Jerzy Laudanski

Identification of the vasodilatory endothelial cannabinoid receptor in the human pulmonary artery.

Background The endocannabinoid anandamide is implicated in the pathogenesis of hypotension in haemorrhagic, endotoxic, and cardiogenic shock. It has been demonstrated in animal, but not in human, vessels that the vasodilatory effects of anandamide and abnormal cannabidiol are partially mediated by an as yet unidentified endothelial cannabinoid receptor. Our study was performed to examine the influence of abnormal cannabidiol on the human pulmonary artery. Methods Isolated human pulmonary arteries were obtained from patients without clinical evidence of pulmonary hypertension during resection of lung carcinoma. Vasodilatory effects of abnormal cannabidiol were examined on endothelium-intact vessels preconstricted with serotonin or potassium chloride. Results Anandamide and abnormal cannabidiol relaxed serotonin-preconstricted vessels concentration-dependently. The effect of abnormal cannabidiol was reduced by endothelium denudation, pertussis toxin and two antagonists of the novel endothelial receptor, cannabidiol and O-1918, but not by the nitric oxide synthase inhibitor L-NAME given together with the cyclooxygenase inhibitor indomethacin. It was also diminished by blockade of calcium-activated potassium channels by the nonselective blocker tetraethylammonium or by combination of selective blockers of small (apamin) and intermediate and large (charybdotoxin) conductance calcium-activated potassium channels. The potency of abnormal cannabidiol to relax vessels was lower in potassium chloride than in serotonin-preconstriced preparations. Conclusions Abnormal cannabidiol relaxes human pulmonary arteries in an endothelium-independent and endothelium-dependent manner. The latter component is probably mediated via the putative endothelial cannabinoid receptor, activation of which may release endothelium-derived hyperpolarizing factor, which in turn acts via calcium-activated potassium channels. Abnormal cannabidiol is behaviourally inactive; it may have a therapeutic implication in vascular diseases, especially in the treatment of pulmonary hypertension.

Prognostic significance of p53 and bcl-2 abnormalities in operable nonsmall cell lung cancer.

The association of p53 abnormalities and bcl-2 protein expression with clinical data and prognosis in 102 patients with resected nonsmall cell lung cancer (NSCLC) was investigated. Deoxyribonucleic acid analysis of exons 5-8 of the p53 gene showed mutations (p53-M) in 47% of resected NSCLC, serum p53 antibodies (p53-Abs) were detected in 25%, p53 protein overexpression (p53-PE) in 54%, and bcl-2 protein overexpression (bcl-2-PE) in 48%. A statistically significant association was found between p53-PE, serum p53-Abs and the presence of a p53 gene alteration. No significant associations were found between results of the p53-M, p53-Abs, bcl-2-PE tests and clinicopathological parameters. In the case of the p53-PE test there were significantly fewer positive results for adenocarcinoma than for squamous cell carcinoma and large cell carcinoma. Survival analysis showed that both p53 abnormalities and negative staining for bcl-2, when analysed separately, were associated with poor overall survival. In a multivariate analysis, only the positive result of the p53-M test remained an independent, statistically significant, unfavourable prognostic factor for survival. When the p53 mutation test was removed from the model, positive results of the p53-PE test and the p53-Abs test became statistically significant, unfavourable prognostic factors. To conclude, among p53 and bcl-2 abnormalities, only p53 gene mutations seem to have a strong and independent effect on prognosis. When deoxyribonucleic acid sequence information is not available, p53 protein expression and the presence of p53 antibodies in serum may be used to obtain important prognostic information.

Virodhamine relaxes the human pulmonary artery through the endothelial cannabinoid receptor and indirectly through a COX product

The endocannabinoid virodhamine is a partial agonist at the cannabinoid CB1 receptor and a full agonist at the CB2 receptor, and relaxes rat mesenteric arteries through endothelial cannabinoid receptors. Its concentration in the periphery exceeds that of the endocannabinoid anandamide. Here, we examined the influence of virodhamine on the human pulmonary artery.

