Photosynthetic photon flux effects on bean response to nitrogen dioxide

Phaseolus vulgaris L. cv. Kinghorn Wax seedlings, supplied with nutrient solution containing either 0 or 5 mM nitrate as sole N source, were exposed to 0.25 μl/l NO₂ for 6 hr each day for 10 days at continuous photosynthetic photon flux (PPF) of 100, 300, 500 or 700 μmol m⁻² sec⁻¹. There was a significant interaction of PPF and nitrate. Shoot and root dry weights increased with increasing PPFs only when nitrate was supplied. The main effects of NO₂ on plant growth were significant; none of the interactions involving NO₂ were significant. Exposure to NO₂ decreased shoot and root dry weight in both the presence and absence of nutrient N and at all PPF levels. All interactions were significant for in vitro leaf nitrate reductase activity (NRA), which increased markedly at PPFs above 100 μmol m⁻² sec⁻¹ when nitrate was supplied. Treatment with NO₂ strongly inhibited enzyme activity in the presence of nitrate, particularly at the 300 μmol m⁻² sec⁻¹ PPF level. These experiments demonstrated that PPF level does not modify the effect of NO₂ on growth but does have a major effect on NRA and on NO₂ effects on NRA in the presence of nutrient nitrate.     

小麦NO3-转运蛋白基因TaNRT2.3的克隆和表达分析

用RT-PCR和RACE技术在NO3-诱导处理的小麦(Triticum aestivum L.)根中克隆到一个硝酸根转运蛋白基因的cDNA,命名为TaNRT2.3(GenBank登录号AY053452).序列分析表明,TaNRT2.3全长1 744 bp,其中含有1 521bp的ORF,编码507个氨基酸,具有12个跨膜区,属于MFS超基因家族中的NNP家族.TaNRT2.3与其他植物中已知的NRT2具有很高的同源性.Northern杂交表明:TaNRT2具有在根中表达的组织特异性,而在叶中未检测到.TaNRT2的表达受NO3-诱导,在含NH4 介质中不表达.NO3-在低浓度(5～200μmol/L)和高浓度(2.0 mmol/L)时均起作用.通过研究小麦在0.2 mmol/LNO3-条件下TaNRT2的表达水平及对NO3-的吸收效率,表明TaNRT2在小麦高效吸收NO3-方面起着重要的作用.分根实验表明植物中N循环本身可以作为吸收N的调节信号.     

Nutrient removal by thermophilic Fischerella (Mastigocladus laminosus) in a simulated algaculture process

A novel nutrient removal/waste heat utilization process was simulated using semicontinuous cultures of the thermophilic cyanobacterium Fischerella. Dissolved inorganic carbon (DIC)-enriched cultures, maintained with 10 mg l⁻¹ daily productivity, diurnally varying temperature (from 55°C to 26–28°C), a 12:12 light cycle (200 μE sec⁻¹ m⁻²) and 50% biomass recycling into heated effluent at the beginning of each light period, removed > 95% of NO₃⁻ + NO₂⁻−N, 71% of NH₃-N, 82% of PO₄³⁻ −P, and 70% of total P from effluent water samples containing approximately 400 μg l⁻¹ combined N and 60 μg l⁻¹ P. Nutrient removal was not severely impaired by an altered temperature gradient, doubled light intensity, or DIC limitation. Recycling 75% of the biomass at the end of each light period resulted in unimpaired NO₃⁻ + NO₂⁻ removal, 38–45% P removal and no net NH₃ removal. Diurnally varying P removal, averaging 50–60%, and nearly constant > 80% N removal, are therefore projected for a full-scale process with continuous biomass recycling.     

Nitrite reduces the effects of HOCl on unsaturated phosphatidylcholines--a MALDI-TOF MS study

The concentration of nitrite (NO₂⁻) increases under inflammatory conditions. However, the physiological role of nitrite is so far controversial discussed: it was reported that effects of HOCl (an important inflammation mediator) on phospholipids (PL) may be enhanced but also reduced in the presence of nitrite.

