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Ferritin and transferrin levels in human breast cyst fluids: relationship with intracystic electrolyte concentrations.

The concentrations of sodium and potassium and the content of ferritin and transferrin, proteins considered as potential markers for identifying cells undergoing divisional activity, were measured in fluid from 30 human breast cysts. On the basis of the relative electrolyte concentrations, two main classes of cysts were defined. When the cyst fluids were subdivided according to their Na+/K+ ratio, a significant difference was found between menstruating vs. menopausal patients. The relationship between the two major iron-binding proteins and the Na+/K+ ratio may indicate the expression of a local higher biosynthetic activity in apocrine cysts associated with higher cancer risk.

Body Fluids↗

[An immunohistochemical study of cutaneous tumors using an antibody to the breast cyst fluid protein (GCDFP-15)].

The antibody against a molecular weight 15,000 protein isolated from the breast cyst fluid (GCDFP-15) was applied to tissue from cutaneous tumors, especially to sweat gland tumors. This protein was purified from the breast cyst fluid; antisera was prepared in rabbits. Ninety six cases of cutaneous tumors were stained by the PAP method. Apocrine hydrocystoma, syringocystadenoma papilliferum, so-called mixed tumor of the skin, extramammary Paget's disease, microcystic adnexal carcinoma, and mucinous carcinoma of the skin were positively stained. Eccrine poroma, eccrine spiradenoma, syringoma, eccrine hydrocystoma, papillary eccrine adenoma, adenoid type of basalioma were negative. These observations indicate that the positively stained tumors have functioning seromucous cells, which are characteristic of apocrine gland cells and eccrine dark cells in the skin.

Animals↗

Enterolactone in breast cyst fluid: correlation with EGF and breast cancer risk.

The purpose of our study was to investigate whether enterolactone does accumulate into breast cyst fluid and whether it correlates with breast cancer risk. We included 258 women who had at least one cyst aspiration and known intracystic cation and epidermal growth factor (EGF) concentration values. For 191 of such women serum aliquots were also available. The median value of serum enterolactone was 17 nM/l (range 1-140 nM/l). The median intracystic level of enterolactone was much higher (63 nM/l, range 0-872 nM/l) and was significantly higher in type I cysts (p = 0.000). This cyst type contained also significantly higher levels of EGF (p = 0.000). A direct relationship was found between serum and cyst fluid enterolactone levels (p = 0.000) and between cyst enterolactone and EGF levels (p = 0.03), the latter correlation being evident especially in type II cysts. Twelve patients in the cohort of women were found to have developed a breast cancer. After univariate analysis breast cancer risk was associated with cyst type and especially with EGF concentration. No association was evident for enterolactone concentration. However, enterolactone concentration appeared to significantly decrease the risk of patients with high EGF concentrations. Our results show that enterolactone does accumulate in breast cysts, and that it modulates the risk related to the intracystic level of EGF, which is confirmed to be a strong predictor of breast cancer risk.

4-Butyrolactone↗

Monoclonal antibodies against ovarian carcinoma-associated antigens, raised by immunization with cyst fluids.

Intrasplenic immunization with ovarian tumour derived cyst fluids was used to generate monoclonal antibodies (Mabs) against human ovarian carcinomas. Seven Mabs were characterized immunocytochemically using indirect immunofluorescence assays (IFA) on frozen sections of gynaecologic tumours (n = 85), non-gynaecologic tumours (n = 50), normal human tissues (n = 66), and on 8 ovarian carcinoma cell lines. The Mabs reacted with 67-96% of 49 ovarian carcinoma tissues tested. Although 6 out of 7 Mabs showed slight cross reactivity with some endometrial, cervical, and oviductal carcinomas, little reactivity with non-gynaecologic tumours or normal tissues was found. Based on immunoelectronmicroscopy, all the antigenic determinants defined by the Mabs appeared to be associated with the tumour cell membrane. Among the seven Mabs, OV-TL 16 (IgG1 subclass) and OV-TL 23 (IgM class) reacted positively with 96% and 82% of the ovarian carcinomas respectively. Staining of 100% of the ovarian carcinomas examined (n = 49) could be achieved by combining OV-TL 16 with 23, as also by other combinations of the Mabs. It is concluded that a panel of these newly developed Mabs may be suitable for diagnostic purposes in histopathology.

