Lipid peroxidation stimulated by iron nitrilotriacetate in rat liver

The complex of ferric iron with nitrilotriacetate (iron-NTA) given i.p. is an unusually potent stimulus for lipid peroxidation (LP) in vivo, as monitored by exhaled alkanes. Localization of 59Fe-labeled NTA radioactivity in mouse liver and accumulation of thiobarbituric acid (TBA)-reacting material in liver after i.p. injection suggested that the effect of i.p. iron-NTA could be primarily hepatic. It was found that 100 microM iron-NTA added to a hepatocyte suspension gassed with air stimulated ethane production (3 +/- 1 pmoles/10(6) cells/min) versus an undetectable control, and at a sensitivity of 0.083 pmole/10(6) cells/min. Under similar conditions, hepatocytes stimulated by iron-NTA generated low level chemiluminescence (CL) in parallel with formation of TBA-reactants; the generation of CL was concentration related. Liver was homogenized and fractionated by ultracentrifugation: iron-NTA stimulated CL in whole liver homogenate as in intact cells. The greater part of this activity localized to the microsomal and mitochondrial fractions where NADH or NADPH was required. Using rat liver microsomes, it was shown that iron-NTA in the presence of NADPH stimulated two phases of CL with an initial phase maximum in 1-2 min (phase 1) which decreased abruptly to be followed by a prolonged rise (phase 2); NADH could replace NADPH. Ferrous iron (as chloride) caused a burst of CL, whereas ferric iron was inactive. However, complex differences exist between CL stimulated by Fe(II) and by iron-NTA in the presence of reducing equivalents. Under conditions resulting in the production of CL, a microsomal system with iron-NTA and reducing equivalent accumulated TBA-reactants in parallel with the stimulated CL and rapid increase in oxygen consumption. Both desferrioxamine and butylated hydroxyanisole were able to strongly inhibit the CL stimulated by iron-NTA. When iron-NTA and iron-ADP were compared in the microsomal system, similar responses were obtained but major differences characterized the effects of these iron chelates on whole cells with the ADP complex being relatively inactive. We conclude that iron-NTA stimulated free radical reactions in liver by undergoing cyclic oxidation and reduction and that these reactions utilized oxygen, generated CL, and formed TBA-reactants and ethane. At a subcellular level, the reactions of iron-NTA resembled those reported for iron-ADP.     

Dopamine: its potential for inducing ischemic left ventricular dysfunction

As an agent potentially capable of inducing ischemia in patients with coronary artery disease, dopamine administered intravenously was evaluated as a pharmacologic stress agent by supine radionuclide angiography, and the results were compared with ergometer exercise. In a preliminary group of 11 subjects (4 normal subjects and 7 patients with coronary disease), dopamine alone was administered in increments of 2.5 micrograms/kg per min to a maximum of 15 micrograms/kg per min. There were significant differences between exercise and dopamine in maximal stress heart rates, 129.3 +/- 30.0 versus 88.0 +/- 35.8 beats/min (p less than 0.05) in normal subjects and 118.9 +/- 21.1 versus 87.6 +/- 22.6 beats/min (p less than 0.05) in patients with coronary disease, as well as in maximal stress rate-pressure products, 213.3 +/- 51.4 versus 155.0 +/- 52.5 mm Hg/min X 10(2) (p less than 0.02) in normal subjects and 216.0 +/- 45.6 versus 161.0 +/- 48.6 mm Hg/min X 10(2) (p less than 0.003) in patients with coronary disease. As a result, in these patients the ejection fraction response was significantly different: -3.3 +/- 4.5% with exercise versus + 6.3 +/- 4.6% with dopamine (p less than 0.05). In a second group of 41 subjects (9 normal subjects and 32 patients with coronary disease), atropine (0.6 mg) was administered intravenously before and after every second dopamine dose increment. This produced statistically similar maximal stress heart rates as compared with exercise in all subjects, rate-pressure products in normal subjects and slightly higher values with dopamine in patients with coronary disease: 200.3 +/- 47.2 versus 183.1 +/- 43.0 (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Quality of life after repair of tetralogy of Fallot

OBJECTIVE: To determine the quality of life in individuals with corrected tetralogy of Fallot. METHODS AND SUBJECTS: Questionnaires concerning quality of life were sent to all 87 surviving patients aged between 16 and 40 years who had undergone intracardiac repair of tetralogy of Fallot and follow-up in the Wessex Cardiothoracic Unit, and to 87 age and sex matched controls, with medically treated haemodynamically insignificant ventricular septal defects. RESULTS: The only significant difference found between the cases and controls was in requirements for schooling, where those with tetralogy of Fallot were more likely to require additional educational help at school (p = 0.044). For all other aspects of quality of life examined by the questionnaire, including social and genetic history, exercise ability, and health related quality of life, no significant differences were found. Different operative techniques, such as transjunctional patching, right ventriculotomy, and previous palliative shunting, did not affect the quality of life of our population with Tetralogy of Fallot, on average twenty years after their surgery, although the range of operative techniques was limited. Neither age at surgery, nor time since surgery, was correlated with measurements of quality of life. CONCLUSIONS: Those who have undergone surgical correction of tetralogy of Fallot have a normal quality of life, with few differences compared to controls.     

