Individual Dose-response Curves Research Articles

Dose Response of Alcuronium and d-Tubocurarine in Infants, Children and Adolescents

Seventy neonatal to adolescent general surgical patients were studied to create an individual dose-response curve for the long-acting neuromuscular blocking agents, alcuronium and d-tubocurarine. The mean (SEM) ED95 of alcuronium was 196 (9), 271 (13) and 243 (8) micrograms/kg in infants, children and adolescents, respectively (P less than 0.01). d-tubocurarine showed a similar age dependent dose-response relationship. ED95 doses were 414 (40), 499 (41) and 445 (31) micrograms/kg, respectively. The onset time (time from intravenous administration to maximal effect) following equipotent dosages was 40-50% shorter in infants than in children or adolescents (1.5 vs 2.7 minutes, P less than 0.05).

Quantification of proliferative and suppressive responses of human T lymphocytes following ConA stimulation

The mitogenic response of human T lymphocytes to graded doses of concanavalin A (ConA) has been measured by means of an MTT tetrazolium dye metabolic assay. Three groups of healthy subjects, representing children, younger adults and elderly persons, were investigated. It was shown that a typical bell-shaped course of the ConA dose-response curve is the result of a proliferative response to suboptimal concentrations of ConA and a toxic action of ConA at supraoptimal concentrations. The ascending part of the response curve reflects in its shape the regulatory interaction of responding cells. A decrease in suppressive functions is accompanied by a shift of this part of the curve to lower concentrations of ConA. By means of a mathematical model derived from enzyme kinetics, an attempt was made to quantify the suppressive functions from the course of the individual dose-response curve. It was found that after suitable data processing, suppression-related shape changes can be assigned to a single parameter. The value of this parameter as a diagnostic tool was tested in a study of the age dependence of human T lymphocyte responses to ConA. While the proliferative response decreased with age, the suppressive functions exhibited their maximum effect in the group of adults. Thus it could be demonstrated that ConA induced proliferative and suppressive responses are due to two different pathways which can be independently extracted from the dose-response curve.

Influence of the duration of inhalation of cold dry air on the resulting bronchoconstriction in asthmatic subjects

Hyperventilation of cold dry air causes bronchoconstriction in asthmatic subjects and has been proposed as a test for assessing bronchial hyperresponsiveness. The influence of the duration of inhalation of unconditioned cold air has not been studied. We have investigated the question in 12 asthmatic subjects in a clinically stable state. Each subject underwent three inhalation tests at a maximum interval of two weeks. On each day, the duration of inhalation was different, being randomly 2, 3 or 4 min depending on the subject. Doubling doses of cold air produced by a freon conditioner were administered, increasing ventilation from 7.5 to 15, 30, 60 l.min-1 and maximum voluntary ventilation (MVV). Forced expiratory volume in one second (FEV1) was assessed after each period of cold air inhalation. The test was stopped when the FEV1 had decreased by 20% or more, or when MVV had been achieved. The dose of cold air expressed as the level of ventilation causing a 20% change in FEV1 (PD20) was interpolated from individual dose-response curves. Dose-response curves shifted to the left when the duration of ventilation was increased. PD20 was significantly lower after 3 min of ventilation than after 2 min (mean +/- SD PD20 of 41.7 +/- 1.4 l.min-1 compared with 53.3 +/- 1.2 l.min-1; p = 0.002). There was a further fall in PD20 after 4 min of ventilation (PD20 = 36.1 +/- 1.5 l.min-1) but the difference compared with the values obtained after 3 min was not significant (p = 0.09), thus suggesting a plateau.(ABSTRACT TRUNCATED AT 250 WORDS)

Two-dose technique to create an individual dose-response curve for atracurium.

While monitoring the thenar EMG response to ulnar nerve stimulation, the authors gave either 105, 150, 210, or 300 micrograms/kg of atracurium to 60 patients. The maximal neuromuscular responses were plotted on a log-probit paper. The individual second dose required to produce 95% neuromuscular block (NMB) was estimated from a graph drawn on the paper. The maximal response following this second dose was then plotted. The mean maximal response following the second atracurium dose was 95.5% (SD range, 92.6-97.4%) NMB. The two dose-response points thus acquired resulted in the individual two-dose dose-response curve. The ED50 and ED95 and slope of the two-dose dose-response curve were compared with the single-dose dose-response curve. The average ED50 and ED95 determined by the two-dose and the single-dose techniques were nearly identical [160 micrograms/kg (SD range, 126-201 micrograms/kg) vs. 164 micrograms/kg (SD range, 150-179 micrograms/kg) and 302 micrograms/kg (SD range, 251-363 micrograms/kg) vs. 336 micrograms/kg (SD range, 274-411 micrograms/kg) respectively]. Also, the slopes of the curves were similar [6.2 (SD range, 5.2-7.2) vs. 5.4 (SD range, 4.5-6.4) probit/log]. It is therefore possible to construct an individual dose-response curve for atracurium within 7-9 min and to determine individual pharmacodynamic characteristics of atracurium from this curve.

