Novel methylation panel for the early detection of colorectal tumors in stool DNA.

BACKGROUND: Previous studies showed that the assessment of promoter hypermethylation of a limited number of genes in tumor biopsies may identify the majority of colorectal tumors. This study aimed to assess the clinical usefulness of a panel of methylation biomarkers in stool DNA in the identification of colorectal tumors, using methylation-specific melting curve analysis (MS-MCA), a technique that simultaneously analyzes all cytosine-phosphate-guanine (CpG) residues within a promoter. MATERIALS AND METHODS: The promoter methylation status of 4 tumor-related genes (RARB2, p16INK4a, MGMT, and APC) was analyzed in DNA stool samples and corresponding tissues in an initial set of 12 patients with newly diagnosed primary colorectal carcinomas and 20 patients with newly diagnosed colorectal adenomas, using methylation-specific polymerase chain reaction. Results were replicated in a set of 82 patients (20 healthy subjects, 16 patients with inflammatory bowel disease (IBD), 20 patients with adenomas, and 26 patients with carcinomas), using MS-MCA analyses. RESULTS: In the initial set, >or= 1 positive methylation marker was detected in the stools of 9 of 12 patients (75%) with carcinomas and 12 of 20 patients (60%) with adenomas, with no false-positive results. Stool analyses missed 7 methylated lesions (25%). In the replication set, stool DNA testing detected 16 of 26 carcinomas (62%) and 8 of 20 adenomas (40%). The MS-MCAs missed 14 methylated tumors (37%). No aberrant methylation was evident in healthy subjects, but the RARB2 marker was positive in 2 of 15 stool samples (13%) of patients with IBD. CONCLUSION: Analysis via MS-MCA of a panel of methylation markers in stool DNA may offer a good alternative in the early, noninvasive detection of colorectal tumors.

[Prediction of prognosis of patients with pancreatic adenocarcinoma with curative intent resection by means of histologic grade and pathologic N stage]. / Predicción del pronóstico de los pacientes con adenocarcinoma pancreático resecado con intención curativa mediante el grado histológico y el estadio N patológico.

BACKGROUND AND OBJECTIVE: Pancreatic cancer has the poorest prognosis of any common gastrointestinal malignancy, with a 5-year overall survival of less than 5%. A better knowledge of prognostic factors related to this neoplasia might help improve the survival of these patients. We evaluated the prognostic significance of different factors in both overall survival and tumor recurrence in patients with pancreatic adenocarcinoma who had undergone pancreatic resection with curative intent. PATIENTS AND METHOD: All patients with pancreatic adenocarcinoma submitted to surgical resection in our unit from January 1995 to February 2005 were evaluated. Twenty-three pre-surgical, therapeutic, and histopathologic variables were analyzed. Univariate (Kaplan-Meier, log-rank test) and multivariate (Cox regression) analyses were performed to select independent prognostic factors. RESULTS: Ninety-four patients were evaluated. The median age of patients was 63 years and 53% were woman. The probability of overall survival was 63% at 1 year, 18% at 3 years, and 8% at 5 years, with a median survival of 18 months. Univariate analysis identified performance of adjuvant therapy, histologic grade, percentage of involved-resected lymph nodes, pathologic N stage, and pathologic TNM stage as variables associated with overall survival. On the other hand, the probability of tumor recurrence was 52% at 1 year, 83% at 3 years, and 91% at 5 years, with a median time to tumor recurrence of 12 months. Predictive variables of tumor recurrence in the univariate analysis were preoperative N stage, preoperative TNM stage, postoperative CA 19.9 serum concentration, histological grade, percentage of involved-resected lymph nodes, pathologic N stage and pathologic TNM stage. Multivariate analysis identified histological grade and pathologic N stage as independent predictive factors of both overall survival (histologic grade: HR=2.341 [CI 95%, 1.342-4.098; p=0.003]; pathologic N stage: HR=2.242 [1.213-4.149; p=0.01]) and tumor recurrence (histological grade: HR=1.742 [CI 95%, 1.121-3.086; p=0.05]; pathologic N stage: HR=2.096 [1.089-4.032; p=0.027]). CONCLUSIONS: The histological grade and pathologic N stage predict the prognosis of patients with pancreatic adenocarcinoma after surgical resection.

