartz). Nine genotypes exhibiting the widest range of salt tolerance were grown in sea-salt amended nutrient solution in a greenhouse. Salinity ranges were 1.1 (EC_w0, control) to 49.7 dS m⁻¹ (EC_w50) based on electrical conductivity of the solution (EC_w). Organic osmolytes most important within seashore paspalum under salinity stress were proline, Gly-betaine, and trigonelline in terms of explaining intraspecific salt tolerance differences and, therefore, should be the focus of biotechnology approaches to enhance these traits. While these osmolytes differed in accumulation with increasing salinity and absolute concentrations among salt tolerant and intolerant genotypes, the magnitude of responses was not sufficiently large to suggest use for salt screening as physiological/biochemical markers. Fructose concentration increased with salinity, especially for salt sensitive ecotypes, and may have potential as a marker. Glucose, sucrose, and myo-inositol tended to increase with salinity, but changes did not relate to intraspecific salt tolerance, while mannitol and sorbitol were not affected by salinity. Proline demonstrated a 20.8-fold increase averaged across genotypes from EC_w0 to EC_w50 salinity. Proline was the primary organic osmolyte for osmotic adjustment accounting for an average of 9.3% to total solute potential (Ψ_s) at EC_w50 and 56% of the organic solute contribution to Ψ_s. In the salt tolerant genotype, SI 93-2, proline and Gly-betaine exhibited greater absolute concentration and accumulation rate relative to the least salt tolerant, Adalayd. The intraspecific role of Gly-betaine did not relate to osmotic adjustment differences, suggesting another role perhaps in protection of the thylakoid membrane.     

Synthesis of L-chiro-inositol 1,4,6-trisphosphorothioate, a potent and selective inhibitor of myo-inositol 1,4,5-trisphosphate 5-phosphatase

L-chiro-inositol 1,4,6-trisphosphate and trisphosphorothioate have been synthesized from L-quebrachitol; the trisphosphorothiate is the most potent inhibitor of Ins(1,4,5)P₃ 5-phosphatase yet discovered.     

Dual effect of formycin a upon the hydrolysis of phosphoinositides in perifused pancreatic islets

Formycin A (1.0 mM) caused a rapid, sustained and rapidly reversible inhibition of effluent radioactivity in rat pancreatic islets prelabelled with myo-[2-³H]inositol and perifused in the presence of 8.3 mM -glucose. This coincided with a progressive decrease in islet ATP content and transient inhibition of insulin release. Theraafter, however, formycin A increased glucose-induced insulin release. Moreover, in islets that were preincubated with myo-[2-³H]inositol and then exposed during perifusion to a rise in -glucose concentration from 2.8 to 16.7 mM, the release of insulin and ³H fractional outflow rate at both the low and high hexose concentrations were much higher when both the preincubation and perifusion were conducted in the presence, rather than absence, of formycin A. It is concluded that formycin A first inhibits and later enhances both the hydrolysis of phosphoinositides and release of insulin, these effects being possibly related to changes in the islet cell content of adenosine and/or formycin A triphosphates.     

Total synthesis of L-2,2-difluoro-2-deoxy-MYO-inositol 1,4,5-trisphosphate, a potent inhibitor of the enzymes of d-MYO-inositol 1,4,5-trisphosphate metabolism

The synthesis of the enantiomers of 2,2-difluoro-2-deoxy-myo-inositol 1,4,5-trisphosphate is reported. L-2,2-difluoro-2-deoxy-myo-inositol 1,4,5-trisphosphate is a potent inhibitor of 3-kinase and 5-phosphatase.     

Synthesis of enantiomerically pure lysophosphatidylinositols and alkylphosphoinositols

A convenient synthesis for enantiomeric pure

Full-size image

,

Full-size image

(or and (or has been described. Starting from myo-inositol, penta-O-acetyl-myo-inositol was made in five steps. Then enantiomeric purification was done by a diastereomeric salts separation method, and the purity of each enantiomer was spectroscopically measured (¹⁹F-NMR). The phosphodiester was made via phosphoramidites. The enantiomeric products (>99% optical purity) of all compounds were easily obtained in large quantities (5–10 g). Synthetic phosphatidylinositol analogues of precisely defined structure and configuration are interesting tools for studying signal transduction mechanism and cell activity modulation.     

