Article

Simultaneous production of carbon monoxide and thiobarbituric acid reactive substances in rat tissue preparations by an iron-ascorbate system.

Department of Pediatrics, Neonatal and Developmental Medicine Laboratory, Stanford University School of Medicine, CA 94305-5208, USA.
Canadian Journal of Physiology and Pharmacology (Impact Factor: 1.55). 01/1999; 76(12):1057-65. DOI: 10.1139/cjpp-76-12-1057
Source: PubMed

ABSTRACT Most of the carbon monoxide (CO) produced by mammals is a product of the heme oxygenase (HO) reaction, the rate-limiting step in the heme degradation pathway leading to the generation of bilirubin in man. However, some CO is derived from other sources. We studied the association of CO production with lipid peroxidation in tissue preparations from adult male Wistar rats. Supernatants, from 20% tissue homogenates in potassium phosphate buffer, centrifuged for 1 min at 13,000 x g, were incubated for 30 min at 37 degrees C in septum-sealed vials in the dark with ascorbate (100 microM) and Fe(II) (6 microM) and (or) Fe(III) (60 microM). Butylated hydroxytoluene (BHT, 100 microM) was added for the blank reaction. CO produced into the headspace was quantitated by gas chromatography. Thiobarbituric acid reactive substances (TBARS), conjugated dienes (CD), and lipid hydroperoxides (LOOH) in the reaction medium were quantitated by spectrophotometry. Of the tissues studied, CO and TBARS formation was greatest for brain, followed by kidney, lung, spleen, and blood, but no CO or TBARS formation was detected for testes, intestine, liver, and heart. Cell fractionation studies indicated that these differences might be due to the presence of endogenous soluble antioxidants in the latter tissues. Furthermore, these studies demonstrated that CO was exclusively generated by subcellular fractions that contained membranes. The magnitude of the rate of product formation in brain supernatants depended on the concentration of Fe(II) and (or) Fe(III). The formation of CO, TBARS, CD, and LOOH increased linearly with time for up to 30 min, but the rates of product formation were different. Product formation was completely inhibited by BHT (100 microM), biliverdin (50 microM), bilirubin (50 microM), citrate (100 microM), and the Fe(II) chelators, desferrioxamine mesylate (100 microM) and diethylenetriaminepentaacetate, but not by 10 microM of the HO inhibitor, zinc deuteroporphyrin bis glycol. We conclude that CO generation is associated with the process of in vitro lipid peroxidation in tissues with limited antioxidant reserves.

0 Followers
 · 
45 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: Hemolytic disease in newborns can result from a number of conditions, which can place such infants at an increased risk for the development of severe hyperbilirubinemia. Because the catabolism of heme produces equimolar amounts of carbon monoxide (CO) and bilirubin, measurements of end-tidal breath CO (corrected for ambient CO) or ETCOc can serve as an index of hemolysis as well as of bilirubin production from any cause. Elevated levels of ETCOc have been correlated with blood carboxyhemoglobin levels and thus hemolysis. However, the detection of hemolysis can be a clinically challenging problem in newborns. Here, we describe the importance of determining ETCOc levels and their application in identifying infants at risk for developing hyperbilirubinemia associated with hemolysis and other causes of increased bilirubin production.Journal of Perinatology advance online publication, 17 April 2014; doi:10.1038/jp.2014.66.
    Journal of perinatology: official journal of the California Perinatal Association 04/2014; 34(8). DOI:10.1038/jp.2014.66 · 2.35 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The exogenous application of carbon monoxide (CO) is a valuable strategy which enables study of the effects under different stress conditions. However, in this experimental model a true endogenous CO production by plants can not be measured. In this work, so as to achieve an elevated sensitivity and to avoid invasive techniques, we quantify the endogenous CO production by tissues in salt-treated soybean plants through gas chromatography coupled to a reduction gas detector. This technique allows short and room temperature incubation of intact tissues and homogenates. We found that a 200 mM NaCl treatment induces total CO production in leaves and roots. The sensitivity of the technique offers no correlation between this increment and heme oxygenase (HO) activity measured as a function of CO production. We also found that untreated soybean plants continue to produce significant CO levels up to 7 days post planting, after which CO content decreases to a third and remains constant in the next days. However, HO activity does not change throughout these days. The data here reported shows that HO activity is not the main source of CO in soybean plants. We discuss alternative sources that could be implicated in this production. Taking our own results and data reported by other colleagues, we propose lipid peroxidation and ureide metabolism as potential sources of CO.
    Environmental and Experimental Botany 06/2014; 102. DOI:10.1016/j.envexpbot.2014.01.012 · 3.00 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Increased levels of end-tidal carbon monoxide (ETCOc) in preterm infants during the first day of life are associated with oxidative stress, inflammatory processes and adverse neurodevelopmental outcome at 2 years of age. Therefore, we hypothesized that early ETCOc levels may also be associated with impaired growth of unmyelinated cerebral white matter. From a cohort of 156 extremely and very preterm infants in which ETCOc was determined within 24 h after birth, in 36 infants 3D-MRI was performed at term-equivalent age to assess cerebral tissue volumes of important brain regions. Linear regression analysis between cerebral ventricular volume, unmyelinated white matter/total brain volume-, and cortical grey matter/total brain volume-ratio and ETCOc showed a positive, negative and positive correlation, respectively. Multivariable analyses showed that solely ETCOc was positively related to cerebral ventricular volume and cortical grey matter/total brain volume ratio, and that solely ETCOc was inversely related to the unmyelinated white matter/total brain volume ratio, suggesting that increased levels of ETCOc, associated with oxidative stress and inflammation, were related with impaired growth of unmyelinated white matter. Increased values of ETCOc, measured within the first 24 hours of life may be indicative of oxidative stress and inflammation in the immediate perinatal period, resulting in impaired growth of the vulnerable unmyelinated white matter of the preterm brain.
    PLoS ONE 03/2014; 9(3):e89061. DOI:10.1371/journal.pone.0089061 · 3.53 Impact Factor