Effect of indomethacin on adenosine-induced bronchoconstriction

The exact mechanism of adenosine-induced bronchoconstriction in patients with asthma is unknown. Adenosine contraction of guinea pig trachea was antagonized by inhibitors of cyclooxygenase. The aim of this study was to investigate the effect of indomethacin (100 mg/day) on adenosine-induced bronchoconstriction in 14 asymptomatic patients with asthma. Airway response was evaluated as FEV 1, and adenosine was administered as an aerosol diluted in 0.9% saline to produce a concentration range of 0.125 to 4 mg/ml. The dose of adenosine producing a 20% change in FEV 1 (PD 20) was calculated from the individual semilogarithmic dose-response curve; the results of PD 20 were converted to log values for statistical analysis (Student's paired t test). The study was performed on 3 separate days. On the first day, the adenosine challenge was performed, and on subsequent days patients were pretreated with either placebo or indomethacin in a randomized, double-blind manner. Inhaled adenosine caused bronchoconstriction with a geometric mean PD 20 of 0.71 mg (95% confidence limits, 0.44 to 1.16). After placebo, a geometric mean PD 20 of 0.91 mg (95% confidence limits, 0.53 to 1.58) was obtained. Indomethacin pretreatment decreased adenosine hyperresponsiveness and shifted the dose-response curves of adenosine challenge to the right with a geometric mean PD 20 of 1.28 mg (95% confidence limits, 0.64 to 2.56). The effect of indomethacin on adenosine bronchoconstriction ( p < 0.01 versus baseline; p < 0.05 versus placebo) suggests an indirect mechanism of adenosine on inducing release of arachidonic acid derivatives. Inflammatory mediators inhibited by indomethacin may be involved in adenosine bronchoconstriction, even if this mechanism is not relevant.

4 Desferrioxamine-induced iron excretion in humans

Iron excretion in response to DF in humans is dependent upon the degree of iron overload, particularly of parenchymal liver cells. However, a number of variables, including ascorbate status, erythroid activity and liver disease, affect both the amount of iron mobilized and the route by which it is excreted. Faecal iron, derived from the bile, appears to arise from intracellular chelation of a transit iron pool related to hepatocyte iron stores, whereas urine iron may be derived from iron capable of exchanging with plasma transferrin at cell membranes of both hepatocytes and macrophages. Faecal iron predominates as iron stores return towards normal on regular chelation therapy. An understanding of the variables which influence iron excretion allows rational planning of long-term therapy with DF in patients with iron-loading anaemias. In young children a dose of 40-50 mg/kg given on five or six nights a week from the age of about 3 years is appropriate for prevention of serious iron loading. In older children the dose must be carefully tailored (by means of an individual urinary iron excretion dose-response curve) to achieve maximum safe chelation of pre-existing iron stores. In patients with slower rates of iron loading from excessive gastrointestinal iron absorption, intermittent use of DF infusions may be sufficient to maintain normal iron stores.

Effect of vasoactive intestinal peptide (VIP) on propranolol-induced bronchoconstriction

There is now considerable evidence in favor of vasoactive intestinal peptide (VIP) as a neurotransmitter of nonadrenergic noncholinergic nerves in the airways. The purpose of our study was to evaluate the influence of inhaled VIP on bronchomotor tone after a β-adrenergic- and cholinergic-receptor blockage. The study was peformed in six patients with asthma in 4 days. On the first day, a propranolol provocative dose producing a 20% change in FEV 1 (PD 20) was determined from the individual semilogarithmic dose-response curve. On the other days, the propranolol challenge was performed after inhalation of ipratropium bromide (40 μg), VIP (70 μg), and both drugs in randomized double-blind order. Statistical analysis was performed by two-way analysis of variance. The results demonstrated that mean propranolol PD 20 was 0.14 mg (geometric mean + SD = 1.22). Ipratropium bromide administration, like VIP administration, significantly raised the PD 20 value. The administration of both drugs elicited a further remarkable increase of mean propranolol PD 20. The results demonstrated that inhaled VIP influences bronchomotor tone and that this effect is independent of the cholinergic blockage.