In this paper a simple model system was used: unsaturated phosphatidylcholine (PC) vesicles were treated with HOCl in the presence of varying NaNO₂ concentrations and the yield of reaction products was determined by MALDI-TOF MS: the extent of chlorohydrin generation was significantly reduced in the presence of NaNO₂ because HOCl is consumed by the oxidation of NO₂⁻ to NO₃⁻.

Similar results were obtained when HOCl was generated by the myeloperoxidase (MPO)/H₂O₂/Cl⁻ system or the experiments were carried out in the presence of a simple peptide. It is concluded that the transient products of the reaction between HOCl and NO₂⁻ do not have a sufficient reactivity to modify PL.     

The interaction between nitrogen oxides and hemoglobin and endothelium-derived relaxing factor

Among nitrogen oxides, NO and NO₂ are free radicals and show a variety of biological effects. NO₂ is a strongly oxidizing toxicant, although NO, not oxidizing as NO₂, is toxic in that it interacts with hemoglobin to form nitrosyl-and methemoglobin. Nitrosylhemoglobin shows a characteristic electron spin resonance (ESR) signal due to an odd electron localized on the nitrogen atom of NO and reacts with oxygen to yield nitrate and methemoglobin, which is rapidly reduced by methemoglobin reductase in red cells. NO was found to inhibit the reductase activity. Part of NO inhaled in the body is oxidized by oxygen to NO₂, which easily dissolves in water and converts to nitrite. The nitrite oxyhemoglobin autocatalytically after a lag. The mechanism of the oxidation, particularly the involvement of superoxide, was controversial. The stoichiometry of the reaction has now been established using nitrate ion electrode and a methemoglobin free radical was detected by ESR during the oxidation. Complete inhibition of the autocatalysis by aniline or aminopyrine suggests that the radical catalyzes conversion of nitrite to NO₂, which oxidizes oxyhemoglobin. Recently NO was shown to be one of endothelium- derived relaxing factors and the relaxation induced by the factor was inhibited by hemoglobin and potentiated by superoxide dismutase.     

Second-sphere coordination of ‘star’-shaped hexakis(N-pyridin-4-one)benzene (HPOB) with hexakis(methanol)nickel(II) nitrate; unusual (NO3)(HPOB)(NO3) π-stacked ‘sandwiching’

It is shown by X-ray studies that the compound Ni(HPOB)(NO₃)₂(MeOH)₉ [where HPOB=hexaxis(N-pyridin-4-one)benzene] contains [Ni(MeOH)₆]²⁺ cations hydrogen-bonded to the oxygen atoms of the pyridone units in HPOB, with the resulting six-connectivity at both metal and HPOB producing a three-dimensional network array essentially topologically equivalent to the -Po structure. The pyridone rings in the HPOB molecules are arranged orthogonally to the central C₆ ring and the nitrate anions form an unusual (NO₃ ⁻)(HPOB)(NO₃ ⁻) ‘sandwich’ by a combination of π-stacking and C---HO hydrogen bonds.     

Perchlorate and nitrate reductase activity in the perchlorate-respiring bacterium perclace

The perchlorate (ClO₄⁻)-respiring organism, strain perc1ace, can grow using nitrate (NO₃⁻) as a terminal electron acceptor. In resting cell suspensions, NO₃⁻ grown cells reduced ClO₄⁻, and ClO₄⁻ grown cells reduced NO₃⁻. Activity assays showed that nitrate reductase (NR) activity was 1.31 μmol min⁻¹ (mg protein)⁻¹ in ClO₄⁻ grown cells, and perchlorate reductase (PR) activity was 4.24 μmol min⁻¹ (mg protein)⁻¹ in NO₃⁻ grown cells. PR activity was detected within the periplasmic space, with activities as high as 14 μmol min⁻¹ (mg protein)⁻¹. The NR had a pH optimum of 9.0 while the PR had an optimum of 8.0. This study suggests that separate terminal reductases are present in strain perclace to reduce NO₃⁻ and ClO₄⁻.     

