The effect of combined antenatal vitamin K and phenobarbital therapy on umbilical blood coagulation studies in infants less than 34 weeks' gestation

OBJECTIVE: To determine if antenatal vitamin K and phenobarbital therapy affect coagulation studies in umbilical blood at birth, and to provide 95% reference ranges for umbilical blood coagulation parameters in premature gestations. METHODS: Patients at imminent risk for spontaneous or indicated premature delivery less than 34 weeks' gestation were randomized to receive either placebo or vitamin K and phenobarbital. Prothrombin time (PT), activated partial thromboplastin time (PTT), functional coagulation factors, and decarboxylated prothrombin assays were performed on umbilical blood specimens. Decarboxylated prothrombin, also known as "protein induced by vitamin K absence-factor II" or precursor prothrombin, is a sensitive marker for vitamin K deficiency. Standardized values of PT and PTT are reported in seconds and standardized values of factor assays in percentage of normal adult functional activity (mean +/- one standard deviation). RESULTS: Newborns in the placebo and treatment groups had similar umbilical blood PT (12.6 +/- 1.2 versus 12.7 +/- 1.4 seconds), PTT (48.8 +/- 13.4 versus 49.6 +/- 13.8 seconds), and functional activity of factor II (40.3 +/- 12.5 versus 42.0 +/- 12.1%), factor VII (67.0 +/- 20.9 versus 66.8 +/- 18.9%), factor IX (27.4 +/- 12.8 versus 25.8 +/- 8.9%), and factor X (47.0 +/- 12.8 versus 49.2 +/- 11.6%). Newborns in the treatment group were about half as likely as those in the placebo group to have detectable decarboxylated prothrombin levels in umbilical blood at birth (gestational age-adjusted odds ratio 0.47, 95% confidence interval 0.22-1.01; P = .05). CONCLUSIONS: Combined maternal therapy with vitamin K and phenobarbital before premature delivery does not affect umbilical blood PT, PTT, or functional activity of vitamin K-dependent coagulation factors II, VII, IX, and X. However, it is associated with the reduced presence of decarboxylated prothrombin in umbilical blood at birth. There is significant improvement in umbilical blood coagulation tests as gestational age advances from 24 to 34 weeks.     

D-dimer, thrombin-antithrombin III-complex (TAT) and prothrombin fragment 1+2 (PTF). Parameters for monitoring therapy with low molecular-weight heparin in coagulation disorders

Two groups of 15 patients each with disseminated intravascular coagulation in association with septic disease were treated with low-molecular-weight heparin (lmw-heparin) in different dosages (group I: 1.5-5 IE/kg body weight (BW) per hour; group II: 8-15 IE/kg BW). We studied the levels of D-dimer, thrombin-antithrombin III complex (TAT), prothrombin fragments 1 and 2 (PTF), and global tests of coagulation like prothrombin time (PT), activated partial thromboplastin time (PTT), thrombin time (TT) and platelet count, plasminogen activation (PA) and fibrinogen concentration to estimate the success of heparin therapy in the two groups. TT and fibrinogen concentration were not suitable to follow the course of the coagulation disorder, PT, PTT, platelet count progressively PA, D-dimer, TAT, and PTF normalised progressively after heparinisation. However, only the last three parameters were sensitive enough to show different effects of variable dosages of lmw-heparin. D-dimer, TAT, and PTF levels declined in proportion with heparin concentrations, and thus appear to be the most useful parameters for monitoring the therapeutic effect of heparin in septic coagulopathies.     

Autotransfused shed mediastinal blood has normal erythrocyte survival

The anticoagulant action of Anisakis simplex larvae on human blood in vitro was examined. Anticoagulant activity was assessed by routine screening tests that evaluate the overall competency of the coagulant mechanism. A slight prolongation of the prothrombin time (PT) was observed with the larval crude extracts. Prolongation of the PT was seen at a concentration of excretory/secretory (ES) products greater than 62.5 micrograms/ml. No prolongation of the activated partial thromboplastin time (PTT) was observed using crude extracts. There was a prolongation of the PTT with ES products at concentrations greater than 62.5 micrograms/ml. ES products of the larvae were able to prolong coagulation times indicating that they contain an inhibitory or anticoagulant property. Preparation of crude extracts of A. simplex showed only minimal anticoagulant activity. The results obtained by measurements of the PT and the PTT suggest a probable alteration of one of the coagulation proteins namely factors Xa, IIa or Va. These findings suggest that the anticoagulant activity demonstrated in the ES products may play an important role during invasion of the gastric or intestinal mucosa by larvae and could have biological significance in infected patients.     