Structural studies on [Mo3S4] and [Mo3S4Cu] complexes with tripodal ligands providing various NxOy (x + Y = 3) donor sets

The interaction of 1,3,5-triamino-1,3,5-trideoxy-cis-inositol (taci) and its N-methylated derivative 1,3,5-trideoxy-1,3,5-tris(dimethylamino)-cis-inositol (tdci) with the incomplete [Mo₃S₄]⁴⁺ cube and the heterometallic [Mo₃S₄Cu]⁴⁺ cube have been investigated by X-ray analysis. The crystal structures of [Mo₃S₄(taci+ rmC₃H₆O-H₂O)₃-4H]·2OH₂O (1a, rhombohedral, space group R32, A = 15.964(3), C = 40.59(1) Å, Z = 6), [Mo₃S₄(tdci)₃]Br₄·9.5EtOH·5H₂O (2a, triclinic, space group

Full-size image

and [CuBrMo₃S₄(tdci)₃]Br₃·11 H₂O·EtOH (3a, monoclinic, space group P2,/n, A = 14.887(3), B = 22.570(4), C = 21.974(5) Å, β = 98.54(2)°, Z = 4) revealed andN-N-O and an N-O-O coordination mode for taci and tdci, respectively. In 1a, taci is coordinated as an anion with deprotonated oxygen and nitrogen donors. In addition, the non-coordinating amino group reacted with one equivalent; of acetone, forming a Schiff base condensation product. For 2a, short Mo---O bonds and high pK_a values (compared to the aqua ion [Mo₃S₄(H₂O)₉]⁴⁺) indicate the formation of a zwitterionic form of the tdci ligand with coordinated alkoxo groups and peripheral dimethylammonium groups. No significant differences were found for the structural properties of the Mo-tdci fragment in 2a and 3a. The coordination modes of taci and tdci, as observed in the solid state, are in agreement with the previously reported solution structures, established by NMR spectroscopy. They are attributed to the specific steric requirements of the two ligands and to a pronounced preference of the [Mo₃(μS)₃(μ₃S)]⁴⁺ core to coordinate a nitrogen donor trans to μ₃S.     

Dual effects of ATP on phosphatidylinositol breakdown in rat hepatocyte membranes

The mechanisms whereby adenosine-5−triphosphate (ATP)_regulated the inositol phospholipid-signalling system were studied in rat hepatocytes. Intact hepatocytes respond to extracellular ATP, adenosine-5′-O-(3-thiotriphosphate) (ATPγS), ADP and weakly to guanosine-5′-triphosphate (GTP), but not to other purine nucleotides (GDP or AMP). This is consistent with the ideal that a P₂ purinergic receptor is coupled to the phosphatidylinositol metabolism in these cells. Partially purified plasma membranes prepared from myo-[³H]inositol prelabelled hepatocytes exhibit a phosphatidylinositol-4,5-bisphosphate phospholipase C activity sensitive to ATP, ATPγS and guanosine-5′-O-(3-thiotriphosphate) (GTPγS). Moreover the GTPγS effect of greatly enhanced by ATP and ATPγS. These potentiating effects differ according to the adenylnucleotide considered. ATP produces (1) an increase in the GTPγS-PLC sensitivity, (2) a potentiation of the phospholipase C (PLC) response induced by maximal dose of GTPγS, and (3) an increase in the inositol lipids pools. At variance, ATPγS, a nonhydrolysable analogue of ATP, only increases the PLC-sensitivity towards GTPγS. These results may signify that ATP stimulates inositol phosphate accumulation via at least two distinct mechanisms (i) a direct activation of a P₂ purinergic receptor coupled to a PLC via a GTP binding protein and (ii) a stimulation of the phosphatidylinositol (PI) and phosphatidyinositol-4-phosphate (PIP) kinases which increased the pool of phospholipase C substrates.     