Effect of nedocromil on bronchospasm induced by inhalation of substance P in asthmatic subjects

Several studies have demonstrated that neuropeptides are present in peptidergic fibres of bronchial tissue. The aim of the present study was to evaluate in vivo the effect of nedocromil sodium (2 x 2 mg) on bronchospasm induced by inhalation of substance P. Six moderate asthmatic patients, mean age 25.17 years, were studied. Airway response was measured as FEV1 and the dose of substance P (using a dose range of 23-736 nmol) producing a 20% decrease in FEV1 (PD20) was calculated from the individual semilogarithmic dose-response curves. Patients were studied on 3 separate days in a randomized, double-blind manner. On the first day a baseline PD20 value was determined. On subsequent days substance P challenge was performed after pretreatment (20 min before challenge) with either placebo or nedocromil sodium. Student's paired t-test and Wilcoxon's test were used for statistical analysis. The results of this study demonstrated that inhalation of substance P causes a dose-dependent bronchoconstriction and that the bronchoconstriction induced by substance P can be prevented by pre-treatment with nedocromil sodium.

Comparison of the dose-response curves obtained by forced oscillation and plethysmography during carbachol inhalation

We compared the cumulative dose-response curves obtained during carbachol inhalation by simultaneous measurements of airway specific conductance (sGaw) and respiratory conductance, in 23 subjects with or without bronchial hyperresponsiveness. The sGaw was measured by a body plethysmograph, whilst the random noise forced oscillation technique (FOT) was used to determine respiratory conductance. The sGaw was compared to respiratory conductance extrapolated to zero frequency (Grs0). Bronchial sensitivity was assessed by the threshold dose of carbachol (TD) that induced a decrease in sGaw and Grs0 of twice the baseline coefficient of variation. Bronchial responsiveness was assessed by the slopes (S) of the individual dose-response curves. The TD and S values obtained by FOT and by plethysmography were closely correlated (p less than 0.001). The carbachol doses inducing a 50% reduction in sGaw were equivalent to those causing a 42% reduction in Grs0 (r = 0.90; p less than 0.001). During bronchial challenge testing, FOT provides comparable information in terms of bronchial sensitivity and responsiveness to that supplied by plethysmography.

Cumulative dose-response curves for assessing combined effects of salbutamol and ipratropium bromide in chronic asthma

We investigated whether salbutamol (S) and ipratropium bromide (IB) exerted a true additive bronchodilator effect in asthma. In fifteen selected chronic asthmatics, individual cumulative dose-response curves to S and IB were performed on two separate days (linear regression of bronchodilator response (delta FEV1) between 20 and 80% of maximal response, versus log dose), and the dose of S equipotent to the IB dose giving the maximal bronchodilator effect (IBopt) was calculated by interpolation of each S curve. On two other days, each patient received IBopt or the equipotent S dose followed by an additional 400 micrograms S. On day 1 or 2, FEV1 reached 220 +/- 410 ml and 2410 +/- 380 ml (p less than 0.05) after the maximal dose of IB and S respectively. On day 3 or 4 after pretreatment by IB or S an additional 400 micrograms S gave a further increase, which was similar in both series (315 and 320 ml, respectively). FEV1 after combination treatment reached 238 +/- 350 ml and was not significantly different from the maximal effect of S (2440 +/- 290 ml). We conclude that S and IB exert a true pharmacological additive effect, since the combination effect is as great as the maximal effect of the most potent drug (S) and greater than the maximal effect of IB, and that the same additional dose of S gives the same increase after equipotent doses of S and IB.

Imparied lymphocyte proliferative response to mitogen in alcoholic patients. Absence of a relation to liver disease activity.

Concanavalin A-induced lymphocyte proliferation was studied in 25 patients with alcoholic hepatitis or compensated alcoholic cirrhosis. Nine alcoholics without evidence of liver disease were also evaluated. A nonlinear correlation equation, which was natural logarithmic, was applied to individual dose-response proliferation curves and permitted comparisons between patient groups and controls. The proliferative response in all patient groups was significantly lower when compared to healthy controls and was independent of the presence or absence of liver disease. This suggests that some changes in immune function observed in alcoholics may be linked to the direct effects of alcohol on the immune system rather than to the associated liver disease.

