[Hormone-dependence and C-ERBB2: relationship between estrogen receptor expression, estrogen-regulated proteins, and C-ERBB2 in breast carcinoma]. / Ormono-dipendenza e C-ERBB2: rapporto tra espressione dei recettori estrogenici, estrogen regulated proteins e C-ERBB2 nel carcinoma della mammella.

We reviewed 1491 consecutive cases of operated breast cancer between 1999-2004, and found that hormone-dependence, particularly if evaluated with functional markers of the estrogen receptor (estrogen regulated proteins, ERP), is inversely proportional to antigen neu expression. Our data confirms that ERP can give additional information on the probability of response to hormonal therapy.

[Predictive variables of lymphatic metastasis in breast carcinoma with a diameter below 2 cm]. / Variabili predittive di metastasi linfonodali nei carcinomi mammari di diametro inferiore a cm 2.

The axillary lymph node status of patients with newly diagnosed breast cancer remains the most important prognostic information available at the moment. However, only a minority of patients presents with such node metastases at diagnoses. We reviewed our database and studied 500 consecutive patients with early breast cancer, and found that age inferior to 50 years, high grade, diameter superior to 1 cm, elevated Ki-67, and expression of oncogene p-53 are all factors associated with lymph node metastases.

Optimisation of isolation of richly pure and homogeneous primary human colonic smooth muscle cells.

BACKGROUND: Inherent properties of gastrointestinal smooth muscle can be assessed using isolated cell suspensions. Currently available isolation techniques, based on short 2-h enzymatic digestion, however, present the disadvantage of low cellular yield with brief viability. These features are an important limiting factor especially in studies in humans in which tissue may not be available daily and mixing of samples is not recommended. AIMS: To optimise the isolation procedure of cells from human colon to obtain a richly pure primary smooth muscle cell preparation. METHODS: Slices of circular muscle layer, obtained from surgical specimens of human colon, were incubated overnight in Dulbecco's modified eagle's medium supplemented with antibiotics, foetal bovine serum, an ATP-regenerating system and collagenase. On the following day, digested muscle strips were suspended in HEPES buffer, and spontaneously dissociated smooth muscle cells were harvested and used either immediately or maintained in suspension for up to 72 h. Cell yield, purity, viability, contractile responses, associated intracellular calcium signals and RNA and protein extraction were evaluated and compared to cell suspensions obtained with the current short digestion protocol. RESULTS AND CONCLUSION: The overnight isolation protocol offers the advantage of obtaining a pure, homogeneous, long-life viable cell suspension that maintains a fully differentiated smooth muscle phenotype unchanged for at least 72 h and that allows multiple functional/biochemical studies and efficient RNA extraction from a single human specimen.