Joël Pincemail

Thromboxane and prostacyclin release in adult respiratory distress syndrome

Plasma thromboxane B2 (TXB2) and 6-ketoprostaglandin F1α (6-keto-PGF1α) were measured in 84 patients at risk of developing adult respiratory distress syndrome (ARDS) (44 patients following multiple trauma, 29 patients following abdominal surgery and 11 patients with acute pancreatitis). Forty-nine of these 84 patients developed an ARDS. High (>140 pg/ml plasma) TXB2 values were found in 52/84 patients. The median values of TXB2 were: 360 pg/ml in multiple injured, 250 pg/ml in abdominal surgery and 410 pg/ml in acute pancreatitis patients. The median TXB2 value was 575 pg/ml in patients developing ARDS and 140 pg/ml in those without this complication: this difference was statistically significant (p<0.05). The median values of 6-keto-PGF1α were 55pg/ml in multiple injured, 25 pg/ml in abdominal surgery and 120 pg/ml in acute pancreatitis patients. The median 6-keto-PGF1α value was 122 pg/ml in ARDS patients and 25 pg/ml in non-ARDS patients (statistically significant: p<0.05). High TXB2 and 6-keto-PGF1α values were particularly related to sepsis in abdominal surgery patients (p<0.05) and in multiple injured patients (p<0.01). No relation could be established between abnormal TXB2 or 6-keto-PGF1α values and death. High TXB2 values often persisted for several days and were observed particularly at the time ARDS diagnostic criteria were fulfilled. An imbalance between TXB2 and 6-keto-PGF1α was observed: 6-keto-PGF1α values were always lower than TXB2 values and did not persist for more than 24 h except in four cases. Our data demonstrate a significant production of prostanoids in ARDS patients particularly in sepsis and indicate a disturbance in balance of the prostacyclin/thromboxane axis.

Equine postanaesthetic myositis: thromboxanes, prostacyclin and prostaglandin E2 production

Arachidonic acid cyclooxygenase metabolites, thromboxane B2 (TXB2), prostaglandin E2 (PGE2) and 6-keto-prostaglandin F1 (6-keto-PGF1) were measured in horses where anaesthesia was maintained with halothane. Two horses suffering from postanaesthetic myositis were compared with four normal horses. TXB2 and PGE2 levels were higher in mixed venous blood drawn from the myopathic horses. An increase of TXB2 and PGE2 levels appeared when myopathic horses were rolled into dorsal recumbency after a prolonged period of lateral recumbency. One hour after the end of anaesthesia, TXB2 had continued to increase whereas PGE2 decreased. By measurements on blood samples drawn from the brachial vein, we have shown that the rising level of TXB2 in mixed venous blood is mainly due to the increase of TXB2 in blood draining the dependent leg. The origin of the rise in PGE2 is not demonstrated in this study. 6-keto-PGF1 did not change during anaesthesia. An explanation of this imbalance between TXB2 and 6-keto-PGF1 production is considered.

Pentane measurement in man as an index of lipoperoxidation

The peroxidation of unsaturated fatty acids, constituents of tissues, is one proposed mechanism of invivo oxidant damage. Several methods are described to quantify in vivo lipid peroxidation: malonaldehyde determination, measurement of conjugated dienes and the detection of volatile products in expired gases. The measurement of expired volatile hydrocarbons is a specific and non-invasive technique for monitoring in vivo lipid peroxidation. We describe here an improved method for detecting and quantifying volatile products of lipid peroxidation in the breath of healthy human volunteers. We also include a discussion of the obtained results and of the advantages and disadvantages of measuring expired hydrocarbon gases.

Fast double antibody radioimmunoassay of human granulocyte myeloperoxidase and its application to plasma

The haem enzyme myeloperoxidase (MPO) (EC 1.11.1.7) with a spectral A430/A280 ratio greater than 0.7 and a specific activity of 125 U/mg was purified from isolated human neutrophils. To obtain a radioimmunoassay (RIA) for this enzyme, a specific antiserum against human neutrophil MPO was raised in rabbits and used at an initial dilution of 1/10,000. MPO labelled with 125iodine by a technique of self-labelling in the presence of H2O2, had a specific activity of 24 mCi/mg. After incubation at room temperature (2 h) and separation by double antibody precipitation in the presence of polyethylene glycol, the sensitivity of the RIA was 21 ng/ml. The RIA showed good precision and accuracy with intra- and interassay coefficients of variation of less than 7% for MPO concentrations ranging from 100 to 800 ng/ml, and satisfactory recoveries of known amounts of exogenous MPO in plasma. For the measurement of MPO in blood, the best sampling technique was to collect blood into EDTA. Rapid centrifugation (within 20 min) was necessary for blood collected into heparin. Mean MPO values in normal individuals were 340 +/- 98 ng/ml in EDTA plasma (n = 152) and 332 +/- 82 ng/ml in heparinized plasma (n = 34). When MPO was measured 12-6 h after injury in critically ill patients high values (above 1000 ng/ml) were found in 6/15 patients with multiple injuries. In patients with sepsis (n = 22), MPO values were always above 1000 ng/ml.