华北二月兰-春玉米轮作体系中土壤硝态氮的时空变化特征

二月兰-春玉米轮作生产体系是近年来为解决华北地区出现的大面积冬闲田而提出的冬绿肥-春玉米生产新模式.本文依托定位试验,研究了该体系从二月兰翻压到玉米收获期间的土壤硝态氮时空变化特征.结果表明：土壤硝态氮含量呈玉米生育前期高后期低的时间变化特征和硝态氮含量峰值随着生育期的推移逐渐下移的空间变化特征,且土壤硝态氮含量随施肥量的增加而显著增加.翻压二月兰对土壤硝态氮含量的时空变化有一定影响,冬春季种植二月兰可降低0～180 cm土壤硝态氮累积量;二月兰翻压后,春玉米苗期与喇叭口期土壤硝态氮规律基本一致,主要集中在0～20 cm土层,0～100 cm土壤剖面为有二月兰处理高于无二月兰处理,100～180 cm土壤剖面则为有二月兰处理低于无二月兰处理;抽雄期以后,土壤硝态氮含量普遍较低,100～180 cm土层土壤硝态氮含量为有二月兰处理略高于无二月兰处理.总体上,翻压二月兰可以增加0～180 cm土层土壤硝态氮保蓄量.     

Kinetics of the reactions of nitrogen dioxide with glutathione,cysteine, and uric acid at physiological pH

Nitrogen dioxide (NO₂^•) is a key biological oxidant. It can be derived from peroxynitrite via the interaction of nitric oxide with superoxide, from nitrite with peroxidases, or from autoxidation of nitric oxide. In this study, submicromolar concentrations of NO₂^• were generated in < 1 μs using pulse radiolysis, and the kinetics of scavenging NO₂^• by glutathione, cysteine, or uric acid were monitored by spectrophotometry. The formation of the urate radical was observed directly, while the production of the oxidizing radical obtained on reaction of NO₂^• with the thiols (the thiyl radical) was monitored via oxidation of 2,2′-azino-bis-(3-ethylthiazoline-6-sulfonic acid). At pH 7.4, rate constants for reaction of NO₂^• with glutathione, cysteine, and urate were estimated as 2 × 10⁷, 5 × 10⁷, and 2 × 10⁷ M⁻¹ s⁻¹, respectively. The variation of these rate constants with pH indicated that thiolate reacted much faster than undissociated thiol. The dissociation of urate also accelerated reaction with NO₂^• at pH > 8. The thiyl radical from GSH reacted with urate with a rate constant of 3 × 10⁷ M⁻¹ s⁻¹. The implications of these values are: (i) the lifetime of NO₂^• in cytosol is < 10 μs; (ii) thiols are the dominant ‘sink’ for NO₂^• in cells/tissue, whereas urate is also a major scavenger in plasma; (iii) the diffusion distance of NO₂^• is 0.2 μm in the cytoplasm and < 0.8 μm in plasma; (iv) urate protects GSH against depletion on oxidative challenge from NO₂^•; and (v) reactions between NO₂^• and thiols/urate severely limit the likelihood of reaction of NO₂^• with NO• to form N₂O₃ in the cytoplasm.     

大气NO2影响植物生长与代谢的研究进展

二氧化氮(NO₂)是大气氮氧化物之一,是大气气溶胶颗粒形成的主要成分,降低大气NO₂浓度可减轻空气中的雾霾.大气NO₂通过干沉降和湿沉降两种方式降落到植物叶片.植物吸收NO₂后主要通过两种代谢途径来降低空气中NO₂浓度: 一是主要在细胞质和叶绿体中利用还原酶的氮代谢途径,二是在质外体和细胞质中的歧化反应.植物吸收NO₂干扰了植物正常的生长和生理代谢,包括: 植物营养和生殖生长,植物体内硝酸还原酶(NaR)活性、亚硝酸还原酶(NiR)活性、氮素吸收、光合等生理代谢过程.对目前国内外有关大气NO₂影响植物生长与代谢的研究进展进行了综述,并对植物吸收NO₂的生理及分子机制的未来研究方向进行了展望.     

