[Genomic profile in high risk neuroblastoma by comparative genomic hybridization]. / Perfil genómico del neuroblastoma de alto riesgo mediante hibridación genómica comparada.

Different subtypes of neuroblastoma (NB) carry associated genetic aberrations that predict their clinical course. Whole chromosome gains are usually associated with early clinical stages and good prognosis, while 1p deletion, 17q gain and MYCN amplification (MNA) are related to advanced stages and poor prognosis. High-risk neuroblastomas (NB-HR) include NB in children aged more than 1 year old, either stage 4 or any stages showing MNA except stage 1. The prognosis of NB-HR patients remains poor, despite aggressive therapy. Only MNA confers poor prognosis. Between January 2000 and February 2005, tumoral specimens from 60 patients with NB-HR were sent to the Spanish Reference Center for NB biological studies. In all cases, MYCN together with 1p36 status was analyzed by fluorescence in situ hybridization (FISH). Comparative genomic hybridization (CGH) was performed in 24 cases. Using FISH we detected 31 MNA cases including 29 with 1p36 deletion; there were 21 cases without MYCN amplification (MNNA) but 7 of these had 1p36 deletion; 8 cases showed MYCN gain (MNG) but 6 of these had 1p36 deletion. CGH showed other chromosomal alterations. Of 11 MNA cases, none had 11q loss and all of them showed 17q gain or 17 disomy. Of the 7 MNNA cases, there were 4 with 11q loss including 2 with 3p loss and all presented 17q gain or 17 disomy. The 6 MNG cases included 4 cases with 11q loss and 5 cases with 17q gain or 17 disomy. Genomic profiling by CGH in NB-HR confirms the interaction among genetic alterations, the prognostic significance of which should be evaluated to establish new treatment criteria.

Perfil genómico del neuroblastoma de alto riesgo mediante hibridación genómica comparada / Genomic profile in high risk neuroblastoma by comparative genomic hybridization

El neuroblastoma presenta alteraciones genéticas que predicen su evolución clínica. Ganancias cromosómicas completas están asociadas a estadios clínicos no avanzados y evolución favorable, mientras que pérdidas de 1p, ganancia de 17q y amplificación del gen MYCN (MNA) son indicativas de estadios clínicos avanzados y pronóstico desfavorable. Son neuroblastomas de alto riesgo (NB-HR) los presentes en niños mayores de un año: estadio 4 o MNA en cualquier estadio de enfermedad, excluyendo estadio 1. El pronóstico de estos enfermos es malo, incluso con tratamientos agresivos. Sólo MNA confiere valor pronóstico negativo. Se remitieron al Centro de Referencia Nacional del neuroblastoma 60 casos de NB-HR. En todos analizamos MYCN y 1p36 con la técnica de hibridación in situ fluorescente (FISH) y en 24 de ellos el perfil genómico con la técnica de hibridación genómica comparada (CGH). Mediante FISH detectamos 31 casos MNA presentando 29 pérdida de 1p36; 21 casos no amplificados (MNNA) con pérdida de 1p36 en 7 casos; y 8 casos con ganancia del gen MYCN (MNG), seis con pérdida de 1p36. Mediante CGH detectamos otros reordenamientos cromosómicos. De 11 casos MNA ninguno presentó pérdida en 11q, todos presentaban disomía del 17 o ganancia 17q. De 7 casos MNNA, cuatro mostraron pérdida de 11q, 2 con pérdida de 3p. Todos mostraron disomía del 17 o ganancia 17q. De 6 casos MNG, cuatro mostraron pérdida de 11q y cinco disomía del 17 o ganancia 17q. El estudio del perfil genómico en NB-HR revela la interacción de alteraciones genéticas cuyo significado pronóstico debe ser evaluado para establecer nuevos criterios terapéuticos

