内源性CO在心血管系统的细胞信使作用

近年的研究发现内源性一氧化碳（ＣＯ）不仅是中枢误字率牟细胞信使,也是心血管扩细胞信使。血红素－ＨＯ－ＣＯ－ＣＧＭＰ系统与Ｌ－ａｒｇ－ＮＯＳ－ＮＯ－ｃＧＭＰ系统及血管活性物质的关系密切,涉及许多生理和病理生理过程。ＣＯ在心血管系统的血管舒张,血压调控和心肌保护中起重要作用。     

Hepatic calcium-binding protein regucalcin is released into the serum of rats administered orally carbon tetrachloride

The change in calcium-binding protein regucalcin, mainly localized in liver, in the liver and serum of rats received a single oral administration of carbon tetrachloride (50%; 1.0 ml/100 g body weight) was investigated. The change of regucalcin mRNA levels in the liver was analyzed by Northern blotting using liver regucalcin cDNA (0.6 kb). At 10 and 24 h after the administration, liver regucalcin mRNA levels were reduced markedly. Moreover, regucalcin concentration in the liver and serum was estimated by enzyme-linked immunoadsorbent assay (ELISA) with rabbit-anti-regucalcin IgG. Administration of carbon tetrachloride (CCl₄) induced a significant decrease in liver regucalcin concentration and a corresponding elevation of serum regucalcin concentration at 24 h after the administration. An appreciable increase in serum regucalcin concentration was seen at 2 h after the administration. Meanwhile, serum transaminases (GOT and GPT) activities were significantly increased by CCl₄ administration, indicating that liver injury is induced. The present study demonstrates that hepatic regucalcin is released into the serum of rats administered orally CCl₄, suggesting that the estimation of serum regucalcin is a useful tool for diagnosis of liver injury.     

Nickel is required for the transfer of electrons from carbon monoxide to the iron-sulfur center(s) of carbon monoxide dehydrogenase from Rhodospirillum rubrum

The role of nickel in CO oxidation and electron flow was investigated in carbon monoxide dehydrogenase from Rhodospirillum rubrum. The Fe-S centers of oxidized, nickel-containing (holo) CO dehydrogenase were completely reduced within 1 min of exposure to CO. The Fe-S centers of oxidized, nickel-deficient (apo) CO dehydrogenase were not reduced during a 35-min incubation in the presence of CO. Apo-CO dehydrogenase Fe-S centers were reduced by dithionite. The Fe-S centers of cyanide-inhibited, holo-CO dehydrogenase were not reduced in the presence of CO but were reduced by dithionite. Treatment of apo-CO dehydrogenase with cobalt(II), zinc(II), and iron(II) resulted in association of these metal ions (0.70, 1.2, and 0.86 mol of M2+/mol, respectively) with the protein but no increase in specific activity. Purified holo-CO dehydrogenase contained 1.1 mol of nickel/mol of protein and could not be further activated upon addition of NiCl2, suggesting the presence of one catalytic nickel site on the enzyme. The M2+-treated enzymes could not be further activated by addition of NiCl2 as opposed to the untreated apoenzyme, whose activity was stimulated 50-100-fold to the level of holoenzyme upon addition of NiCl2. When placed under CO, the Fe-S centers of the cobalt-treated enzyme became reduced over a 35-min time course, as opposed to the zinc- and iron-treated enzymes, which remained oxidized. We conclude that nickel, or an appropriate nickel analogue in the nickel site, mediates electron flow from CO to the Fe-S centers of CO dehydrogenase.     

