Search

To clarify the accuracy of immunocytochemical detection of estrogen receptors (ER) in breast carcinomas using cytological materials, imprint specimens from tumor tissue were compared with frozen tissue sections and tumors analyzed by the dextran coated charcoal (DCC) method and enzyme immunoassay (EIA). Out of 50 cases examined by imprint immunocytochemistry, there were 39 ER positive cases (78.0% positivity). The positivity in the imprint materials agreed with that of the DCC in 36 out of 40 cases (85.0%), with 100% sensitivity and 60.0% specificity. The two methods statistically correlated with each other in their positivity and grade (p less than 0.001). The positivity and grades of imprint and frozen immunocytochemistry as well as those of imprint immunocytochemistry and the EIA agreed almost perfectly with each other. As a result of the present study, we concluded that immunocytochemical detection of ER is indeed reliable, as accurate as other procedures. We recommend that aspiration biopsy cytology (ABC) be used for morphological examination and ER immunocytochemistry when adequate materials are available and that imprint materials be used when ABC materials are inadequate and fresh tissue is available at the time of surgery.

Two cell lines, NF and JoN, derived from human ovarian carcinosarcomas, were established in tissue culture and in nude mice. Both lines, growing in monolayers, showed morphologic features of adenocarcinoma cells (NF being aneuploid with a modal number of 53, and JoN being pseudodiploid with a modal number of 44). Intermediate filaments were demonstrated immunohistochemically; the JoN line expressed keratin, but not vimentin or desmin, whereas the NF line expressed vimentin and desmin, but not keratin. Plasminogen activator activity was found in both lines. It is concluded that both of these lines are potentially useful models for studying the diverse characteristics of malignant mixed Müllerian tumors.

DNA from 24 ovarian tumors, including 16 carcinomas, was examined for amplification of the proto-oncogenes c-myc, int-2, and rc-erbB-2. All cases of carcinoma were also examined by flow cytometry for DNA ploidy and cell cycle analysis, and eight cases of carcinoma were examined for estrogen and progesterone receptors. Protooncogene amplification was not detected in the DNA of benign ovarian neoplasms, or of ovarian carcinomas with low malignant potential. Amplification of c-myc was detected in six of 12 cases of invasive carcinoma, int-2 amplification was present in one case, and c-erbB-2 amplification was not detected in any case. Among the seven cases evidencing protooncogene amplification, three cases showed aneuploidy in tumor DNA, while four showed diploidy. Two cases which showed aneuploidy in tumor DNA did not demonstrate any degree of protooncogene amplification. Protooncogene amplification was frequently associated with morphologic nuclear anaplasia and high mitotic count. Six of the seven cases demonstrating c-myc or int-2 were of the serous type or showed some degree of serous differentiation, while none of the four cases of purely mucinous carcinoma had evidence of amplification. While the total number of cases in the study was limited, it would appear from the trend demonstrated by the data that protooncogene amplification (particularly c-myc) may be involved in the pathogenesis of aggressive common epithelial tumors of the ovary.

Four tumor markers (CA125, CA19-9, CEA, TPA) were analysed between the localizations and the serum data in the 36 ovarian cancers. The positive rates of each tumor markers were; 23 cases (63.9%) on CA125, 15 cases (41.7%) on CA19-9, 12 cases (33.3%) on CEA, and 27 cases (75.0%) on TPA. CA125 and CA19-9 were observed in the luminar borders and cellular membranes on serous adenocarcinomas and in the cytoplasms on mucinous adenocarcinomas. Both CEA and TPA were stained in cytoplasms. The correlation in the localization was observed between CA19-9s and CEAs (Kendall's rank correlation = 0.5303 greater than 0.5). The correlations between the localization and the serum data were observed on CA19-9 and CA125.