Bronchospasm Induced by Inhalation of Substance P: Effect of Sodium Cromoglycate

The aim of this study was to evaluate in vivo the effect of inhaled substance P (SP) and to determine the effect of sodium cromoglycate (SCG) on bronchospasm induced by its inhalation in 6 asthmatic patients. At the beginning of the study, all patients were asymptomatic, with an FEV1 value not less than 20%. SP was administered as aerosol, prepared in 0.9% saline to produce a dose range of 0.03-1 mg. Airway response was measured as FEV1 using a pulmonary system 47120A Hewlett-Packard instrument. The dose of SP producing a 20% change in FEV1 was calculated from the individual semi-logarithmic dose-response curve (PD20). After administration of the placebo, SP produced a dose-related bronchoconstriction with a geometric mean PD20 of 0.15 mg. After SCG, the mean PD20 value was of 0.64 mg (p less than 0.01). These results confirmed the bronchospasm induced by inhalation of SP and demonstrated that SCG is able to prevent this effect although it is impossible to define the exact mechanism of the drug.

Histamine H2-receptor of human and rabbit parietal cells.

The histamine H2-receptor on the human parietal cell has been characterized by using dose-response curves and the negative logarithm of the molar concentration of an antagonist (pA2) analyses of cimetidine antagonism of betazole, histamine, and impromidine stimulation in isolated human and rabbit gastric glands. To evaluate the in vitro results, betazole-stimulated gastric acid secretion with and without cimetidine was also studied in healthy subjects. In the in vivo model, individual dose-response curves were shifted to the right with increasing cimetidine concentrations, but this was counteracted by increasing betazole doses, indicating competitive, reversible antagonism. The pA2 values ranged from 6.1 to 6.3. In isolated human gastric glands, impromidine was shown to be eight times more potent than histamine, indicating higher receptor affinity, but the maximally stimulated aminopyrine accumulation was the same as for histamine, and the pA2 values for cimetidine antagonism did not differ significantly, i.e., 5.7 (histamine) and 6.1 (impromidine). In isolated rabbit gastric glands, cimetidine inhibited the histamine- and impromidine-stimulated response with pA2 values of 6.0 and 7.3, respectively. Impromidine was shown to be approximately 100 times more potent than in human gastric glands, whereas histamine had the same potency. This confirms the role of the histamine H2-receptor and suggests a difference between the species concerning receptor affinity.

Insulin deficiency and insulin resistance in type 2 (non-insulin-dependent) diabetes: quantitative contributions of pancreatic and peripheral responses to glucose homeostasis.

A non-steady state dose-response study was designed to quantitate peripheral sensitivity to insulin and pancreatic responsiveness to glucose, and to assess their relative contribution to glucose intolerance in Type 2 diabetes (Type 2 DM, non-insulin-dependent). Eleven lean and eleven obese patients with mild diabetes (fasting plasma glucose, FPG, 10.3 +/- 1.0 and 9.4 +/- 0.6 mmol l-1, respectively) were examined; twenty-six lean and twelve weight-matched obese subjects served as controls. Pancreatic response was measured by sequential injection of 0.1, 0.3 and 0.9 g kg-1 glucose; peripheral sensitivity to insulin was determined from the rate of clearance (Kgluc) of 0.3 g glucose injected sequentially together with 25, 50 and 100 mU insulin kg-1 or with 0, 12.5 and 50 mU kg-1, under somatostatin infusion. The mean dose-response curve describing glucose-induced insulin release showed increased maximal capacity to secrete insulin in obese controls, while the responses of lean as well as obese Type 2 DM were reduced by more than 80%. The mean dose-response curves relating plasma exogenous insulin levels to Kgluc were similar in lean diabetics and lean controls. The curves of both obese controls and obese diabetics were shifted to the right, demonstrating similar insulin resistance. In four lean controls, sensitivity to insulin was tested also during a hyperglycemic clamp set at 10.3 +/- 0.6 mmol l-1. Hyperglycemia reduced the Kgluc at all insulin levels. Individual dose-response curves were transformed to single weighted numerical pancreatic responsiveness scores [PRS], and peripheral sensitivity scores [PSS].(ABSTRACT TRUNCATED AT 250 WORDS)