Predicción del pronóstico de los pacientes con adenocarcinoma pancreático resecado con intención curativa mediante el grado histológico y el estadio N patológico / Prediction of prognosis of patients with pancreatic adenocarcinoma with curative intent resection by means of histologic grade and pathologic N stage

Fundamento y objetivo: el cáncer de páncreas tiene el peor pronóstico de los tumores gastrointestinales, con una supervivencia media a los 5 años de menos del 5%. Un mejor conocimiento de los factores pronósticos de esta neoplasia podría ayudarnos a mejorar la supervivencia de estos pacientes. El objetivo de este estudio fue evaluar el significado pronóstico de diferentes factores en relación tanto con la supervivencia general como con la recurrencia tumoral en pacientes con adenocarcinoma pancreático intervenidos con intención curativa. Pacientes y método: se ha evaluado a todos los pacientes con adenocarcinoma pancreático intervenidos en nuestra unidad desde enero de 1995 a febrero de 2005. Se analizaron 23 variables prequirúrgicas, terapéuticas e histopatológicas. Para seleccionar los factores pronósticos independientes se realizó un análisis univariable (con curvas de Kaplan-Meier y prueba de rangos logarítmicos) y otro multivariable (regresión de Cox). Resultados: se evaluó a 94 pacientes. La mediana de edad de los pacientes fue 63 años y el 53% eran mujeres. La supervivencia general fue del 63% al año, del 18% a los 3 años y del 8% a los 5 años, con una mediana de supervivencia de 18 meses. El análisis univariable identificó como variables relacionadas con la supervivencia general el grado histológico, el porcentaje de ganglios afectados de los que fueron resecados, el estadio N patológico y el estadio TNM patológico. Por otro lado, la probabilidad de recurrencia tumoral fue del 52% al año, del 83% a los 3 años y del 91% a los 5 años, con una mediana para la recurrencia tumoral de 12 meses. Las variables predictivas de recurrencia tumoral en el análisis univariable fueron el estadio N preoperatorio, el estadio TNM preoperatorio, la concentración sérica postoperatoria de CA19.9, el grado histológico, el porcentaje de ganglios afectos de los que fueron resecados, el estadio N patológico y el estadio TNM patológico. El análisis multivariable identificó el grado histológico y el estadio N patológico como los factores predictivos independientes tanto de la supervivencia general (grado histológico: hazard ratio [HR]=2,341; intervalo de confianza[IC] del 95%, 1,342¿4,098; p=0,003; estadio N patológico: HR=2,242; IC del 95%, 1,213¿4,149; p=0,01) como de la recurrencia tumoral (grado histológico: HR=1,742; IC del 95%, 1,021¿3,086; p=0,05; estadio N patológico: HR=2,096; IC del 95%, 1,089¿4,032; p=0,027). Conclusiones: el grado histológico y el estadio N patológico predicen el pronóstico de los pacientes con adenocarcinoma pancreático después de la resección quirúrgica (AU)

Background and objective: Pancreatic cancer has the poorest prognosis of any common gastrointestinal malignancy, with a 5-year overall survival of less than 5%. A better knowledge of prognostic factors related to this neoplasia might help improve the survival of these patients. We evaluated the prognostic significance of different factors in both overall survival and tumor recurrence in patients with pancreatic adenocarcinoma who had undergone pancreatic resection with curative intent. Patients and method: All patients with pancreatic adenocarcinoma submitted to surgical resection in our unit from January 1995 to February 2005 were evaluated. Twenty-three pre-surgical, therapeutic, and histopathologic variables were analyzed. Univariate (Kaplan-Meier, log-rank test) and multivariate (Cox regression) analyses were performed to select independent prognostic factors. Results: Ninety-four patients were evaluated. The median age of patients was 63 years and 53% were woman. The probability of overall survival was 63% at 1 year, 18% at 3 years, and 8% at 5 years, with a median survival of 18 months. Univariate analysis identified performance of adjuvant therapy, histologic grade, percentage of involved-resected lymph nodes, pathologic N stage, and pathologic TNM stage as variables associated with overall survival. On the other hand, the probability of tumor recurrence was 52% at 1 year, 83% at 3 years, and 91% at 5 years, with a median time to tumor recurrence of 12 months. Predictive variables of tumor recurrence in the univariate analysis were preoperative N stage, preoperative TNM stage, postoperative CA 19.9 serum concentration, histological grade, percentage of involved-resected lymph nodes, pathologic N stage and pathologic TNM stage. Multivariate analysis identified histological grade and pathologic N stage as independent predictive factors of both overall survival (histologic grade: HR=2.341 [CI 95%, 1.342¿4.098; p=0.003]; pathologic N stage: HR=2.242 [1.213¿4.149; p=0.01]) and tumor recurrence (histological grade: HR=1.742 [CI 95%, 1.121¿3.086; p=0.05]; pathologic N stage: HR=2.096 [1.089¿4.032; p=0.027]). Conclusions: The histological grade and pathologic N stage predict the prognosis of patients with pancreatic adenocarcinoma after surgical resection (AU)

