Miguel Morell

Overexpression of Clusterin in Human Breast Carcinoma

Clusterin has been implicated in numerous processes including active cell death, immune regulation, cell adhesion and morphological transformation. The purpose of this study was to examine clusterin expression in a large series of breast carcinomas by immunohistochemistry and in situ hybridization. The study included 40 samples of non-neoplastic glandular epithelia, 42 benign lesions, 15 atypical intraductal hyperplasias, 35 carcinomas in situ, 114 invasive carcinomas, and lymph node metastases from 40 patients. Epithelial normal cells were always negative for clusterin expression and only 19% of the benign lesions presented positive staining. In contrast to the benign lesions, however, the frequency of clusterin positive samples increased in atypical hyperplasias (47%, P = 0.08), intraductal carcinomas (49%, P = 0.01) and invasive carcinomas (53%, P < 0.001). Positive staining presented a cytoplasmic pattern, except in 3 cases of invasive carcinomas which had nuclear staining. Clusterin mRNA by in situ hybridization confirmed the specific cellular pattern of clusterin expression by immunohistochemistry. Clusterin expression was associated with large tumor size (P = 0.04), estrogen and progesterone receptor negative status (P = 0.02 and P = 0.001, respectively) and with the progression from primary carcinoma to metastatic carcinoma in lymph nodes (80% metastatic nodes had positive expression) (P = 0.004). Ten of 15 (67%) primary carcinomas without clusterin expression became positive in lymph node metastases, while most (22 of 25, 88%) of the clusterin-positive primary carcinomas were also immunoreactive in metastases. In survival analysis, clusterin expression did not represent a prognostic indicator by uni- or multivariate analysis. The increased clusterin expression in breast carcinomas tended to correlate inversely with the apoptotic index (P = 0.09) which indicates that clusterin gene expression is not a prerequisite to cellular death by apoptosis. From these results, we suggest that clusterin may have a role in tumorigenesis and progression of human breast carcinomas.

Expression of the Antiapoptotic Proteins Clusterin and Bcl-2 in Laryngeal Squamous Cell Carcinomas

Bcl-2 and clusterin genes have been related to the inhibition of apoptosis, an event that plays a key role in malignant transformation and in invasive disease. In this work, we determine the significance of clusterin and bcl-2 expression in a large series of laryngeal carcinomas. We used immunohistochemical methods and in situ hybridization to examine the expression of these proteins. Nontumoral epithelial laryngeal tissues did not express clusterin and bcl-2 proteins. However, 9% (14 out of 154) and 25% of these tumors (39 of 154) had positive clusterin and bcl-2 staining, respectively. Clusterin expression was significantly related to the degree of local invasion and higher bcl-2 expression was found in these clusterin-positive tumors (p < 0.05). Bcl-2 expression was significantly correlated with supraglottic localization, nodal metastases, invasion in depth, and poorly differentiated tumors. However, by multivariate analysis, bcl-2 was shown to be an independent predictor of good prognosis in these tumors (OR = 0.12, 95% CI = 0.02–0.91). These findings indicate that clusterin and bcl-2 are upregulated in laryngeal carcinomas and their expression is related to the invasiveness of these tumors.

Expression of bax and p53 Proteins in the Tumorigenesis and Progression of Breast Carcinomas

Objectives: Dysregulation of normal programmed cell death mechanisms plays an important role in the pathogenesis of breast cancer. The purpose of this study was to investigate the role of bax and p53 expression in tumorigenesis and progression of breast carcinoma as well as their relationship with proliferative and apoptotic activity. Methods: We used immunohistochemical methods and in situ detection of apoptotic cells to examine 30 carcinomas in situ (CIS), 131 invasive breast carcinomas and 45 lymph node metastases. Results: In 25% (33 of 131) of invasive breast carcinomas examined, bax expression was absent, while p53 accumulation was present in 37% (49 of 131). Interestingly, p53 accumulation and loss of bax expression occur in breast CIS as frequently as in invasive breast carcinoma. Thus, in 17% (5 of 30) of CIS bax expression was absent, and 30% (9 of 30) presented nuclear expression of p53. p53 accumulation was related to apoptosis and proliferative activity. However, the protein level of bax was unrelated to all parameters studied, including proliferation and apoptosis of tumor cells. A multivariate analysis of disease-free survival demonstrated that p53 accumulation and bax expression are not significant independent indicators of prognosis in operable breast carcinoma. Our results also show that the proportion of bax- and p53-positive cells does not vary between primary and metastatic tumors. Conclusions: p53 accumulation and loss of bax expression influence the acquisition of a malignant phenotype but seem to have no further impact on tumor progression.

