ABSTRACT
BACKGROUND: Previous studies have indicated that alterations in blood glucose and/or insulin levels modify the inflammatory response. The purpose of this study was to elucidate whether increased levels of glucose and/or insulin influence the activation pattern of blood leucocytes and their production of cytokines in vitro. METHODS: Venous blood was obtained from eight healthy male volunteers after an overnight fast. Glucose and/or insulin were added to aliquots of whole blood to increase the blood glucose concentration by 5 or 20 mmol/l and/or the insulin concentration by 6 or 30 nmol/l, respectively, before stimulation with E. coli lipopolysaccharide (LPS) at concentrations of 10, 100 or 1000 ng/ml. The samples were subsequently incubated at 37 degrees C for 6 h before cytokine measurements. After centrifugation the levels of interleukins (IL)-1beta, IL-6, IL-8, IL-10 and tumour necrosis factor (TNF)-alpha were measured in plasma using enzyme-linked immuno-sorbent assays. The results were compared with cytokine levels in parallel control samples to which only identical amounts of LPS were added. RESULTS: The LPS-stimulated production of IL-1beta was significantly reduced by on average 26% in samples to which glucose 20 mmol/l was added; addition of insulin and/or glucose 5 mmol/l had no apparent effect on the IL-1beta production at any LPS concentration. The levels of IL-6, IL-8, IL-10 and TNF-alpha were not manifestly altered by addition of glucose and/or insulin at any LPS concentration. CONCLUSION: A substantial increase in blood glucose concentration changed the IL-1beta production, but not the production of other cytokines, in response to LPS stimulation.
Subject(s)
Glucose/pharmacology , Insulin/pharmacology , Interleukins/blood , Leukocytes/drug effects , Lipopolysaccharides/pharmacology , Tumor Necrosis Factor-alpha/blood , Humans , In Vitro Techniques , Interleukins/metabolism , Leukocytes/metabolism , Male , Tumor Necrosis Factor-alpha/metabolismABSTRACT
BACKGROUND: Fibrinolysis in blood is mainly reflected by the activities of tissue plasminogen activator (tPA) and of plasminogen activator inhibitor-1 (PAI-1). The effect of myocardial ischemia on their activities in the coronary circulation is, however, not established. OBJECTIVES: With an improved experimental model, we therefore examined the effect of a brief period of myocardial ischemia on their activities. Furthermore, the consequences of repeated periods of ischemia, mimicking the situations in patients with unstable angina, were investigated. METHODS: In six anesthetized pigs, we occluded the distal left anterior descending coronary artery (LAD) four times for 10 min with 40 min intervals and determined the activities of tPA and PAI-1 in arterial and coronary venous blood. By simultaneously recording LAD flow, we could estimate cardiac release of these factors at baseline conditions and during reperfusion. RESULTS: Neither net cardiac release of PAI-1 nor alterations in plasma PAI-1 levels were demonstrated during the experiment. However, a significant net release of tPA activity of 10.4 +/- 3.2 IU mL(-1) (P < 0.005) was recorded during baseline conditions. During reperfusion following the first period of ischemia, the cardiac release of tPA activity increased to a peak of 103 +/- 30-fold baseline release, but declined progressively after repeated periods of ischemia. After the fourth period, tPA release did not exceed an estimated baseline accumulation during ischemia and early reperfusion. CONCLUSIONS: In this porcine model, a substantial local increase in fibrinolytic capacity was observed after brief periods of ischemia, but declined subsequently by repeated periods of ischemia.
Subject(s)
Coronary Circulation , Fibrinolysis , Myocardial Ischemia/physiopathology , Angina Pectoris , Animals , Coronary Vessels , Female , Male , Models, Animal , Myocardial Reperfusion Injury , Plasminogen Activator Inhibitor 1/blood , Plasminogen Activator Inhibitor 1/metabolism , Recurrence , Swine , Tissue Plasminogen Activator/blood , Tissue Plasminogen Activator/metabolismABSTRACT
In a randomized trial on the effect of dalteparin for 5 weeks after HRS we evaluated hemostatic variables in plasma sampled before and 1, 6 and 35 days postoperatively. In 218 patients we found that prothrombin fragment 1 + 2 (F1 + 2), thrombin-antithrombin complexes (TAT), d-dimer and fibrinogen were significantly higher on day 35 as compared with baseline values in the placebo group (P < 0.001 for all). The same pattern was found in the dalteparin group, but with significantly lower values for F1 + 2, TAT and d-dimer. In patients in the placebo group with venographically proven deep vein thrombosis (DVT) on day 35 (33%), significantly higher values were found for F1 + 2, TAT and d-dimer than in patients without DVT. Patients in the highest quartile of d-dimer (>2850 ng mL-1) had an odds ratio for the presence of DVT of 24.0 when compared with patients in the lowest quartile (<1625 ng mL-1). It is concluded that a substantial hypercoagulability is sustained until day 35 after HRS, significantly reduced with prolonged administration of dalteparin.