Positive VEGF immunostaining independently predicts poor prognosis in curatively resected gastric cancer patients: results of a study assessing a panel of angiogenic markers.

Angiopoietin-2 (Ang-2) and vascular endothelial growth factor (VEGF) contribute to gastric cancer aggressiveness by up-regulating the expression of proteases. We evaluated the expression and the prognostic significance of angiogenic factors and proteases in 148 patients with R0-resected gastric cancer. Expression of VEGF, Ang-2, cyclooxygenase-2 (COX-2), urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1, matrix metalloproteinases (MMP)-1 and -9 were assayed by immunohistochemistry. After a mean of 63 +/- 4 months, 81 out of 148 patients had died due to disease. The probability of being free of recurrence was 62, 48, and 42% at 2, 5, and 10 years, respectively. Single bivariate analysis identified VEGF, Ang-2, COX-2, PAI-1, and MMP-9 expression, along with several clinicopathological parameters (grade of curability, lymph node ratio, pTNM, pT, pN), as variables associated with both decreased disease-specific survival and recurrence. On multivariate analysis, after adjusting for significant clinical covariables, positive VEGF immunostaining was the primary prognostic factor, and no other tumor marker variable could add any significant improvement for the prediction, for both disease-specific survival (p = 0.001; HR, 3.27; 95% CI, 1.76 to 6.10) and tumor recurrence (p = 0.002; HR, 2.81; 95% CI, 1.48 to 5.35). Our study suggests that VEGF alone may be clinically useful for establishing therapeutic decisions in gastric cancer patients.

[Current status of treatment of inflammatory bowel disease]. / Estado actual del tratamiento de la enfermedad inflamatoria intestinal.

Inflammatory bowel disease encompasses a group of diseases with poorly defined etiology that affect the digestive tract. These diseases are characterized by their chronic course and by periods of disease activity, of variable severity, that alternate with periods of clinical remission. In the last few years, inflammatory bowel disease has been the object of intense research, which has increased knowledge of the physiopathogenic mechanisms involved. This has enabled the development of a new generation of biotechnological drugs effective in patients previously considered to be refractory to medical treatment and has allowed the accumulated corticosteroid dose to be reduced and the indications for surgery and hospital admissions to be decreased, thus improving quality of life. In addition, some classical drugs have been demonstrated to be effective in recurrence prevention after surgery for Crohn's disease and in the prevention of dysplasia and colorectal cancer in inflammatory bowel disease.

Estado actual del tratamiento de la enfermedad inflamatoria intestinal / Current status of treatment of inflammatory bowel disease

La enfermedad inflamatoria intestinal engloba un grupo de enfermedades del tracto digestivo de etiología todavía no bien conocida, que se caracterizan por su curso crónico y por la alternancia de períodos de actividad, de gravedad variable, con períodos de remisión clínica. Durante los últimos años, la enfermedad inflamatoria intestinal ha sido objeto de una intensa investigación, circunstancia que ha promovido un mejor conocimiento de sus mecanismos fisiopatogénicos. Este hecho ha permitido el desarrollo de una nueva generación de fármacos biotecnológicos capaces de controlar a pacientes considerados previamente refrac-tarios al tratamiento médico, reducir la dosis acumulada de corticoides, y disminuir las indicaciones de cirugía y los ingresos hospitalarios, con un incremento de la calidad de vida. Por otro lado, se ha demostrado la eficacia de determinados fármacos clásicos en la prevención de la recurrencia tras la cirugía de la enfermedad de Crohn, así como su utilidad en la prevención de la displasia y el cáncer colorrectal en los pacientes con enfermedad inflamatoria intestinal (AU)

