Reduction of reperfusion injury of human myocardium by allopurinol: a clinical study.

To determine the possibility of myocardial protection against reperfusion injury by allopurinol, 22 aortocoronary bypass patients were studied. Eight patients received allopurinol (200 mg during induction of anesthesia and 100 mg after starting extracorporeal circulation) during surgery (group B), and 14 patients served as a control (group A). Blood samples and myocardial biopsies were taken before and 10 min after aortic cross-clamping. No statistically significant difference between the two groups was observed considering gender, age, prior myocardial infarction, left ventricular end diastolic pressure (LVEDP), and aortic cross-clamp time. Preservation of cardiac tissue was assessed by the measurement of quantitative birefringence (QBR) changes upon the addition of adenosine 5'-triphosphate (ATP) plus calcium in biopsies and the need for postoperative inotropes. The synthesis of peroxides was estimated by the measurement of leukotriene B4 and C4 (LTB4, LTC4). LTB4 was below the level of detection (< 1.5 ng/l) before and after cross-clamping in both groups, while the LTC4 level for group A increased from < 1.5 to 27 +/- 17 ng/l compared to an increase of < 1.5 to 11 +/- 8 ng/l for group B after 10 min of reperfusion (p = .036). The decrease in QBR value in group A was 1.26 +/- 0.28 and 0.35 +/- 0.23 for group B (p < .003). Postoperatively, 11 out of 14 patients in group A needed inotropic support (dopamine or dobutamine), whereas two patients out of eight did so in group B.(ABSTRACT TRUNCATED AT 250 WORDS)

Lysine-binding heterogeneity of Lp(a): consequences for fibrin binding and inhibition of plasminogen activation.

Lipoprotein(a) [Lp(a)] is recognized as an independent risk factor for atherosclerosis. Lp(a) consists of a LDL-like moiety with an additional glycoprotein, apo(a), linked to apolipoprotein B-100. Apo(a) has a high homology with plasminogen (Pg). In vivo, Pg is activated on a fibrin surface by tissue Pg activator (tPA). We prepared Lp(a) from plasma by sequential ultracentrifugation followed by lysine-sepharose affinity chromatography. We found that a changing (donor dependent) fraction of the Lp(a) did not bind to lysine-sepharose. This fraction, designated Lp(a)lys-, was further purified using gel filtration. Bound Lp(a) [Lp(a)lys+] was eluted with 0.2 M EACA. Apo(a) isoforms in both fractions were identical. In contrast Lp(a)lys+ inhibited Pg activation by tPA in vitro (IC50% 20 mg/l), whereas Lp(a)lys- did not. In addition Lp(a)lys- did not bind to CNBr-digested fibrinogen whereas Lp(a)lys+ did (Kd, app = 0.2 nM). Therefore we conclude that a changing donor dependent fraction of human plasma Lp(a) does not inhibit Pg activation in vitro and does not bind to CNBr-digested fibrinogen.

Monitoring rejection after heart transplantation: cytoimmunological monitoring on blood cells and quantitative birefringence measurements on endomyocardial biopsy specimens.

Cytoimmunological monitoring and quantitative birefringence measurements were used as potential aids in diagnosing acute rejection after heart transplantation instead of histopathological assessment of the endomyocardial biopsy specimen alone. Cytoimmunological monitoring was based on morphological inspection and quantitation of mononuclear cells, particularly activated lymphoid cells. Quantitative birefringence measurements comprise a variable for myocyte contractile function. Its read out is the ratio of the degree of birefringence before contraction to that after. Cytoimmunological monitoring indicated significantly higher concentrations of activated lymphocytes in moderate or severe acute rejection, and quantitative birefringence measurements indicated decreased myocyte function during severe and resolved or resolving rejection. Cytoimmunological monitoring and quantitative birefringence measurements were diagnostically most useful in terms of sensitivity, specificity, and predictive value, when only data gathered before the first episode of acute rejection were considered. For cytoimmunological monitoring, diagnostic relevance was optimal when the data were expressed as relative proportions of activated lymphocytes. The quantitative birefringence measurements correlated best with analysis of the endomyocardial biopsy specimen when a cut off value of 1.25 was used. When both methods for diagnosing acute rejection were analysed together, no improvement in sensitivity (value 0.44) was found, but the specificity increased to 0.98 and the predictive value to about 0.80. It is concluded that cytoimmunological monitoring is a useful, non-invasive additional method for diagnosing the first period of acute rejection after heart transplantation and that quantitative birefringence measurements give valuable information on the extent of myocyte damage.

