Impact of the fibrinolytic enzyme system on prognosis and survival associated with non-small cell lung carcinoma.

Comprehensive studies of fibrinolysis in non-small cell lung carcinoma have been limited, and assignment of patients to high/low prognosis groups based on arbitrary cut-offs utilizing fibrinolytic measurements is unstandardized. This study was performed in 166 patients to examine the effects of cut-off values determined in three ways. Model 1 assigned patients to one of three equal groups (low, medium, high) based on fibrinolytic measurements made at diagnosis, Model 2 divided patients into low/high groups using median values, and Model 3 grouped according to the parameter being above/below normal range. In model 1, raised plasma fibrinogen, D-dimer and soluble fibrin were positively associated with poorer survival. In model 2, tissue plasminogen activator antigen was additionally related to poorer prognosis. Model 3 identified seven parameters as significantly related to survival, two not identified by the other models becoming significant [plasmin-antiplasmin, tissue plasminogen activator inhibitor-1 (PAI-1) antigen]. Using multivariate analysis, plasma fibrinogen level was the most uniformly significant parameter. Relative risk estimates indicated that raised plasma fibrinogen, soluble fibrin and D-dimer were associated with increased risk of death. Use of the normal/above normal cut-off is recommended to provide the maximum number of significant parameters relating to prognosis, and increased plasma D-dimer, PAI-1 antigen and fibrinogen were most closely related to survival/prognosis.

Alterations to the fibrinolytic enzyme system in patients with non-small cell lung carcinoma.

Although fibrinolysis has been implicated in the progression and metastasis of lung cancer, no detailed study has been carried out on components measured in samples from both plasma and tumour. This study thus provides the first comprehensive data obtained from 166 patients diagnosed with non-small cell lung carcinoma. Plasma samples were obtained at diagnosis and tumour samples during surgical resection. Appropriate control samples were obtained from normal subjects and patients with chronic obstructive airways disease (plasma) and from organ donors (normal lung tissue). Assays were performed on plasma and tissue extracts for tissue plasminogen activator, urokinase-like activator and plasminogen activator inhibitor (activity and antigen in all cases), together with plasmin-antiplasmin complex, soluble fibrin, D-dimer and thrombin-antithrombin complex. Levels of D-dimer, thrombin-antithrombin complex and plasmin-antiplasmin complex were all significantly higher in plasma from patients, whereas urokinase-like activator activity was reduced. Only two parameters were significantly altered in both the core and periphery of tumour tissue: levels of D-dimer were increased and tissue-type plasminogen activator activity was reduced. Interestingly, significant differences in levels of other fibrinolytic parameters were detected in the core and periphery of tumours. Significant activation of fibrinolysis was indicated in patients, although the origin of this could not be related consistently to changes in levels of plasminogen activator and inhibitor.

Preoperative platelet dysfunction increases the benefit of aprotinin in cardiopulmonary bypass.

BACKGROUND: This study was designed to determine the benefit of aprotinin therapy in reducing bleeding during and after cardiopulmonary bypass in patients with preoperative platelet dysfunction. Platelet function involvement in the mechanism by which aprotinin acts was also investigated. METHODS: In a double-blind, randomized study, patients received high-dose aprotinin (n = 54) or placebo (n = 52). Whole blood aggregation was measured preoperatively. Platelet function and activation in both groups were assessed intraoperatively and postoperatively at five times. RESULTS: Aprotinin significantly reduced perioperative bleeding and postoperative blood transfusion. Placebo-treated patients with reduced preoperative platelet aggregation bled more postoperatively, but aprotinin reduced the bleeding in patients with normal or reduced platelet function to similar levels. Any cardiopulmonary bypass-induced changes in platelet aggregation, platelet activation as measured by P-selectin expression, and von Willebrand factor antigen and function were similar in aprotinin-treated and placebo-treated groups. CONCLUSIONS: The mechanism by which aprotinin reduced bleeding was independent of any effect on platelet function. However, aprotinin produced a greater reduction in bleeding among patients whose condition was hemostatically compromised by preoperative platelet dysfunction.

Changes in plasminogen activator inhibitor-1 levels in non-small cell lung cancer.