Clotting times and antithrombin III activity in cats with naturally developing diseases: 85 cases (1984-1994)

OBJECTIVE: To determine the prevalence of abnormalities of in vitro prothrombin time (PT) and activated partial thromboplastin time (APTT), or antithrombin III (ATIII) activity or all 3 variables in cats; and the association of abnormalities of these variables with naturally developing diseases or disorders. DESIGN: Retrospective study. ANIMALS: 85 cats from which blood had been obtained for measurement of a coagulation profile (PT, APTT, and ATIII activity) and concentration of fibrin degradation products. PROCEDURE: Medical records from the Texas A&M College of Veterinary Medicine were reviewed to determine clinical diagnosis, results of CBC and coagulation profile, and clinical evidence of abnormal bleeding or thrombotic disease. RESULTS: 38 cats had one or more abnormality in the coagulation profile; most had multiple abnormalities. Twenty of these 38 cats had concurrent thrombocytopenia. Thrombocytopenia was identified in 9 of 47 cats in which results of the coagulation profile were normal. Most cats did not have clinical evidence of a coagulation disorder, and testing had been requested as part of a diagnostic work-up or before surgery. Diseases commonly associated with laboratory evidence of a coagulation disorder, either singly or in combination, included hepatic disease, neoplasia, and systemic infections. CLINICAL IMPLICATIONS: On the basis of laboratory evidence, hemostatic disorders develop more commonly in cats than clinical signs would suggest. Coagulation profiles may be warranted in high-risk cats to alert clinicians to potential problems.     

Correlation between the leak point pressure and the clinical grade of incontinence

Twenty-two cats with liver disease were evaluated for coagulation abnormalities including alterations in prothrombin time, activated partial thromboplastin time, thrombin time, factor VII activity, and platelet count. The purpose of the study was to determine the prevalence of coagulation abnormalities in this population of cats, classify abnormalities according to underlying pathogenesis, and determine if serum biochemical parameters typically used as indicators of liver disease showed any correlation with the coagulation abnormalities present. Study results indicated that at least 1 coagulation abnormality was present in 82% of the cats. Prolongation of prothrombin time was most common (16/22 cats) and factor VII activity was below reference range (< 60%) in 15 cats. When classified according to underlying pathogenesis, vitamin K deficiency was the most common abnormality found (11/22). Other abnormalities were less common and included hepatic synthetic failure (3/22), indeterminate (3/22), and disseminated intravascular coagulation (1/22). Increase in alkaline phosphatase (ALP) activity was the only biochemical abnormality that showed statistically significant correlation with coagulation abnormalities (P = .023). Cats with marked increases in ALP activity were more likely to have coagulation abnormalities than those with only mild increases in ALP activity.     

Persistent airway colonization with Alcaligenes xylosoxidans in two brothers with cystic fibrosis

Thirty-six patients, 9 males and 27 females, average age 13.2 years, undergoing posterior fusion and segmental instrumentation surgery for idiopathic adolescent scoliosis were studied for abnormalities of platelet count (PC), prothrombin time (PT) and partial thromboplastin time (PTT). Intraoperative and immediate postoperative values were obtained at 2, 4, 6, 12, 24, and 48 hours. All patients showed diminution of platelet counts during the study period, but mean values were reduced below normals only at 4 hours after operation. The PT was significantly elevated in all patients intraoperatively and for the first 24 hours postoperatively, with return to normal by 48 hours. The PTT levels were essentially unaffected at any time.     

Retinoblastoma protein expression in lung cancer: an immunohistochemical analysis