Cyclic changes in the concentration of glucose and fructose in human cervical mucus.

OBJECTIVE: To determine a possible cyclic change in the concentration of glucose and fructose in the aqueous phase of human cervical mucus (CM). DESIGN: Concentrations of glucose and fructose were longitudinally determined in the aqueous phase of CM of normal cycling women using enzymatic techniques, modified for small quantities. SETTING: Patients visiting a fertility clinic were selected. PATIENTS: Nine healthy women with regular menstrual cycles of 28 +/- 3 days that appeared to be ovulatory, demonstrated by sonographic follicle immaging and serum progesterone (P) measurements. INTERVENTIONS: Cervical mucus samples were longitudinally collected preovulatory, postovulatory, and premenstrual in ovulatory cycles, monitored by ultrasound and blood estradiol and P measurements. MAIN OUTCOME MEASURES: The study was designed to measure glucose and fructose longitudinally on three different points during one cycle. RESULTS: The preovulatory glucose concentrations in CM were lower than postovulatory and premenstrual. The preovulatory fructose concentrations were lower than premenstrual. The glucose concentration correlated with the blood P level. CONCLUSION: There is a consistent change in the glucose concentration measured in human CM in three phases of the menstrual cycle. The preovulatory and premenstrual fructose concentrations differ significantly. Knowledge of the carbohydrate metabolism in human cervical mucus may contribute in illuminating the possible role of the carbohydrate metabolism in sperm migration at midcycle and implantation in the luteal phase.

The origin of plasma creatine kinase 1, detected in patients during and following open heart surgery.

Plasma creatine kinase 1 (CK-1) was detected intra-operatively in 6 out of 6 patients and postoperatively in 15 out of 22 patients, undergoing cardiac surgery. A transient increase in plasma levels of creatine kinase 2 (CK-2) and total creatine kinase (CK-tot.) activity was observed in all patients. The disappearance rates for the 2 isoenzymes in the circulation were CK-1: Kd = 4.7 X 10(-3) min-1, and CK-2: Kd = 0.60 X 10(-1) min-3. Analysis of vessel and heart tissue showed that the saphenous vein contained mainly CK-1; high activities of all three isoenzymes were found in the parts of the heart investigated. Most probably, both CK-1 and CK-2 are liberated from injured cardiac tissue.

Modified creatine kinase-MB plots as a tool to detect perioperative myocardial infarction. Evaluation of sensitivity and specificity as compared with electrocardiographic changes.

Based on electrocardiographic criteria, coronary artery bypass grafting patients were divided into two groups, one with and one without perioperative myocardial infarction. Serial total-creatine kinase activity did not discriminate between the two groups; however, serial creatine kinase-MB activity showed a consistent difference. Patients with perioperative myocardial infarction showed an increase, whereas patients without perioperative myocardial infarction showed a decrease during the postoperative period. The creatine kinase-MB plots showed a sensitivity of 0.92 and a specificity of 0.98 as compared with the electrocardiograms.

Reference interval for the bilirubin excess in cerebrospinal fluid by derivative spectrophotometry.

The value of the bilirubin excess can be a useful aid for recognizing blood from haemorrhage in cerebrospinal fluid. One of the parameters needed for the calculation of the bilirubin excess is the total bilirubin concentration in CSF. A method for measuring total bilirubin in cerebrospinal fluid is presented, based on diazotization of bilirubin according to Jendrassik-Gróf, combined with multiwavelength first derivative spectrophotometry. This bilirubin assay allows determination of total bilirubin concentrations as low as 0.045 +/- 0.002 mumols/l. This method also enables a correction for oxyhaemoglobin interference. The value of the bilirubin excess was calculated for patients (n = 66) not showing any neurological disorder. A reference interval of 0.07 +/- 0.06 mumols/l was calculated for the bilirubin excess.

Postprandial decrease in HDL cholesterol and HDL apo A-I in normal subjects in relation to triglyceride metabolism.