Subject(s)
Arthroplasty, Replacement, Hip/adverse effects , Thrombophilia/drug therapy , Thrombosis/prevention & control , Aged , Anticoagulants/pharmacology , Anticoagulants/therapeutic use , Blood Coagulation Factors/analysis , Blood Coagulation Factors/drug effects , Dalteparin/pharmacology , Dalteparin/therapeutic use , Female , Humans , Male , Phlebography , Postoperative Complications/drug therapy , Postoperative Complications/prevention & control , Thrombophilia/etiology , Thrombophilia/prevention & control , Thrombosis/drug therapy , Thrombosis/etiology , Venous Thrombosis/diagnosis , Venous Thrombosis/prevention & controlABSTRACT
The purpose of these studies was to examine if perfluorochemical (PFC) liquids stimulate blood leukocytes to secrete nitric oxide (NO) and/or endothelin-1 (ET-1). As such, NO and ET-1 may modulate broncho- and vascular dilatation and constriction, respectively, and thereby influence the clinical condition of a patient in respiratory distress with persistent pulmonary hypertension. Blood leukocytes in their natural habitat (whole blood) were incubated in the presence of two different perfluorochemicals (perflubron and perfluorodecalin). The overall response in ET-1 or NO (indirectly measured as nitrite/nitrate) production was examined at increasing PFC percentages (wt/vol) of PFC/whole blood. The lowest proportion used, 0.001% (wt/vol), was relevant to serum concentrations of PFC observed in liquid-ventilated individuals, whereas the highest proportion PFC, 50% (wt/vol), would mimic a situation where leukocytes are presented to PFC-filled airways. Plasma levels of freshly drawn blood, similar to levels of incubated (6 h) non-PFC-supplemented cultures, were ET-1 0.59 +/- 0.07 pg/ml (6 h, mean +/- SEM) and NO(-2)/NO(-3) 50 +/- 9 microM (6 h). Perflubron or perfluorodecalin did not induce significant differences in ET-1 or NO(-2)/NO(-3) levels as function of PFC type or dose. In conclusion, PFC liquids do not stimulate production in leukocytes in vitro of substances that may modulate constriction or dilatation in the vascular and respiratory tract systems.
Subject(s)
Endothelin-1/biosynthesis , Fluorocarbons/pharmacology , Leukocytes/drug effects , Leukocytes/metabolism , Nitric Oxide/biosynthesis , Humans , Hydrocarbons, Brominated , Nitrates/metabolism , Nitrites/metabolism , SolutionsABSTRACT
To elucidate the possible roles of nitric oxide (NO), endothelin-1 (ET-1), and reactive oxygen species (ROS) in the pathophysiology of serogroup A streptococcal (GAS) peritoneal sepsis, we investigated the effects of aminoethylisothiourea (AE-ITU), an inducible NO synthase (iNOS) inhibitor, and a ROS scavenger, and the ET-1 receptor antagonist bosentan. In rats, live GAS inocula, 3 x 10(8) and 1 x 10(9) cfu/kg, entailed a 24-h mortality of 10% and 90%, respectively. GAS caused increases in tissue iNOS activity (9 h), in serum nitrite/nitrate (9-24 h), and in intracellular leukocyte ROS levels (3-6 h). These changes were all prevented by the pre-treatment with AE-ITU. A novel finding was that AE-ITU also prevented the GAS-induced marked increase in plasma ET-1 at 6 h. Short-term (7-h) survival was improved by both AE-ITU and by bosentan. The mechanism(s) for the beneficial effects of AE-ITU may possibly be a combined mode of action; iNOS inhibition, ROS scavenging, and inhibition of the increase in plasma ET-1 caused by GAS.