Increased urokinase plasminogen activator (uPA) levels are increased in a number of malignancies and have been correlated with decreased disease-free interval and decreased overall survival. We have, therefore, examined components of this plasminogen activating system in patients with Non-Small Cell Lung Cancer (NSCLC). Levels of uPA, urokinase-plasminogen activator receptor (uPAR) and plasminogen activator inhibitor-1 (PAI-1) were measured semiquantitatively in paraffin sections of tumours from 147 patients with NSCLC. Immunohistochemically stained sections of tumour were allocated a score for stain intensity and results correlated to: survival; tumour stage(T); nodal stage(N); stage grouping (I to IIIb), survival status and sex. Increased levels of PAI-1 were associated with a decreased survival in squamous cell carcinoma (SCC) X2 = 5.72, p = 0.017 (n = 74). There was a significant positive relationship between PAI-1 levels and N-stage (p = < 0.05), presence of nodal metastases (p = < 0.05), stage grouping (p = < 0.01) and extent of disease (p = < 0.05) in the total group and the SCC subgroup, but not adenocarcinoma. There was a significant positive relationship between PAI-1 levels and T-stage (p = < 0.05) in the total group, and survival status (p = < 0.05) in the SCC subgroup alone. uPA and uPAR levels were not significantly associated with tumour staging or survival. We conclude that increased PAI-1 antigen levels may be associated with a decreased survival in patients with SCC.

A proposed model to monitor heparin therapy using the concentrated thrombin time which allows standardisation of reagents and improved estimation of heparin concentrations.

The concentrated thrombin time (CTT), a thrombin time performed with a high concentration of thrombin, was evaluated as an alternative to the activated partial thromboplastin time (APTT) for monitoring of heparin therapy. Forty-nine plasmas from patients receiving unfractionated heparin therapy were tested. It was first demonstrated that CTTs using three commercial reagents could be standardised against CTTs performed with a reference reagent, MRC reagent 66/305. For comparison, APTTs were performed on the plasmas. As a benchmark of the degree of heparinisation, the heparin concentration of the plasmas was determined by chromogenic anti-IIa heparin assays, therapeutic range being 0.2-0.4 units/ml. The optimal relationships of the CTTs and APTT with the heparin concentration were established. These were used to predict the heparin concentrations of the plasmas from the results of the APTT, CTT performed with the reference reagent, and transformed CTT performed with each of the three commercial reagents. In predicting the assayed plasma heparin concentrations, the accuracy of the APTT was only 53%, while the CTT was from 78 to 82%. The CTT can be standardised and, subject to results of clinical trials, could provide an improved method of monitoring heparin therapy.

The role of von Willebrand factor in haemostasis and blood loss during and after cardiopulmonary bypass surgery.

Excessive perioperative and postoperative bleeding continue to complicate cardiopulmonary bypass surgery (CPB). In this study we measured the von Willebrand factor antigen (vWF:Ag), collagen binding assay (CBA) and ristocetin cofactor assay (RiCo) in 52 patients undergoing CPB. The collagen binding assay employs the affinity of the high-molecular-weight multimers of vWF to collagen and was used in this study to demonstrate differences in the multimeric composition of vWF during and after CPB. The observed values for the vWF:Ag, CBA and RiCo were correlated to the amount of postoperative bleeding. Both the preoperative vWF:Ag and the CBA showed significant negative correlation with postoperative blood loss (r = -0.3046, P < 0.05 and r = -0.3228, P < 0.05 respectively). Higher blood loss figures correlated with lower vWF:Ag, CBA and RiCo during and after surgery with the strongest correlation 1 h post-op between both vWF:Ag and CBA and actual blood loss (r = -0.5061, P < 0.001 and r = -0.4942, P < 0.001 respectively). The strong negative correlations between vWF:Ag, CBA and RiCo before, during and after CPB and blood loss data verify the important role of vWF in primary haemostasis. Of particular note, the negative correlations between preoperative levels of vWF:Ag, CBA and postoperative blood loss provide valuable insight into another possible mechanism of excessive bleeding post CPB.

Relationship of fibrinolysis and platelet function to bleeding after cardiopulmonary bypass.

In order to study the effects of cardiopulmonary bypass (CPB) on fibrinolysis and platelet function and the possible relationship of these effects on post-operative blood loss, 127 patients undergoing CPB were examined. There was a significant reduction in the median levels of fibrinogen, plasminogen, alpha 2-antiplasmin, fibrinolytic potential and platelet aggregation during CPB (P < or = 0.001). Median levels of soluble fibrin, fibrinogen degradation products, D-dimer and PAI-1 were increased, while the level of t-PA activity remained constant. Post-CPB levels of fibrinogen and plasminogen correlated negatively with blood loss (P = 0.003 and P < 0.001, respectively) and interestingly, lower levels of alpha 2-antiplasmin and higher levels of t-PA activity before CPB were associated with greater blood loss after CPB (P < 0.001 and P = 0.004, respectively). Better pre-CPB platelet function correlated with lower levels of D-dimer before and after CPB. As expected, haemodilution had a significant effect on fibrinolytic and coagulation parameters post-CPB; the greater the haemodilution, the more the concentration of fibrinogen, plasminogen and alpha 2-antiplasmin fell post-CPB and the greater the blood loss. The increase in PAI-1 levels intra-CPB appeared to result in mean t-PA activity remaining unchanged 1 h post-CPB. Post-CPB increases in soluble fibrin were paralleled by increases in fibrinogen degradation products and D-dimer, suggesting that intra-operative contact activation is related to activation of the fibrinolytic system. The present findings indicate the greater the fibrinolytic activation, the greater the post-CPB blood loss.

