Recombinant human activated protein C attenuates endotoxin-induced lung injury in awake sheep.

INTRODUCTION: Acute lung injury often complicates severe sepsis. In gram-negative sepsis, bacterial endotoxin activates both coagulation and inflammation. Enhanced lung vascular pressures and permeability, increased extravascular lung water content and deteriorated gas exchange characterize ovine endotoxin-induced lung injury, a frequently used model of acute lung injury. Recombinant human activated protein C (rhAPC), with its anticoagulant, anti-inflammatory, fibrinolytic and antiapoptotic effects, reportedly reduces the respirator-dependent days and the mortality of patients with severe sepsis. We speculate whether rhAPC antagonizes endotoxin-induced lung injury in sheep. METHODS: Two groups of sheep were exposed to Escherichia coli endotoxin (lipopolysaccharide) 15 ng/kg/minute intravenously from 0 to 24 hours; one group received only lipopolysaccharide throughout (n = 8), and the other group received lipopolysaccharide in combination with rhAPC 24 microg/kg/hour from 4 to 24 hours (n = 9). In addition, one group received rhAPC as above as the only intervention (n = 4), and four sham-operated sheep were used for determination of the alpha and epsilon isoforms of protein kinase C in pulmonary tissue. Data were assessed by one-way analysis of variance for repeated measurements. Biochemical data were analyzed using Student's t test, or using the Mann-Whitney U test when appropriate. RESULTS: Infusion of endotoxin caused lung injury, manifested by increments in pulmonary artery pressure, in pulmonary micro-occlusion pressure, in pulmonary vascular downstream resistance, in pulmonary vascular permeability index, in extravascular lung water index and in deterioration of oxygenation that were all attenuated by infusion of rhAPC. Endotoxemia led to changes in inflammation and coagulation, including pulmonary neutrophil accumulation paralleled by increased TNFalpha and decreased protein C and fibrinogen in animal plasma, which all improved following infusion of rhAPC. Moreover, rhAPC prevented the translocation of protein kinase C alpha and epsilon isoforms from the cytosolic fraction of lung tissue extracts. CONCLUSION: In awake sheep, rhAPC alleviates endotoxin-induced lung injury--as characterized by improvements of oxygenation, coagulation and inflammation, as well as by reversal of pulmonary hemodynamic and volumetric changes.

Significant contribution of liver nonparenchymal cells to metabolism of ammonia and lactate and cocultivation augments the functions of a bioartificial liver.

A bioartificial liver (BAL) will bridge patients with acute liver failure (ALF) to either spontaneous regeneration or liver transplantation. The nitrogen metabolism is important in ALF, and the metabolism of nonparenchymal liver cells (NPCs) is poorly understood. The scope of this study was to investigate whether cocultivation of hepatocytes with NPCs would augment the functions of a BAL (HN-BAL) compared with a BAL equipped with only hepatocytes (H-BAL). In addition, NPCs were similarly cultivated alone. The cells were cultivated for 8 days in simulated microgravity with serum-free growth medium. With NPCs, initial ammonia and lactate production were fivefold and over twofold higher compared with later time periods despite sufficient oxygen supply. Initial lactate production and glutamine consumption were threefold higher in HN-BAL than in H-BAL. With NPCs, initial glutamine consumption was two- to threefold higher compared with later time periods, whereas initial ornithine production and arginine consumption were over four- and eightfold higher compared with later time periods. In NPCs, the conversion of glutamine to glutamate and ammonia can be explained by the presence of glutaminase, as revealed by PCR analysis. Drug metabolism and clearance of aggregated gamma globulin, probes administered to test functions of hepatocytes and NPCs, respectively, were higher in HN-BAL than in H-BAL. In conclusion, NPCs produce ammonia by hydrolysis of amino acids and may contribute to the pathogenesis of ALF. High amounts of lactate are produced by NPCs under nonhypoxic conditions. Cocultivation augments differentiated functions such as drug metabolism and clearance of aggregated gamma-globulin.

Analysis of protein S-100B in serum: a methodological study.

BACKGROUND: Dysfunction and damage of the human central nervous system can be detected with biochemical markers, and protein S-100B is the best-established such marker. The aim of this study was to evaluate whether the protein is stable during long-term storage, to establish reference values for the new Elecsys S100 test and to compare this new method with the Liaison Sangtec 100 test. METHODS: We analysed blood samples from 118 blood donors and 196 patients with subarachnoid haemorrhage or head injury. The long-term stability of S-100B in frozen serum samples was evaluated with repeated analysis in 1997 and 2003 using an immunoradiometric assay. Method comparison between the Liaison Sangtec 100 and Elecsys S100 tests was performed using Bland-Altman difference plots. RESULTS: Serum concentrations increased significantly during long-term storage (mean difference 0.15 microg/L; +/-2 SD, 0.55 microg/L). Serum measurements using the Elecsys S100 method in 118 healthy blood donors showed S-100B levels between 0.02 and 0.08 microg/L (mean 0.05). The 95th percentile was 0.07 microg/L. The Liaison Sangtec 100 test usually measured higher concentrations than the Elecsys S100 method, and the difference between the two methods increased with increasing concentrations. The mean difference between the methods was 0.14 microg/L (+/-2 SD, 0.39 microg/L). CONCLUSIONS: Protein S-100B is not stable during long-term storage and the two analytical methods are not interchangeable.

