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A simple, one-step fractionation technique suitable for the finest quantitation of type V collagen from acidic mixtures of interstitial collagens is reported. The technique is based on the higher solubility of this collagen type in alkaline KCl-phosphate solution, and the purity of the sample has been monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electron microscopic analyses. The latter reveal that the majority of the molecules are endowed with a thread portion and a terminal knob, and show a low tendency to originate ordered and reproducible polymers at low temperature.

Amniotic fluid was analyzed for the presence of the 15,000 monomer molecular size glycoprotein found in human breast gross cystic disease fluid (GCDFP-15). From 24 weeks' gestation a log-linear increase in levels of GCDFP-15 was noted. The levels of GCDFP-15 doubled every 16 to 28 days, and the highest value recorded was 7,200 ng/ml. At delivery, levels of GCDFP-15 in cord blood plasma were in a background range (mean, 8 ng/ml). Maternal plasma levels of GCDFP-15 were one to tenfold higher in the third trimester of pregnancy when compared to those in nonpregnant women. Since saliva is known to contain high concentrations of GCDFP-15 (10 to 70 micrograms/ml), it is proposed that the levels in amniotic fluid originate from saliva and tracheobronchial secretions.

We assayed Type 1 (high K+) and Type 2 (high Na+) human breast cyst fluids for DHEA-S. When an antibody specific for the 3-sulfoconjugate end of DHEA-S was used, Type 1 cyst fluids (n = 18) showed a content of 114 +/- 68 micrograms/mL (mean +/- sigma) and Type 2 cyst fluids (n = 14) of 35 +/- 17 micrograms/mL (P less than 0.01). Using an antibody specific for the D-ring, the results were 151 +/- 91 micrograms/mL and 51 +/- 32 micrograms/mL, respectively (P less than 0.01). The apparent concentrations of DHEA-S were statistically different, even though both assays gave equal results in serum from normal adults. The presence of other compounds in individual cyst fluid samples was examined by extraction and chromatography. DHEA-S immunoreactivity was found in both early and late eluting fractions in Type 1 cyst fluids and in late eluting fractions from Type 2 cyst fluids. Only the late eluting fraction from Type 2 fluids had approximately equal immunoreactivity with both antibodies. In addition to authentic DHEA-S, breast cyst fluids contain other materials that react with DHEA-S antibodies. Radioimmunoassays for DHEA-S in cyst fluid must be specifically validated because of the presence of these compounds.

The presence of CEA was studied in paraffin sections of 100 benign breast lesions using an immunoperoxidase technique. In fibroadenomas, CEA was found in 25% of cases as opposed to 64% with cystic disease with or without epitheliosis. Our results demonstrate a tendency for an increased frequency of CEA tissue positivity in association with a more prominent form of epithelial hyperplasia with cystic disease. The observed higher positivity of tissue CEA may reflect a higher 'cellular activity' in the latter lesions compared with fibroadenomas.

A previous study reported pH levels in human breast cyst fluid ranging from 7.00 to 7.80 with the cut off between Type I (high K+, low Cl-) and Type II (high Na+, high Cl-) cysts at pH 7.40. Our data indicate a wider range of pH levels in human breast cyst fluids with Type I cysts consistently having acidic levels below pH 7.0 while Type II cysts had pH levels greater than 7.0. Type I acidic cysts characteristically also contain greater amounts of organic acids than Type II cysts. Thus, the pH of breast cyst fluid appears to be related to the electrolyte profile.

The aims of this paper are to demonstrate the stainability of estrogen, CEA, and ferritin in breast carcinomas, fibroadenomas, and fibrocystic diseases; to examine whether the findings of endogenous estrogen using the immunohistochemical detection method are related to estrogen receptor (ER) assays; and to determine whether the stainability of estrogen, CEA, and ferritin were related to the prognosis of breast carcinomas. In breast cancer, the stainability of estrogen using the peroxidase-antiperoxidase (PAP) method was positively correlated with the dextran-coated charcoal (DCC) assay for ER. In breast cancers, the percentage of positive staining was 46% for estrogen, 48% for CEA, and 47% for ferritin. With all three stains, significant differences were observed between cancer and benign diseases. Cases that were both positive for estrogen staining and negative for CEA showed a good prognosis after the recurrence of disease. Our data suggest that the immunohistochemical staining of estrogen, CEA, and ferritin might predict the biological behavior of breast carcinomas and be a prognostically useful indicator of breast cancer patients.

A simple and reliable radioimmunoassay for the determination of 5-androstene-3 beta,17 beta-diol (5-Adiol) in peripheral plasma and in breast cyst fluid, after chromatography on Celite microcolum has been described and evaluated. The antiserum used was raised in the rabbit injected with dehydroepiandrosterone-15 alpha-carboxymethyl-bovine serum albumin. In men below 40 years of age the levels ranged from 0.85 to 2.80 ng/ml (mean +/- SEM: 1.52 +/- 0.11; n = 24) and from 0.50 to 2.20 ng/ml (mean: 0.93 +/- 0.09; n = 20) in men aged between 41 and 62 years. The mean level was significantly different (p less than 0.001) between the two groups. A significant correlation (r = -0.56 p less than 0.01) was demonstrated between age and all male levels. In women the mean plasma level was in the follicular phase: 0.81 +/- 0.07 ng/ml (range: 0.40 - 1.50; n = 17; age: 19-41 years) and in the luteal phase: 0.83 +/- 0.05 ng/ml (range: 0.40 - 1.30; n = 29; age: 18-43 years). No cyclical change and no correlation with age could be evidenced. A significant difference (p less than 0.001) was shown between females and the young male group. In breast cyst fluid the levels of unconjugated 5-Adiol ranged from 0.05 to 13.70 ng/ml (mean: 2.21 +/- 0.73; n = 23) whereas the sulfate concentrations ranged from 75 to 7,500 ng/ml (mean: 1,973 +/- 543; n = 18), thus demonstrating very wide inter-individual variations.