几种旱地农作物在农田N2O释放中的作用及环境因素的影响

用封闭箱法原位观测几种旱田N₂O的排放通量,并与裸地N₂O通量比较,评价植物在农田N₂O释放中的作用。田间观测与室内模拟实验结合,考察环境因子对N₂O通量的影响。结果表明1d内大豆田N₂O通量有两个释放高峰,而菠菜田和春小麦田只有1个释放高峰。种植大豆较大地提高了农田N₂O的排放通量。农田裸地为一较弱的N₂O释放源,且在1年的一定时期内表现为大气N₂O的汇。光照变化对植物N₂O通量影响很大,在较弱的光照条件下,植物释放N₂O的通量较高。     

Gill microsomal (Na+,K+)-ATPase from the blue crab Callinectes danae: Interactions at cationic sites

Euryhaline crustaceans tolerate exposure to a wide range of dilute media, using compensatory, ion regulatory mechanisms. However, data on molecular interactions occurring at cationic sites on the crustacean gill (Na⁺,K⁺)-ATPase, a key enzyme in this hyperosmoregulatory process, are unavailable. We report that Na⁺ binding at the activating site leads to cooperative, heterotropic interactions that are insensitive to K⁺. The binding of K⁺ ions to their high affinity sites displaces Na⁺ ions from their sites. The increase in Na⁺ ion concentrations increases heterotropic interactions with the K⁺ ions, with no changes in K_0.5 for K⁺ ion activation at the extracellular sites. Differently from mammalian (Na⁺,K⁺)-ATPases, that from C. danae exhibits additional NH₄⁺ ion binding sites that synergistically activate the enzyme at saturating concentrations of Na⁺ and K⁺ ions. NH₄⁺ binding is cooperative, and heterotropic NH₄⁺ ion interactions are insensitive to Na⁺ ions, but Na⁺ ions displace NH₄⁺ ions from their sites. NH₄⁺ ions also displace Na⁺ ions from their sites. Mg²⁺ ions modulate enzyme stimulation by NH₄⁺ ions, displacing NH₄⁺ ion from its sites. These interactions may modulate NH₄⁺ ion excretion and Na⁺ ion uptake by the gill epithelium in euryhaline crustaceans that confront hyposmotic media.     

Isolation, characterization, and nitrification potential of a methylotroph and two heterotrophic bacteria from a consortium showing methane-dependent nitrification

Abstract A methanotrophic nitrifying consortium was previously obtained from a humisol which showed CH₄-dependent nitrification. Although the methanotroph could not be obtained in pure culture, three other members of the consortium have been isolated: An obligately methylotrophic Methylobacillus (Is-1) which grows only on CH₃OH and does not nitrify; a Pseudomonas (Is-2) which grows on Is-1 culture filtrate and produces NO₂⁻, NO₃⁻ and N₂O from NH₂OH, and NO₃⁻ from NO₂⁻; and a second Pseudomonas (Is-3) which produces NO₃⁻ from NH₄⁺ or NO₂⁻, and N₂O from NH₂OH. A model is proposed for the trophic relations and nitrogen transformations in the consortium which may apply to some natural systems.     

The activation by Ca of the ATPase of extracted muscle fibrils with variation of ionic strength, pH and concentration of MgATP

1. The alteration of the Ca²⁺ requirements of the ATPase activity of fibrils from rabbits and crabs at varying ionic strength, pH and concentration of MgATP (i.e. MgATP²⁻ + MgHATP⁻) was investigated.

2. Under physiological conditions, it was found that the ATPase activity of rabbit and crab fibrils after an initial increase decreased steeply when the Ca²⁺ concentration is raised above 1×10⁻⁴ M. This is a primary effect of the over-optimal Ca²⁺ concentration and not a secondary one caused by the influence of accompanying ions.

3. The Ca²⁺ requirements for ATP splitting by rabbit fibrils remain constant at an ionic strength from 0.1 to 0.2 and for a MgATP concentration in the range from 0.5 to 10 mM. At I = 0.05 it is about 5 times smaller than at 0.1. When the pH is decreased from 8 to 7, the Ca²⁺ requirements are increased some 10 times but only 3 times when the pH is varied between 7 and 6.

4. In crab fibrils, there is no alteration of the Ca²⁺ requirements when the ionic strength is varied between 0.05 and 0.2, but a reduction of the pH from 8.0 to 6.0 raises the Ca²⁺ requirements for half activation and for threshold by a factor of 10. Changing the MgATP concentration increases the Ca²⁺ requirements only in the range from 1 to 5 mM, while the concentration required in 0.5 mM is identical with that at 1 mM, and 10 mM corresponds to 5 mM.