Changes in haemoglobin levels according to changes in body mass index and smoking habits, a 20-year follow-up of a male cohort: the Tromsø Study 1974-1995.

Haemoglobin level declines with increasing age in cross sectional studies. Little is known about the longitudinal changes of haemoglobin. Because both high or low haemoglobin levels increase mortality and morbidity we examined how changes in lifestyle factors like body mass index (BMI) and smoking habits influence cohort changes in haemoglobin level. In all, 4159 men aged 20-49 years at baseline were examined in 1974 and 1994-1995 in a longitudinal, population-based study from the municipality of Tromsø, Northern Norway. Mean haemoglobin was 148 g/l. There was no difference in mean haemoglobin after 20 years in any strata of age. Mean BMI increased 2.1 kg/m(2). The prevalence of smokers decreased 20.1 percentage points. In a multiple regression analysis increase in BMI was associated with increased haemoglobin change. Smoking cessation lowered mean haemoglobin 1.6 g/l compared to never smokers. Haemoglobin increased 0.8 g/l in smoking quitters whose BMI increased >2.5 kg/m(2) compared to a decrease of 6.7 g/l in weight reducers. There was a positive dose-response relationship between changes in cigarettes smoked per day and change in haemoglobin among consistent smokers. In conclusion, in contrast to cross sectional studies, mean haemoglobin did not change during 20 years ageing of relatively young men. This could be explained by higher BMI and less smoking. The increase in BMI affected haemoglobin change to such an extent that the reduction in haemoglobin due to smoking cessation was counteracted. Prospective studies are needed to address the health implications.

Purpura fulminans as a sequel to erythema nodosum in a child with homozygous Leiden mutation and acquired protein S deficiency.

UNLABELLED: A 6-y-old boy presented with generalized, bruise-like swelling of both legs. Three weeks later, he developed purpura fulminans in one of the affected feet. Histology of the leg swelling was in accordance with erythema nodosum. The boy proved to be homozygous for the Factor V Leiden mutation and to have acquired protein S deficiency. He recovered, with partial loss of two toes. CONCLUSION: In contrast to what is often stated, erythema nodosum is not always a benign condition. On the basis of this report, we suggest that if extensive erythema nodosum develops in an individual without any known thrombophilic disorder, investigations with respect to the latter should be performed.

Haemoglobin and anaemia in a gender perspective: the Tromsø Study.

OBJECTIVES: To examine the gender-specific distribution of haemoglobin (Hb) and the World Health Organization (WHO) criteria for anaemia compared with the 2.5 percentile for Hb. METHODS: A population-based study from Tromsø, Northern Norway. All inhabitants above 24 yr were invited. In total, 26 530 (75%) had their Hb analysed. RESULTS: The 2.5-97.5 percentile of Hb was 129-166 and 114-152 g/L for all men and women, respectively. In men, mean Hb decreased from 148 to 137 g/L between 55-64 and 85+ yr. In women, mean Hb increased from 132 to 137 g/L between 35-44 and 65-74 yr and then decreased to 131 g/L among the oldest. Using the WHO criteria for anaemia (Hb: <130 and <120 g/L, men and women respectively), the prevalence of anaemia in men increased with age from 0.6% aged 25-34 to 29.6% aged 85+. For women, the prevalence of anaemia varied from 9.1%, 2.2% and 16.5% in the age groups of 35-44, 55-64 and 85+ yr, respectively. The WHO criteria gave a two to three times higher prevalence of anaemia compared with the 2.5 percentile of Hb in women, but the difference was small in men. Poor self-rated health was not associated with low values of Hb in women. In men, there was an association in some age groups. CONCLUSION: The WHO criteria for anaemia and the 2.5 percentile for Hb corresponded well for men, but not for women. The WHO criteria of anaemia may result in medicalization of healthy women.

Dietary supplementation with bean extract improves lipid profile in overweight and obese subjects.

The aim of this study was to evaluate the long-term effects of a dietary supplementation of bean extract on serum lipids, nutritional parameters, and fat excretion in feces.Sixty-two overweight and obese (body mass index > 25 kg/m(2)) volunteers were randomized to receive the dietary supplement (n = 31, supplement group) or the placebo (n = 31, placebo group). There were 41 women and 21 men, ages 22 to 66 years. Two capsules of a dietary supplement or a placebo were administered three times daily for 3 mo. The supplement group was then invited to participate in an open-label study for 9 mo. Twenty-four subjects (7 men and 17 women) were randomized to receive two or four capsules of the supplement three times daily. Lipids and nutritional blood parameters were measured at baseline, after 3 mo, and at 12 mo. Excretion of fat in feces was measured. At 3 mo, reduction in serum concentration of cholesterol was found in the supplement group but not in the placebo group. High-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and triacylglycerols in serum did not change in either group. In the 9-mo open-label study, no further reduction in serum cholesterol was observed. Low-density lipoprotein and the ratio of low- to high-density lipoprotein decreased, whereas triacylglycerols remained unchanged. Serum vitamin B12 and folic acid decreased but remained within the normal range. Ferritin and albumin in serum remained unchanged. No differences were observed in serum lipids and nutritional parameters between groups. The bean extract significantly increased fat excretion in feces. In conclusion, this dietary supplementation improved lipoprotein profile and enhanced fat excretion in feces in overweight and obese subjects.