Surgically resected signet-ring cell carcinomas of the stomach have been clinicopathologically investigated. Although this type of carcinoma was found to be widely spread in the propria mucosa, a deeper invasion beyond the submucosa appeared more slowly than in other types of carcinomas. For example, the larger the early carcinoma in which the invasion was restricted to within the submucosa, the greater the incidence of a signet-ring cell carcinoma increase, especially in cases involving the mucosa. The incidence of a nodal metastasis was found to be lower than in cases of a moderately and poorly-differentiated adenocarcinoma in the early stage. However, when the signet-ring cell carcinomas invaded beyond the submucosa, the tumor cells spread rapidly and widely in the wall with a subsequent abdominal implantation, causing ascites and peritonitis carcinomatosa. It seemed likely that this deep invasion was accelerated by cellular change, such as the enlargement of nucleus, cellular atypy, and a decreased mucin production. As signet-ring cell carcinoma arise from the neck of glands and infiltrate the propria mucosae under the superficial epithelium, diagnosis by the barium enema and an endoscopic examination is very difficult in the case of small-size lesions. In lesions greater than 1 cm in diameter, these carcinomas usually showed an erosive and/or ulcerated appearance.

Regional ploidy variations within individual tumors were analyzed by in-situ cytofluorometry of metaphase cells in Feulgen-stained paraffin sections, using 45 resected stomachs with early and advanced signet ring cell carcinomas. Aneuploid cells were found in one of 30 early cancers and in eight of 15 advanced cancers, and were almost always accompanied by diploid cancer cells in the mucosal part of the cancers. The diploid and the aneuploid cells were generally found to be distributed in different territories in the mucosa, and aneuploid foci were often included in the diploid area. These findings suggest the diploid origin of signet ring cell carcinomas and the occurrence of aneuploidy during the tumor development. Moreover, the aneuploid cells appeared to infiltrate beyond the mucosa more readily than the diploid cells; most of the aneuploid populations already invaded the extramucosal tissue, and the cancer cells infiltrating in the extramucosal tissue were predominantly aneuploid in six of the nine cancers with aneuploidy. Thus, it appears that the occurrence of aneuploid clones may accelerate the progression of signet ring cell carcinomas from early to advanced stages.

Adenosine deaminase complexing protein (ADCP) immunoreactivity was investigated in 40 ovarian tumors and correlated with clinicopathologic parameters, including tumor steroid receptor content. Ten (29%) of 34 common epithelial ovarian carcinomas showed ADCP reactivity. Reactivity for ADCP was seen more frequently in mucinous (100%; p less than 0.001), well-differentiated (73%; p less than 0.001) and Stage I (56%; p less than 0.05) ovarian carcinomas. Furthermore, tumors that contained high levels of androgen receptors and tumors that did not contain estrogen receptors were more frequently ADCP positive (p less than 0.05). However, after stratifying for histologic grade, no correlation between ADCP reactivity and receptor status was found. Determination of ADCP reactivity appears to be of limited value in ovarian cancer.

Only 69% (42 of 61) of the documented ovarian tumor specimens received between September 1981 and August 1982 contained 75% or more viable tumor. Of the 42 measured for estrogen receptors (ER) and progesterone receptors (PR), 34 (81%) had estrogen-binding titers. greater than or equal to I fmol/mg of cytosol protein, while 18 (43%) had progesterone-binding titers greater than or equal to 5 fmol/mg of cytosol protein. ER and PR titers were found to be independent of histologic grade, clinical stage, patient age, age at first pregnancy, age at menarche, age at menopause and number of pregnancies. The presence of ER and PR in malignant ovarian tumors suggests that assays positive for these hormones could aid in the selection of patients suitable for hormonal therapy.