Prognostic value of serum p53 antibodies in patients with resected non-small cell lung cancer

BACKGROUND Serum antibodies against p53 protein (p53-Abs) have been detected in some cancer patients. The significance and use of p53-Abs as a marker of the clinical behavior of lung cancer is currently under investigation. PURPOSE In this study, we measured the serum p53-Abs in 84 patients with operable non-small cell lung cancer (NSCLC) and evaluated potential association between the presence of these antibodies and prognosis. METHODS Enzyme-linked immunosorbent assay (ELISA) was used to detect p53-Abs in serum. Survival and disease-free survival curves related to initial p53-Abs status were estimated using the Kaplan-Meier method. RESULTS At the time of diagnosis 19 (22.6%) of 84 analyzed patients had positive result from the serum p53 antibodies (p53-Abs) test. No association was found between p53-Abs, histological types of tumors and clinical stage of disease. We found that patients with a positive result from the p53-Abs test had lower probability of overall and disease-free survival. The unfavorable effect was significant both in the univariate analysis, as well as in the multivariate analysis (after adjustment for sex, histopathological type of tumor. TNM stage). CONCLUSION The results of the present study indicate that serum p53 antibodies may be an independent prognostic factor in NSCLC, especially in the squamous cell carcinoma (SqCC) patients and may be useful in identifying resected lung cancer patients at high risk for treatment failure.

Preoperative CYFRA 21-1 level as a prognostic indicator in resected nonsmall cell lung cancer

Cytokeratin 19 is particularly abundant in carcinoma of the lung. The CYFRA 21-1 assay has recently been developed for detection of a cytokeratin 19 fragment in serum. In the current study, the prognostic information provided by the CYFRA 21-1 assay in operable nonsmall cell lung cancer (NSCLC) was analysed. Serum levels of CYFRA 21-1 were measured using an immunoradiometric assay (DiaSorin) in 94 patients with operable NSCLC. Survival and disease-free survival curves related to initial levels of this marker were estimated using the Kaplan-Meier method. Elevated preoperative CYFRA 21-1 levels were identified in 42% of patients with NSCLC. The number of patients with elevated levels of this marker increased with tumour node metastasis (TNM) stage (p=0.02). In univariate analysis elevated levels of CYFRA 21-1 were significantly associated with poor overall survival (p<0.001) and with disease-free survival (p<0.001). The results remained significant when the comparisons were adjusted, using the stratified log-rank test, for patients TNM stage (p<0.001 for both overall and disease-free survival). Elevated preoperative levels of CYFRA 21-1 decreased the probability of survival or surviving without recurrence 15 months or more after the operation. This was confirmed by the results of the multivariate analysis. In conclusion, CYFRA 21-1 may be an independent prognostic parameter of survival and tumour relapse in nonsmall cell lung cancer and may be useful in identifying resected cancer patients at high risk for treatment failure.

Serum vascular endothelial growth factors C and D in patients with oesophageal cancer.