大气NO2影响植物生长与代谢的研究进展

二氧化氮(NO₂)是大气氮氧化物之一,是大气气溶胶颗粒形成的主要成分,降低大气NO₂浓度可减轻空气中的雾霾.大气NO₂通过干沉降和湿沉降两种方式降落到植物叶片.植物吸收NO₂后主要通过两种代谢途径来降低空气中NO₂浓度: 一是主要在细胞质和叶绿体中利用还原酶的氮代谢途径,二是在质外体和细胞质中的歧化反应.植物吸收NO₂干扰了植物正常的生长和生理代谢,包括: 植物营养和生殖生长,植物体内硝酸还原酶(NaR)活性、亚硝酸还原酶(NiR)活性、氮素吸收、光合等生理代谢过程.对目前国内外有关大气NO₂影响植物生长与代谢的研究进展进行了综述,并对植物吸收NO₂的生理及分子机制的未来研究方向进行了展望.     

Characterization of the particulate nitrite oxidase and its component activities from the chemoautotroph Nitrobacter agilis

1. Difference spectra, at room and liquid N₂ temperatures, of S₂O₄²⁻-, and NO₂⁻-reduced intact cells and cell-free preparations of Nitrobacter agilis demonstrated the presence of cytochromes of the c- and a-types. Reduction of cytochromes by succinate, and to a limited extent, by NADPH also occurred, provided KCN (0.1 mM) was also present.

2. A particulate, heat-labile nitrite oxidase having an absolute requirement for O₂ was prepared from N. agilis cells using sonic oscillation and differential centrifugation. The particles also possessed NADH oxidase, succinoxidase, formate oxidase and traces of NADPH oxidase activity. The stoichiometry of the nitrite oxidase reaction approached the theoretical value of 2 moles of NO₂⁻ consumed per mole of O₂ consumed. The pH optimum of the nitrite oxidase system shifted to progressively more alkaline values as the NO₂⁻ concentration was increased, changing from a pH value of 6.8 at 0.6 mM KNO₂ to pH 8.0 at 0.01 M KNO₂ with apparent K_m's of 0.2 and 1.2 mM NO₂⁻, respectively. Computations of the HNO₂ concentrations present under the above conditions showed an approx. 500-fold greater affinity for HNO₂ which was independent of pH, suggesting the involvement of HNO₂ as both a substrate and an inhibitor (at higher concentrations) of the nitrite oxidase system. The marked inhibition by NaN₃, NaCN and Na₂S, as well the light-reversible inhibition by CO, indicated the presence of cytochrome oxidase which was subsequently characterized. NO₂⁻ proved to be a competitive inhibitor of the nitrite oxidase system.

3. The particulate preparation also possessed a heat-labile nitrite-cytochrome c reductase activity which was energy independent and routinely measured under anaerobic conditions. As in the case of nitrite oxidase, the affinity of the enzyme for NO₃⁻ increased as the pH was lowered, but the pH optimum remained unaffected. In terms of calculated HNO₂ concentration an approximately constant K_m of about 0.2 μM was estimated at the several pH's examined. The inhibition by NO₃⁻ was shown to be competitive. The marked sensitivity of the reductase to several metal-binding agents implicated a metal component in the electron transport chain at the site prior to cytochrome c.

4. The membrane-like composition of the nitrite oxidase system is indicated.     

A nitrite reductase with cytochrome oxidase activity from Micrococcus denitrificans

The formation of nitrite reductase and cytochrome c in Micrococcus denitrificans was repressed by O₂. The purified nitrite reductase utilized reduced forms of cytochrome c, phenazine methosulphate, benzyl viologen and methyl viologen, respectively, as electron donors. The enzyme was inhibited by KCN, NaN₃ and NH₂OH each at 1 mM, whereas CO and bathocuproin, diethyl dithiocarbamate, o-phenanthroline and ,'-dipyridyl at 1 mM concentrations were relatively ineffective. The purified enzyme contains cytochromes, probably of the c and a₂ types, in one complex. A K_m of 46 μM for NO₂⁻ and a pH optimum of 6.7 were recorded for the enzyme. The molecular weight of the enzyme was estimated to be around 130000, and its anodic mobility was 6.8·10⁻⁶ cm²·sec⁻¹·V⁻¹ at pH 4.55.