From 1963 to 1986, 1126 cases (1300 lesions) of early gastric carcinoma were treated at Fukui Prefectural Hospital. Mucinous adenocarcinomas comprised 16 lesions in this series. These cases of mucinous adenocarcinoma were younger than all the other early carcinomas of the stomach and they often were locted in the lower third of the stomach. Macroscopically, most were types "Iia + IIc". Dividing them into 3 grouping: papillary, tubular, and signet ring cell types, depending on the microscopic appearance of the intramucosal carcinoma, the signet ring cell group differed from the other two as to age, macroscopic appearance, and region it occupied. An immunochemical study showed that CA19-9 stained positively in a mucous lake, much like a carcinoma cell, but that CEA stained little.

By affinity chromatography on peanut agglutinin-agarose, receptors for peanut agglutinin were isolated from three lines of human malignant cells labeled with [3H]-galactose. The cell lines examined were a Burkitt lymphoma line Daudi, a lung squamous carcinoma QG-56 and a gastric signet ring carcinoma KATO-III. The isolated receptors from the three sources had similar properties and behaved as glycoproteins with apparent molecular weights of 160,000 to 280,000 upon sodium dodecyl sulfate gel electrophoresis. However, they showed molecular weights of only around 70,000 upon gel filtration, suggesting that they contained large amounts of carbohydrates. Pronase digestion depolymerized the glycoproteins, though significant amounts of the products were large enough to be excluded from a column of Sephadex G-50. The large molecular weight products were not detected when the receptors were treated with mild alkali and NaBH4 prior to pronase digestion. Major components of the radioactive oligosaccharides and glycopeptides released in the latter case had molecular weights in the range from 3,500 to less than 1,000. These results indicate that the glycoprotein receptors had clusters of oligosaccharide chains of small and medium sizes.

Long-term observation on transplanted tumor of human intestinal mucoid adenocarcinoma in nude mice is reported. The transplanted tumor which has been passed 57 passages in 151 nude mice with transplant survival rate of 99.34% in more than 5 years is the tumor with the utmost number of passages domestically undertaken as yet. During the long-term observation, the transplanted tumor was not only identical to the primary tumor in histopathology, ultrastructure and chromosome pattern, but also constant in biological characteristics, such as secretion of mucin and production of carcinoembryonic antigen. These tumor cells were able to combine with CEA mcAb specifically by obviously showing the scanning image of transplanted tumor of human intestinal mucoid adenocarcinoma in nude mice with 131I-CEAmcAb. The roles of the transplanted tumor model in cancer research are discussed.

Glucocorticoid receptors (GR) were studied in tumor cytosols from 140 breast cancer patients. The level of GR depended upon tumor histology. The highest GR level was found in medullary cancer, while the lowest--in a mucinous one. The GR level in ductal-invasive cancer was significantly higher than in the lobular infiltrative type. The frequency of GR in tumors of the 1st grade of malignancy was significantly lower than in those of the 2nd and 3rd grade of malignancy. The 1st grade of malignancy tumors which contained GR had estrogen and progesterone receptors, too. The increasing malignancy was matched by the rising percentage of such tumors which was as high as 47% among 3rd grade of malignancy tumors.

The DNA stemline of 45 mucinous breast carcinomas was determined by flow cytometry using paraffin embedded archival tissue sections. The material consisted of 26 pure mucinous and 19 mixed mucinous carcinomas. The patients were followed up for at least 15 years or until death. Nearly all pure mucinous carcinomas had a normal DNA stemline (25 of 26) with only one aneuploid tumour. Mixed mucinous carcinomas had a DNA content resembling that of common ductal carcinoma with 11 aneuploid tumours. Aneuploid tumours tended to be of higher grade and stage than diploid tumours. The survival of patients with pure mucinous carcinoma was better than that of patients with mixed mucinous carcinoma. Mucinous carcinoma should be classified as such only if it is a pure mucinous carcinoma.