OBJECTIVE Lymph node metastasis is a characteristic of malignant cancers and is observed more frequently in oesophageal cancer than in other digestive tract cancers, making it one of the most important prognostic factors. Vascular endothelial growth factors C (VEGF-C) and D (VEGF-D) are important lymphangiogenic factors in human cancers and lymphangiogenesis is associated with lymph node metastasis. The aim of the study was to determine the correlation between pre-treatment serum levels of VEGF-C (sVEGF-C) and VEGF-D (sVEGF-D) and clinicopathologic features in patients with oesophageal cancer. METHODS Serum VEGF-C and sVEGF-D were measured by enzyme-linked immunoadsorbent assay (ELISA) on 149 patients with oesophageal cancer, 29 patients with benign oesophageal diseases and 30 healthy controls. RESULTS Serum VEGF-C and sVEGF-D levels were significantly higher in patients with oesophageal carcinoma than in the control group (p<0.001 and p=0.001, respectively) or in the benign oesophageal diseases group (p=0.04 and p=0.03, respectively). Subgroup analysis showed that lymph node metastasis (p=0.001), stage (p=0.001), tumour depth (p=0.006), resectability (p=0.002), tumour size (p=0.01), distant metastases (p=0.01) and histological grading (p=0.04) were correlated with an elevated level of sVEGF-C. Elevated levels of sVEGF-D were associated with tumour depth (p=0.002), stage (p=0.01) and lymph node metastasis (p=0.02). Among the patients (n=83) who underwent potentially curative surgery, the overall survival time (p=0.008) was shorter for patients with a high level (>8667 pg ml(-1)) of sVEGF-C than for those with a low level (<8667 pg ml(-1)), when the cut-off value was determined on the basis of the median value in oesophageal cancer patients. On univariate regression analysis, tumour size, tumour depth, stage, lymph node metastases, distant metastases, resectability and sVEGF-C were found to be significant prognostic factors. CONCLUSIONS These results suggest that pre-treatment levels of sVEGF-C and sVEGF-D reflect lymph node metastases and advanced stage of oesophageal cancer. Serum VEGF-C may be useful in predicting poor outcome for patients undergoing a potentially curative oesophagectomy.

The preoperative study of mediastinal lymph nodes metastasis in lung cancer by endoscopic ultrasonography (EUS) and helical computed tomography (CT)

OBJECTIVE Accurate staging of mediastinal lymph nodes in patients with lung cancer is fundamental for their treatment and prognosis. The aim of this study was to compare the value of EUS and CT staging in patients with non-small-cell lung cancer (NSCLC) with postsurgical stage. METHODS Ninety two patients with NSCLC underwent EUS and CT for preoperative detection of metastases to the mediastinal lymph nodes. EUS examinations were done with the ultrasonic linear array scanning echoendoscope (FG 32 UA, Hitachi/Pentax), CT-Toshiba Exvision GX scanner, with 24-s spiral acquisition, pitch 1:1 (7 mm collimation, 4 mm reconstruction index), during i.v. administration of non-ionic iodinated contrast media. RESULTS The frequency of mediastinal involvement was 22.7%. The regions most accessible by EUS evaluation were subaortic, subcarinal and paraoesophageal lymph nodes. On a per-patient basis, EUS and CT results were: sensitivity 70.0 and 60.0%, specificity 80.6 and 72.6%, accuracy 77.2 and 68.5%. On a per-sites basis, the sensitivity of EUS evaluation was 78.8%, specificity 89.9%, accuracy 87.7%, comparing with CT-63.6, 84.0, 79.9%, respectively. When the EUS and CT images were analysed in combination, the sensitivity increased to 86.4%. CONCLUSION We believe that EUS and CT should be used together for preoperative non-invasive staging of mediastinal lymph nodes in patients with NSCLC.

Ligands at β2-, β3-, and the Low-affinity State of β1-adrenoceptors Block the α1-adrenoceptor-mediated Constriction in Human Pulmonary and Rat Mesenteric Arteries

We examined whether the β2-adrenoceptor agonists fenoterol and salbutamol, the β3-adrenoceptor agonists CL 316243 and ZD 2079, and the agonists of the low-affinity state of β-adrenoceptors, cyanopindolol and CGP 12177 block α1-adrenoceptors in that concentration range in which they relax the human pulmonary and rat mesenteric arteries preconstricted with phenylephrine 10 μM and 1 μM, respectively. For quantification of vasodilatation pEC25 values and for the antagonism toward α1-adrenoceptors, pA2 values were determined. We found that in the rat mesenteric artery, (1) the pEC25 values of the β-adrenoceptor ligands resemble their respective pA2 values (difference ≤ 0.9 log units), and (2) the order of potencies is the same for both parameters, ie, cyanopindolol ∼ fenoterol > CGP 12177 > salbutamol > ZD 2079 > CL 316243. In the human pulmonary artery, (1) the pEC25 values are slightly lower (by 0.6-1.3 log units) than their respective pA2 values, and (2) the rank order of potencies is the same for both parameters. In conclusion, the present study suggests that ligands of β2-adrenoceptors and of non-β1-non-β2-adrenoceptors relax rat and human vessels preconstricted with phenylephrine or norepinephrine mainly through their α1-adrenolytic effects. Hence, for the investigation of the role of β-adrenoceptors in vessels, the constrictor agent should be chosen with great caution.