5. It can be deduced from the experimental results that at a pH above 6.0 maximal activation is always obtained if the Ca²⁺ concentration is 5×10⁻⁵ M. By contrast, relaxation is only achieved when the Ca²⁺ concentration is below 1×10⁻⁷ M for pH 7.0 and I > 0.1 or below 1×10⁻⁸ for pH > 7.0 or I < 0.1.

6. To achieve complete relaxation, an ethyleneglycoldiaminotetraacetate (EGTA) concentration of 1 mM is sufficient, even when there is a large degree of contamination by Ca²⁺ as long as the pH stays above 6.5.     

On two photoreactions in System II of plant photosynthesis

Light-induced absorbance changes of cytochrome b₅₅₉ and C₅₅₀ in chloroplasts indicate that noncyclic electron transport from water to ferredoxin (Fd)-NADP⁺ is carried out solely by System II and includes not one but two photoreactions (IIa and IIb) that proceed effectively only in short-wavelength light. (C₅₅₀ is a new chloroplast component identified by spectral evidence and distinct from cytochromes.) The evidence suggests that the two short-wavelength light reactions operate in series, being joined by a System II chain of electron carriers that includes (but is not limited to) C₅₅₀, cytochrome b₅₅₉, and plastocyanin (PC).

H₂O → IIb_hv → C₅₅₀ → cyt. b₅₅₉ → PC → IIa_hv → Fd → NADP⁺

Photoreaction IIb involves an electron transfer from water to C₅₅₀ that does not require plastocyanin and is the first known System II photoreaction resistant to inhibition by 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU) and o-phenanthroline. Cytochrome b₅₅₉ is reduced by C₅₅₀ in a reaction that is readily inhibited by DCMU or o-phenanthroline. Thus, the site of DCMU (and o-phenanthroline) inhibition of System II appears to lie between C₅₅₀ and cytochrome b₅₅₉. Photoreaction IIa involves an electron transfer from cytochrome b₅₅₉ and plastocyanin to ferredoxin-NADP⁺.     

A functional site of Ca in the oxygen-evolving photosystem II preparation from Synechococcus sp.

Treatment of the oxygen-evolving photosystem II preparation from the thermophilic cyanobacterium Synechococcus sp. with EDTA inhibited electron flow from Z to P680 and consequently induced a back electron flow from Q⁻_a to P680⁺. The inhibition was reversed fully by Ca²⁺and partially by Mn²⁺ and Mg²⁺ when EDTA-treated preparations had been incubated with respective divalent metal cations for several minutes, whereas diphenylcarbazide had no effect on the recombination between q⁻_a and P680⁺ in EDTA-treated preparations. It is concluded that Ca²⁺ is essential for electron transport from Z to P680.

Oxygen evolution Electron transport Photosystem II Ca²⁺ Thermophilic cyanobacterium     

田间大豆植株N2O通量的测定及光照的影响

采用封闭式箱法,在田间自然状况下对大豆植株N₂O通量进行了测定.结果表明,在主要生育期内,大豆植株N₂O通量有2个释放高峰,分别位于苗期和开花结荚期.大豆植株N₂O通量的昼间变化模式基本上为上午有1个释放高峰,而下午有一个释放低谷.施肥和对照小区N₂O平均通量分别为2.27和1.28μgN₂Om^-2·h^-1.在较强的光照条件下(10⁴lx数量级),大豆植株N₂O通量较低,甚至可吸收大气中的N₂O,而在较弱光照条件下(10³～10²lx数量级),大豆植株N₂O通量较高.     

Coordination of a disubstituted alkene in a chelated d-metal-alkyl-alkene complex. Evidence for equilibrium between complexes with coordinated and free alkenes

Addition of (Cp*₂YH)₂ (4) to 2-methyl-1,4-pentadiene produced the yttrium-alkyl-alkene chelate complex Cp*₂YCH₂CH₂CH₂C(CH₃)=CH₂ (2) in which a disubstituted alkene is complexed to the metal center. Evidence for coordination of the alkene unit of 2 comes from the ¹H and ¹³C NMR chemical shifts of the vinyl units and from observation of nOe effects between Cp* protons and vinyl hydrogens. The disubstituted alkene ligand of 2 is weakly bound, and evidence for an equilibrium with substantial amounts of complex 3 with a free alkene was obtained from variable temperature ¹H NMR spectroscopy.     