Assessment of retinoid-induced differentiation of F9 embryonal carcinoma cells with an enzyme-linked immunoadsorbent assay for laminin: Statistical comparison of dose-response curves

A convenient procedure, using enzyme-linked immunoadsorbent assay of laminin, to measure retinoid-induced F9-cell differentiation into parietal endoderm was developed. Dose-response curves were fitted with the Allfit program, a statistical method for the analysis and simultaneous comparison of sigmoidal curves, which has been modified for use with a microcomputer. The procedure was standardized with respect to time of retinoid incubation, time-course of laminin production, effects of dibutyryl cAMP, and nature of individual dose-response curves. Retinoic acid produced a half-maximal response at 1.3 n m. Retinol was 175-fold less potent than retinoic acid and required 72 h to effect a maximum response, in contrast to 48 h for retinoic acid. Six oxidized and/or isomerized metabolites of retinoic acid, including 13- cis-retinoic acid, were less potent than retinoic acid, but were more potent than retinol. The dose-response curves had identical slopes with the exception of those obtained with 13- cis-4-oxo- and 4-oxo-16-hydroxyretinoic acids, the only metabolites tested with two structural alterations relative to retinoic acid. Multiple functional group alterations were synergistic in deactivating retinoic acid. The synthetic retinoids 13- cis-N-ethylretinamide and 4-hydroxyphenylretinamide and the steroid hormone 1,25-dihydroxycholecalciferol were inactive.

Activation of neutrophil chemotaxis by leukotriene B4 and 5-hydroxyeicosatetraenoic acid in chronic inflammatory bowel disease.

Circulating neutrophils were investigated in 15 patients with Crohn's disease (CD), 15 with ulcerative colitis (UC), and 15 healthy volunteers. Dose-response curves for chemotaxis in Boyden chambers were analysed for sensitivity to leukotriene B4 (LTB4), its 20-hydroxy-LTB4 (20-OH-LTB4) and 20-carboxy-LTB4 (20-COOH-LTB4) catabolites, and 5- and 15-hydroxyeicosatetraenoic acids (HETEs). Positive controls included: complement 5a (C5a), formy-L-methionyl-L-leucyl-L-phenylalanine (f-Met-Leu-Phe), and casein. Control chemotaxis test were performed at concentrations yielding optimal responses in leucocytes of healthy volunteers. Chemotaxis to suboptimal concentrations of LTB4 1.0 and 3.2 nmol/l, and 5-HETE 316 nmol/l, was markedly depressed in patients with chronic inflammatory bowel disease (CIBD). Analyses of individual dose-response curves revealed an underlying decreased sensitivity to LTB4 in 11 out of 30 patients, to 5-HETE in 10 out of 30 patients with a corresponding decrease of median sensitivity to LTB4 and 5-HETE in both CD and UC. Peak responses to LTB4, 5-HETE, f-Met-Leu-Phe, and casein were identical in the three groups tested, whereas the C5a values were significantly depressed in both groups of patients (p less than 0.05). The potency of LTB4 exceeded that of 5-HETE by a factor of approximately 100 whereas 20-OH-LTB4 was nearly as potent as LTB4. 20-COOH-LTB4 and 15-HETE did not activate chemotaxis of human neutrophils. These findings are suggestive of a competitive inhibition of receptors with heterogeneity for LTB4 and 5-HETE.(ABSTRACT TRUNCATED AT 250 WORDS)

Phorbol myristate acetate lung injury and airway responsiveness to aerosol histamine in awake sheep

To test the hypothesis that phorbol myristate acetate (PMA), a potent granulocyte activator, would increase airway responsiveness to aerosol histamine, we quantitated airway responsiveness to aerosol histamine in nine awake sheep before and 4 1 2 hours after intravenous PMA, 5 μg/kg. A dose-response curve to aerosol histamine was constructed by administering histamine aerosols of 0.1, 0.3, 1.0, 3.0, 10.0, and 30.0 mg/ml sequentially until dynamic lung compliance (C dyn) decreased to ⩽65% baseline. The dose of histamine that caused a reduction of C dyn to 65% of baseline (ED 65C dyn) was determined by linear interpolation. We then administered PMA, defined a new baseline C dyn 4 1 2 hours later, and administered aerosol histamine as before. Control experiments were performed on another day in eight of the nine sheep by administering aerosol histamine 4 1 2 hours apart without infusing PMA. Histamine responsiveness did not change significantly during 4 1 2 hours in the control experiments. Control (preaerosol histamine) C dyn decreased from 0.090 ± 0.010 L/cm H 2O before PMA to 0.048 ± 0.004 L/cm H 2 4 1 2 hours after PMA. The mean pre-PMA ED 65 C dyn for the nine sheep was 3.80 ± 0.87 mg/ml, and the mean post-PMA ED 65C dyn was 2.13 ± 0.98 mglml ( p < 0.05). When individual doseresponse curves are examined, the small statistical increase in aerosol histamine responsiveness does not appear important, especially when the increase is compared to the changes previously reported after endotoxemia in unanesthetized sheep.