Subject(s)
Endothelin-1/blood , Enzyme Inhibitors/pharmacology , Shock, Septic/drug therapy , Streptococcal Infections/blood , Streptococcal Infections/drug therapy , Streptococcus pyogenes , Sulfonamides/pharmacology , Thiourea/analogs & derivatives , Thiourea/pharmacology , Animals , Antihypertensive Agents/pharmacology , Blood Pressure/drug effects , Bosentan , Endothelin Receptor Antagonists , Male , Nitric Oxide Synthase/antagonists & inhibitors , Nitric Oxide Synthase Type II , Rats , Rats, Sprague-Dawley , Receptor, Endothelin A , Shock, Septic/blood , Survival Rate , Time FactorsABSTRACT
OBJECTIVE: To test the effect of a continuous infusion of the nitric oxide (NO) synthase (S) inhibitor aminoethyl-isothiourea (AE-ITU) on survival time, hemodynamics, and oxygen transport in a porcine model of live group A streptococcal (GAS) sepsis. Furthermore, to examine the role of endothelin-1, histamine, and reactive oxygen species (ROS) in streptococcal shock. DESIGN: Prospective, randomized trial. SETTING: Laboratory at a university hospital. SUBJECTS: Twenty-eight pigs with an average weight of 25 kg. INTERVENTIONS: Sixteen animals received a continuous infusion of live Streptococcus pyogenes 1.3 x 10(10) colony forming units/hr: eight received fluids only, and the other eight received an intravenous infusion of AE-ITU 10 mg/kg/hr starting 30 mins before the GAS challenge. Six control pigs received AE-ITU 10 mg/kg/hr iv for 5 hrs. Another six animals received half the dose of GAS over 5 hrs. MEASUREMENTS AND MAIN RESULTS: GAS infusion caused a rapid increase in pulmonary, hepatic, and systemic vascular resistance, followed by hypotension with a 90% lethality at 4 hrs. Treatment with AE-ITU increased 4-hr survival in septic animals from 1/8 to 8/8 and 5-hr survival from 0/8 to 5/8, prevented hypotension, and increased urine output. AE-ITU attenuated the decrease in cardiac output, liver blood flow, and oxygen delivery, and hepatic arterial blood flow as a fraction of cardiac output increased (all p < .05). Plasma nitrate/nitrite levels decreased in all animals. Inducible NOS and endothelial constitutive NOS activities in liver, gut, and lung were not increased during sepsis, nor were they decreased after AE-ITU. Plasma levels of endothelin-1 and methylhistamine increased in all septic animals and were not modified by AE-ITU. AE-ITU prevented the increase in monocyte ROS production caused by GAS. In control animals, AE-ITU caused an increase in mean arterial pressure, liver blood flow, and oxygen delivery. CONCLUSIONS: In this model of porcine GAS-induced septic shock, which was not associated with enhanced NO production, infusion of the NOS inhibitor AE-ITU prolonged survival, prevented hypotension, and improved cardiac contractility, organ perfusion, and tissue oxygenation. These beneficial effects of AE-ITU might be a result of the combined effect of ROS scavenging and modulation of local NO production, thus improving the balance of vasodilator and vasoconstrictor forces and reducing oxidative stress.