Lymphatic vessel invasion detected by the endothelial lymphatic marker D2-40 (podoplanin) is predictive of regional lymph node status and an independent prognostic factor in patients with resected esophageal cancer.

The discovery of markers to lymphatic endothelial cells and the development of novel antibodies to these markers have brought increasing attention to the lymphatics and progress in the understanding of lymphangiogenesis and cancer metastasis. In this study, we investigate the presence of lymphatic vessel invasion (LVI) detected by D2-40 immunohistochemical staining in resected esophageal cancer and correlated with clinicopathologic data and patient survival. Sixty nine patients, who had a primary resection of esophageal cancer, were analyzed by univariate and multivariate logistic regression, and univariate and multivariate survival analysis. The total rate of LVI was 72% (50/69). Positive LVI was significantly correlated with lymph node metastasis (p < 0.001), tumor size (p < 0.001), histological grading (p = 0.017), tumor depth (p = 0.001), and stage (p < 0.001). Multivariate logistic analysis identified LVI (p = 0.036) as a predictor of regional lymph node metastasis. On univariate survival analysis, patients with LVI had a significantly shorter disease-free survival, cancer-specific survival and overall survival. Multivariate analysis proved that LVI diagnosed by D2-40 is an independent prognostic factor of both disease-free survival (p = 0.04) and overall survival (p = 0.032) in resected esophageal cancer. These results show that LVI assessment identifies patients at high risk for regional lymph node metastasis and that LVI is an independent prognostic factor in patients with esophageal cancer.

Clinical significance of serum macrophage-colony stimulating factor (M-CSF) in esophageal cancer patients and its comparison with classical tumor markers.

Abstract Background: Hematopoietic growth factors (HGFs), such as macrophage-colony stimulating factor (M-CSF) are produced aberrantly in many malignancies. Esophageal cancer (EC) is characterized by late diagnosis, rapid progression and poor survival. The purpose of the present study was to determine the clinical usefulness of M-CSF in the diagnosis of EC and its histological type – squamous cell cancer of the esophagus (ESCC). Methods: The study included 80 patients with EC (54 with ESCC and 26 with adenocarcinoma of the esophagus) and 30 healthy subjects. The serum concentrations of M-CSF, carcinoembryonic antigen (CEA) and squamous cell cancer antigen (SCC-Ag) were determined using immunoenzyme assays. We defined the percentage of increased concentrations of proteins tested and the areas under the receiver-operating characteristics (ROC) curves. Results: In EC and ESCC patients, serum concentrations of M-CSF, CEA and SCC-Ag were significantly higher compared with healthy subjects. Concentrations of M-CSF in both groups analyzed (EC and ESCC) showed a tendency for increases with depth of tumor invasion, the presence of lymph node and distant metastases. The combined use of M-CSF with SCC-Ag increased their percentage of increased concentrations, and this value was higher than for classical tumor markers in both groups analyzed. The area under ROC curve for M-CSF was larger than for CEA in EC patients. Conclusions: Our findings suggest the potential usefulness of serum M-CSF concentrations as a tumor marker for EC, especially in combination with SCC-Ag. However, the diagnostic value of this cytokine may be limited because of its non-specific nature. Clin Chem Lab Med 2010;48:1467–73.