The most highly purified nitrite reductase still exhibited cytochrome c oxidase activity with a K_m of 27 μM for O₂. This activity was also inhibited by KCN, NaN₃ and NH₂OH and by NO₂⁻.

A constitutive cytochrome oxidase associated with membranes was also isolated from cells grown anaerobically with NO₂⁻. It was inhibited by smaller amounts of KCN, NaN₃ and NH₂OH than the cytochrome oxidase activity of the nitrite reductase enzyme and also differed in having a pH optimum of about 8 and a K_m for O₂ of less than 0.1 μM. Spectroscopically, cytochromes b and c were found to be associated with the constitutive oxidase in the particulate preparation. Its activity was also inhibited by NO₂⁻.

The physiological role of the cytochrome oxidase activity associated with the purified nitrite reductase is likely to be of secondary importance for the following reasons: (a) it accounts for less than 10% of total cytochrome c oxidase activity of cell extracts; (b) the constitutive cytochrome c oxidase has a smaller K_m for O₂ and would therefore be expected to function more efficiently especially at low concentrations of O₂.     

不同浓度硝态氮供应下小麦生长、硝态氮累积及根系钙信号特征

以小麦品种‘石麦15’和‘衡观35’为材料进行营养液水培试验,研究不同浓度硝态氮供应对小麦苗期根系形态、钙离子流特征及钙调蛋白(CaM)含量的影响。结果表明,与适宜浓度硝态氮处理(2.5mmol/L)相比,无外源硝态氮供应时小麦地上部鲜重、硝态氮含量均降低,侧根数量显著减少;高浓度硝态氮处理(50mmol/L)下两个小麦品种地上部硝态氮含量升高,根系总长度降低,‘石麦15’侧根数量减少。无硝态氮和高浓度硝态氮处理下,根系中钙调蛋白含量降低,且‘衡观35’的降低幅度大于‘石麦15’。无外源硝态氮供应时小麦根尖表现出较为明显的钙离子外流特征;与适宜浓度硝态氮处理相比,高硝态氮处理下小麦根尖Ca2+的内流速度显著下降。说明硝态氮供应不足和高浓度硝态氮供应会影响小麦根系生长,根系Ca2+外流或Ca2+内流速度下降,CaM含量减少,Ca2+/CaM可能介导硝态氮调控小麦根系生长发育。     

外源γ-氨基丁酸对Ca(NO3)2胁迫下甜瓜幼苗NO3--N同化的影响

以盐敏感型甜瓜品种‘一品天下208’为试材,用80 mmol·L^-1 Ca(NO₃)₂模拟设施土壤盐渍化,采用深液流水培,研究外源 γ-氨基丁酸(GABA)对Ca(NO₃)₂胁迫下甜瓜幼苗硝态氮(NO₃^--N)同化的影响.结果表明: Ca(NO₃)₂胁迫显著降低了甜瓜幼苗体内硝酸还原酶(NR)、谷氨酰胺合酶(GS)和谷氨酸合酶(GOGAT)活性,增强了谷氨酸脱氢酶(GDH)、谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性,导致铵态氮(NH₄⁺-N)和游离氨基酸含量增加,NO₃^--N和可溶性蛋白质含量下降,植株生长和光合作用受到严重抑制.Ca(NO₃)₂胁迫下,外源喷施GABA有效促进了甜瓜根系对NO₃^--N的吸收及其向地上部的转运,并通过增强NR、GS和GOGAT活性提高了甜瓜幼苗对NH₄⁺的同化力;通过抑制GDH脱氨作用减少了甜瓜幼苗体内NH₄⁺的释放量,从而缓解了盐诱导产生的NH₄⁺-N积累所造成的氨毒害作用;外源喷施GABA也能调节甜瓜组织中氨基酸代谢途径,促进蛋白质的合成.表明外源GABA能增强甜瓜幼苗对NO₃^--N的同化能力,调控氨基酸代谢,进而有效缓解Ca(NO₃)₂胁迫对甜瓜幼苗的盐伤害作用.     