The postprandial lipoprotein metabolism is important since it determines the circulation of potentially atherogenic particles and influences the metabolism of high-density lipoproteins (HDL) in a complex manner that is at present not completely understood. Therefore, the short-term (24-h) changes in postprandial lipoprotein metabolism, including retinyl palmitate (RP), apolipoprotein A-I (apo A-I), and apolipoprotein B, were studied in relation to postheparin lipolytic activities in six healthy normolipidemic men after an oral RP fat tolerance test. The fat load (98 g) was cleared in 7 h, because the triglyceride (TG) concentrations had returned to initial values (0.72 +/- 0.31 mmol/l) at that time. RP showed a peak in plasma at 4 and 5 h but remained present in chylomicron (remnants) in low concentrations after 8 and 24 h. After the fat load, HDL cholesterol and HDL-associated apo A-I showed a significant decrease in concentration of 35 and 29%, respectively. The decrease coincided with the increase in chylomicron remnants and the transient appearance of TG-enriched HDL. Hepatic lipase was correlated to both the initial HDL cholesterol concentration as well as the peak concentration of TG in chylomicron remnants, suggesting that it could be one of the regulating common physiological pathways in postprandial HDL and TG metabolism. In the subjects studied, the atherogenic potential of plasma increased in response to an oral fat load, characterized by a decrease in HDL cholesterol and HDL-associated apo A-I.

Cell genetic studies on propionyl-CoA carboxylase deficient cell lines.

Thirteen propionyl-CoA carboxylase (PCC)-deficient fibroblast lines were analysed for complementation by examining the incorporation of [14C]propionate into trichloroacetic-acid insoluble cellular macromolecules of polyethylene-glycol-dimethyl-sulphoxide induced heterokaryons formed by fusion of mutant cell lines. Corrections for variations in the rate of protein synthesis of the heterokaryons and individual cell lines was made by measuring the incorporation of [3H]lysine, added simultaneously with the labelled propionate. Two distinct complementation groups were found in this sample of PCC-deficient cell lines. Evidence for intragenic complementation was not obtained.

Prolone metabolism in isolated rat liver cells.

The metabolism of proline was studied in liver cells isolated from starved rats. The following observations were made. 1. Consumption of proline could be largely accounted for by production of glucose, urea, glutamate and glutamine. 2. At least 50% of the total consumption of oxygen was used for proline catabolism. 3. Ureogenesis and gluconeogenesis from proline could be stimulated by partial uncoupling of oxidative phosphorylation. 4. Addition of ethanol had little effect on either proline uptake or oxygen consumption, but strongly inhibited the production of both urea and glucose and caused further accumulation of glutamate and lactate. Accumulation of glutamine was not affected by ethanol. 5. The effects of ethanol could be overcome by partial uncoupling of oxidative phosphorylation. 6. The apparent K(m) values of argininosuccinate synthetase (EC 6.3.4.5) for aspartate and citrulline in the intact hepatocyte are higher than those reported for the isolated enzyme. 7. 3-Mercaptopicolinate, an inhibitor of phosphoenolpyruvate carboxykinase (EC 4.1.1.32), greatly enhanced cytosolic aspartate accumulation during proline metabolism, but inhibited urea synthesis. 8. It is concluded that when proline is provided as a source of nitrogen to liver cells, production of ammonia by oxidative deamination of glutamate is inhibited by the highly reduced state of the nicotinamide nucleotides within the mitochondria. 9. Conversion of proline into glucose and urea is a net-energy-yielding process, and the high state of reduction of the nicotinamide nucleotides is presumably maintained by a high phosphorylation potential. Thus when proline is present as sole substrate, the further oxidation of glutamate by glutamate dehydrogenase (EC 1.4.1.3) is limited by the rate of energy expenditure of the cell.

Role of anion translocation across the mitochondrial membrane in the regulation of urea synthesis from ammonia by isolated rat hepatocytes.

The regulation of urea synthesis from ammonia was investigated using isolated hepatocytes from fasted rats. Addition of ammonia alone produced only a small increase of urea formation, which was stimulated 2-fold by ornithine in conjunction with a fall of ATP levels and an accumulation of citrulline. Further addition of oleate or beta-hydroxybutyrate produced an additional 2-fold stimulation of urea formation to approximately 200 mumol/g dry weight/hour. The presence of oleate also protected against the inhibitory effect of 2,4-dinitrophenol on urea synthesis and the cellular ATP content. The data suggest that both the rate of of energy production and the rate of generation of reducing equivalents from endogensou substrates are insufficient to meet the requirements for optimal rates of urea synthesis. Urea formation from NH3 in the presence of ornithine and oleate, but iin the absence of gluconeogenic precursors, was inhibited by butylmalonate, a known inhibitor of malate-phosphate exchange across the mitochondrial membrane, and stimulated by theaddition of malate and other dicarboxylic acids and amino acids to the cell suspension...