Expression of cathepsin D and pS2 in imprint smears of breast carcinoma.

The aim of this study was to add to existing information on the effects of certain tumour markers expressed by breast cancers on tumour malignancy as evidenced by size of primary and occurrence of lymph node invasion. One hundred freshly resected breast cancers were examined by immunocytochemical staining of imprint smears for Cathepsin D and pS2. Oestrogen receptor (ER) and progesterone receptor (PR) were tested for by dextrose-coated charcoal (DCC) assay and the results correlated with tumour size, histology and presence or absence of lymph node metastases at the time of surgery using chi(2) analysis. A significant positive correlation was demonstrated between Cathepsin D positivity and ER, PR and pS2 positivity. In tumours < 2 cm in diameter at surgery a positive correlation was observed between Cathepsin D positivity and the presence of lymph node metastases. The findings support the hypothesis that Cathepsin D may promote early metastasis, possibly by its proteolytic activity.

Antigen expression of alveolar macrophages in smokers and patients with lung diseases.

Alveolar macrophage function was studied immunocytochemically using three monoclonal antibodies--macrophage CD 68 KP 1 (M), protein CD 11C (P), and anti-elastin (EL)--and three polyclonal antibodies--lysozyme (LZ), alpha-1-antitrypsin (AAT), and alpha-1-antichymotrypsin (AACT). The material for study was smears obtained from bronchial washings from 15 healthy persons and 60 patients with respiratory infections or primary or secondary malignant lung infiltration. Eight of the healthy group and 40 of the patient group were smokers (SM). The percentage of cells obtained from the washings which were macrophages was also measured. The intensity of staining reactions for each of the six antigens was noted and in general more intense staining was noted in smokers than in non-smokers. More intense staining was observed in patients with pulmonary infections (group II PI) and metastatic pulmonary infiltrations (group IV MP Ca) than in controls (group IC), while patients with primary lung cancer (group III PP Ca) had highly reduced staining reactions. The number of macrophages was similarly increased in all groups in comparison with the IC group for non-smokers and in all groups except III PP Ca for smokers. It is concluded that smoking, pulmonary infections, and metastatic infiltration of the lung are associated with an increase in the number and activity of alveolar macrophages, while patients with primary lung cancer have an increase in the number of macrophages which are functionally incompetent.

Detection of c-Ha-ras oncogene expression in pleural and peritoneal smear effusions by in situ hybridization.

This study was undertaken to determine the presence of the c-Ha-ras protooncogene in pleural and peritoneal effusions. Cells from 40 pleural and peritoneal fluids from patients with malignant and nonmalignant diseases were studied by in situ DNA hybridization utilizing a biotinylated probe. The results were correlated with those obtained by conventional cytological methods. A cytological diagnosis of metastatic carcinoma was established in 67% of the patients with malignancy and effusion, and in 6% of the patients with benign disease accompanied by effusion. On the other hand, a relatively high proportion of ras expression was observed in 38% of the cases representing benign and reactive effusions and as high as 88% in the cases with malignant tumor and effusion. These results suggest that the ras oncogene may have an important role in differentially diagnosing benign and reactive mesothelial cells from malignant cells.

Cytological evaluation of the effect of Tamoxifen in premenopausal and post-menopausal women with primary breast cancer by analysis of the karyopyknotic indices of vaginal smears.

The influence of Tamoxifen on the vaginal epithelium of 33 premenopausal and 99 post-menopausal women with primary advanced breast cancer was investigated. The karyopyknotic index (KPI) values were assessed before starting therapy and at monthly intervals during therapy with Tamoxifen. A decrease in the KPI in menstruating women and a slight but definite increase in KPI values in post-menopausal women were observed.

Presence of epidermal growth factor receptor in breast smears of cyst fluids: relationship to electrolyte ratios and pH concentration.