Relationship of platelet aggregation to bleeding after cardiopulmonary bypass.

Excessive bleeding after cardiopulmonary bypass operations is a persistent problem. This study assessed the influence of platelet function on blood loss for 134 patients undergoing cardiopulmonary bypass. Platelet function was measured by platelet aggregation in platelet-rich plasma and whole blood using collagen as the agonist. Adenosine triphosphate release was assessed concurrently. Measurements were made 1 day before operation and 1 hour after the cessation of cardiopulmonary bypass. Three important findings were made. First, statistically significant correlations were shown between preoperative and postoperative platelet aggregation and blood drainage for the first 3 hours postoperatively. Second, correlations were greatest when preoperative measurement was performed on whole blood and postoperative measurement was performed on platelet-rich plasma. Third, patients with reduced postoperative platelet aggregation in platelet-rich plasma had significantly greater transfusion requirements in the first 24 hours postoperatively. In defining the 16 patients who bled excessively among the 134 patients studied, the preoperative aggregation in whole blood had a sensitivity of 62%, specificity of 75%, positive predictive value of 26%, and negative predictive value of 94%. The postoperative aggregation in platelet-rich plasma had a sensitivity of 86%, specificity of 69%, positive predictive value of 28%, and negative predictive value of 97%. These results indicate that preoperative and postoperative measurement of platelet aggregation may provide a rationale for the prophylaxis or treatment of patients to reduce blood loss after cardiopulmonary bypass.

The effect of oral anticoagulant therapy on APTT results from a bedside coagulation monitor.

OBJECTIVE: The Ciba Corning 512 coagulation monitor (CC512) can be used to monitor heparin therapy by performing an activated partial thromboplastin time (APTT) at the patient's bedside. This study was designed to compare the CC512 results to results using the laboratory system. The relative sensitivities of both systems to the effect of oral anticoagulant therapy also was investigated. METHODS: Activated partial thromboplastin times were performed with both the CC512 and laboratory system on 74 specimens from patients receiving i.v. heparin therapy, and on 14 specimens from patients on warfarin only. Heparin assays were performed on 43 of the specimens from the heparinized patients. RESULTS: When a patient was receiving heparin only, the APTT results of the CC512 proved to be similar to existing laboratory methods. The CC512 APTT results of patients on warfarin only were markedly prolonged, whereas the laboratory APTTs were only slightly affected. CONCLUSION: The CC512 results were comparable to the laboratory system. However, the CC512 APTT was more sensitive to the effect of warfarin than the laboratory APTT system used in this study. CC512 APTT results on a patient receiving both oral and intravenous anticoagulation could be misleading.

A low volume specimen container suitable for monitoring the aPTT of heparinized patients.

Differences in the activated partial thromboplastin time (aPTT) were shown when blood taken from patients receiving intravenous heparin therapy was collected into 5 ml and 1 ml citrate containers. Mean aPTTs were 27% shorter with the plasma from the 1 ml citrate containers (n = 23). These results were paralleled by a 37% reduction in the mean heparin concentration (n = 11) and a 77% increase in the mean platelet factor 4 (PF4) concentration (n = 7). This phenomenon is due to increased platelet activation and subsequent increased heparin neutralization in the 1 ml citrate container. In an attempt to overcome this, the citrate was removed from a 1 ml container and replaced with a buffered tri-sodium citrate solution containing theophylline, adenosine and dipyridamole anticoagulant (CTAD). Blood from heparinized patients taken into both 5 ml citrate and 1 ml CTAD showed a correction of the shortening artefact in the low volume container. The mean aPTT of plasmas from the 1 ml CTAD container showed an increase of 10% compared with the 5 ml citrate. There was no significant difference in the mean heparin or PF4 concentrations of blood taken into either container. The 1 ml CTAD tube described is a suitable collection container for monitoring heparin in neonates or patients who are difficult to venepuncture and overcomes the neutralization of heparin in part filled low volume containers.