The topomer-sampling model of protein folding

Clearly, a protein cannot sample all of its conformations (e.g., approximately 3(100) approximately 10(48) for a 100 residue protein) on an in vivo folding timescale (<1 s). To investigate how the conformational dynamics of a protein can accommodate subsecond folding time scales, we introduce the concept of the native topomer, which is the set of all structures similar to the native structure (obtainable from the native structure through local backbone coordinate transformations that do not disrupt the covalent bonding of the peptide backbone). We have developed a computational procedure for estimating the number of distinct topomers required to span all conformations (compact and semicompact) for a polypeptide of a given length. For 100 residues, we find approximately 3 x 10(7) distinct topomers. Based on the distance calculated between different topomers, we estimate that a 100-residue polypeptide diffusively samples one topomer every approximately 3 ns. Hence, a 100-residue protein can find its native topomer by random sampling in just approximately 100 ms. These results suggest that subsecond folding of modest-sized, single-domain proteins can be accomplished by a two-stage process of (i) topomer diffusion: random, diffusive sampling of the 3 x 10(7) distinct topomers to find the native topomer ( approximately 0.1 s), followed by (ii) intratopomer ordering: nonrandom, local conformational rearrangements within the native topomer to settle into the precise native state.     

Effect of luminal acid on intracellular pH in oxynticopeptic cells in intact frog gastric mucosa.

The effect of changes in luminal [H+] on intracellular pH in oxynticopeptic cells was examined using intact sheets of frog (Rana catesbeiana) gastric mucosa in which oxynticopeptic cells were selectively loaded with the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). The serosal solution was buffered with either HCO3- or N-2-hydroxymethylpiperazine-N'-2-ethanesulfonic acid (HEPES). Luminal pH was decreased from 7.2 to 1.5 and changed back to 7.2. In stimulated (forskolin-treated) tissues, intracellular pH decreased at luminal pH 1.5 only in HEPES, with complete recovery at 7.2. In resting (omeprazole-treated) tissues, intracellular pH began to decrease at luminal pH 2.0 in HEPES and at 1.5 in HCO3-, with complete recovery at 7.2 in both. In resting tissues bathed in Cl(-)-free HEPES, the recovery of intracellular pH at luminal pH 7.2 was completely prevented by serosal amiloride (1 mmol/L) but was not affected by serosal 4,4'-diisothiocyanatodihydrostilbene-2-2'-disulfonic acid (H2-DIDS; 0.5 mmol/L). In resting tissues bathed in Cl(-)-free HCO3-, the recovery of intracellular pH at luminal pH 7.2 was not affected by amiloride but was prevented partially by H2-DIDS and completely by combination of H2-DIDS and amiloride or by removal of ambient Na+. These results suggest that during exposure to high luminal [H+]: (a) stimulated oxynticopeptic cells maintain a steady intracellular pH more readily than resting cells; (b) serosal HCO3- protects oxynticopeptic cells from intracellular acidosis; and (c) both Na+/H+ exchange and Na(+)-HCO3- cotransport are involved in the recovery from intracellular acidosis in resting oxynticopeptic cells.     

The immunomodulatory effects of thalidomide on human immunodeficiency virus-infected children

The safety and immune effects of low-dose thalidomide treatment (3 mg/kg/day for 28 days) were evaluated in a study involving 8 South African human immunodeficiency virus (HIV)-infected children. The children were 7-69 months old and in disease stages A1-C3. Thalidomide therapy did not affect virus load, even though none of the children was receiving antiretroviral therapy. Thalidomide stimulated CD8+ T cells in peripheral blood, which increased expression of the activation markers CD38 and human leukocyte antigen DR and of the memory cell marker CD45RO. The frequency of HIV gag-specific CD8+ T cells in peripheral blood increased in 3 of 4 children who were evaluated during treatment with thalidomide. Clinical adverse events were mild. In this study, thalidomide was found to be safe and well tolerated and caused significant immunomodulation at a low dose. This is the first report describing use of an oral drug that may enhance HIV-specific CD8+ T cell function in HIV-infected children.