Extramammary Paget's disease--evidence for an apocrine origin. An immunoperoxidase study of gross cystic disease fluid protein-15, carcinoembryonic antigen, and keratin proteins.

The histogenesis of extramammary Paget's disease has long remained unresolved and controversial. In an attempt to delineate the origin of the neoplastic cells in this disease, the immunoperoxidase localization of gross cystic disease fluid protein (GCDFP-15), a marker of apocrine epithelium, carcinoembryonic antigen (CEA), and keratin proteins, was determined for seven cases of extramammary Paget's disease (five vulvar, one anogenital, and one axillary). Immunoreactivity for GCDFP-15 was localized within Paget cells in six of our seven cases, including five cases from the vulva and one case from the axilla. CEA was present in the Paget cells in all seven cases. None of the Paget cells exhibited immunoreactivity for keratin proteins. Within normal skin, eccrine glands were immunoreactive for both keratin and CEA, whereas GCDFP-15 localized only to apocrine ducts and glands. Our findings strongly support an apocrine cell derivation for extramammary Paget's disease.

Immunoperoxidase localization of GCDFP-15 with mouse monoclonal antibodies versus rabbit antiserum.

Five monoclonal antibodies (Mabs) raised against separate determinants on a breast gross cystic disease fluid protein of 15 KD (GCDFP-15) were compared to one another and to a rabbit antiserum (Rb) against GCDFP-15 by radioimmunoassay (RIA) and by immunoperoxidase localization in paraffin-embedded tissues. All five Mabs and the Rb were equivalent in recognition of GCDFP-15 in solution, as determined by RIA. However, two of the Mabs (A5, B15) showed only minimal binding to GCDFP-15 in paraffin-embedded tissues, whereas the other three Mabs (B1, B4, D6) were equivalent to the Rb in staining intensity. These latter three Mabs and the Rb were evaluated by the immunoperoxidase technique on a variety of benign and malignant neoplasms as well as normal tissues (150 specimens) for staining specificity. Immunoperoxidase staining by the three Mabs vs the Rb was equivalent in apocrine glands, metaplastic apocrine epithelium of breast, and breast carcinomas with apocrine features. No staining of the Mabs or Rb was seen in the other tissue specimens.

Intranuclear androgen and cytosolic receptor concentrations in the axillary skin of osmidrosis.

5 alpha-Dihydrotestosterone (DHT) and testosterone were measured by radioimmunoassay in the crude nuclear and cytoplasmic fractions of the axillary skin of both male and female patients with osmidrosis and the levels compared with those of nongenital skin. The intranuclear levels of DHT were 1.44 +/- 0.22 and 1.77 +/- 0.38 pg/micrograms DNA in men and women, respectively. Those of testosterone were about 10% of DHT levels. In the skin of nontarget regions nuclear DHT was much scarcer or undetectable. Cytosolic androgen receptors in isolated apocrine glands were also measured using 3H-R1881 as a ligand. Typical androgen receptors were present in all of eight patients (KD = 1.32 +/- 0.24 X 10(-9)M, Bmax = 10.3 +/- 0.51 fmol/mg protein). Neither the intranuclear androgen concentrations nor the cytosolic androgen receptor levels were significantly different between the two sexes. These data indicate clearly that the apocrine gland of patients with osmidrosis is a typical androgen target organ, irrespective of sex, and suggest that nuclear DHT in the axillary skin of women is derived from not only testosterone but also other precursors, especially in consideration of the very low serum concentrations of testosterone in females.

Immunohistochemical identification of epithelial membrane antigen in sweat gland tumors by the use of a monoclonal antibody.

A total of 78 cases of adnexal tumors of the skin were examined with the use of monoclonal antibody against epithelial membrane antigen (EMA). The EMA reaction was confined to luminal surfaces and lateral borders of sweat glands, both eccrine and apocrine, being usually absent in ductal segments. Neoplastic lesions of all the adenomatous tumours and mixed tumours of sweat glands showed specifically positive EMA staining of luminal surfaces and lateral borders of tubular, duct-like, and cystic structures. However, solid foci of those tumours were negative for EMA. Tumours of ductal origin, e.g. syringomas and poromas, exhibited positive EMA staining in their plasma membrane, although normal intact keratinocytes did not stain for EMA. Immunohistochemical distribution of EMA in skin adnexal tumours was classified into two types: one in which the luminal surfaces and lateral outer borders were positive, similar to that of the normal secretory coil, and the other in which the plasma membrane of neoplastic cells of ductal origin was positive.

A chemist's view of the analysis of human hair for trace elements.