Subject(s)
Isothiuronium/analogs & derivatives , Nitric Oxide/antagonists & inhibitors , Shock, Septic/drug therapy , Shock, Septic/mortality , Streptococcal Infections/drug therapy , Streptococcal Infections/mortality , Thiourea/analogs & derivatives , Animals , Disease Models, Animal , Female , Hemodynamics , Isothiuronium/therapeutic use , Male , Random Allocation , Shock, Septic/physiopathology , Streptococcal Infections/physiopathology , Streptococcus pyogenes , Survival Rate , Swine , Thiourea/pharmacologyABSTRACT
In this study, we have established a pig model that can combine extensive hemodynamic monitoring with simultaneous repetitive (serial) blood sampling for the study of multiple variables related to the hemostatic system. Sixteen healthy young pigs were studied to evaluate the influence of continuous endotoxin infusion on hemodynamic patterns and activation of coagulation and fibrinolysis. The chief aim of the study was to investigate the applicability of analytical methods primarily developed for use with human plasma samples in quantification of factors and reaction products of the porcine coagulation and fibrinolytic systems, and further, to use these methods to study the longitudinal changes in the plasma levels of these hemostatic variables as a consequence of endotoxin infusion. We found that acute, controlled endotoxemia induced a hemodynamic state of shock and reduced pulmonary gas exchange. Simultaneously, a gradual increase in peripheral blood mononuclear cell tissue factor activity was demonstrated, and increased maximally 5.5-fold 4 hours after onset of endotoxin infusion. Thrombin-antithrombin complexes increased in plasma to maximum levels after 3 hours, accompanied by an ethanol gelation test that was regularly positive after 1 to 2 hours, and fibrin monomer levels that gradually increased maximally 3.8-fold after 6 hours. These changes were followed by gradual decreases of both fibrinogen and factor VII levels, mainly due to consumption. Plasma levels of tissue type plasminogen activator activity peaked at 1.5 hours (11.3-fold increase), whereas the peak of plasminogen activator inhibitor-1 activity (14-fold increase at 4.5 hours) was delayed compared to tissue plasminogen activator and completely extinguished plasma tissue plasminogen activator activity. The sequential activation of coagulation and fibrinolysis established a procoagulant state favoring disseminated intravascular coagulation and microthrombus formation, potentially leading to multiple organ dysfunction.
Subject(s)
Blood Coagulation/drug effects , Endotoxemia/physiopathology , Fibrinolysis/drug effects , Hemodynamics/drug effects , Animals , Antifibrinolytic Agents/metabolism , Blood Coagulation Factors/drug effects , Blood Coagulation Factors/metabolism , Blood Pressure/drug effects , Cardiac Output/drug effects , Endotoxemia/metabolism , Endotoxins/administration & dosage , Endotoxins/pharmacology , Leukocyte Count/drug effects , Platelet Count/drug effects , Swine , Vascular Resistance/drug effectsABSTRACT
To elucidate the pathophysiology of acute shock caused by serogroup A streptococci (GAS), GAS were given intravenously to 25 pigs. Short-time infusions of GAS (n=11) caused variable and unpredictable responses. A continuous infusion of 5x108 cfu/kg/h (n=8) caused pulmonary hypertension, arterial hypotension, and reduced cardiac output and liver perfusion, progressing to circulatory shock within 2-4 h. Halving the infusion rate (n=6) induced a more gradual development of shock and doubled the mean survival time from 2.1 to 4.0 h. Mean tumor necrosis factor-alpha levels (+/-SE) increased from 25+/-1 to 40+/-3 pg/mL. Only slight signs of organ dysfunction were observed, which indicates that this is primarily a model of acute septic shock. Light microscopy revealed moderate inflammatory reactions in lung, liver, and gut biopsy samples, although high numbers of viable, M-typeable GAS were recovered from tissues. The present model may be useful to study mechanisms involved in acute septic shock as well as therapeutic interventions.
Subject(s)
Shock, Septic/physiopathology , Streptococcal Infections/physiopathology , Streptococcus pyogenes/pathogenicity , Animals , Blood/microbiology , Blood Gas Analysis , Disease Models, Animal , Embolism, Air/etiology , Female , Hypertension, Pulmonary/etiology , Hypotension/etiology , Male , Multiple Organ Failure/etiology , Shock/etiology , Shock, Septic/metabolism , Streptococcal Infections/metabolism , Streptococcus pyogenes/growth & development , Swine , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Increased nitrogen monoxide (NO) concentrations change leukocyte function under a multitude of experimental conditions. NO inhalation is an experimental treatment for lung failure and exposes leukocytes to increased NO concentrations during passage through the lungs. To investigate whether short-term NO inhalation induces lasting changes in the function of circulating human leukocytes, venous blood samples were drawn from eight healthy male volunteers before and at the end of a 35-min period of breathing 40 ppm NO in 30% O(2). The leukocytes in the samples were subsequently analyzed for NO-induced changes in expression of cell surface molecules, generation of reactive oxygen species (ROS), and cytokine production by flow cytometry and ELISA techniques. The results were (1) NO inhalation changed neither the baseline nor the Escherichia coli lipopolysaccharide (LPS)-induced expression of the cell adhesion molecules CD11a, CD11b, CD11c, and CD62L (l-selectin) on neutrophilic granulocytes (PMN) or monocytes (Mo). The expression of CD14 and HLA-DR was also unchanged. (2) The generation of ROS in response to activation with phorbol myristate acetate increased in PMN after NO inhalation; an increase in Mo did not reach significance. (3) Baseline and LPS-stimulated production of IL-1beta decreased after NO inhalation, while the LPS-stimulated production of TNF-alpha increased. No changes in IL-6 production were detected.