Several analyses of the retinoblastoma (RB) gene in lung cancer at the DNA, mRNA and protein levels have recently been reported. In particular, small cell lung carcinoma shows a high incidence of RB gene abnormalities, suggesting that alterations of this gene may participate in tumor development. In the present study, we used an immunohistochemical technique with a monoclonal antibody raised against RB protein (PMG3-245) to detect its expression in representative paraffin sections of tissues obtained from 108 patients with various types of lung cancer treated by surgical resection of the primary tumor. While deletion of RB protein expression was observed in 7 (88%) of small cell lung carcinomas, only 17 (17%) of 100 non-small cell lung carcinomas showed decreased RB protein levels and 6 (6%) showed no RB protein expression. This low incidence of RB protein expression abnormalities in non-small cell lung carcinomas was significant (p < 0.0001). Thus, in contrast to small cell lung carcinoma, abnormalities in RB protein expression may be minor events in non-small cell lung carcinoma. In addition, no significant correlation was found between abnormalities in RB protein expression and clinical factors such as stage, tumor size, and nodal involvement in non-small cell lung carcinoma. However, abnormalities in RB protein expression in squamous cell carcinoma were observed only in the less differentiated types (p = 0.144), and there was a weak but not statistically significant association in non-small cell lung carcinoma between RB protein status and prognosis (p = 0.09). Therefore, in non-small cell lung carcinoma, although abnormalities in RB protein appear not to be closely associated with tumor development, further studies on a larger scale and with a longer-term follow-up are required to determine the clinicopathological significance of RB gene abnormalities, in particular the relationship between abnormalities of RB protein and differentiation or prognosis.     

Two genes in the rat homologous to human NKG2

OBJECTIVES: The purpose of this study was to investigate the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) genotype and endothelial cell dysfunction or hypercoagulable state in elderly hypertensive patients. BACKGROUND: Angiotensin-converting enzyme (ACE) insertion/ deletion (I/D) polymorphism was recently reported to be associated with various cardiovascular diseases. However, the precise mechanism of this association remains unknown, and some confounding factors might also affect the association. Endothelial cell dysfunction and coagulation activation play important roles in both the atherosclerotic process and the onset of cardiovascular events. METHODS: We identified the ACE I/D genotype and measured the plasma levels of markers of endothelial cell damage (von Willebrand factor [vWF] and thrombomodulin) and of coagulation activation (prothrombin fragment F1 + 2 [F1 + 2]) in 318 asymptomatic elderly patients with hypertension, aged 59-93 years. RESULTS: The vWF level was significantly higher in those with the DD genotype (n = 54) than in those with the II genotype (n = 131, p < 0.0001) or with the ID genotype (n = 133, p < 0.0001). The TM levels were also higher in patients with the ID genotype (p < 0.005) and the DD genotype (p < 0.01) than in those with the II genotype. There were no differences in F1 + 2 level among the groups. Positive correlations of systolic blood pressure with levels of both vWF and thrombomodulin were found predominantly in patients with the II genotype (both p < 0.001), but no correlation was noted in those with the DD genotype. CONCLUSIONS: Considering the increased plasma levels of both endothelial cell-derived markers in the hypertensive patients with ACE DD genotype, we speculate that the ACE D allele is a risk factor for the development of hypertensive cardiovascular disease associated with endothelial cell damage.     

Correlation between P450 CYP1A1 inducibility, MspI genotype and lung cancer incidence

The aim of this study was to verify a possible correlation between CYP1A1 induction, MspI genotype and lung cancer incidence. A case-control study was performed on 48 lung cancer patients and 81 healthy subjects to test the existence of a correlation, within a European population. The hyperinducible group exhibited a significantly higher risk of lung cancer (odds ratio = 3.41; P = 0.036), especially for adenocarcinoma (odds ratio = 5.29; P = 0.033). In contrast with the situation observed in Asian populations, the frequency of the M2 allele did not differ significantly in the total lung cancer population (7.82%) and the group of healthy subjects (10.71%). The median inducibility value was slightly higher among cancer patients with one or two M2 alleles than among patients homozygous for the wild-type allele (P = 0.09). However, the percentage of individuals possessing at least one mutated allele was not significantly higher among hyperinducible patients (37.5%) than among non-hyperinducible patients (16.0%). No significant correlation could be found between M2 allele and lung cancer or between M2 allele and CYP1A1 inducibility; the only positive correlation found was between CYP1A1 hyperinducibility and lung cancer incidence. Our observations do not support the view that the presence of the M2 allele at the MspI site of the CYP1A1 gene constitutes a significant lung cancer risk in Caucasians.     