Characterization of insulin resistance in type I diabetes.

Insulin sensitivity was assessed using the euglycaemic clamp technique in eight type I diabetic patients (after overnight blood glucose normalization with an artificial pancreas) and in six healthy subjects. Basal insulin concentrations were higher in diabetic patients (25 +/- 4 microU/ml) than in control subjects (17 +/- 1 microU/ml; P less than 0.05). Insulin infusion of 0.5, 1.0, 2.0 and 5.0 mU/kg per min during subsequent 2-h periods resulted in similar mean steady-state insulin concentrations in both groups. The mean dextrose requirements during the last 40 min of each period were nevertheless decreased in diabetic patients (1.6 +/- 0.5, 3.5 +/- 0.8, 6.5 +/- 0.7, 10.2 +/- 0.7 mg/kg per min) as compared with control subjects (4.7 +/- 0.3, 8.2 +/- 0.9, 10.2 +/- 0.9, 12.4 +/- 0.9 mg/kg per min). At low insulin concentrations dextrose requirements were diminished in all diabetic subjects. At the highest insulin levels, individual dose-response curves from only four patients were within the normal range. Under basal conditions, the monocyte receptor number was significantly reduced in diabetic patients (17,500 +/- 2,800 sites/cell) as compared with control subjects (26,700 +/- 2,500 sites/cell; P less than 0.05), whereas there were no differences regarding empty site affinities. Receptor data did not differ in patients with normal and decreased maximal dextrose requirements. Insulin resistance is apparently a common feature of type I diabetes at serum insulin concentrations of approximately 100 microU/ml. Normalization of the insulin effect by higher insulin concentrations is not possible in all patients. Insulin antibodies at concentrations observed in this study (less than 0.16 mU/ml) do not contribute significantly to insulin resistance; receptor and postreceptor defects are possibly more important.

Histamine release from whole blood induced by anti-IgE: relationship to patient age, age at onset and serum IgE levels.

Anti-IgE-induced histamine release from basophils was examined in 46 asthmatic subjects using a whole blood method. Basophils from subjects less than 30 years old released more histamine than those from subjects aged 41 to 50. The age at onset of the disease also affected the reactivity of basophils to anti-IgE: basophils showed a high response in subjects whose age at onset was between 0 and 10 years, and low response in the subjects whose age at onset was between 41 and 50 years. There was a correlation between histamine release and serum IgE levels. However, individual dose-response curves of histamine release varied greatly in whom serum IgE levels were low.

The value of activated coagulation time in monitoring heparin therapy during extracorporeal circulation.

The anticoagulant effect of heparin was studied in 20 patients undergoing aortocoronary bypass surgery. The protamine dose necessary to reverse heparin after extracorporeal circulation (ECC) was assessed in ten patients from individual heparin dose-response curves (HDR group). The other ten patients received protamine according to a routine protocol (control group). The protamine administration was followed in both groups by injection of 3-5 g tranexamic acid (Cyklokapron). A wide range of sensitivity to heparin was shown by the patients. Although almost twice as much protamine was given to the control group as to the HDR group, the effect on heparin reversal was similar. The variability of protamine dose did not influence the post-bypass levels of fibrinogen, AT-III, activated coagulation time (ACT) or Cephotest, and fibrinolysis was not observed in either of the groups. During ECC there was poor correlation between ACT and plasma heparin levels, and the use of heparin dose-response curves was grossly misleading in regard to true heparin concentration. The postoperative bleeding was not related to the levels of heparin or coagulation parameters after heparin reversal. The concentrations of fibrinogen and AT-III showed variations dependent on the changes in haematocrit. A number of factors other than heparin that influence ACT are discussed.