Initial chemotherapy followed by surgery in malignant thymoma.

Initial combination chemotherapy with cisplatinum, adriamycin and cyclophosphamide was given prior to surgery in a young woman with an unresectable malignant thymoma. Complete remission was achieved following three cycles of chemotherapy. Subsequent thymectomy revealed no evidence of residual malignancy. Prolonged remissions following cisplatin-based chemotherapy have recently been achieved in metastatic thymoma. Initial chemotherapy should now be considered prior to surgery or radiotherapy in those patients presenting with very bulky or unresectable disease.

Cisplatinum-based chemotherapy in malignant mediastinal teratoma.

Five patients with primary malignant mediastinal non-seminomatous germ cell tumours have been treated with a multimodality approach, including cisplatinum-containing chemotherapy, at this institution over the past 7 years. All patients had bulky disease (greater than 10 cm maximum diameter) at presentation and showed raised serum concentrations of human chorionic gonadotrophin or alpha-fetoprotein. Two patients are alive with no evidence of disease at 22 months and 6 years, respectively, from initial diagnosis; two patients have died from progressive disease and one from acute non-lymphocytic leukaemia without evidence of residual germ cell tumour. Long-term survival is achievable for these poor risk patients with a combined modality approach.

A comparison of two APTT reagents which use silica activators.

The Ortho activated partial thromboplastin time (APTT) reagent, Thrombosil 1 (TS), was compared to the General Diagnostics automated APTT reagent (GD). TS produced more precise results over a 38-day period of testing a normal control plasma, indicating that the upper limit of the normal range could be more precisely set with TS. This normal range was better represented if the normal values with both reagents were logarithmically transformed before calculating the mean +/- 2 SD. TS was more sensitive to plasma which had been heparinized in vitro. This was also demonstrated in vivo by the testing of 100 plasmas from heparinized patients. On testing of in-vitro dilutions of normal plasma with factor-deficient plasmas, TS was more sensitive to decreasing levels of factors VIII, IX and XI but less sensitive to decreasing factor XII. This was demonstrable in vivo in 71% of cases with plasmas from factor-deficient patients. GD was more sensitive to the lupus anticoagulant in most cases.

Ploidy as a prognostic determinant in surgically treated lung cancer.

The usefulness of tumour ploidy as a prognostic determinant in lung cancer was evaluated in a group of 100 surgically treated patients. Archival paraffin sections of the tumours were analysed by flow cytometry. 45% of tumours were aneuploid and 55% were diploid. Overall, patients with aneuploid tumours had significantly shorter survival (p less than 0.0005) than those with diploid tumours. The subset of patients without nodal involvement at operation and with diploid tumours had a particularly long survival rate. Of these 45 patients 41 (91%) were alive at 2 years compared with only 16 (55%) of the 29 with aneuploid tumours (p less than 0.05). A group with such a favourable prognosis has not previously been recognised except when staging is based on total mediastinal nodal clearance. Ploidy was found to be independent of age, sex, type of operation, site of primary tumour, histology, or TNM category. On Cox multivariate analysis ploidy was the most important and independent prognostic determinant. Therefore, in patients with operable lung cancer, ploidy should be taken into account in planning of management, in estimation of prognosis, and in stratification for treatment trials.

Prolonged survival with conventional cisplatinum/vinblastine/bleomycin therapy alone for mediastinal endodermal sinus tumour.

Combination chemotherapy with cisplatin, vinblastine, and bleomycin (PVB) is considered to be the best treatment for disseminated non-seminomatous germ cell tumours, with a 70% long term disease-free survival rate. For one subgroup of patients, those with primary mediastinal endodermal sinus tumours, results with PVB have been disappointing and more intensive regimens with major toxicity have recently been proposed. We report a patient who, following standard PVB chemotherapy, is one of the longest reported survivors of mediastinal endodermal sinus tumours. Optimal chemotherapy for endodermal sinus tumours is yet to be determined.

Acute pancytopenia due to megaloblastic arrest in association with co-trimoxazole.

Three patients who were taking co-trimoxazole developed acute pancytopenia due to megaloblastic arrest, and two of the three died while pancytopenic. One patient had a pre-existing megaloblastic anaemia. In the other two patients, there was no macrocytosis or neutrophil hypersegmentation despite the severe megaloblastosis. Because specific treatment is needed urgently, it is important to distinguish megaloblastic arrest from drug-induced hypoplasia.