恶性淋巴瘤患者凝血纤溶指标变化的意义

目的 探讨恶性淋巴瘤患者凝血纤溶指标的变化,提高对恶性淋巴瘤患者合并血栓的认识方法应用全自动血凝分析仪对20名健康对照及71例恶性淋巴瘤患者的血浆凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、纤维蛋白原定量(FIB)及凝血酶时间(TT)进行检测,用免疫散射比浊法测定两组血浆D-二聚体(D-DI)的含量.结果 71例恶性淋巴瘤患者APTT(30.44±1.43)s、FIB(3.28±0.20)g/L、D-DI(297.05±56.59)μg/L均高于健康对照组的(23.72±0.76)s、(2.57±0.22)g/L、(94.50±26.07)μg/L,差异有统计学意义(P＜0.05).Ⅰ、Ⅱ期淋巴瘤患者各项凝血纤溶指标较健康对照组差异无统计学意义(P＞0.05).Ⅲ、Ⅳ期淋巴瘤患者APTT延长,FIB增高,与健康对照组及Ⅰ、Ⅱ期组比较差异有统计学意义(P＜0.05).合并静脉血栓患者7例,该组与Ⅰ、Ⅱ期患者相比FIB及D-DI均高,与Ⅲ、Ⅳ期患者相比D-DI明显升高.结论 恶性淋巴瘤患者存在凝血纤溶指标的异常,需定期动态检测,化疗后易发生静脉血栓,需及早预防,延长患者生存期.     

Prothrombin time derived fibrinogen determination on Sysmex CA-6000

AIM: To evaluate PT derived fibrinogen determinations with reference to the Clauss fibrinogen assay using a Sysmex CA-6000 random access coagulation analyser. METHODS: Samples were analysed from normal subjects (n = 20), patients with renal or liver dysfunction (n = 25), critically ill patients (n = 25), patients receiving oral anticoagulant treatment (n = 50), and patients with a haemoglobinopathy (n = 127). Prothrombin times were performed using two thromboplastins: one derived from rabbit brain (Dade: Thromboplastin IS) and the other from recombinant human tissue factor (Dade: Innovin). Fibrinogen was assayed by the Clauss method using a commercial kit (Dade: Fibrinogen). RESULTS: The relation between Clauss fibrinogen and PT derived fibrinogen was found to be dependent on the patient's clinical group and source of the thromboplastin used. When the data from the above sample groups were pooled there was still a significant difference (p < 0.001) between Clauss fibrinogen and PT derived fibrinogen, irrespective of thromboplastin used. CONCLUSIONS: It is unsafe to use the PT derived fibrinogen for patient monitoring owing to non-uniform variability in response to clinical status and reagent employed; however, it may prove to be a useful screening test in a research environment for estimating fibrinogen levels among defined patient groups.     

Presence of serum anti-p53 antibodies is associated with pleural effusion and poor prognosis in lung cancer patients

This study was designed prospectively to evaluate the development of anti-p53 antibodies (Abs) in lung cancer patients in relation to their clinical outcome. Sera, derived from 125 lung cancer patients, consisting of 14 small cell lung cancers (SCLC) and 111 non-SCLCs (NSCLC), were surveyed. The p53-null human NSCLC cell line, NCI-H1299, transfected with a human mutant p53 gene was prepared as the source of p53 antigen for immunoblotting analyses to detect the presence of serum anti-p53 Abs. The control group included sera from 10 healthy adults and 14 patients with benign pulmonary diseases. Clinical data including staging and survival were recorded for statistical analyses. The anti-p53 Abs were found in 8% (10 of 125) of the lung cancer patients studied (8.1% of NSCLC versus 7.1% of SCLC patients), whereas none of the control sera had detectable anti-p53 Abs. The presence of anti-p53 Abs was closely associated with malignant pleural effusions (P = 0.001). The p53 Ab-positive patients had a worse prognosis than the p53 Ab-negative patients (P < 0.02; median survival, 20 versus 41 weeks). In both univariate and multivariate analyses, the tumor extension and probably the presence of anti-p53 Abs were significant predictors for cancer death. The development of anti-p53 Abs (n = 9) was also a predictor for poor survival in patients with malignant effusions (n = 51). In conclusion, the presence of serum anti-p53 Abs is closely associated with malignant pleural effusions in lung cancer patients. It may serve as a negative prognostic factor for survival independent of malignant pleural effusions and tumor staging.     

