The effect of rapid cooling to 20-24 degrees C of whole blood immediately after collection, using 'cooling units' with butane-1,4-diol and prolonged storage up to 24 h at ambient temperature was investigated in the whole blood and the subsequently prepared plasma, buffy coat and buffy-coat-poor red cell concentrate (BC-poor RCC) in saline-adenine-glucose-mannitol (SAG M) solution. Factor VIII:C content of the plasma (n = 10), after 24 h storage was 80 +/- 3% of the initial value. In routine procedures factor VIII:C content in the plasma (n = 129 pools of 20 donor units plasma) was 0.77 +/- 0.078 IU/ml, after storage of the whole blood for 16-20 h. In whole blood (n = 10), the 2,3-diphosphoglycerate (2,3-DPG) content of the red cells decreased from 4.36 +/- 0.55 to 1.47 +/- 0.6 mumol/ml red cells after 24 h storage at 20-24 degrees C. After storage of the BC-poor RCC (n = 10) at 2-6 degrees C for 1 week, the 2,3-DPG had dropped to 0.76 +/- 0.46 mumol/ml red cells. During the first 24 h of storage of whole blood, the adenine triphosphate (ATP) levels of the red cells remained stable. A mean increase of 20% of the initial value was observed after addition of SAG M solution. In the BC-poor RCC the ATP slowly decreased to 81 +/- 5% after 5 weeks and to 68 +/- 6.6% of the initial value after 6 weeks storage.(ABSTRACT TRUNCATED AT 250 WORDS)

Background: Interleukin 10 (IL-10) exerts anti-inflammatory actions by counteracting many biological effects of interferon γ (IFN-γ). Aims: To investigate this in humans, we studied the effects of human recombinant IL-10 administration on IFN-γ production by patient leucocytes. Furthermore, we assessed the IFN-γ inducible molecule neopterin and nitrite/nitrate serum levels, which are indicative of endogenous nitric oxide formation. Methods: As part of two placebo controlled double blind studies, we analysed patients with chronic active Crohn's disease (CACD) who received either subcutaneous recombinant human IL-10 (n=44) or placebo (n=10) daily for 28 days, and patients with mild to moderate Crohn's disease (MCD) treated with either subcutaneous IL-10 (n=52) or placebo (n=16) daily for 28 days. Neopterin and nitrite/nitrate concentrations were measured in serum, and ex vivo IFN-γ formation by lipopolysaccharide or phytohaemagglutinin (PHA) stimulated whole blood cells were investigated before, during, and after IL-10 therapy. Results: In patients with CACD, the highest dose of 20 µg/kg IL-10 caused a significant increase in serum neopterin on days +15 and +29 of therapy compared with pretreatment levels. No changes were observed for nitrite/nitrate levels under either condition. In MCD, treatment with 20 µg/kg IL-10 resulted in a significant increase in PHA induced IFN-γ production. Conclusions: High doses of IL-10 upregulate the production of IFN-γ and neopterin. This phenomenon may be responsible for the lack of efficacy of high doses of IL-10 in the treatment of CACD and MCD.

Lipopolysaccharide (LPS), the major glycolipid component of gram-negative bacterial outer membranes, is a potent endotoxin responsible for pathophysiological symptoms characteristic of infection. The observation that the majority of LPS is found in association with plasma lipoproteins has prompted the suggestion that sequestering of LPS by lipid particles may form an integral part of a humoral detoxification mechanism. Previous studies on the biological properties of isolated lipoproteins used differential ultracentrifugation to separate the major subclasses. To preserve the integrity of the lipoproteins, we have analyzed the LPS distribution, specificity, binding capacity, and kinetics of binding to lipoproteins in human whole blood or plasma by using high-performance gel permeation chromatography and fluorescent LPS of three different chemotypes. The average distribution of O111:B4, J5, or Re595 LPS in whole blood from 10 human volunteers was 60% (؎8%) high-density lipoprotein (HDL), 25% (؎7%) low-density lipoprotein, and 12% (؎5%) very low density lipoprotein. The saturation capacity of lipoproteins for all three LPS chemotypes was in excess of 200 g/ml. Kinetic analysis however, revealed a strict chemotype dependence. The binding of Re595 or J5 LPS was essentially complete within 10 min, and subsequent redistribution among the lipoprotein subclasses occurred to attain similar distributions as O111:B4 LPS at 40 min. We conclude that under simulated physiological conditions, the binding of LPS to lipoproteins is highly specific, HDL has the highest binding capacity for LPS, the saturation capacity of lipoproteins for endotoxin far exceeds the LPS concentrations measured in clinical situations, and the kinetics of LPS association with lipoproteins display chemotype-dependent differences.The lipopolysaccharide (LPS) components of gram-negative bacterial outer membranes are potent endotoxins responsible for hemodynamic, hematological, and metabolic changes observed during severe infection. Activation of responsive cells of the host immune system by low concentrations of LPS results in the production of high levels of endogenous mediators of inflammation such as tumor necrosis factor alpha (TNF-␣) and interleukin-1␤ (IL-1␤), IL-6, and IL-8, which are capable of sustaining the inflammatory state. Among the observed metabolic changes in patients are profound disturbances in plasma lipid profiles (1,5,14) that may also be induced in experimental animals by LPS challenge (12). Lipid metabolism appears to be extensively regulated during the host response to infection; however, increased cytokine levels do not solely appear to be responsible for the characteristic alterations in plasma lipid profiles. It has recently been suggested that disturbances in lipid metabolism may, in fact, form part of the host defense because the immune response is tightly linked to the metabolic response (15). LPS binding protein (LBP) is an acute-phase protein (29) that plays a central role in the attenuation of the cellular response to end...