Inflammatory bowel disease encompasses a group of diseases with poorly defined etiology that affect the digestive tract. These diseases are characterized by their chronic course and by periods of disease activity, of variable severity, that alternate with periods of clinical remission. In the last few years, inflammatory bowel disease has been the object of intense research, which has increased knowledge of the physiopathogenic mechanisms involved. This has enabled the development of a new generation of biotechnological drugs effective in patients previously considered to be refractory to medical treatment and has allowed the accumulated corticosteroid dose to be reduced and the indications for surgery and hospital admissions to be decreased, thus improving quality of life. In addition, some classical drugs have been demonstrated to be effective in recurrence prevention after surgery for Crohn's disease and in the prevention of dysplasia and colorectal cancer in inflammatory bowel disease (AU)

Effect of cyclosporin A on cell adhesion molecules and leukocyte-endothelial cell interactions in experimental colitis.

BACKGROUND: Cyclosporin A (CsA) is an immunosuppressive agent that is believed to act primarily through effects on T-helper lymphocyte function and proliferation. The aim of this study was to investigate whether modulation of leukocyte recruitment and expression of cell adhesion molecules contribute to the therapeutic efficacy of CsA in a model of experimental colitis. METHODS: The therapeutic effects of CsA were assessed in mice with dextran sulfate sodium-induced colitis. Leukocyte-endothelial cell interactions were determined in colonic venules by intravital microscopy. The expression of cell adhesion molecules intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and mucosal addressin cell adhesion molecule 1 (MAd-CAM-1) was measured by the radiolabeled antibody technique. RESULTS: Treatment with CsA (4 mg/kg/day) significantly improved the clinical course of colitis, decreasing weight loss, diarrhea, rectal bleeding, disease activity index, colon weight, and colonic shortening. Microscopic damage score, myeloperoxidase activity, tumor necrosis factor alpha (TNF-alpha), and interleukin-6 in colonic tissue were significantly diminished by CsA. CsA also significantly reduced ICAM-1 and VCAM-1, but not MAdCAM-1, expression in colitic mice. TNF-alpha-induced ICAM-1 and VCAM-1 expression in primary cultures of human umbilical vein endothelial cells was reduced by co-incubation with CsA. The reduction in adhesion molecule expression was followed by a marked decrease in leukocyte adhesion in colonic venules of colitic mice. CONCLUSIONS: CsA ameliorates experimental colitis in mice. Reduced adhesion molecule expression resulting from diminished pro-inflammatory cytokine production and from a direct effect of CsA in endothelial cells decreases leukocyte recruitment into the inflamed intestine, contributing to this protective effect.

Trefoil peptide TFF2 treatment reduces VCAM-1 expression and leukocyte recruitment in experimental intestinal inflammation.

There is evidence for a beneficial effect of trefoil peptides in animal models of gastric damage and intestinal inflammation, but the optimal treatment strategy and the mechanistic basis have not been explored thoroughly. It has been suggested that these proteins may modulate the inflammatory response. The aims of this study were to compare the protective and curative value of systemic and topical trefoil factor family (TFF)2 administration in dextran sulfate sodium-induced experimental colitis and to investigate the relationship between the therapeutic effects of TFF2 and modulation of leukocyte recruitment and expression of cell adhesion molecules. Clinical and morphologic severity of colitis was evaluated at the end of the study (Day 10). Leukocyte-endothelial cell interactions were determined in colonic venules by fluorescence intravital microscopy. The expression of cell adhesion molecules vascular cell adhesion molecule 1 (VCAM-1) and mucosal addressin cell adhesion molecule 1 (MAdCAM-1) was measured by the dual radiolabeled monoclonal antibody technique. Pretreatment with TFF2 by subcutaneous or intracolonic (ic) route ameliorated the clinical course of colitis, and the luminal route had a significantly superior effect. This beneficial effect was correlated with significant reductions in endothelial VCAM-1 but not MAdCAM-1 expression and leukocyte adhesion to intestinal venules, which returned to levels similar to those of controls. In established colitis, ic TFF2 treatment did not modify the severity of colonic lesions. In conclusion, TFF2 is useful in the treatment of colitis, and topical administration is superior to the systemic route. Reduction in adhesion molecule expression and leukocyte recruitment into the inflamed intestine contributes to the beneficial effect of this treatment.