Antibodies, Monoclonal↗

Serum antibody response following parenteral immunization with hydatid cyst fluid in sheep infected with Echinococcus granulosus.

Production of specific serum antibodies following immunization with hydatid cyst fluid antigens was investigated in sheep with Echinococcus granulosus infection and in noninfected controls. Six of 10 infected animals responded to intramuscular injection of antigen by rapid production of antibodies detected in indirect hemagglutination assays. Similar responses did not occur in any of 10 noninfected controls. It is suggested that differences in the rate of response to immunization with cyst fluid antigens between groups of sheep could be exploited in serodiagnosis of E. granulosus infection in sheep. The results also suggest that low levels of antibody found in the serum of sheep infected with E. granulosus are not the result of immunosuppression or immunological tolerance, but are due to sequestration of antigen from the immune system of the host.

Animals↗

Bile acids in human breast cyst fluid: the identification of lithocholic acid.

Breast cyst fluid (BCF) aspirated from 12 women with fibrocystic disease of the breast and sera obtained simultaneously were analyzed for bile acids. Analysis was performed by gas-liquid chromatography of the acetoxy methyl esters of the bile acids prepared after alkaline hydrolysis of the bile salts. An internal standard served to correct for methodological losses. Low levels of bile acids were found in serum samples, precluding overt hepatobiliary complications. Deoxycholic acid (17-160 mumol/L), chenodeoxycholic acid (18-305 mumol/L), and cholic acid (3-119 mumol/L) were detected in 11 of 12 samples of BCF. In 2 cases, chosen at random, the identities of the bile acids were verified by mass spectrometry. Lithocholic acid (9-23 mumol/L), a reported cocarcinogen, was detected in 6 of the 12 samples of BCF. This is the first report of the presence of lithocholic acid in BCF with confirmation by Mass spectrometry. There was no correlation between the levels of individual bile acids and those of potassium ion, Na+/K+, estriol-3-sulfate, or 16 alpha-hydroxyandrogen sulfates that had been quantified previously in these samples. There was borderline correlation between concentrations of total bile acids and K+ (P less than 0.06) and Na+/K+ (P less than 0.07). Yet to be elucidated are the mechanism of accumulation of bile acids in BCF and whether levels of particular bile acids in BCF may serve to identify that small subset of women with fibrocystic disease at risk for developing breast cancer.

Adult↗

DNA ploidy of ovarian and adnexal cyst fluid. A useful adjunct to cytology.

It is increasingly common for cytology laboratories to receive ovarian, adnexal and pelvic cyst fluids obtained via sonographically directed aspiration and laparoscopic techniques, especially from women who are desirous of preserving fertility or who are undergoing in vitro fertilization (IVF). Accurate characterization of such cysts is a worthwhile goal, given the superior prognosis for ovarian carcinomas that are diagnosed at an early stage. In an effort to improve upon the false-negative diagnosis rate associated with cytology, we evaluated DNA ploidy as a possible adjunctive criterion. We examined 55 benign, 3 borderline and 6 malignant aspirates received by our cytopathology laboratory; 35 were aspirated directly from the patient from clinically and ultrasonographically benign cysts, and 29 were aspirated from surgically removed benign (20) and malignant (9) cysts. Adjunctive DNA ploidy and cell cycle analysis was performed using the Cell Analysis Systems CAS-200 on Feulgen-stained cytologic smears of the 64 cyst fluids. Adequate material for DNA analysis was obtained from 33/35 in situ aspirated cysts and from 19/29 surgical specimen cysts. Forty-seven of 52 cytologically benign cysts were diploid. Of the 5 nondiploid benign cysts, 3 were follicle cysts (2 from hormonally stimulated IVF patients and 1 from a postpartum patient), and 1 was a benign cystic teratoma. Their nondiploid DNA pattern or tetraploidy may be due to a high proliferative index. The fifth nondiploid benign aspirate was from a resected benign epithelium-lined cyst; its DNA histogram contained a conspicuous tetraploid population. All 9 malignant cysts were cytologically malignant. Of the 3 borderline cysts, 1 was nondiploid, and 2 were diploid. All 6 fully malignant cysts were nondiploid; 2 of them were tetraploid. Based on our results, we conclude that DNA ploidy analysis of cells derived from ovarian and adnexal cyst aspirates is feasible (in 95% of cases) and relatively specific (90%) and has a relatively high negative predictive value (92%). The results are not sufficiently predictive of the histology of the lesion to warrant therapeutic intervention based on ploidy alone (sensitivity of nondiploid results, 78%; positive predictive value of nondiploid results, 58%). Nondiploidy should suggest consideration, but is not conclusive, of a malignancy diagnosis. There may even be prognostic implications to the ploidy pattern, particularly in borderline tumors, in which nondiploidy portends a poor prognosis.