Different subtypes of neuroblastoma (NB) carry associated genetic aberrations that predict their clinical course. Whole chromosome gains are usually associated with early clinical stages and good prognosis, while 1p deletion, 17q gain and MYCN amplification (MNA) are related to advanced stages and poor prognosis. High-risk neuroblastomas (NB-HR) include NB in children aged more than 1 year old, either stage 4 or any stages showing MNA except stage 1. The prognosis of NB-HR patients remains poor, despite aggressive therapy. Only MNA confers poor prognosis. Between January 2000 and February 2005, tumoral specimens from 60 patients with NB-HR were sent to the Spanish Reference Center for NB biological studies. In all cases, MYCN together with 1p36 status was analyzed by fluorescence in situ hybridization (FISH). Comparative genomic hybridization (CGH) was performed in 24 cases. Using FISH we detected 31 MNA cases including 29 with 1p36 deletion; there were 21 cases without MYCN amplification (MNNA) but 7 of these had 1p36 deletion; 8 cases showed MYCN gain (MNG) but 6 of these had 1p36 deletion. CGH showed other chromosomal alterations. Of 11 MNA cases, none had 11q loss and all of them showed 17q gain or 17 disomy. Of the 7 MNNA cases, there were 4 with 11q loss including 2 with 3p loss and all presented 17q gain or 17 disomy. The 6 MNG cases included 4 cases with 11q loss and 5 cases with 17q gain or 17 disomy. Genomic profiling by CGH in NB-HR confirms the interaction among genetic alterations, the prognostic significance of which should be evaluated to establish new treatment criteria

Characterization of a new rat cell line established from 2'AAF-induced combined hepatocellular cholangiocellular carcinoma.

A rat cell line-nominated CC-62 derived from a combined hepatocellular and cholangiocellular carcinoma obtained by administration of 2-acetylaminofluorene to male Wistar rats, has been established. Using light and electron microscopy it was determined that morphologically the tumor consisted of a mixed population of hepatocytes and cholangiolar neoplastic cells, intermingled with small, undifferentiated oval-like cells. The CC-62 line has been maintained through 90 passages in culture adopting a paving stone arrangement. Doubling time at the 12th passage was 23 h. Immunostaining with a panel of antisera was performed to identify the cytological profiles of the cell line. There was no k-ras or p53 expression by immunohistochemistry, and molecular biology failed to detect mutations. Molecular analysis by reverse transcriptase-polymerase chain reaction revealed transcripts for c-met but no expression of HGF messenger ribonucleic acid. Three cell lines cloned from CC-62 showed the same immunohistochemical and molecular pattern as the parental line. Cytogenetic analysis revealed a chromosome number ranging from 74 to 82 with a modal number of 79 but no clonal structural abnormalities were found. Deoxyribonucleic acid ploidy analysis showed an aneuploid peak. CC-62 caused tumors 1 mo after subcutaneous transplantation into nude mice, with morphological patterns of mucosecretory solid and spindle-shaped carcinoma. This cell line is the first established from a primary rat combined hepatocellular and cholangiocellular neoplasm. The resulting cells expressed biological and morphological markers of hepatocytes and cholangiolar cells. Therefore this cell line may contribute to a better understanding of the histogenesis of liver cancer.

Tratamiento del cáncer colorrectal: encuesta de la Sociedad Valenciana de Cirugía / Management of colorectal cancer: a survey by the Surgical Society of Valencia