DD genotype of the angiotensin-converting enzyme gene is a risk factor for left ventricular hypertrophy

BACKGROUND: The cardiac renin-angiotensin system has been suggested to be involved in the development of left ventricular hypertrophy. In humans, a strong correlation has been found between plasma angiotensin I-converting enzyme (ACE) activity and the insertion/deletion (I/D) polymorphism of the ACE gene, which has been reported to be associated with myocardial infarction, ischemic and idiopathic dilated cardiomyopathy, sudden death in hypertrophic cardiomyopathy, and restenosis after percutaneous transluminal coronary angioplasty. In the present study, we examined the possibility that the genotype of the ACE gene might influence the development of left ventricular hypertrophy. METHODS AND RESULTS: The study population consisted of 268 subjects randomly selected from our outpatient clinic. In 142 subjects, left ventricular mass (LVM) was determined by echocardiogram. The genotype of the ACE gene was determined by the polymerase chain reaction. ANCOVA revealed that the genotype of the ACE gene had no effect on blood pressure. The percentage of the explained variance of LVM with variables including diastolic blood pressure (DBP, P = .0001), body mass index (BMI, P = .0001), sex (P = .0009), and the genotype of the ACE gene (P = .0017) was 34.61%. Significant differences in the effects of the genotype of the ACE gene on LVM were observed between the II and DD (P = .0004) and between the ID and DD (P = .0077) genotypes. The percentage of the explained variance of the LVM/ht ratio with variables including sex (P = .134), age (P = .3655), the genotype of the ACE gene (P = .0014), BMI (P = .0001), and DBP (P = .0001) was 31.25%. Significant differences in the effects of the genotype of the ACE gene on LVM/ht were observed between the II and DD genotypes (P = .0003) and between the ID and DD genotypes (P = .0091). CONCLUSIONS: In addition to BMI and DBP, the genotype of the ACE gene was a significant predictor of LVM and LVM/ht in our study population.     

Tumour necrosis factor-alpha (TNF-alpha) in patients who have asbestosis and develop cancer

OBJECTIVES: Concentrations of tumour necrosis factor-alpha (TNF-alpha) were assayed by radioimmunoassay in serum samples collected between 1981 and 1987 from 111 patients with asbestosis who were at a high risk of cancer. Follow up of these patients until 1993 showed that 38 had developed cancer (27 lung, three mesotheliomas, and eight diverse malignancies). RESULTS: The mean serum concentrations of TNF-alpha given in fmol/100 microliters serum in all the cases with cancer (14.1) and the cases with lung cancer (13.6) were significantly higher (P < 0.05) than the mean concentrations in the exposed controls (10.5). A positive increase was considered to be any value that was > 2 SDs above the mean of the exposed controls. 22% (six of 27) of the cases with lung cancer were positive compared with 4% (three of 73) of the exposed controls, a significant difference (P < 0.001). The serum concentrations of TNF-alpha correlated moderately with cancer (r = 0.3), lung cancer (r = 0.3), and Neu oncoproteins and epidermal growth factor receptor (EGFR) (r = 0.3, 0.5 respectively). Also, there was a significant correlation between development of cancer and severity or progression of asbestosis. There was no correlation between the concentrations of TNF-alpha and severity or progression of asbestosis. CONCLUSIONS: These results showed high concentrations of TNF-alpha in the patients who had cancer. TNF-alpha may offer an auxiliary method in early diagnosis of cancers related to asbestosis.     

The insertion allele of the ACE gene I/D polymorphism. A candidate gene for insulin resistance?

BACKGROUND: The insertion/deletion (ID) polymorphism of the angiotensin-converting enzyme (ACE) gene has been associated with increased coronary heart disease (CHD), although the mechanism of this association is not apparent. We tested the hypothesis that the deletion allele of the ACE gene is associated with insulin resistance. METHODS AND RESULTS: We related ACE genotype to components of the insulin-resistance syndrome in 103 non-insulin-dependent diabetic (NIDDM) and 533 nondiabetic white subjects. NIDDM subjects with the DD genotype had significantly lower levels of specific insulin (DD 38.6, ID 57.1, and II 87.4 pmol.L-1 by ANOVA, P = .011). Non-insulin-treated subjects with the DD genotype had increased insulin sensitivity by HOMA % (DD 56.4%, II 29.4%, P = .027) and lower levels of des 31,32 proinsulin (DD 3.3, II 7.6 pmol.L-1, P = .012) compared with II subjects. There were no differences in prevalence of CHD or levels of blood pressure, serum lipids, or plasminogen activator inhibitor-1 (PAI-1) activity between the three ACE genotypes. In nondiabetic subjects there were no differences in insulin sensitivity, levels of insulin-like molecules, blood pressure, PAI-1, serum lipids, or CHD prevalence between the three ACE genotypes. CONCLUSIONS: We conclude that increased cardiovascular risk of the DD genotype is not mediated through insulin resistance or abnormalities in fibrinolysis. Conversely, we report an increased sensitivity in NIDDM subjects with the ACE DD genotype.     