The dissociation of carbon monoxide from hemoglobin intermediate

To investigate the mechanism of allosteric switching in human hemoglobin, we have studied the dissociation of the ligand (CO) from several intermediate ligation states by a stopped-flow kinetic technique that utilizes competitive binding of CO by microperoxidase. The hemoglobin species investigated include Hb(CO)4, the diliganded symmetrical species (alpha beta-CO)2 and (alpha-CO beta)2, and the di- and monoliganded asymmetrical species (alpha-CO beta-CO)(alpha beta), (alpha-CO beta)(alpha beta-CO), (alpha beta-CO) (alpha beta), and (alpha-CO beta)(alpha beta). They were obtained by rapid reduction with dithionite of the corresponding valence intermediates that in turn were obtained by chromatography or by hybridization. The nature and concentration of the intermediates were determined by isoelectric focusing at -25 degrees C. The study was performed at varying hemoglobin concentrations (0.1, 0.02, and 0.001 mM [heme]), pH (6.0, 7.0, 8.0), with and without inositol hexaphosphate. The results indicate that: (a) hemoglobin concentration in the 0.1-0.02 mM range does not significantly affect the kinetic rates; (b) the alpha chains dissociate CO faster than the beta chains; (c) the symmetrical diliganded intermediates show cooperativity with respect to ligand dissociation that disappears in the presence of inositol hexaphosphate; (d) the monoliganded intermediates dissociate CO faster than the diliganded intermediates; (e) the asymmetrical diliganded intermediates are functionally different from the symmetrical species.     

Estimation of amount of stationary pulmonary blood from carbon monoxide uptake measurements

A mathematical model of CO uptake from a single alveolus is modified to include stationary pulmonary blood arising from a pulmonary vascular obstruction. From this model an estimator model is developed that produces simultaneous estimations of the diffusing capacity of the lung for CO and the fraction of the pulmonary capillary blood that is stationary. The estimator model was tested using simulated data from uniform and non-uniform simulators and found to be only mildly sensitive to noise and incorrect values for the pulmonary capillary blood volume. Both the estimator model and breath-to-breath changes in the diffusing capacity of the lung for CO (exhaled) were found to be greatly affected by inhomogeneity of diffusing capacity and ventilation. At times both returned false positive results that limit their use as a screening test for stationary pulmonary blood. Although changes in CO uptake may at times indicate the presence of stationary pulmonary blood, the confounding effects of inhomogeneity of ventilation and diffusing capacity make the use of such changes impractical under most circumstances.     

The reticuloplasmin calreticulun is released into the medium by carrot cell cultures

We report here the presence of a 58-kDa protein in the cells of Daucus carota L. cultivated in vitro. Two lines of carrot cells are used: wild-type line (wt) and mutant line (ts11). We describe here also presence of this protein in the media of cultured cells. Strong reaction of this intracellular and extracellular protein with an anti-calreticulin antiserum indicates that it is a major high capacity, low affinity Ca²⁺-binding reticuloplasmin–calreticulin. No differences in biochemical characterization is found between calreticulin purified from the wild-type line and the mutant line. Moreover molecular mass, type of glycosylation and the ability of extracellular protein to bind calcium is found to be indistinguishable from those of the purified intracellular calreticulin. Calreticulin release is attributed to some stress imposed on cultured cells by growth conditions. It is shown that this process can be also induced in CR-non-releasing systems such as carrot somatic embryos by applying a high-cell-density stress.     

The rate of entry of dioxygen and carbon monoxide into myoglobin.

The model for carbon monoxide or dioxygen recombination with heme proteins developed by the group at the University of Illinois is reexamined. We propose that the carbon monoxide or dioxygen molecule enters the protein at essentially a diffusion-limited rate determined by the solvent viscosity and that the protein offers no important barriers to this entry. The viscosity dependence of the entry rate k(ED), its magnitude (1 x 10(10) M(-1)s(-1), and the rate of quenching of triplet states of protoprophyrin IX in apomyoglobin by dioxygen are used as supporting evidence. Comparison is made to the model of a fluctuating protein developed by G. Weber.     