With the comparatively recent development of analytical techniques of great power and sensitivity, the significance of the levels of trace elements in human hair has attracted the attention of many disciplines including the environmental sciences. This paper presents the view that an agreed basis for the chemical analysis of trace elements in hair has not been established by the many workers in the field; a chemical basis is proposed here. Levels of 37 trace elements found in human hair are tabulated. Endogenous and exogenous sources of such trace elements are described and discussed. An extended review of the many pre-analysis treatments of hair (for the removal of exogenous elements) is presented. Twenty-four representative treatments are tabulated. Some of these treatments clearly removed significant fractions of endogenous elements along with exogenous elements. It is clear that method of cleaning have frequently been chosen without knowing enough about the basic chemistry and behaviour of the hair shaft. The significance of the results obtained cannot therefore be reliably assessed. A collation of recent literature reports leads to the tentative conclusion that disulphide bonds in the cuticular proteins of hair are major sites both for the deposition of metals during formation of hair and for interaction with exogenous elements. The feasibility of a holistic, no-wash policy for hair analysis is outlined and supported.

The nature of pigment in pigmented apocrine hidrocystoma.

Apocrine hidrocystomas are often pigmented clinically. The cause of this pigmentation is not known. A case of pigmented apocrine hidrocystoma is presented with evidence of melanin as the underlying mechanism of the pigmentation. Review of an additional 150 cases suggests that this is rare. The Tyndall effect, analogous to that seen in blue dome cysts of fibrocystic disease of the breast may be the likely explanation of the pigmentation seen clinically in most cases of apocrine hidrocystomas.

The polymorphic apocrine nevus: a study of a unique tumor including carcinoembryonic antigen staining.

The apocrine nevus is a rare tumor. We report a 32-year-old man with a neck nodule, histologically an apocrine nevus displaying mature and immature apocrine structures in a distinctive pattern. The carcinoembryonic antigen staining was positive intracellularly only in the smaller luminal structures. We have named this unique tumor a polymorphic apocrine nevus.

The isolation of human sebaceous glands and apocrine sweat glands by shearing.

A new method of isolating human sebaceous and apocrine sweat glands by the repeated dissection of skin biopsies with scissors is described. The success of the technique is attributed to a line of weakness between the investing capsule and the surrounding connective tissue which parts under shear forces. The glands are judged to be viable by: (i) light and electron microscopy; (ii) ATP, ADP and AMP contents of 148.8 +/- 30.3, 30.6 +/- 4.7 and 14.9 +/- 4.7 pmol (mean +/- s.e.m.) for sebaceous glands and 310.2 +/- 34.1, 90.35 +/- 16.3 and 40.1 +/- 11.8 pmol (mean +/- s.e.m.) for apocrine sweat glands, which gave energy charges of 0.84 and 0.81, respectively; and (iii) a rate of sebaceous gland lipogenesis of 39.7 +/- 3.7 pmol glucose incorporated into lipid/gland/h (mean +/- s.e.m.).

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.

Nature of so-called "metaplasia of the apocrine epithelium". Macrophages attack apocrine epithelium". Macrophages attack apocrine epithelium.

Immunohistochemical and electron-microscopic studies were performed on the lesion termed "metaplasia of the apocrine epithelium." which was seen in association with an apocrine adenocarcinoma. The cells of this so-called "metaplasia" lacked cytokeratin, which was present in the apocrine epithelium. Surprisingly, the lesion ultrastructurally consisted mainly of terminally differentiated macrophages, the cytoplasms of which were filled with numerous phagosomes and lipid droplets. The cells lacked a desmosomal connection at their borders and some had Langerhans granule-like structures in the cytoplasm. The luminal wall was often infiltrated with macrophages and lymphocytes, and, in some portions, was replaced by macrophages with a large cytoplasm filled with numerous phagosomes. The immunohistochemical and ultrastructural findings indicate that the lesion is not composed of cells of the apocrine epithelial origin, but of macrophages that have ingested apocrine epithelium.

Mixed tumors of the skin. A histological and immunohistochemical study.

Sixty-four specimens of mixed tumors of the skin were studied by conventional microscopy. Sections from all 64 specimens were stained by hematoxylin and eosin, and sections from 18 of those specimens were stained by immunoperoxidase techniques for the presence of S-100 protein, carcino-embryonic antigen (CEA), keratin, actin, vimentin, epithelial membrane antigen (EMA), and gross cystic disease fluid protein-15 (GCDFP-15). Two distinctive histopathological patterns of mixed tumors of the skin became apparent, namely, apocrine and eccrine. Mixed tumors with apocrine features are by far the most common. Immunoperoxidase techniques, in our experience, do not enable differentiation between apocrine and eccrine types of mixed tumors.

A source of cutaneous maternal semiochemicals in the mink?

Unique hypertrophic apocrine sweat glands are described in the neck, perineal and inguinal skin of mink kits. These glands enlarge after birth, only to regress rapidly and become vestigal by weaning. No similar phenomenon has been recognized before in mammals. Behavioral studies indicate a possible role for the glandular secretion in maternal recognition of the young.

Lectin-binding sites in Paget's disease.