Subject(s)
Cell Adhesion Molecules/metabolism , Cytokines/metabolism , Leukocytes, Mononuclear/drug effects , Nitric Oxide/pharmacology , Reactive Oxygen Species/metabolism , Administration, Inhalation , Adult , Humans , Leukocytes, Mononuclear/metabolism , Male , Methemoglobin/metabolism , Nitrates/metabolism , Nitrites/metabolismABSTRACT
Elevated levels of endothelin-1 (ET-1) and von Willebrand factor (vWF), both markers indicative of endothelial function, are associated with hypertension. In a randomized open study we investigated the effect of antihypertensive treatment with the alpha-blocker doxazosin (n = 23) or the beta-blocker atenolol (n = 22) for 22 weeks on circulating levels of ET-1 and vWF in middle-aged men with essential hypertension. Blood pressure reduction was satisfactorily achieved with both drugs, although the decrease in the atenolol group was larger than that in the doxazosin group. A reduction in the levels of vWF occurred in both groups, being more pronounced in the alpha-blocker group compared with the decrease on beta blockers, p = 0.004 and p = 0.056, respectively. In the alpha-blocker group, there was a significant correlation (r = 0.50, p = 0.022) between the reduction in diastolic blood pressure and the decline in vWF. A highly significant decrease in plasma ET-1 was obtained during beta blockade (p = 0.007), whereas no significant change occurred within the alpha-blocker group. There was, however, no correlation between the decrease in blood pressure and the reduction in ET-1. The different favorable effects of alpha and beta blockers on endothelial function expressed as vWF and ET-1, could indicate that the effects are probably related not only to the blood pressure per se, but also to the different pharmacologic mechanisms of the drugs.
Subject(s)
Atenolol/pharmacology , Doxazosin/pharmacology , Endothelin-1/blood , Hypertension/pathology , von Willebrand Factor/metabolism , Adrenergic alpha-Antagonists/pharmacology , Adrenergic beta-Antagonists/pharmacology , Adult , Antihypertensive Agents/pharmacology , Biomarkers , Blood Pressure/drug effects , Endothelium/drug effects , Humans , Male , Middle Aged , Single-Blind MethodABSTRACT
A prospective open-labeled clinical study was carried out to compare the safety of dextran 70 and low molecular weight heparin (dalteparin; DD group) versus dextran 70 alone (D group) in patients subjected to elective hip replacement surgery. Dalteparin, 5,000 IU/day and dextran 70, 500 ml during surgery and on the first postoperative day were administered to 214 patients. Dextran 70 alone was infused in 44 patients, 500 ml during surgery and on the 1st, 3rd and 5th postoperative day. Mean total blood loss during the operation and until the 2nd postoperative day was 1,708 ml in the DD group and 1,712 ml in the D group (p = 0.79). During the 1st postoperative week, no group differences were found in the relative number of patients that received packed red blood cells (p = 0.95), the amount of transfused packed red blood cells (p = 1.0) and changes in hemoglobin concentrations (p = 0.69). The present results suggest that dextran 70 and dalteparin can be combined in recommended doses without significantly increasing perioperative bleeding in patients undergoing hip replacement surgery. Bone traumatization and insufficient plugging of surgical traumatized bone surfaces with bone cement favor bleeding. Further well-designed studies are needed to evaluate the safety and efficacy of this regimen.