Adnexal Diseases↗

Cyst fluid cytologic analysis in the differential diagnosis of pancreatic cystic lesions.

Pancreatic cystic lesions include inflammatory pseudocysts, benign serous cystadenomas, and mucinous neoplasms, some of which are malignant. Cytologic analysis of cyst fluid has been proposed to diagnose pancreatic cysts before definitive therapy. The authors report an analysis of 31 pancreatic cyst aspirates: 9 pseudocysts, 5 serous cystadenomas, 8 mucinous cystic neoplasms, 4 mucinous cystadenocarcinomas, 2 papillary cystadenocarcinomas, 1 mucinous ductal adenocarcinoma with cystic degeneration, and 2 cystic islet cell tumors. All pseudocysts were correctly classified as probable inflammatory lesions, because of the presence of abundant acute inflammation and histiocytes and the absence of glandular epithelium. Three of five serous cystadenomas were correctly classified, based on the presence of small cuboidal cells in clusters with microvesicular cytoplasm containing glycogen. Eleven of 12 mucinous tumors contained round cells with large cytoplasmic mucin vacuoles or columnar cells containing cytoplasmic mucin. Malignancy was diagnosed in 5 of 7 carcinomas, 1 case was classified as suspicious for malignancy, and 1 case was nondiagnostic because of the absence of a cellular component. The authors concluded that pancreatic cyst fluid cytologic analysis is useful in differentiating mucinous from nonmucinous pancreatic cysts and may provide definitive evidence of malignancy. In some cases, serous cystadenoma can be diagnosed based on cytologic analysis. An inflammatory smear without epithelial cells suggests a pseudocyst, but these findings are nonspecific, as a similar pattern may occur when a cystic neoplasm undergoes degenerative changes. Therefore, pseudocyst remains a diagnosis of exclusion.

Biopsy, Needle↗

Prostate-specific antigen found in type I breast cyst fluids is a secretory product of the apocrine cells lining breast gross cysts.

Prostate-specific antigen (PSA), a serine protease thought to be exclusively produced by the prostate epithelial cells, has been recently found in human breast tissues and fluids. PSA in breast cancer is associated with the presence of steroid-hormones and receptors, and its presence seems to be a favourable prognostic indicator. In order to clarify whether the cells lining breast cysts may represent the source of PSA found in human breast cyst fluid, we performed an ultrastructural immunolocalization of PSA in the cells surrounding Type I breast cysts, obtained from breast cyst fluids of women affected by breast gross cystic disease, the most commonly occurring benign breast lesions associated with increased cancer risk. These apocrine cells show morphological features typical of actively synthesizing and secreting cells, and a PSA labelling distributed on free ribosomes, RER cisternae, and secretory granules, indicating that the metabolically active apocrine cells lining the Type I cysts are responsible for the production and secretion of PSA in Type I breast cyst fluids. The synthesis and intracystic accumulation of this serine protease in biosynthetically active apocrine Type I cysts can play an important role in the natural history of breast gross cystic disease as well as in the mechanism of cyst evolution.