The purpose of this study was to evaluate and correlate different high risk factors in breast cysts to produce a more accurate prognosis. We considered the cytology of the lining epithelium (apocrine or flattened), the Na+/K+ ratio (less than 3; greater than or equal to 3), the pH (lower, equal, or higher than 7.30), and the EGFr (present or not) in an attempt to reach better prognoses and diagnoses by evaluating more than one risk factor. Our material of 40 macrocysts consisted of 23 simple cysts with flattened epithelium and 17 complex cysts with apocrine or hyperplastic epithelium. In the simple cysts, the Na+/K+ ratio was greater than or equal to 3, while in the complex cysts the Na+/K+ ratio was, in all cases but one, less than 3. The pH was not significantly lower than neutral in the complex cysts. The EGFr was detected in 5 of 23 simple cysts and in 12 of 17 complex cysts. In conclusion, reaction of EGFr in smears of cyst fluids with the low intracystic electrolyte ratios may provide an explanation of why women with apocrine metaplasia or epithelial hyperplasia may be at higher risk of breast cancer than women with cysts which are lined by flattened epithelium.

Immunodetection of neuron-specific enolase and keratin in cytological preparations as an aid in the differential diagnosis of lung cancer.

The results of neuron-specific enolase (NSE) and keratin immunodetection in cytological specimens of sputum secured from 41 patients with lung cancer are presented. All 19 cases of small-cell carcinoma showed intense immunoreactivity for NSE. No such immunoreactivity was found in 21 of 22 cases of non-small-cell carcinoma. The single positive result was from a case of large-cell undifferentiated carcinoma. All 10 cases of squamous-cell carcinoma showed immunoreactivity for keratin. The 19 cases of small-cell carcinoma showed no such reactivity. Our findings indicate that immunostaining for NSE and keratin is a valuable aid when a definite diagnosis of small-cell carcinoma of the lung can not be made on the basis of conventional cytologic features.

Detection of benign or malignant origin of ascites with combined indirect immunoperoxidase assays of carcinoembryonic antigen and lysozyme.

The indirect immunoperoxidase method was used to study the presence of the intracellular carcinoembryonic antigen (CEA) and lysozyme (LZ) in alcohol-fixed cytologic smears of peritoneal fluids from 2 patients with chronic active hepatitis, 31 patients with liver cirrhosis and 7 patients with malignant liver disease. In the two patients with hepatitis, LZ was positive in both CEA was positive in one and negative in the other. Of the 31 patients with liver cirrhosis, 21 (67.5%) were LZ positive, 27 (87%) were CEA negative and only 4 (13%) were CEA positive. Of the seven patients with malignant disease, six were CEA positive and six were LZ negative. It is of interest that 23 of 24 (96%) LZ-positive results and 28 of 29 (97%) CEA-negative results corresponded to negative cytologic diagnoses for malignancy. Cytologic diagnosis of "reactive mesothelial cells" seemed to correlate better (71%) with CEA-negative and LZ-positive results. The data suggest that the investigation of CEA and LZ in the cells of peritoneal fluids appears to have promise as an adjunct to cytology in differentiating benign from malignant origins of the fluid.

The cytologic application of carcinoembryonic antigen for the discrimination of malignant from benign serous effusions.

In an attempt to diminish false cytologic diagnosis in discriminating between benign and malignant effusions, the value of intracellular carcinoembryonic antigen (CEA) was estimated in cytologic smears of effusions using a modified immunoperoxidase technique. CEA serum level was estimated by radioimmunoassay technique. In this study we included a group of 42 patients, 23 with a malignant and 19 with a benign effusion. CEA in cells of effusions was positive and useful for confirming malignancy in 65% of malignant effusions. The percentage increased to 83% with the combination of the results of positive CEA in cells and serum. Cytology was positive for malignancy in 16 of the 23 (70%) malignant cases. CEA in cells of benign effusions was negative in 84% and cytology in 100%. It is concluded that intracellular CEA estimation, when combined with CEA estimation in serum, improves the discriminating range of cytology for differentiating malignant and benign effusions.