Oxidant Stress and Glomerular Prostanoid Production: Influence of Angiotensin Converting Enzyme Inhibition

1. The effect of H₂O₂ (4.7 × 10^-9 4.7 × 10^-3M) on prostanoid production by isolated glomeruli from normotensive (WKY) and, spontaneously hypertensive rats (SHR) has been studied.

2. Oxidant stress significantly increased synthesis of prostaglandin E₂(PGE₂), I₂(PGI₂)and thromboxane A₂ (TxA₂) by glomeruli from both strains whereas the ratio (PGE₂ + PGI₂)/TxA₂ increased in only SHR.

3. Pre-incubation of glomeruli with the angiotensin converting enzyme inhibitors captopril or lisinopril, had virtually no effect on H₂O₂-induced synthesis of individual prostanoids nor on the ratio (PGE₂ + PGI₂)/TA₂ by glomeruli from either WKY or SHR.

4. The findings suggest that H₂O₂-induced changes in glomerular function may be mediated, in part, by PGs but fail to support the suggestion that the ability of ACEI to protect glomeruli from H₂O₂-induced damage is determined by PGs.     

广州地区晚稻田 CH4和N2O 的排放通量及其影响因素

用密闭箱法同时研究了广州地区晚稻田CH₄和N₂O的排放通量。结果表明,连续淹水、常规连作和水旱轮作等3种处理的CH₄平均排放通量分别为1.763、2.84和0.36mg·m^-2·h^-1,而N₂O的平均排放通量分别为6.74、11.69和55.07μgN₂O-N·m^-2·h^-1,表明稻田连续淹水显著增加CH₄的排放而降低N₂O的排放。水旱轮作降低CH₄排放而提高N₂O的排放,说明稻田CH₄和N₂O排放之间存在着消长关系。讨论了这2种温室气体排放的影响因素,并初步分析了它们对温室效应的相对贡献。     

Denitrification in sediment determined from nitrogen isotope pairing

Abstract A new method for accurate and easy measurement of denitrification in sediments is presented. The water overlying intact sediment cores was enriched with ¹⁵NO₃⁻ which mixed with the ¹⁴NO₃⁻ of the natural sources of NO₃⁻. The formation by denitrification of single-labeled (¹⁴N¹⁵N) and double-labeled (¹⁵N¹⁵N) dinitrogen pairs was measured by mass spectrometry after a few hours incubation. Total denitrification including the formation of unlabeled (¹⁴N¹⁴N) dinitrogen could be calculated assuming random isotope pairing by denitrification of the uniformly mixed NO₃⁻ species. In contrast to previous approaches, by this method it is possible to measure denitrification of both NO₃⁻ diffusing from the overlying water and NO₃⁻ from nitrification within the sediment.     

Changes in the Activities of H+, K+-ATPase and Na+, K+-ATPase in Cultured Cells Derived from Micromeres of Sea Urchin Embryos with Special Reference to Their Roles in Spicule Rod Formation

In cultured cells derived from micromeres isolated at the 16-cell stage of sea urchin embryos, the activity of H⁺, K⁺-ATPase became detectable after 15 hr of culture, when the cells started to form spicules, and then increased reaching a plateau from 25 hr of culture. The Na⁺, K⁺-ATPase activity of isolated micromeres increased to a maximum at 20 hr of culture and thereafter decreased gradually. Allylisothiocyanate, an inhibitor of H⁺, K⁺-ATPase, caused a decrease in intracellular pH (pHi) accompanied by blockage of ⁴⁵Ca deposition in spicule rods in spicule-forming cells at 30 hr of culture. Ouabain and amiloride had scarcely any effect on the pHi or ⁴⁵, deposition. In cultured cells exposed to nifedipine, which blocked ⁴⁵Ca deposition in spicule rods, allylisothiocyanate did not cause any decrease in pHi. These results show that H⁺, which is generated in the overall reaction to produce CaCO₃ from Ca²⁺ and HCO₃⁻, is probably released from the cells mainly in the reaction catalyzed by H⁺, K⁺-ATPase to maintain successive production of CaCO₃.