Introducción. Motivados por conocer la situación actual del tratamiento del cáncer colorrectal en los hospitales de la Comunidad Valenciana, y por encargo de la Sociedad Valenciana de Cirugía, se elaboró una encuesta dirigida a todos los Servicios de Cirugía General y Aparato Digestivo de estos centros, cuyo resultado exponemos. Material y método. Se realizó un análisis retrospectivo mediante encuesta, y se obtuvieron los datos desde el año 1997 hacia atrás, agrupándolos por años naturales. La encuesta abordó siete apartados del tratamiento del cáncer colorrectal (diagnóstico, cirugía programada, cirugía de urgencias, terapéutica adyuvante, enfermedad avanzada, seguimiento y anatomía patológica), y se estudiaron tanto parámetros de estructura como de proceso, así como los resultados de los mismos. Resultados. Se remitieron un total de 20 cuestionarios obteniéndose 17 respuestas (85 por ciento). Sólo 2 hospitales (11,7 por ciento) disponen de ecografía endorrectal. Ninguno realiza por sistema enema de doble contraste. Únicamente en 3 hospitales (17,64 por ciento), el cáncer rectal es tratado por un grupo determinado de cirujanos. Cinco hospitales (29,4 por ciento) realizan con asiduidad el lavado colónico intraoperatorio en la cirugía del cáncer obstructivo. Siete centros (41,1 por ciento) llevan a cabo algún tipo de terapéutica adyuvante preoperatoria en el cáncer rectal, siendo la cifra total de recidivas locales del 11,58 por ciento a los 2 años de seguimiento. No existe ningún protocolo establecido de seguimiento postoperatorio de estos pacientes en 3 hospitales (17,64 por ciento). El número medio de ganglios aislados por pieza quirúrgica es de nueve, y sólo 2 centros (11,7 por ciento) reflejan en sus informes anatomopatológicos la afectación del margen circunferencial. Conclusiones. De los resultados obtenidos en esta encuesta y su posterior comparación con la bibliografía concluimos: a) en el aspecto diagnóstico, se debe mejorar el porcentaje de colonoscopias completas; los enemas opacos, cuando se realicen, deberían llevarse a cabo sistemáticamente mediante la técnica de doble contraste; sería aconsejable implantar la ecografía endorrectal como exploración de rutina para la correcta estadificación del cáncer rectal con el fin de realizar una correcta selección de los pacientes candidatos a terapéutica adyuvante preoperatoria; b) en la cirugía electiva, dado que la cirugía del cáncer rectal depende del cirujano, creemos que debería ser realizada por personal especialmente entrenado; c) respecto a la cirugía del cáncer colorrectal obstructivo, si las condiciones del paciente lo permiten, debería tratarse de aumentar el porcentaje de resecciones con anastomosis primaria, entrenando al equipo quirúrgico en la realización del lavado intraoperatorio; d) se necesitan estudios prospectivos para valorar el régimen terapéutico adyuvante preoperatorio más adecuado; e) es recomendable que los distintos hospitales dispongan de protocolos de seguimiento postoperatorio homogéneos, con la finalidad de uniformizar el control de estos pacientes, además de prestarles un apoyo psicológico y servir de auditoría de sus propios resultados, y f) debe tratarse de que en los informes anatomopatológicos se especifique el margen circunferencial, así como intensificar el aislamiento de ganglios linfáticos a fin de evitar la infraestadificación tumoral (AU)

[Multiple analysis of morphologic factors with prognostic value in transitional cell papillary carcinoma of the bladder. Retrospective study of 571 cases]. / Análisis múltiple de factores morfológicos con valor pronóstico en el carcinoma papilar de células transicionales de vejiga. Un estudio retrospectivo de 571 casos.

A 7-year retrospective study of 571 cases of transitional cell carcinoma of the bladder (1990-1996) was made in order to demonstrate, statistically, the existence of a close relation between the tumoral staging of Jewett and the cytological grading of Mostofi (p < 0.05). From the results obtained we deduce that grades 1 and 2 are usually associated with stage A tumours, whereas grade 3 is associated with stage B. Thereafter all stage B tumours (184 cases) were selected to determine whether or not cytological grading was related to any histological parameter in order to differentiate between superficial (stage B1) and deeply invasive tumours (stage B2). According to results obtained, the existence of a close relation between grading and the way of muscular infiltration in a focal or diffuse manner could be suggested. These data will be correlated to prognosis in a subsequent clinical follow-up. Therefore, we conclude that grades 1 and 2 usually infiltrate the muscular layer of the bladder in a focal manner with better prognosis, while grade 3 tumors infiltrate in a diffuse way with a poor prognosis.