A 62-year-old male was admitted to our clinic with the complaints of gross-hematuria and miction pain. Cystoscopic examination revealed non-papillary tumor around the orifice of the left ureter and the left wall. Histopathological findings of the biopsy specimen demonstrated signet-ring cell carcinoma, and the specimen was stained positively by the peroxidase-antiperoxidase technique. No malignant findings in any other organs including gastrointestinal tract and prostate were detected. This patient underwent total cystectomy with ileal conduit and histopathological staging was pT3bNOMO. He was followed with no evidence of local recurrence or metastasis for 29 months after operation. The 45 reported cases with primary signet-ring cell carcinoma of the urinary bladder including our case are reviewed and some characteristics of this entry are discussed.

The expression of the ras gene product p21 in normal gastric mucosa, early gastric carcinoma of diffuse (gastric) and intestinal types, and in adjacent mucosal abnormalities is reported. The analysis was performed on paraffin sections by an immunohistochemical assay using the mouse monoclonal antibody RAP-5 and the rat monoclonal antibody Y13-259. Expression of ras p21 was assessed by staining intensity and percentage of positively stained cells. In comparison to normal gastric mucosa of non-cancer patients, p21 was overexpressed in nearly all early carcinomas of both types and in the dysplastic and/or metaplastic mucosal alterations accompanying intestinal type of gastric cancer. Increased p21 expression was also observed in the normal-appearing mucosa adjacent to early carcinomas of diffuse type, but not in the morphologically normal gastric epithelium adjacent to the intestinal type. The results of this investigation suggest that ras p21 overexpression may be related to early events of human gastric carcinogenesis. The study supports the notion of different pathways in the development of diffuse (gastric) and intestinal types of gastric carcinomas.

Estrogen (ER) and progesterone (PR) receptors in 20 primary carcinomas of the ovary were studied by the customary DCC method. One of the receptors examined was found in 12 patients (60%) and both of them in 5 patients (25%). Serous types of tumour revealed a higher concentration of receptors than mucinous ones. The study of this kind is considered useful for the selection of patients in whom the adjuvant hormonal therapy could be applied.

A detailed histopathologic analysis of 399 primary breast carcinomas was performed, and several morphologic features were correlated with the estrogen receptor (ER) status. In all cases ER status was determined immunocytochemically by estrogen receptor immunocytochemical assay (ER-ICA). In 359 carcinomas ER status was also biochemically determined. Invasive lobular, mucoid, and tubular carcinomas rather than ductal carcinomas were ER-positive more frequently in ER-ICA. Medullary and papillary carcinomas had corresponding lower or higher ER positivity, respectively, by both methods. The correlation of histologic grade and its single factors with ER status was statistically significant by both methods. Lymphocytic reaction to tumor showed a significant inverse relationship to ER status by both methods. A statistically significant higher number of ER-positive carcinomas in ER-ICA and dextran-coated charcoal assay (DCC) occurred when elastic tissue was present. Different associations were found between stromal content, tumor diameter, and ER status in DCC and ER-ICA, respectively.

Two hundred two human, mucinous breast carcinomas were investigated for the presence of argyrophilic granules, and these granules were found in 25% of the cases. The granules were located in the cytoplasm and were heterogeneously distributed within the tumors. Tumors with granules were otherwise morphologically indistinguishable from those tumors without granules. The recurrence-free survival was independent of the presence of granules, and no relation was found to other clinical or histopathologic factors. Tumors with granules were found to be estrogen-receptor positive, and they appear to have a slightly less aggressive growth pattern than tumors without granules, but the difference is far from being statistically significant. It is concluded that there is no convincing evidence that this group of primary breast carcinomas with argyrophilia originates from APUD cells.

Tissues from 12 patients with gastric ulcers, 50 patients with gastric carcinoma, and 2 normal subjects who died suddenly were examined immunohistochemically with antiserum against the beta-subunit of human chorionic gonadotropin. Human chorionic gonadotropin-beta-immunoreactive cells were present in the neck portion of the pyloric and pseudopyloric glands of normal and inflammatory gastric tissue. Gastric cancers located in the antrum-pylorus and the cardia were frequently associated with human chorionic gonadotropin immunoreactivity. These findings indicate that human chorionic gonadotropin-producing cells normally exist in the gastric mucosa, especially in the pylorus. It is speculated that normal human chorionic gonadotropin-producing cells may give rise to cancer in certain cases.