Síntesis de colágeno e insuficiencia cardíaca

Se desconoce el metabolismo del colageno en la insuficiencia cardiaca con y sin disfuncion sistolica ventricular izquierda. Estudiamos las concentraciones de los marcadores de sintesis de colageno tipo I (peptido C-terminal del procolageno tipo I [PIP]) y de degradacion (telopeptido del colageno tipo I [CITP]) en un grupo de 70 pacientes tras un ingreso por insuficiencia cardiaca (35 con funcion ventricular izquierda deprimida y 35 con funcion ventricular conservada), asi como en 30 individuos sanos. Excluimos a los pacientes con insuficiencia renal, enfermedad hepatica, enfermedad autoinmunitaria o del metabolismo oseo, traumatismo mayor y cirugia reciente (

Collagen Synthesis and Heart Failure

Little is known about collagen metabolism in heart failure, with or without left ventricular systolic dysfunction. We studied serum concentrations of the carboxy-terminal propeptide of procollagen type I (PIP), a marker of collagen type-I synthesis, and of the carboxy-terminal telopeptide of collagen type I (ICTP), a marker of collagen type-I degradation, in 70 patients admitted for heart failure (35 with depressed left ventricular systolic function and 35 with preserved left ventricular systolic function) and in 30 control subjects. Patients with kidney failure, liver disease, metabolic bone disease, rheumatic disease, recent (within 3 months) major trauma or surgery, or serious wounds were excluded. The concentration of the collagen synthesis marker, PIP, was higher in heart failure patients than control subjects, at 140+/-56.38 mg/L vs 113.66+/-36.6 microg/L, respectively (P=.01). However, there was no difference in the concentration of the collagen degradation marker, ICTP, between heart failure patients and control subjects, at 2.89+/-2.37 mg/L vs 2.26+/-1.7 microg/l, respectively. In heart failure patients, left ventricular systolic function had nonsignificant effect on the PIP or ICTP concentration.

El análisis computarizado cromático es un predictor precoz de riesgo cardiovascular asociado a hipercolesterolemia

Introduccion y objetivos La deteccion precoz de la enfermedad cardiovascular es el mayor objetivo de la medicina contemporanea para la prevencion de la enfermedad coronaria. El objetivo de este estudio fue buscar un numero de cambios que pudieran ser detectados en las neuronas del area 19 de Brodman, por medio del analisis computarizado cromatico (ACC), como consecuencia de una disfuncion neurobiologica, que tradujera un fallo en la percepcion del color y que ese fallo, cuantificado numericamente, expresara la existencia de una hipercolesterolemia y, por ende, de riesgo cardiovascular. Pacientes y metodo Estudiamos a 208 pacientes (grupo 1) (153 varones y 55 mujeres) con valores de colesterol total por encima de 200 mg/dl. El grupo control (grupo 2) estaba formado por 208 sujetos (153 varones y 55 mujeres), pero con concentraciones de colesterol total por debajo de 200 mg/dl. Todos los pacientes fueron examinados mediante ACC, descartando antes cualquier enfermedad sistemica u oftalmologica. Resultados Todos los indices de la prueba fueron altamente significativos en ambos grupos. La relacion directa entre la concentracion de colesterol y el area 19 alcanzo una correlacion canonica de 0,825, con una sensibilidad del 90% y una especificidad del 93%. Los resultados de la regresion multiple con el colesterol total, como variable dependiente, y el mas significativo parametro de ACC, como variable independiente, fueron de R = 0,89 (p Conclusiones El estudio determina una fuerte asociacion entre la hipercolesterolemia y las areas visuales de la corteza cerebral.

Estudio comparativo del impacto de la dieta frente a pravastatina en los centros de análisis cromático del área 19 de Brodman, mediante el análisis computarizado cromático (Estudio CARDIOCOLOUR)