Immunoreactive trypsin in the adult respiratory distress syndrome

With the purpose of studying the role of proteinases in the development of ARDS, plasma levels of immunoreactive trypsin (IRT) and amylase were measured in 43 intensive care patients at risk of developing ARDS (22 polytrauma, seven abdominal surgery, four burns, two DIC and eight pancreatitis). Twenty four of these 43 patients developed ARDS and 31 presented abnormal IRT values (above 70 μg/L). Twenty-one of these 31 patients had ARDS; a significant correlation thus appeared between ARDS and abnormal IRT values. In nine patients, IRT values were higher than 800 μg/L and remained high for 3 to 4 days. A statistically significant correlation also appeared between abnormal IRT and septic phenomena: 20 patients with high IRT values presented septic problems. When IRT values were high, amylase values were often also abnormal: 12 of 23 patients with high IRT had abnormal amylase levels (the eight patients with documented pancreatitis were excluded); no other clinical signs or symptoms of pancreatitis were present in these patients. IRT could be one of the mediators of ARDS in septic patients. It is not clear that the pancreas is the origin of IRT in all cases.

Tocopherol mobilization during dynamic exercise after beta-adrenergic blockade

This study addresses the question of whether tocopherol mobilization during exercise could be explained by a lipolysis effect. Nine healthy male subjects were submitted to dynamic exercise of graded intensity (45, 60, 75% VO2max) on a cycle ergometer after ingestion of either a placebo or 40 mg propranolol as beta-blocker. Plasma tocopherol concentration increased toward a peak value reached during or at the end of exercise. The magnitude of this increase did not differ in the two experimental conditions while plasma free fatty acids concentration was lowered under beta-adrenergic blockade by propranolol. From these results, we conclude that tocopherol mobilization during dynamic exercise does not depend on lipolysis.

Plasma vitamin E, total lipids and myeloperoxidase levels during spinal surgery. A comparison between two anesthetic agents: propofol and isoflurane

Plasma levels of vitamin E (Vit. E), total lipids (TL), Vit. E to TL ratio and myeloperoxidase (MPO) were studied in 20 patients undergoing lumbar spinal surgery and randomly allocated to two anesthetic groups: propofol (bolus dose + continuous infusion) and thiopental/isoflurane. Peripheral blood samples were withdrawn prior to induction, each hour during anesthesia and 1 h after the end of surgery. Mean Vit. E and TL levels as well as mean Vit. E to TL ratios remained in the normal range over the entire period of study whatever the anesthetic regimen. MPO levels rose significantly in the post‐operative period only, but without statistical difference between the two groups. Therefore, anesthesia with propofol or thiopental/isoflurane modifies neither total lipid concentrations nor plasma Vit. E, which is a potent endogenous inhibitor of lipid peroxidation bound to lipoproteins. The rise of plasma MPO suggests a moderate post‐operative neutrophil activation which is not influenced by anesthetic techniques.

Self-Labeling of Human Polymorphonuclear Leucocyte Myeloperoxidase with 125iodine

In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with125Iodine: chloramine T, lactoperoxidase, and an original technique of ‘self labeling’ based on the ability of the enzyme to oxidize and bind125I in the presence of H2O2. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 μCi/gmg MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (≤3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits.

Plasma myeloperoxidase and vitamin E levels in head injury: preliminary results related to outcome.

This preliminary study was designed to assess a possible role of neutrophil activation and to determine the prognostic value of plasma myeloperoxidase (MPO) and vitamin E (Vit. E) levels in severe head injury. Plasma MPO and Vit. E levels were measured in nine severely head-injured patients (Glasgow Coma Score </=8) (ages 12-80 years) 6, 12. 18, 24, and 30 h after trauma. Patients were classified into two groups according to outcome after discharge from the ICU: group D (death; n = 5) and group S (survival; n = 4). Plasma MPO levels were increased immediately after trauma and then decreased. The MPO peak observed after 6 h was significantly higher (p < 0.05) in group D (mean +/- SEM: 1,237 +/- 122 ng/ml) than in group S (mean +/- SEM: 543 +/- 148 ng/ml). Plasma Vit. E levels were lower than normal values and decreased over time. They were always significantly lower (p < 0.05) in group D than in group S, except for the first sample. These differences cannot be explained entirely by total plasma lipid (TL) values since no statistical difference in TL concentrations was found between the two groups during the course of study. The ratio of Vit. E to TL. considered as the best index of Vit. E status, was lower in group D than in group S. and the difference reached statistical significance (p < 0.05) 12 h after trauma. In conclusion, in spite of the limited number of patients included in this study, it appears that severe head injury is associated with an increase in MPO and a decrease in Vit. E levels. These biochemical changes are of greater magnitude in group D than in group S; they suggest neutrophil activation and lipoperoxidation processes. Finally, plasma MPO and Vit. E seem to be new discriminant factors of outcome in head-injured patients.

Trypsin-like activity and thromboxane release in adult respiratory distress syndrome.

Plasmatic immunoreactive trypsin (IRT), thromboxane and trypsin-like enzymatic activity were measured in 117 patients at risk of developing adult respiratory distress syndrome (ARDS) (53 multiple injury, 30 abdominal surgery, 17 acute pancreatitis, 12 burnt and 5 disseminated intravascular coagulation patients). 69 of these patients developed ARDS. Immunoreactive trypsin and thromboxane were measured by radio-immuno-assay and trypsin-like enzymatic activity by spectrophotometry, using a specific chromogenic substrate. Mean IRT value was 675 ng/ml in ARDS and 265 ng/ml in non ARDS patients (p less than 0.05). Mean IRT value was 685 ng/ml in septic and 170 ng/ml in non septic patients (p less than 0.01). An abnormal trypsin-like enzymatic activity was measured in 26 ARDS patients. In 60 patients (37 ARDS and 23 non ARDS), thromboxane appeared in plasma simultaneously or about 24 hours after the beginning of IRT release. The importance of thromboxane release parallels the intensity of IRT. Originating from pancreas, trypsin can appear in plasma either by absorption from gastrointestinal tract or after pancreatic ischemia.