Summary Lumbar bone mineral density (BMD) determination by dual photon absorptiometry was used to study the influence of adjuvant chemotherapy for premenopausal breast cancer on the risk of premature osteoporosis. Six cycles of combination chemotherapy caused ovarian failure in 31 of 44 (71 %) women, amenorrhoea mostly already beginning during treatment. In contrast, only seven of 44 (16%) women, who were pair-matched for age and year of breast cancer surgery and had not been treated with chemotherapy, were post-menopausal at the time of measurement. The mean interval after breast surgery was 3.5 years. The significantly decreased BMD in the treated group (1.17 compared to 1.29 g cm-2) could only be explained by the high incidence of menopause in these women, which on average occurred 10 years prematurely. Extrapolation of these findings suggests that adjuvant chemotherapy may precipitate osteoporotic fractures by some 10 years in a considerable proportion of women cured of premenopausal breast cancer.Gradual loss of bone matrix and mineral is a consequence of ageing leading to osteoporosis. In general, women start off with less peak adult bone mass than men and consequently develop more symptoms from osteoporosis. As a result a considerable proportion of women suffer from spontaneous fractures in later life (Gordan, 1978;Nordin, 1980; NIH Consensus Statement, 1984). Various data suggest that in osteoporosis there is a disproportionately greater loss of trabecular bone from the axial skeleton compared to cortical bone from appendicular sites (Riggs et al., 1981).The average annual loss of bone mass measured by bone mineral density (BMD) in women before menopause amounts to 1-2%, but may rise to some 6-8% during the first 2-5 years after menopause (Krolner & Pors Nielsen, 1982;Genant et al., 1982). This dramatic temporary increase in the rate of bone loss around natural menopause or after bilateral ovariectomy has been ascribed to a ceasing protection from bone loss by ovarian hormones (Riggs et al., 1981(Riggs et al., , 1982(Riggs et al., , 1986Genant et al., 1982;Johnston et al., 1985;Richelson et al., 1984). Oestrogen replacement therapy effectively prevents osteoporosis in post-menopausal women if started within a few years after menopause (Riis, 1987;Lindsay et al., 1976;Christiansen et al., 1980;Recker et al., 1977).Adjuvant chemotherapy in premenopausal women treated for breast cancer frequently leads to diminished ovarian function and premature menopause (Henderson, 1987). Since early menopause is one of the strongest predictors of osteoporosis (Richelson et al., 1984), we anticipated that the widely used adjuvant chemotherapy regimen of cyclophosphamide, methotrexate and 5-fluorouracil (CMF), might seriously precipitate osteoporosis.To investigate this possibility we have compared the bone mineral density of the lumbar spine between women who had been treated with adjuvant CMF and women, matched for age and time of premenopausal breast cancer surgery, who received no such treatment. Patients were consi...

The safety, tolerability, and efficacy of a 12-wk treatment with pravastatin, 5, 10, and 20 mg/d, was evaluated in 72 children with heterozygous familial hypercholesterolemia (FH) in a double-blind, randomized and placebo-controlled study. The results show that pravastatin was well tolerated and that adverse events were mild and equally distributed among the three treatment groups. Plasma total and LDL cholesterol levels were significantly reduced in all pravastatin treatment groups, in comparison with the control group; -24.6% (-28.1 to 21.0) and -32.9% (-37.0 to -28.6), for mean change and 95% confidence interval, respectively. In four children plasma LDL cholesterol levels were reduced within normal limits for sex and age. HDL cholesterol increased in the pravastatin 20-mg group, +10.8% (+3.4 to +18.8), whereas plasma apo B100 and very LDL (VLDL) cholesterol levels were reduced within all pravastatin-treated groups -26.8% (-31.2 [corrected] to -21.7) and -24.5% (-35.0 to -12.3). These data show that short-term pravastatin treatment of children with FH is safe and effective, although long-term dose titration studies with 3-hydroxy-3-methylglutaryl-CoA reductase inhibitors need to be performed, to reduce plasma LDL cholesterol levels below a predefined level. The results of these studies have to be awaited before new treatment strategies are to be considered in these children.