Predictions of carbon monoxide and blood carboxyhemoglobin levels from motor vehicles exhaust emissions

This study aims at predicting Carbon monoxide (CO) emission rate as well as carboxyhaemoglobin (HbCO) levelsin Al-Ain City, United Arab Emirates (UAE). Also, several mathematical models are proposed for estimating both CO emission and HbCO levels. These models yield excellent agreements between observed and predicted data and can be easily coupled with atmospheric models. Carbon monoxide exposure from motor vehicles were measured over a 3-month period from September to December 1995 at three locations in the UAE. The study is based on a sample of 148 cars. The data revealed that the major characteristics of the cars surveyed were as follows: 69.5% of the cars were Japanese, 74.3% of the cars were 5 years old; 41% of the car was range rover or four-wheel-drive car, which indicates a higher proportion of powerful cars; 58.8% was used special fuel; 51.4% had regular maintenance for their cars and 52% of the cars had 4 cylinder engine capacity. Carboxyhaemoglobin (HbCO) levels in the UAE were predicted for 1-hour (1.720 ppm) and 8-hour (8.106 ppm) exposure times. Also, on an average driving of 80 km hr–1 per day, the CO emission rate was found to be 8.729 ppm. These findings indicate that the mean 1-hour and 8-hour Carboxyhaemoglobin (HbCO) levels do not exceed the permissible standards recommended by the WHO. Additionally, the results of stepwise multiple regression analyses revealed that the factors, such as, type of car, size of cylinder, size of car, and maintaining service of the car exerted the greatest influence on the amount and concentration of pollutant emission produced by car exhaust.     

一氧化碳在低氧性肺血管收缩反应中的作用

目的和方法 :观察外源性一氧化碳 (CO)对大鼠离体肺动脉环低氧性收缩反应 (HPV)的影响 ,并通过观察血红素氧化酶抑制剂ZnPPIX对HPV的影响 ,探讨内源性一氧化碳在HPV中的作用及机制。结果 :低氧可使苯肾上腺素 (PE)预收缩的肺动脉环出现明显的收缩反应 ,肺动脉cGMP含量下降 ;用ZnPPIX孵育后 ,低氧后的肺动脉cGMP含量增加 ,低氧性肺血管收缩反应 (HPV)受抑 ;外源性CO可明显增加肺动脉cGMP含量 ,HPV明显受抑。结论 :外源性CO及ZnPPIX可增加低氧后的肺动脉cGMP含量 ,抑制HPV ,内源性CO减少导致cGMP含量下降可能是HPV的原因之一     

The effect of carbon monoxide on haem-facilitated oxygen diffusion

The aim of this paper is to quantify the effect of small quantities of carbon monoxide on the facilitated diffusion of oxygen by haemoglobin in the steady state. It is the first phase in the study of a mathematical model for carbon monoxide poisoning. Here we extend the Wyman model for facilitated diffusion to the case in which there are two ligands. The equations are solved using an asymptotic technique developed by Murray. We obtain accurate analytic approximations for the biologically important quantities of the problem for various percentages of carbon monoxide. These are the concentrations of free oxygen, haemoglobin, oxyhaemoglobin and carboxyhaemoglobin, and hence the saturation of the protein and the facilitated oxygen flux. The major effect of very small quantities of carbon monoxide on the oxygen flux is shown.     

Luminescence from the carbon monoxide derivative of Agaricus bispora tyrosinase

The luminescence of the CO adduct of two isozymic tyrosinases isolated from Agaricus bispora, an edible white mushroom, has been studied. At room temperature the emission appears as a single smooth peak centered at 530 nm with FWHM of 2700 cm-1 and a lifetime of 36 microseconds. The lifetime and wavelength of the emission are virtually unchanged on lowering the temperature from 298 to 77 degrees K. Solvent composition affects the wavelength of emission minimally. The emission is quenched by oxygen but not by a series of substrate analogs, inhibitors, or Lewis bases. The emission further appeared independent of aggregation state of the enzyme or isozyme type. A comparison of these data is made with those obtained by other researchers for the tyrosinase from Neurospora crassa and for several hemocyanins. The comparison supports the hypothesis that regulation of enzymatic activity does not take place within the coordination sphere of the copper atom observed. In addition, it suggests that the 550- to 560-nm emissions previously observed may not be considered characteristic of all CO derivatives of coupled binuclear copper proteins.