Introduccion y objetivos La hipercolesterolemia origina importantes cambios neurodegenerativos en el cortex cerebral, lo que se traduce en un fallo en la vision cromatica, en las neuronas del area 19 de Brodman. Numerosos estudios de intervencion terapeutica para la prevencion primaria y secundaria de la enfermedad coronaria apoyan una reduccion de los acontecimientos cardiacos con la reduccion en el colesterol total o el cLDL. Nuestro objetivo fue elucidar los efectos de la dieta y un inhibidor de la HMG-CoA (pravastatina) sobre la vision de los colores, pero tambien sobre las cifras plasmaticas de colesterol, y correlacionar ambos efectos, mediante el analisis computarizado cromatico (ACC). Pacientes y metodos Estudiamos a 191 pacientes normotensos (133 varones y 58 mujeres), con valores de colesterol total plasmatico por encima de 200 mg/dl. Un total de 70 de estos pacientes fue tratado con la dieta paso II de la American Heart Associaton durante 6 meses. Los restantes 121 fueron tratados con pravastatina, 61 a dosis de 10 mg y los otros 60 con 40 mg. Fueron examinados mediante ACC, descartando previamente cualquier enfermedad sistemica u oftalmologica. Resultados Se logro una recuperacion de la vision cromatica de hasta el 23% con dieta, del 38% con 10 mg/dia de pravastatina y del 92% con 40 mg/dia de pravastatina (p Conclusiones El estudio determina una fuerte asociacion entre la intervencion terapeutica, con dieta o con pravastatina y la mejoria de la vision cromatica.

Serum activity of the key gluconeogenic enzymes in carbon-tetrachloride-induced experimental hepatotoxicity.

Serum activity has been measured in three of the key enzymes in the gluconeogenic pathway in rats subjected to experimental hepatotoxicity after intraperitoneal administration of carbon tetrachloride. The levels of phosphoenolpyruvate carboxykinase (PEPCK) and fructose-1,6-biphosphatase (FBPase) showed a similar behavior to the transaminase (AST and ALT), increasing markedly with respect to the controls at 12 h after administration of the poison, reaching their maximum peak of activity at between 24 and 36 h, and returning to normal values at 96 h. The activity of glucose-6-phosphatase was not significantly modified throughout the treatment. These results seem to demonstrate that the determination of the serum activity of PEPCK and FBPase could be a sensitive and specific marker of hepatic cytolysis.

Estabilidad de NT-proBNP en pacientes con insuficiencia cardiaca sistólica

Los peptidos natriureticos tienen un alto valor diagnostico y pronostico en pacientes con insuficiencia cardiaca, pero desconocemos la estabilidad de sus valores. Determinamos en 2 visitas ambulatorias los valores de N terminal pro-BNP (NT-proBNP) de 30 pacientes consecutivos con insuficiencia cardiaca estable [New York Heart Association (NYHA) II-III] (test 6 min) por disfuncion sistolica [funcion ventricular izquierda deprimida (FEVI)Natriuretic peptides are extremely useful in the diagnosis and prognosis of patients with heart failure. However, it is not clear whether their values are stable. We carried out a prospective study of 30 consecutive ambulatory patients (mean age, 62.6 [12.2] years) with stable systolic heart failure, as determined by the 6-minute walk test, who were in New York Heart Association class II or III and who had a left ventricular ejection fraction <30% (mean ejection fraction, 24.2% [6.68%]). At baseline, the mean N-terminal pro-brain natriuretic peptide (NT-proBNP) level and the mean distance walked in 6 minutes were 2237.3 pg/mL and 348.26 m, respectively. At 3-month follow-up, the corresponding values were 2096.2 pg/mL and 372.05 m, respectively. No significant difference was observed in NT-proBNP level or in distance walked in 6 minutes between baseline and 3 months (P=.8). Overall, there was a good correlation (r=0.94; P< .001) between the plasma NT-proBNP level at baseline and at 3 months in patients with stable chronic heart failure due to systolic dysfunction in New York Heart Association class II or III.

A sensitive enzyme immunoassay for angiotensin II in serum

A sensitive and specific enzyme immunoassay for measuring angiotensin II (AII) has been developed as a convenient alternative to a radioimmunoassay. An antiserum to AII was prepared using AII conjugated by carbodi-imide to rabbit serum albumin, and coated on to microwell plates. The labelled antigen was prepared from AII and horseradish peroxidase using the periodate method. This enzyme immunoassay was a simple two-step procedure: 0.1 ml of AII-extracted plasma was incubated for 1 h at 37 degrees C; and 1 ml of labeled AII was incubated for 1 h at 37 degrees C. Bound horseradish peroxidase activity was then determined using o-phenylenediamine as chromogen by measuring the absorbance at 492 nm. The lower detection limit of the assay was 3.5 pmol l-1. Between- and within-assay RSD values were 8.8-18.3% and 6.9-17%, respectively, for concentrations of 10-40 pmol l-1. The accuracy of the assay, determined by recovery and linearity experiments, was 89-106% for recovery and 91-126% for parallelism. The results obtained by the present ELISA method were well correlated with those obtained by an established radioimmunoassay (n = 10, r = 0.96, intercept = 0.9 and slope = 1.02). This assay is easy to perform, rapid and does not require radioisotopes; thus it could be widely applied in clinical laboratories.