Subject(s)
Blood Loss, Surgical , Dalteparin/administration & dosage , Dextrans/administration & dosage , Orthopedic Procedures/adverse effects , Aged , Analysis of Variance , Anticoagulants/administration & dosage , Anticoagulants/pharmacology , Arthroplasty, Replacement, Hip/adverse effects , Blood Loss, Surgical/prevention & control , Blood Volume , Dalteparin/pharmacology , Dextrans/pharmacology , Drug Therapy, Combination , Erythrocyte Transfusion , Female , Fibrinolytic Agents/administration & dosage , Fibrinolytic Agents/pharmacology , Hemoglobins/analysis , Humans , Male , Plasma Substitutes/administration & dosage , Prospective StudiesABSTRACT
Insulin and angiotensin II (Ang II) are involved in the regulation of endothelin-1 (ET-1). This study investigates their possible influence on plasma levels of ET-1 in humans. Twenty patients with essential hypertension were included in a randomized, double-blind, placebo-controlled crossover study of 4 weeks' treatment with losartan, a selective type 1 angiotensin (AT1) receptor antagonist. The effect was evaluated in the fasting state and during acute hyperinsulinemia physiologically induced by oral glucose ingestion (OGTT) and by euglycemic glucose clamp. Losartan lowered blood pressure significantly, but did not influence plasma levels of ET-1 in the fasting condition (5.2 +/- 0.2 fmol/mL on placebo and 5.6 +/- 0.3 fmol/mL after losartan treatment). During both models of acute hyperinsulinemia, there was a significant decrease in plasma ET-1. In the OGTT the mean values after placebo treatment decreased from 5.2 +/- 0.2 fmol/mL at time 0 to 4.7 +/- 0.4 (P = .001) and 4.0 +/- 0.5 (P = .001) at 60 and 120 minutes, respectively. During the clamp the mean ET-1 values decreased from 5.7 +/- 0.4 fmol/mL at time 0 to 4.6 +/- 0.2 (P < .001) and 4.3 +/- 0.3 (P = .006) at 60 and 120 minutes, respectively. No differences in these profiles occurred after losartan treatment. Significant inverse correlation between fasting levels of ET-1 and insulin sensitivity index was found, r = -.51, P = .003. In conclusion, losartan did not influence the circulating levels of ET-1 in basal condition or during acute hyperinsulinemia, whereas a significant decrease in plasma ET-1 occurred during acute hyperinsulinemia. A significant inverse correlation demonstrated between basal levels of plasma ET-1 and the insulin-stimulated glucose uptake could point to a possible regulatory influence of ET-1 production on glucose metabolism or vice versa.
Subject(s)
Antihypertensive Agents/therapeutic use , Endothelin-1/blood , Hyperinsulinism/blood , Hypertension/drug therapy , Losartan/therapeutic use , Adult , Aged , Cross-Over Studies , Double-Blind Method , Fasting , Female , Glucose Clamp Technique , Glucose Tolerance Test , Humans , Hyperinsulinism/complications , Hypertension/blood , Hypertension/complications , Kinetics , Male , Middle Aged , PlacebosABSTRACT
Discontinuation of thromboprophylaxis a few days after surgery may unmask delayed hypercoagulability and contribute to late formation of deep venous thrombosis (DVT). To investigate whether thromboprophylaxis should be prolonged beyond the hospital stay, a prospective, double-blind randomised study was conducted in 308 patients. All patients received initial thromboprophylaxis with dalteparin, dextran and graded elastic stockings. On day 7, patients were randomised to receive dalteparin (Fragmin) 5000 i.u. once daily, or placebo, for 4 weeks. All patients were subjected to bilateral venography, perfusion ventilation scintigraphy and chest X-ray on days 7 and 35. Patients with venographically verified proximal DVT on day 7 were withdrawn from the randomised study to receive anticoagulant treatment. The overall prevalence of DVT on day 7 was 15.9%. On day 35, the prevalence of DVT was 31.7% in placebo-treated patients compared with 19.3% in dalteparin-treated patients (p = 0.034). The incidence of DVT from day 7 to day 35 was 25.8% in the placebo-treated group versus 11.8% in the dalteparin-treated group (p = 0.017). The incidence of symptomatic pulmonary embolism (PE) from day 7 to day 35 was 2.8% in the placebo-treated group compared with zero in the dalteparin-treated group. This included one patient who died from PE. No patients experienced serious complications related to the injections of dalteparin or placebo. This study shows that prolonged thromboprophylaxis with dalteparin. 5000 IU, once daily for 35 days significantly reduces the frequency of DVT and should be recommended for 5 weeks after hip replacement surgery.