The presence and distribution of lectin-binding sites on neoplastic cells of Paget's disease was studied using fluorescein isothiocyanate (FITC)-conjugated peanut agglutinin (PNA), and FITC-conjugated wheatgerm agglutinin (WGA), and compared with such lectin-binding sites on keratinocytes, and cells of eccrine glands, apocrine glands, and mammary glands. Neoplastic cells of both mammary and extramammary Paget's disease showed cytoplasmic staining with both lectins. There were however fewer stained cells in mammary Paget's disease than in extramammary Paget's disease. The cytoplasmic staining of lectin-binding sites in cells of apocrine glands was in sharp contrast to the cell-surface staining seen on keratinocytes, or cells of eccrine glands or mammary glands. These results indicate that the lectin-binding sites of neoplastic cells of Paget's disease more closely resemble those of cells of apocrine glands than of keratinocytes, cells of eccrine glands or cells of mammary 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.

Immunohistochemistry of gross cystic disease fluid protein (GCDFP-15) in 65 benign sweat gland tumors of the skin.

Sixty-five cases of benign sweat gland tumors of the skin were studied for the expression and localization of gross cystic disease fluid protein-15 (GCDFP-15) by immunoperoxidase methods. There was positive staining of tumors of probable apocrine differentiation in 10 of 11 cases of apocrine hidrocystoma and five of five cases of hidradenoma papilliferum. There was no immunoreactivity for GCDFP-15 for tumors of probable eccrine differentiation, including five cases of eccrine hidrocystoma, five cases of eccrine poroma, five cases of eccrine spiradenoma, 10 cases of clear cell hidradenoma, and nine cases of syringoma. There was variable positive staining of tumors of more uncertain histogenesis, including eight of eight cases of syringocystadenoma papilliferum, one of four cases of cylindroma, and two of two cases of chondroid syringoma (mixed tumor). The above data support a functional differentiation of the expression of GCDFP-15 by eccrine compared to apocrine glandular epithelium with benign tumor development.

Immunohistochemistry of a gross cystic disease fluid protein (GCDFP-15) of the breast. A marker of apocrine epithelium and breast carcinomas with apocrine features.

Gross cystic disease fluid is a pathologic secretion from breast composed of several glycoproteins, including a unique 15,000-dalton monomer protein, GCDFP-15. By the immunoperoxidase technique, GCDFP-15 was localized in the apocrine metaplastic epithelium lining breast cysts and in apocrine glands in the axilla, vulva, eyelid, and ear canal. In normal breast tissue, a few individual epithelial cells within lobules and small ducts were focally positive for GCDFP-15. Fourteen of 30 breast carcinomas stained positively for GCDFP-15. Of 16 carcinomas with apocrine features, 12 stained positively. Benign and malignant lesions from other tissues, including lung, colon, ovary, endometrium, stomach, prostate, liver, esophagus, and kidney, revealed no immunoreactivity. The only cells of "non-apocrine" tissues that contained GCDFP-15 were serous cells of the submandibular salivary gland, submucosal glands of the bronchi, and accessory lacrimal glands. Phylogenetically, these tissues have biologic features in common with apocrine glands. This report is the first to characterize GCDFP-15 as a specific tissue marker of apocrine epithelium.

Immunohistochemistry of a component protein of the breast cystic disease fluid with mol. wt 15,000.

A specific protein from the liquid of a mammary cyst with a molecular weight of 15,000 (GCDFP 15) was studied in normal and pathological mammary tissue using an immunohistochemical method (peroxidase-anti-peroxidase complex). An immunoreactivity of the GCDFP type was found in normal idrosadenoid glands having an apocrine secretion. Histologically normal mammary tissue was not immunoreactive. In benign breast tissue the GCDFP was found particularly in epithelium undergoing apocrine metaplasia (55/55) and in atypical lobular epithelial hyperplasia (8/10). Of the adenocarcinomas of the breast 136/161 (84%) were immunoreactive, especially lobular carcinoma (13/13). The proportion of tumors with a high percentage of immunoreactive cells (76-100%) was greater for Bloom's grade I (1/29: 34%) than for grade III (10/66: 15%). A significant correlation was found between the percentage of immunoreactive cells and the cytosolic concentration of progesterone receptors. The morphological intracellular identification of GCDFP (due to its greater sensitivity) and its correlation with progesterone receptors allowed a more precise evaluation of the functional state and the hormonal dependency of the breast cells by underlining the heterogeneity of the tumoral cell population.

[Apocrine carcinoma of the breast--a morphologic comparison of the apocrine sweat glands and the apocrine metaplastic epithelia in mastopathy].

An operatively removed apocrine carcinoma of the breast from a 62-year-old Japanese lady has been observed by the ABC method, using the monoclonal antibody 115D8. The cancer cells and metaplastic epithelia exhibited similar ultrastructural findings (an apical snout, apocrine granules, etc.) as the apocrine sweat gland cells, although no evidence of apocrine secretion could be detected. The immunohistochemical testing, using monoclonal antibody, 115D8, showed an apical, linear, dot-like, staining in the supranuclear regions on the apocrine sweat gland cells and on the apocrine metaplastic cells of the mammary gland. Similar stainability also was observed in the well-differentiated area of the apocrine carcinoma, while a heterogeneity in staining, such as unstained cells and diffuse cytoplasmic-stained cells were found in the poorly-differentiated areas. These abnormal staining patterns indicate the malignant changes of the apocrine cells.