Estrogen and progesterone receptors were measured in cystosols prepared from 32 normal ovaries and 25 benign and 49 malignant ovarian tumors. In normal ovarian tissue, estrogen and progesterone receptors were detected in 22 and 75% of specimens, respectively. Estrogen receptors were present in low concentrations ranging from 2 to 9 fmol/mg cytosol protein. The estrogen receptor content and distribution were similar in benign tumors (20%), but progesterone receptors were significantly decreased in 16% of specimens (P less than 0.001). In malignant ovarian tissues, estrogen receptors were present in 57% of specimens in concentration ranging from 1 to 132 fmol/mg cytosol protein. Of these, 72% of tissues had estrogen-receptor concentrations greater than 10 fmol/mg cytosol protein. The presence of estrogen receptors in ovarian cancer was significantly different from normal ovaries and benign tumor tissues (P less than 0.01). Progesterone receptors were detectable in 29% of ovarian cancer specimens. Estrogen and progesterone receptors were present alone or in combination, in 65% of ovarian cancers. The similarity in sex steroid content between ovarian and breast cancer warrants prospective studies of sex steroid receptor content in ovarian malignancies as a possible predictive index of survival and response to hormone therapy.

Gastric carcinomas have been assayed for the presence of villin and for the small intestinal hydrolases aminopeptidase N and sucrase isomaltase. These proteins seem not to be present in normal stomach epithelium. However intestinal metaplasia in stomach, and tumour cells in the glandular patterns of gastric carcinoma were positive for all three markers, showing characteristic apical positivity. In contrast, in diffuse gastric carcinomas the percentage of signet ring cells positive for these markers varied from 10-100% with each marker showing a similar percentage of positive cells. Testing of gastric carcinomas with antibodies specific for different keratin polypeptides showed that while all 7 tumours were positive for keratins 8 and 18.2 were also positive for keratin 7. In the keratin 7 positive tumours all tumour cells were keratin 7 positive. The keratin 8 antibody also reacted on routinely fixed specimens. Thus gastric carcinomas reveal different degrees of gastric and intestinal differentiation.

Immunohistochemical distribution of S-100 protein was evaluated in 129 tumors from major and minor salivary glands. Also, two sensitive immunoperoxidase avidin-biotin methods using either overnight incubation with primary antibody or pretreatment trypsin digestion and half-hour incubation were compared. Tumors with S-100 protein immunoreactivity were demonstrated in numerous benign and malignant histologic categories. Adenoid cystic carcinomas, carcinomas ex pleomorphic adenoma, clear cell carcinomas, and adenocarcinomas NOS showed inconsistent positive staining, whereas all monomorphic and pleomorphic adenomas and polymorphous low grade adenocarcinomas examined stained positively. No staining was observed in mucoepidermoid carcinomas or acinic cell carcinomas. Mesenchymal-like tumor cells with positive immunostaining were seen only in pleomorphic adenomas and trabecular-tubular adenomas. Equivalent results were found with both overnight and same-day digestion techniques. The consistent S-100 protein staining in some histologic tumor categories (pleomorphic and monomorphic adenoma and polymorphous low grade adenocarcinoma) compared to mucoepidermoid carcinoma that is devoid of S-100 protein immunoreactivity has application to some microscopic differential diagnostic situations. Inconsistent staining of adenoid cystic carcinomas and adenocarcinomas did not allow discrimination from other benign and malignant salivary gland tumors with similar histomorphology.

Reported in this paper are studies conducted into oestrogen receptor levels of 103 patients with mammary carcinoma, with the dextran-carbon method being used. The findings thus recorded are discussed for their clinical relevance. Recorded were correlations between quantitative oestrogen receptor levels, on the one hand, and menopausal status, grading, histological typing, and tumour size, on the other.