Expression of type I interferon receptor in renal cell carcinoma.

Immunohistochemical analysis of INF-R was performed on 110 renal tumors, 25 peritumoral kidney tissues and 10 lymph node metastases. Pathological material was previously studied and classified according to predominant cell type, stage and grade. A statistical analysis was made in order to determine to what extent the immunoexpression of INF-R differed in relation to the histological variables studied. All peritumoral kidney sections, 89/110 tumors and 9/10 metastases proved positive. Membranous expression was related to clear cell carcinomas. Type I INF-R is expressed in RCC, independent of tumor stage and grade, as well as sex, age and survival. INF-R is widely expressed in RCC in any tumoral type, and its expression is preserved in metastatic disease, which may help to target those patients who could benefit from INF therapy.

Cytogenetic study of a spindle-cell rhabdomyosarcoma of the parotid gland.

The cytogenetic analysis of a spindle-cell rhabdomyosarcoma of the parotid gland in a 6-year-old boy is reported. The tumor cells showed an abnormal karyotype with a hypotriploid modal chromosome number and clonal structural rearrangements affecting chromosomes 1, 8, 12, 21, and 22. The tumor karyotype was: 59, XY, -1, -3, -4, -5, -6, +8, +8, +del(8)(q22q24), -9, -10, del(12)(q13), -15, -16, -17, -18, der(21)t(12;21)(p11;p11), -22, der(22)t(1;22)(q12;p11).

[Neuroblastoma in children under than 1 year of age]. / Neuroblastoma en niños menores de 1 año.

OBJECTIVE: Our objective was to carry out a prospective multicenter study of neuroblastoma patients diagnosed between 0 and 12 months of age. PATIENTS AND METHODS: Diagnostic procedures included histology, catecholamine excretion, bone marrow cytology and MIBG-scan. Staging was evaluated according to the INSS classification. After 1992, Simada criteria were used and also N-myc amplification, DNA index and P-glycoprotein determinations in tumoral tissue. The surgical technique employed and complications derived from it were also evaluated. The patients were treated according to stage with multicenter Spanish protocols N-I-87 and N-II-92. Overall survival and event free survival were calculated by actuarial methods. RESULTS: Between October 1987 and June 1992, a total of 140 infants less than one year of age were registered and diagnosed of neuroblastoma, representing 40% of all neuroblastoma cases. Median age was 0.3 years and 73% were less than 6 months of age at diagnosis. The most frequent stage was 1 (35%) followed by 4-S (20%). The frequency of unfavorable prognostic factors was the following: LDH (21%), NSE (14%), ferritin (18%), Shimada (7%), DNA (35%), NMA (3%), TrakA (23%), P-glycoprotein (19%). Surgery was performed in 133 children: total resection was reported in 94 and > 90% in another 22 cases. Complications attributed to surgery occurred in 12% of the cases. Chemotherapy was given in 73 cases and radiotherapy in 7. The five year total survival is 91% and the event free survival 88%. Survival by stages: Stage 1 = 91%, stage 2A = 88%, stage 2B = 100%, stage 3 = 84%, stage 4 = 56% and stage 4-S = 100%. CONCLUSIONS: 1) The majority of neuroblastoma cases in infants less than one year old are diagnosed before six months of age. 2) For this age group stages 1 and 4-S are the most frequently observed. 3) Unfavorable biological factors are less frequent than for children over one year of age and are associated with disseminated disease (advanced stage). 4) The outcome is excellent, except for stage 4 patients. The cases in stage 1 and 2 may be treated by surgery alone. Chemotherapy may be of benefit for stage 3 patients.

[A glandular-type inverted papilloma of the bladder: apropos a case with an evident chronic irritative factor]. / Papiloma invertido de vejiga tipo glandular: a propósito de un caso con claro factor irritativo crónico.