Subject(s)
Anticoagulants/administration & dosage , Dalteparin/administration & dosage , Hip Prosthesis/adverse effects , Hip/surgery , Postoperative Complications/prevention & control , Thrombosis/prevention & control , Adult , Aged , Double-Blind Method , Female , Humans , Male , Middle Aged , Prospective Studies , Thrombosis/etiology , Treatment OutcomeABSTRACT
In severe trauma, sepsis or during surgery, bacterial lipopolysaccharide (LPS) frequently enters the circulation. Persons with high levels of high-density lipoprotein (HDL) have previously demonstrated higher monocyte procoagulant activity (PCA) when whole blood is challenged with LPS. The aim of the study was to investigate the distribution of radiolabelled LPS (125I-LPS) in plasma from six persons with high (2.14-2.82 mmol l-1) and six persons with low (0.54-1.04 nmol l-1) HDL, subjecting plasma to fast protein liquid chromatography (FPLC), or agarose electrophoresis followed by quantitative autoradiography. In heparin plasma 125I-LPS was located mainly in parts of plasma containing low-density lipoprotein (LDL) or very low-density lipoprotein (VLDL) and the immunoglobulins, and located to a lesser extent in HDL. However, persons with high HDL showed significantly higher binding of 125I-LPS to HDL and the immunoglobulins, probably to IgG, and significantly lower binding to LDL/VLDL. In calcium-depleted plasma (EDTA) 125I-LPS demonstrated a sharp increase in the binding to HDL, combined with a persistently high binding to LDL/VLDL and binding to the immunoglobulins was almost eliminated in all subjects investigated. Likewise, the binding of 125I-LPS to HDL in EDTA plasma was also significantly higher and to LDL/VLDL significantly lower in persons with high HDL. This study demonstrates that the distribution of 125I-LPS in heparin and EDTA plasma from persons with high or low HDL is different, which is presumed to be of importance concerning the various bioactivities of LPS.
Subject(s)
Blood Proteins/metabolism , Lipopolysaccharides/metabolism , Lipoproteins, HDL/blood , Phosphorus Radioisotopes/metabolism , Autoradiography , Cholesterol, HDL/blood , Cholesterol, HDL/pharmacology , Chromatography, Affinity , Edetic Acid , Electrophoresis, Agar Gel , Female , Heparin , Humans , Lipopolysaccharides/blood , Lipoproteins, HDL/pharmacology , Male , Protein Binding/drug effects , Protein Binding/physiology , SepharoseABSTRACT
Endotoxin interacts with several plasma protein systems and blood cells, causing release of a multitude of endogenous mediators that contribute to the pathophysiological process of sepsis. Binding of 125I-labelled lipopolysaccharide, LPS, to human blood in vitro showed that the major part of the 125I-LPS was recovered in plasma, whereas only small amounts were retained in washed suspensions of granulocytes, erythrocytes, monocytes and lymphocytes, respectively. Whole leukocyte preparations or isolated subpopulations incubated with 125I-LPS or fluorescein-conjugated LPS followed by autoradiography, flow cytometry or immunofluorescence microscopy showed unequivocally that monocytes bound much more LPS than did granulocytes and lymphocytes. Lipoprotein electrophoresis followed by autoradiography showed that 125I-LPS bound to all the purified lipoprotein fractions, which was also confirmed by gel filtration chromatography. These findings demonstrate that monocytes represent the most important blood cell for LPS binding and that radiolabelled LPS is able to bind to lipoproteins as well as to other serum constituents.