Adult↗

An immunohistochemical study of the tissue distribution of the breast cyst fluid protein, zinc alpha 2 glycoprotein.

Zinc alpha 2 glycoprotein is one of the proteins present in breast cyst fluids, being found at levels 30-50 times its plasma concentration. Using an immunoperoxidase technique the distribution of this glycoprotein has been studied in a range of non-mammary tissues and carcinomas, as well as in normal, benign and malignant breast specimens. The breast cyst fluid protein was detected in all apocrine cells of skin and in the apocrine metaplastic epithelium lining of breast cysts. A progression was apparent from normal to hyperplastic breast in the number of cells reacting, particularly of cystically dilated acini, to a final consistent staining of apocrine-lined cysts. Zinc alpha 2 glycoprotein was demonstrated in 16 of 33 invasive carcinomas, 15 of which were eosinophilic on haematoxylin and eosin staining, and in one of three non-invasive carcinomas. No staining was apparent in other non-mammary tissues and carcinomas apart from weak reactivity of serous cells of the parotid gland. Zinc alpha 2 glycoprotein is, therefore, a reliable immunohistochemical marker of apocrine cell differentiation.

Apocrine Glands↗

[An immunohistochemical study of normal sweat glands using an antibody to the breast cyst fluid protein (GCDFP-15)].

In this study, we applied the antibody against the molecular weight 15,000 protein purified from breast cyst fluid (GCDFP-15) to cutaneous tissue, especially to sweat glands. Breast cyst fluid was obtained by needle aspiration from patients with gross cystic disease. DEAE-cellulose column chromatography was performed and followed by SDS-polyacrylamide gel electrophoresis. Antisera against the purified protein were prepared in rabbits. We stained specimens of normal skin by the PAP method. The normal apocrine gland cells were strong positive and eccrine gland dark cells were positive. Eccrine gland clear cells were weak positive or negative. Duct cells of both glands were negative. From these observations, we suggest that this antibody is useful for detecting seromucous cells of sweat glands, including both the apocrine gland cells and eccrine dark cells in the skin.

Animals↗

Calcium oxalate crystals in benign breast cyst fluid.

We have recently observed a strongly birefringent material of varying shapes and sizes in a benign breast cyst fluid specimen from a 52-yr-old woman with suspicious mammographic microcalcifications. The finding of calcium oxalate crystals in the breast cyst fluid should be regarded as a recognition of a particular type of calcification, easily overlooked with the conventional light microscopy. To date, we are unaware of any cytologic reports of this type of calcification in the breast.

Body Fluids↗

The value of routine cytologic examination of breast cyst fluids.

The results of the cytologic examination of 6,782 consecutive breast cyst fluids aspirated from 4,105 women during the period 1976 to 1983 were reviewed to assess the value of the routine cytologic study of such specimens. Cases in which cancer had been suspected by physical examination and/or mammography before aspiration of the cyst were excluded from the evaluation. Five clinically and radiologically inapparent intracystic papillomas were detected overall (0.1%). All cases of intracystic papilloma produced a blood-stained fluid and showed an intracystic mass at pneumocystography. Cytology was negative in two of these cases and falsely positive in one case while correctly identifying the papilloma in two cases. One incidental case of occult in situ lobular carcinoma was also detected. Routine cytologic examination of all breast fluids is thus not recommended as a cost-effective practice. Cytology should be used only when a blood-stained fluid is obtained (2% in the present series) since its indiscriminate application to all cyst fluids does not affect the rate of detection of intracystic lesions, most of which are suspected before the aspiration.

Adult↗

Lipid-associated sialic acid levels in human breast cyst fluids.

Benign mammary gross cystic disease is the most common breast lesion. Women with apocrine changes of epithelium lining the cysts are at higher risk for developing breast cancer than the normal female population. Sialic acid has drawn considerable interest because of carbohydrate aberrations in malignant cells. The current investigation determined the concentrations of lipid-associated sialic acid (LASA) in 62 breast cyst fluids and sera. Data analyses show a significant increase in the mean values of LASA in metabolically active apocrine cysts when compared to the cysts with Na+/K+ > 3 (flattened cysts) (p < 0.001). The greater LASA levels in cyst fluids with lower intracystic Na+/K+ ratios could represent an altered expression of biosynthetic activity of the surrounding apocrine cell surface sialoglycolipid metabolism, providing a possible explanation of why women with apocrine cysts may be at greater cancer risk and being useful in further studies on functional stage changes in the cysts and their relationship to breast cancer.