DNA content in the tumour cell nuclei was studied in 16 cases of early stomach carcinoma and in the adjacent areas of the foveolar epithelium dysplasia and metaplastic intestinal epithelium. One wave microspectrophotometry was used; the slides were stained with gallocyanin-chrome alum. The number of DNA synthesizing diploid cells was increasing with the increase of the dysplasia degree. The number of tetraploid cells was higher and the signs of aneuploidy appeared in the areas of noninvasive carcinoma. The stem line in early invasive carcinoma was represented by the diploid cells. The increase of cell atypia in tubular and papillary adenocarcinoma is followed by increase of the tetraploid cell number and by the appearance of polyploidy and aneuploidy. Both undifferentiated and goblet-cell carcinomas have similar parameters of the DNA histogram. DNA content allows an objective judgement on the degree of an early carcinoma cell atypia and differentiation of the advanced dysplasia and noninvasive carcinoma.

Of 534 resected colorectal adenocarcinomas, 165 (31%) contained some mucinous components; these represented the main part of the tumour in 67 (13%). Of the mucin containing tumours, 63 (38%) were in the right colon compared with 50 (13%) of the non-mucinous ones (p less than 0.001). Patients with predominantly mucinous tumours were significantly older than those with non-mucinous tumours, and they tended to present with tumours at a more advanced stage. A multivariate analysis did not show any significant independent prognostic influence of the mucinous component except when this had a predominantly signet ring cell pattern.

The authors report two cases of mucoargyrophilic carcinomas of the breast, occurring in 73 and 81 year old women. Both tumors consist of a central mucoid carcinoma surrounded by solid areas with lobular or endocrinoid features. Most cells showed an abundant eosinophilic granular cytoplasm or a mucinous appearance. The histochemical reactions (Grimelius-Alcian Blue, Grimelius-Mucicarmine) and the ultrastructural study of the peripheral areas disclosed several cell types: poorly differentiated or mucinous cells were present in both cases; but most of the cells were argyrophilic and contained in their hyaloplasm numerous dense core "neurosecretory" granules; such cells revealed many microfilaments arranged in bundles or in whorls. Amphicrine cells were easily demonstrated by histochemical staining and their cytoplasm showed both endocrine granules and mucinous vesicles. These features suggest a possible relationship to other argyrophilic tumors of the breast. The histogenesis of these tumors is still little known.

Monoclonal antibody LICR-LON-M18 is a marker of normal human breast epithelial cell differentiation. The epitope recognized by LICR-LON-M18 is a prominent component of luminal plasma membranes of nonneoplastic resting and lactating human breast epithelial cells but is rarely expressed by human breast carcinomas. With the use of competitive binding-inhibition studies, the immunodominant portion of the LICR-LON-M18 epitope was shown to be the following oligosaccharide sequence [with galactose (Gal) and N-acetylglucosamine (GlcNAc)]: Gal beta 1----4GlcNAc beta 1----. This structure was distinct from Gal beta 1----3GalNac, which was bound by peanut agglutinin (PNA) and was not recognized by LICR-LON-M18 [corrected]. With the use of biochemical techniques, the present data not only confirmed sialylation and consequent "masking" of the LICR-LON-M18 epitope and PNA determinants in human breast carcinomas but also identified the particular groups of glycoproteins involved in this process. These studies provided additional support for the thesis that sialylation of human breast carcinoma glycoproteins represented an enhancement of specific differentiation events normally regulated in the morphogenesis of nonneoplastic human breast epithelium and that specific glycoproteins became masked during the genesis of primary human breast cancer.