Subject(s)
Blood Proteins/pharmacology , Leukocytes, Mononuclear/drug effects , Lipopolysaccharides/pharmacology , Blood Proteins/metabolism , Cell Communication/drug effects , Fluorescent Dyes/metabolism , Humans , Iodine Radioisotopes/metabolism , Leukocytes, Mononuclear/metabolism , Lipopolysaccharides/metabolism , Protein Binding/drug effectsABSTRACT
Hip arthroplasty is associated with a high frequency of postoperative solitary proximal deep vein thrombosis which seems most frequently observed when bone cement is used for prosthesis fixation. Eighteen pigs underwent hemiarthroplasty, eight with cement-fixed prostheses and eight with non-cement prosthesis installation. Levels of thrombin-antithrombin (TAT) complexes, tissue plasminogen activator (t-PA) activity and plasminogen activator inhibitor 1 (PAI-1) activity were determined in femoral vein blood from both limbs during and after surgery. On the operated side, TAT increased during bone traumatization followed by a substantial rise in t-PA activity and a gradual decline in PAI-1 activity. This indicates a local per- and post-operative sequential activation of coagulation and fibrinolysis followed by a fibrinolytic shutdown, all reflected in femoral vein blood on the operated side. In the animals receiving noncemented hip prostheses, the same pattern of activation of coagulation and fibrinolysis occurred on the operated side. This was, however, less marked than with the cement-fixed prostheses. Postoperative scanning electron microscopic (SEM) examination of the femoral veins showed thrombi on the operated side in 62% of the animals in the cement group and 25% in the non-cement group. In an additional study with eight animals undergoing cement-anchored hip prosthesis operations the levels of TAT, t-PA and PAI-I were analysed in femoral vein blood, mixed venous blood and arterial blood. Significantly higher levels were found in femoral vein blood compared with mixed venous blood while no significant change was found in arterial blood compared with mixed venous blood. The hyperthermia induced by curing bone cement was effectively conducted by the implanted prosthesis and did not seem to exert major influence on the activation of coagulation. Extreme rotation of the limbs during surgery did not in itself induce visible vein wall damage as judged by SEM. These studies indicate that traumatization of bone marrow during hip surgery induce a marked local activation of coagulation and a high incidence of deep vein thrombosis in proximal veins, in particular if bone cement is used for prosthesis fixation.
Subject(s)
Antithrombin III/metabolism , Femoral Vein/metabolism , Hip Prosthesis/methods , Plasminogen Activator Inhibitor 1/blood , Thrombin/metabolism , Tissue Plasminogen Activator/blood , Animals , Bone and Bones/injuries , Female , Femoral Vein/ultrastructure , Hematocrit , Male , Swine , Thrombosis/blood , Thrombosis/metabolismABSTRACT
Hip replacement surgery (HRS) is associated with a high frequency of deep vein thrombosis (DVT). At the same time there is a substantial systemic and local activation of coagulation. This study indicates that discontinuation of thromboprophylaxis one week after surgery may allow a second wave of coagulation and fibrinolysis activation to occur. An almost parallel increase in plasma TAT and D-dimer levels between the 6th and the 35th postoperative day may indicate late DVT formation. Repeated bilateral ascending venography is though to be necessary to evaluate the suitability of using selected activation markers of the coagulation and fibrinolytic systems as indices of DVT formation.
Subject(s)
Antithrombin III/metabolism , Hip Prosthesis/adverse effects , Peptide Hydrolases/metabolism , Plasminogen Activator Inhibitor 1/blood , Postoperative Complications/blood , Thrombophlebitis/blood , Tissue Plasminogen Activator/blood , Aged , Anticoagulants/pharmacology , Dalteparin/pharmacology , Female , Humans , Male , Phlebography , Postoperative Complications/diagnostic imaging , Postoperative Complications/prevention & control , Thrombophlebitis/diagnostic imaging , Thrombophlebitis/etiology , Thrombophlebitis/prevention & controlABSTRACT
Endothelin is a novel, potent, endogenous vasoconstrictor derived predominantly from endothelium and macrophages. Release of endothelin-1 (ET-1) into biological fluids was determined by radioimmunoassay in pigs undergoing either a hemorrhagic (3 h) or superior mesenteric artery (SMA) occlusion (5 h) shock followed by reperfusion (90 min) or a control group which was observed for 8 h. After surgery, there was a significant increase in ET-1 in jugular and carotid plasma, lymph, and ascitic fluid in all three models. The portal plasma ET-1 level was significantly increased (p < .05, assessed by the Spearman rank coefficient rho) in both shock models, but no significant increase was noted in the control group. In the SMA occlusion shock model, four pigs died within 30 min of reperfusion, and these animals had a much higher level of portal ET-1 (22.3 +/- 5.5 fmol/mL) than the two pigs that were alive by the end of the observation period (11.5 +/- 1.3 fmol/mL). Reperfusion in the SMA occlusion shock model induced a critical form of circulatory shock characterized by hypotension, decreased cardiac output, and decreased left and right ventricular stroke work index, and death occurred usually within 90 min. Reperfusion of the shed blood in the hemorrhagic shock model almost normalized the hemodynamic derangements caused by the hypovolemia (with the exception of RVSWI), and the portal plasma and ascitic ET-1 levels decreased. These results indicate that ET-1 is released from the gut in response to both general hypoperfusion and selective intestinal ischemia and reperfusion.(ABSTRACT TRUNCATED AT 250 WORDS)