Adult↗

[Tumour markers in human breast cyst fluid in gross cystic breast disease (GCBD)]

AIM: Parallel measurements of tumour markers in the serum and breast cyst fluid in a high risk group (GCBD) of breast cancer. The identification of individuals belonging to this group and their follow-up. MATERIALS AND METHODS: In the breast cyst fluid of 108 patients with GCBD (mean age 47 years) we measured the levels of CA 15-3, TPA, CEA and beta HCG completed by PCT determinations. Simultaneously, the serum CA 15-3 and TPA concentrations were also measured using the Luminescent Immunoassay techniques. RESULTS: Strikingly high TPA values were found in 98% of the patients. The CA 15-3 levels, however, were pathological only in 24%of them. The CEA and beta HCG levels showed hardly any rise and the PCT concentration remained normal. CONCLUSIONS: The lack of any rise in PCT concentration precludes the inflammatory origin of the cystic fluid and the normal serum arker levels exclude ultrafiltration. The increased TPA concentration in the breast cystic fluid and the occurrence of pathological CA 15-3 level in the above percentage of the cases suggest that GCBD represents not only a high risk group but possibly a precancerous state, too.

Journal Article↗

Differential serodiagnosis for cystic and alveolar echinococcosis using fractions of Echinococcus granulosus cyst fluid (antigen B) and E. multilocularis protoscolex (EM18).

Echinococcus granulosus cyst fluid and E. multilocularis protoscolex extract were fractionated by a single step of preparative isoelectric focusing, resulting in an antigen B-rich fraction (8-kD) and an Em18-rich fraction, respectively. The usefulness of both fractions for differential serodiagnosis of cystic (CE) and alveolar (AE) echinococcosis was evaluated by a large-scale immunoblot analysis on a battery of 354 serum samples. These included 66 from AE patients originating from four different endemic areas, 173 from CE patients originating from seven different endemic areas, 71 from patients with other parasitic diseases, 15 from patients with hepatomas, and 29 from healthy individuals. In an immunoblot with the antigen B-rich fraction, 92% (158 of 173) of the CE sera as well as 79% (52 of 66) of the AE sera reacted with the 8-kD subunit. No cross-reactivity occurred with any sera from patients with cysticercosis, other parasitic diseases, or with hepatomas, or from healthy controls. In an immunoblot with the Em18-rich fraction, all but two sera from AE patients (64 of 66, 97%) recognized Em18, and only nine of 34 CE sera from China reacted with it. All other (139) CE sera from six other countries were negative as were all (115) other non-echinococcosis sera. These findings indicate that antigen B (8-kD) is not species-specific for E. granulosus but is genus-specific for Echinococcus, and that the Em18 antigen is a reliable serologic marker for species-specific differentiation of AE from CE.

Animals↗

Presence of laminin fragments in cyst fluid from patients with autosomal dominant polycystic kidney disease (ADPKD): role in proliferation of tubular epithelial cells.

Cyst fluid samples obtained from eight patients with autosomal dominant polycystic kidney disease (ADPKD) were analysed for the presence of the basement membrane component laminin and its breakdown products, using ELISA and immunoblotting techniques. Whole laminin was not detected, whereas laminin fragments of 270, 155, 87, 56, and 14 kDa were detected at a mean total value of approximately 0.5 microgram/ml. The laminin fragments were assessed for their effect on cultured normal and ADPKD epithelial cells. Both cell types showed accelerated growth under these conditions. These findings suggest that basement membrane degradative fragments present in cyst fluid may contribute to cystic epithelial cell proliferation and may therefore be important in the pathogenesis of ADPKD.

Adult↗