Mucin in normal, metaplastic and neoplastic tissue specimens of the human urinary bladder was examined histochemically by the periodic acid-borohydride/potassium hydroxide/periodic acid-Schiff (PB/KOH/PAS) reaction and paradoxical concanavalin A (Con A) staining, both of which have been reported to detect mucins that are specific for the lower gastrointestinal tract. Normal mucosa, Brunn's nest, squamous metaplasia, transitional cell carcinoma, and squamous cell carcinoma produced only small amount of mucin and were negative for both PB/KOH/PAS and labile class III by Con A staining. However, several sections of mucin-producing metaplastic bladder lesions and many samples of adenocarcinoma arising from vesical mucosa and urachus contained PB/KOH/PAS-positive and/or labile class III mucin. Thus the colonic type mucin appeared in some materials of both benign and malignant mucin-producing lesions of the urinary bladder. The findings would be related to the embryogenesis of this organ.

An immunoperoxidase staining technique was used for detecting three major iron binding proteins (ferritin, transferrin and lactoferrin) in 40 breast carcinoma cases and six benign breast proliferative lesions. Ferritin staining was detected mainly in connectival stroma and in histiocytes surrounding neoplastic cells. Few and faint ferritin positivities were also detected in neoplastic cells of 20 carcinoma cases. Transferrin was found inconsistently in myoepithelial cells surrounding normal ductules, or around neoplastic ducts of ductal in situ carcinoma. In eight carcinoma cases, transferrin staining was also positive in neoplastic cells. Lactoferrin was detected only in normal breast epithelial cells and in benign breast proliferative lesions. These immunohistochemical findings may suggest that raised serum ferritin concentrations in breast carcinoma patients might be attributed to stromal reaction rather than to tumour synthesis. Transferrin staining of neoplastic cells in these carcinoma cases appears to be very intriguing, particularly since transferrin is considered an obligate requirement for growing cells, and transferrin receptors have been demonstrated only in dividing cells. On the basis of the immunohistochemical data, lactoferrin might be used as a pointer to benign lesions.

Immunohistochemical determinations of carcinoembryonic antigen, alpha-fetoprotein, human chorionic gonadotropin (hCG), and human placental lactogen (hPL) were performed on tissue sections from 137 epithelial ovarian cancers. Fewer than 25% of serous cystadenocarcinomas contained detectable amounts of any marker. Carcinoembryonic antigen was present in over 50% of tumors, and was noted most frequently in mucinous, endometrioid, and clear cell carcinomas. hPL was demonstrated in 30% of endometrioid carcinomas but was rarely present in other cell types. Both alpha-fetoprotein and hCG were noted in fewer than 10% of all major cell types of epithelial ovarian cancer. Forty-five patients had serial determinations of plasma levels of carcinoembryonic antigen at the time of regular follow-up visits. Serial plasma levels of carcinoembryonic antigen accurately predicated recurrence in nine of 16 patients whose tumors contained carcinoembryonic antigen, in contrast to two of 16 patients whose tumors were devoid of antigen.

An immunoperoxidase method was used to compare the distribution of fibronectin and laminin between superficially spreading and deeply infiltrating parts of signet-ring-cell carcinoma of the stomach. In both parts, laminin-containing basement membranes were generally scarce, but they were observed on some of the cancer cells which had differentiated to glandular cells. Intramucosal invasion fronts of superficially spreading cancers often showed a layered structure, consisting of a middle zone of small cancer cells together with a superficial and a deep zone of signet-ring cells. In this structure, linear fibronectin and laminin deposits were common on the cancer cells in the deep zone, but rare in the superficial zone. However, fibrillar fibronectin deposits in the stroma were not considerably larger in either zone. At extramucosal invasion fronts of deeply infiltrating (advanced) cancers, a stromal remodelling with an increased amount of fibrillar fibronectin deposits was often observed around the cancer cells, whose cell surface fibronectin was largely lost even from some basement membranes. These findings suggest that invasive activity of signet-ring-cell carcinoma may not be related to the mere presence or absence of cell-surface fibronectin and laminin but to the amount of stromal fibronectin, which could reflect a cell-stroma interaction. Signet-ring-cell carcinomas have a stage of intramucosal growth in which cancer cells may live in dependence upon the pre-existing stroma and form the layered structure, while in advanced stages cancer cells seemed to have acquired an ability to elicit their own "tumor stroma".