Rendu-Osler-Weber disease. Report of a clinical case

Rendu-Osler-Weber disease is an hereditary disorder characterized by cutaneo-mucous telangiectasis and vascular abnormalities in several organs. Bleeding, especially epistaxis, represents the most important clinical feature. Pulmonary arteriovenous fistulae can cause hypoxaemia, haemoptysis, polycythaemia and clubbing. Diagnosis is based on family and personal history, teleangiectasis, laboratory (haemochrome, fibrinogen, PT, PTT) and instrumental findings (endoscopy and/or roentgen). Therapy depends on symptoms. Embolization of pulmonary arteriovenous fistulae and laser treatment of intestinal vascular abnormalities have been successful. Danazol treatment yielded controversial results. We report the case of a patient admitted for arterial hypertension and recurrent epistaxis. Rendu-Osler-Weber disease diagnosis was made based on positivity at family and personal history, clinical examination, laboratory and instrumental findings. In conclusion we underline the pivotal role of anamnesis and clinical examination in the differential diagnosis of hereditary bleeding disorders and emphasize the importance of early diagnosis for the correct therapeutic approach.     

Aneuploidy index in blood: a potential marker for early onset, androgen response, and metastasis in human prostate cancer

OBJECTIVES: To investigate whether the frequency of chromosome abnormalities in peripheral blood lymphocytes defined as the aneuploidy index in blood (AnIB) can be used as a clinical marker of early age onset, androgen response, and metastasis in human prostate cancer. METHODS: Peripheral blood samples were collected from 80 patients with prostate cancer, and chromosome preparations were made from 72-hour cultures after mitotic block. The AnIB of 59 informative cases was compared with several parameters, including age at disease onset, Gleason grade of tumor, clinical stage of tumor, metastasis, and prostate-specific antigen (PSA) level. RESULTS: Patients with AnIB levels greater than 3 had a significantly higher incidence of metastasis (P = 0.022), androgen-independent disease (P = 0.002), and early age at disease onset (age at diagnosis less than 65 years) (P = 0.002) compared with the patients with lower AnIB (less than 3) levels. In addition, patients with AnIB levels greater than 5 had higher PSA levels (greater than 20 ng/mL) (P = 0.029) than patients with AnIB levels less than 5. CONCLUSIONS: Chromosome abnormalities can be detected in the peripheral lymphocytes of patients with prostate cancer, and AnIB can be used as an early diagnostic and predictive marker for prostate cancer metastasis and androgen-independent disease.     

Determinants of CPT-11 and SN-38 activities in human lung cancer cells

Irinotecan (CPT-11) is a semisynthetic camptothecin derivative with a broad spectrum of anti-tumour activity. Carboxylesterase (CE) catalyses the conversion of CPT-11 to SN-38 (7-ethyl-10-hydroxycamptothecin), the active form of CPT-11. The antiproliferative effects of CPT-11 and SN-38, CE-activity and topoisomerase I protein expression were investigated in five human small-cell lung cancer (SCLC) cell lines and four human non-small-cell lung cancer (NSCLC) cell lines. Antiproliferative activity, expressed as IC50 values, was determined using the MTT assay. CPT-11 was significantly more active in SCLC than in NSCLC cell lines (P = 0.0036), whereas no significant difference between histological types was observed with SN-38. A significant correlation (r2 = 0.52, P = 0.028) was observed between CE activity and chemosensitivity to CPT-11 but not to SN-38, and significantly higher CE activity was observed in SCLC compared with NSCLC cell lines (P = 0.025). Western blotting experiments showed topoisomerase I protein expressions within a factor of 2, and a granular nuclear staining was detectable in all cell lines by immunocytochemistry of cytospins. No correlation was observed between protein expression and sensitivity to CPT-11 or SN-38. Cellular and medium concentrations of CPT-11 and SN-38 were measured by high-performance liquid chromatography (HPLC) in one SCLC cell line with high CE activity and high sensitivity to CPT-11, and one NSCLC cell line with low sensitivity to CPT-11 and CE activity. Intracellular concentrations of CPT-11 and SN-38 were higher in the SCLC cell line, and this was associated with an increase in cellular uptake of CPT-11 compared with the medium, and an increased intracellular formation of SN-38. In conclusion, CE activity appears to be associated with higher sensitivity to CPT-11 in human lung cancer cell lines and may partly explain the difference in the in vitro sensitivity to CPT-11 between SCLC and NSCLC cells. The assessment of CE activity in clinical material of lung cancer patients undergoing treatment with CPT-11 may be warranted. However, other mechanisms may influence sensitivity to CPT-11, possibly including drug transport.     