OBJECTIVES: Inverted papilloma of the bladder is a rare condition that presents in the middle-aged. Its histological features and prognosis are unique and its diagnosis is always by anatomopathology. To date, its histogenesis has not been elucidated. The present study reports an additional case of this uncommon disease entity, which is even more so due to its glandular anatomopathological morphology. METHODS: A case of inverted papilloma of the bladder in a 52-year-old man with a permanent bladder catheter is described. The tumor was located in the posterosuperior bladder wall and had been successfully resected. RESULTS: The histological diagnosis was that of glandular inverted papilloma of the bladder. The patient is asymptomatic at two years follow up. CONCLUSIONS: In our view, inverted papilloma of the bladder results from a previous metaplasia in which the chronic irritative factor plays an important role.

Immunocytochemical typification of mesothelial cells in effusions: in vivo and in vitro models.

We have performed immunocytochemical, immunoelectron microscopy, Western blot, and culture techniques using monoclonal antibodies against cytokeratin, vimentin, and desmin on 17 benign and 20 malignant effusions of pleural and ascitic origin. Triple coexpression of these three antigens was observed in benign reactive mesothelial cells as well as in one case of mesothelioma. All metastatic adenocarcinoma cells were consistently negative to desmin and positive to cytokeratin and vimentin. Present results were helpful to distinguish reactive and malignant mesothelioma from metastatic carcinoma cells in effusions.

[The determination of tissue CEA in colorectal adenocarcinomas: an immunohistochemical study]. / Determinación del CEA tisular en adenocarcinomas colorrectales: un estudio inmunohistoquímico.

We carried out a CEA immunohistochemical study on 80 colorectal carcinomas, using the PAP methodology. Also we studied peritumoral "normal" mucosa on 69 cases. All tumoral (80/80) and the major part of normal colonic mucosa (68/69) cases stained positively. We present our experience with this immunohistochemical staining using a qualitative semi-quantitative evaluation, as an easy and reliable procedure. This permits to obtain, by immunostaining, a better homogeneous group of tumours (Apical, Mixed and Cytoplasmic, or Weak and Strong), necessary to further correlation with various tumoral parameters. On the basis of this evaluation, we found: A staining of weak intensity (65/68) of Apical type (55/68) in the vast majority of the normal, peritumoral mucosa. In tumors we found a prevalence of strong intensity (67/80), in relation to its major content of CEA. With respect to the type of immunostaining, although Apical staining (32/80) exists the Cytoplasmic (34/80) is predominant together with the Mixed type (14/80). This is expression of the alterations in secretion and distribution of the tissue CEA. We analyze the difficulties of such classification caused by the tumoral heterogeneity and we include other data whose significance is not always clear.

[A correlation between serum levels of preoperative CEA and CEA immunohistochemical staining in colorectal carcinoma]. / Correlación entre niveles séricos de CEA preoperatorios y tinción inmunohistoquímica del CEA en el carcinoma colorrectal.

We analyzed the distribution of tissue CEA in 80 colorectal adenocarcinomas with the PAP immunohistochemical technique. We used a qualitative method with a double grading criterion--topography and intensity of staining--as well as a semiquantitative method in the immunostaining interpretation. We applied a pattern of immunostaining: apical, cytoplasmic or mixed, to each tumor. Likewise, we obtained the pre-operatory serum levels of CEA. The normal value in our laboratory is less than 10 ng/ml. We correlated the immunostaining pattern with the serum levels of CEA, obtaining a global statistical significant correlation (p < 0.01), as well as apical versus cytoplasmic correlation (p = 0,0,3). The apical staining pattern agreed with this CEA levels < 10 ng/ml, whereas the cytoplasmic staining was associated with high frequency with CEA levels > 10 ng/ml. In conclusion the immunohistochemical staining for tissular CEA permits to improve the prognostic efficiency of serum CEA levels.