外源γ-氨基丁酸对Ca(NO3)2胁迫下甜瓜幼苗NO3——N同化的影响

以盐敏感型甜瓜品种‘一品天下208’为试材,用80 mmol·L^-1 Ca(NO₃)₂模拟设施土壤盐渍化,采用深液流水培,研究外源 γ-氨基丁酸(GABA)对Ca(NO₃)₂胁迫下甜瓜幼苗硝态氮(NO₃^--N)同化的影响.结果表明: Ca(NO₃)₂胁迫显著降低了甜瓜幼苗体内硝酸还原酶(NR)、谷氨酰胺合酶(GS)和谷氨酸合酶(GOGAT)活性,增强了谷氨酸脱氢酶(GDH)、谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性,导致铵态氮(NH₄⁺-N)和游离氨基酸含量增加,NO₃^--N和可溶性蛋白质含量下降,植株生长和光合作用受到严重抑制.Ca(NO₃)₂胁迫下,外源喷施GABA有效促进了甜瓜根系对NO₃^--N的吸收及其向地上部的转运,并通过增强NR、GS和GOGAT活性提高了甜瓜幼苗对NH₄⁺的同化力;通过抑制GDH脱氨作用减少了甜瓜幼苗体内NH₄⁺的释放量,从而缓解了盐诱导产生的NH₄⁺-N积累所造成的氨毒害作用;外源喷施GABA也能调节甜瓜组织中氨基酸代谢途径,促进蛋白质的合成.表明外源GABA能增强甜瓜幼苗对NO₃^--N的同化能力,调控氨基酸代谢,进而有效缓解Ca(NO₃)₂胁迫对甜瓜幼苗的盐伤害作用.     

保护性耕作措施对旱地春玉米土壤水分和硝态氮淋溶累积的影响

通过渭北旱塬春玉米15年定位试验,研究6种保护性耕作方式,即传统翻耕(CT)、免耕(NT)、免耕配施生物炭(NB)、免耕秸秆覆盖(NS)、免耕地膜覆盖(NF)和免耕秸秆地膜覆盖(NSF)对土壤水分与硝态氮淋溶的影响,探索适宜于该区可持续发展的农田管理措施.结果表明: 与NT相比,第一次水分补给时期0～100 cm土层,CT处理水分补给量无显著差异,NS、NB、NSF和NF处理显著降低;在100～300 cm土层,NS、NB、NF和NSF处理显著增加了水分补给量,CT显著低于NT处理.第二次水分补给时期,水分补给主要集中在0～100 cm土层,且各处理与NT水分补给量无显著差异.水分消耗时期,各处理与NT处理在0～100 cm土层消耗量无显著差异,NF和NSF在100～300 cm土层水分消耗量显著增加,分别增加了33.9%和59.9%.在0～200 cm土层,与NT相比,CT处理显著增加了硝态氮累积量,是NT的2.2倍,NS、NB、NF和NSF处理显著减少了硝态氮累积量,分别减少了44.6%、61.5%、69.2%和69.8%;而在200～300 cm土层,NS处理显著增加了硝态氮累积量,CT与NT处理无显著差异,NB、NF和NSF处理显著减少.土壤水分运动能显著影响硝态氮在土壤剖面的分布,NB、NF和NSF处理硝态氮主要分布在0～40 cm土层,NS主要分布在0～100 cm和200～300 cm土层,而NT和CT处理硝态氮在整个剖面大量分布,其中NS、NT和CT处理出现两个硝态氮累积峰.不同农田管理措施通过对水分的调控减少硝态氮淋溶,进而提高氮素利用效率,其中免耕基础上秸秆地膜覆盖能有效调控土壤水分运动和减少硝态氮淋溶累积,是旱塬区改善农田水肥状况,增加作物产量的可行措施.     