Recovery of post-tetanic and train-of-four responses at the first dorsal interosseous and adductor pollicis muscles in patients receiving vecuronium

PURPOSE: To compare recovery of accelographical responses to post-tetanic twitch (PTT) and train-of-four (TOF) stimuli obtained at the first dorsal interosseous muscle (DI) with those at the adductor pollicis muscle (AP) after administration of vecuronium 70 micrograms.kg-1. METHODS: Sixty adult patients were randomly assigned to one of four groups: PTT-DI (n = 15), PTT-AP (n = 15), TOF-DI (n = 15), or TOF-AP (n = 15) group. In PTT-DI and PTT-AP groups, responses to PTT were measured accelographically at the DI and at the AP, respectively. In TOF-DI and TOF-AP groups, responses to TOF were measured at the DI and at the AP, respectively. RESULTS: The T1/T0 (T0 = control) was greater in the TOF-DI than in TOF-AP group throughout recovery (P < 0.05), and the T4/T1 was greater in the TOF-DI than in TOF-AP group during the 30-40 min after vecuronium injection (P < 0.05). Time to the return of the first response to PTT (post-tetanic count(1), PTC1) was less in the PTT-DI than in the PTT-AP group (17.7 +/- 4.2 vs 21.7 +/- 5.6 min, mean +/- SD, P = 0.0341). The post-tetanic count PTC (number of single twitch stimuli in response to PTT) was greater in the PTT-DI than in the PTT-AP group during the 10-30 min after vecuronium (P < 0.05). Time to the return of T1 was less in the TOF-DI than in the TOF-AP group (23.1 +/- 6.0 vs 27.6 +/- 4.9 min, P = 0.0334). CONCLUSION: Recovery of responses to PTT and TOF stimuli occurred earlier at the DI than at the AP.     

Polymorphism within the cyclin D1 gene is associated with prognosis in patients with squamous cell carcinoma of the head and neck

We have examined the correlation of a frequent A/G polymorphism within exon 4 of the cyclin D1 gene (CCND1) with genetic susceptibility and clinical outcome in 384 patients with squamous cell carcinoma (SCC) of the head and neck. CCND1 genotype frequencies were similar in the cases and 191 controls. Furthermore, the CCND1 genotype was not associated with susceptibility to SCC of the larynx, pharynx, or oral cavity. The influence of the CCND1 genotype on clinical outcome was also assessed. We found no correlation between genotype and tumor size (T1-T4), the involvement of nodes at presentation, or patient age and gender. However, the distribution of CCND1 genotypes in cases with poorly differentiated tumors was significantly different to that in patients with well-/moderately differentiated tumors (P = 0.016; chi2(2) = 8.71). Homozygosity for CCND1*G (GG genotype) was associated with poorly differentiated tumors (G3). We used Cox's proportional hazards model to investigate the influence of the CCND1 genotype on disease-free interval. CCND1 GG was associated with reduced disease-free interval [P = 0.001; hazard ratio (HR) = 2.95; 95% confidence interval (CI) = 1.54-5.63]. This remained significant after correction for tumor differentiation (P = 0.013; HR = 2.34; 95% CI = 1.2-4.6) and tumor stage (P = 0.005; HR = 2.64; 95% CI = 1.34-5.19). Analysis of the data from patients with tumors at different sites showed that the CCND1 GG genotype was associated with reduced disease-free interval in laryngeal (P = 0.004; HR = 3.63; 95% CI = 1.44-8.83) and pharyngeal (P = 0.006; HR = 3.48; 95% CI = 1.43-8.46) tumors. This is the first report of an association between CCND1 polymorphism and prognosis in SCC of the head and neck. These data show that the CCND1 GG genotype is an independent prognostic indicator of disease-free interval and supports initial observations in non-small cell lung cancer, that polymorphism within CCND1 influences tumor behavior.