土壤中反硝化酶活性变化与N2O排放的关系

研究施肥条件下,土壤反硝化酶活性硝酸还原酶(NR)活性、亚硝酸还原酶(NiR)活性及羟胺还原酶(HyR)活性在玉米生长季节中的变化及其与土壤含水量、硝态氮含量、N₂O排放之间的关系。结果表明,3种还原酶都有明显的季节变化规律并受土壤水分含量及施肥的影响。通过研究3种反硝化酶活性与土壤含水量及N₂O排放量之间的关系后指出,反硝化酶活性变化可作为一个区分旱田N₂O产生途径的指标.     

Particulate formate oxidase from Nitrobacter agilis

1. The nitrite oxidase particles obtained by sonic oscillation of Nitrobacter agilis cells also possessed appreciable formate oxidase activity, ranging from about 25 to 50% of the nitrite oxidase activity depending upon the N. agilis strain. Both activities distributed themselves in the same pattern and proportions during differential centrifugation, and resided solely in the pellet resulting from high-speed centrifugation.

2. Difference spectra of formate-reduced particles or intact cells demonstrated the presence of cytochromes of the c- and a-types like those of the NO₂⁻-reduced material. Under anaerobic conditions NO₃⁻ or fumarate acted as an alternate electron acceptor in place of O₂ in formate oxidation. Under aerobic conditions increasing NO₃⁻ concentrations resulted in (a) an increased role of NO₃⁻ as a terminal electron acceptor compared to O₂, (b) a greater total enzymatic transfer of electrons from formate than if O₂ were the sole electron acceptor, and (c) a partial inhibition of O₂ uptake suggestive of a competition for electrons by the two acceptors. The formate oxidase system failed to catalyze consistently the transfer of electrons to either added mammalian cytochrome c or Fe(CN)₆³⁻. The marked sensitivity of the system to certain inhibitors implicated cytochrome oxidase as an integral part of the formate oxidase. The system was also inhibited significantly by a variety of chelating agents, indicating a metal component in the formate dehydrogenase or early portion of the electron transfer sequence.

3. The stoichiometry of the formate oxidase system was shown to approach the theoretical value of 2 moles of CO₂ evolved per mole of O₂ or per 2 moles of formate consumed.

4. To a limited extent, phosphorylation occurred concomittantly with the oxidation of formate in the presence of the cell-free particulate system.     

Low-dose intravenous nitrite improves hemodynamics in a canine model of acute pulmonary thromboembolism

Acute pulmonary thomboembolism (APT)-induced pulmonary hypertension can be counteracted by activating the nitric oxide (NO)-cGMP pathway. Recent studies have demonstrated that the naturally occurring anion nitrite (NO₂^-) is a bioactive storage reservoir for NO, and is reduced to NO under conditions of hypoxia and acidosis. We hypothesized that nitrite infused intravenously could attenuate the hemodynamic changes associated with APT. APT was induced with autologous blood clots injected into the right atrium in mongrel dogs. After APT (or saline), the dogs received an intravenous nitrite (or saline) infusion (6.75 μmol/kg over 15 min and then 0.28 μmol/kg/min) and hemodynamic evaluations were carried out for 2 h. Plasma nitrite concentrations were measured using ozone-based reductive chemiluminescence methodologies. APT decreased cardiac index (CI) and increased pulmonary vascular resistance index (PVRI); these effects were improved during infusions of sodium nitrite. Accordingly, nitrite infusion increased cardiac index by 28%, reduced the PVRI by 48%, and the systemic vascular resistance index (SVRI) by 21% in embolized dogs, suggesting a greater effect on the ischemic embolized vascular system than the systemic circulation following embolization. Interestingly, in nonembolized control dogs the same nitrite infusion decreased MAP and CI (all P < 0.05). The nitrite infusion increased plasma nitrite concentrations by approximately 2 μM, and produced dose-dependent effects on PVRI, MAP, and SVRI. Remarkably, blood levels of nitrite as low as 500 nM decreased PVRI and SVRI in this model, suggesting a potential role of nitrite in physiological blood flow regulation. These results suggest that a low-dose nitrite infusion produces beneficial hemodynamic effects in a dog model of APT. These findings suggest a new therapeutic application for nitrite and support emerging evidence for a surprisingly potent and potentially physiological vasoactivity of nitrite.