Four cases of primary signet-ring cell adenocarcinoma of the urinary bladder are described. All patients were men, with ages ranging from 46 to 81 years. The number of signet-ring cells varied and showed a direct correlation with poor prognosis. PAP analyses of acid phosphatase and prostate-specific antigen were negative. The results show that signet-ring cell carcinoma has a poor prognosis. This cell type should be sought in all bladder carcinomas since its presence is likely to worsen prognosis.

An immunohistological study of 15 chordomas, six chondrosarcomas, four liposarcomas and seven carcinomas on paraffin embedded samples using anti-cytokeratin, anti-epithelial membrane antigen (EMA), anti-S100 protein, anti-vimentin and anti-neurofilaments showed that chordomas has a characteristic immuno-staining, i.e. positive for cytokeratin, EMA, S100 protein and vimentin; and negative for neurofilaments. This immuno-staining allows a clear distinction of chordomas from other tumours.

The evaluation of serial plasma carcinoembryonic antigen (CEA) levels is one of the most important parameter used to establish the prognosis of surgically cured colorectal cancer patients. Carcinoembryonic antigen is particularly useful in the identification of recurrences and metastasis. However, to improve the usefulness of this assay, it would be helpful to accurately determine, if possible, those patients whose cancers produce CEA. The evaluation of the presence of CEA in these cancer specimens by means of immunoperoxidase staining technique does seem to improve the sensitivity of the CEA test. Fifty-seven patients with colorectal cancer who underwent surgical treatment were studied. Tissue CEA evaluation was correlated with the plasma CEA levels, the pathologic stage and grade, and histologic type of the cancers. Results demonstrate that 66.6% of Dukes' B cancers, 78.9% of Dukes' C, and 77.7% of Dukes' D cancers stained positively for CEA by immunoperoxidase. Thirty of 57 patients with preoperative pathologic plasma CEA levels had positive tissue CEA, whereas 8/57 patients did not. Of patients with a well-differentiated cancer (G1), 81.4% had positive tissue CEA versus the 64% of G2 and 60% of G3 cancers. The authors conclude that the use of the immunoperoxidase stain to measure CEA in tissue, so that the CEA serum assay may be used in those patients known to produce CEA, results in a major increase in the sensitivity of the test.

A combination of reverse phase and normal phase high pressure liquid chromatography has been used to separate the reduced oligosaccharides produced by alkaline borohydride degradation of a blood group A ovarian cyst mucin glycoproteins. Fourteen compounds, ranging in size from a monosaccharide to a decasaccharide, have been isolated preparatively using a Zorbax C-18 reverse phase column eluted with water and a MicroPak AX-5 normal phase column eluted with aqueous acetonitrile. The purity of the products and their structures were determined from the fully assigned high field proton NMR spectra. The resonances of exchangeable amide protons, observed by the Redfield selective pulse sequence in H2O, were assigned by decoupling to the resonances of H2 of the 2-acetamido sugars. Nuclear Overhauser effects were used to establish the relationship of the anomeric protons and those of the aglycone. In exception to earlier proposals that nuclear Overhauser effect on irradiation of the anomeric proton should always be observed at the proton attached to the aglycone carbon, we find that for the linkage of GalNAcp(1----3)Gal, nuclear Overhauser effect on irradiation of the alpha-anomeric proton resonance is observed not at H3 but at H4 of galactose. A combination of NMR methods and enzymatic degradation was employed to determine the structures of 13 different oligosaccharides of which seven have not previously been reported. These oligosaccharides, which terminate with beta-Gal, alpha-Fuc, beta-GlcNAc, and alpha-GalNAc, account for 75% of the total glycoprotein carbohydrate, the remainder being isolated as a mixture of glycopeptides and a high molecular weight polysaccharide whose NMR spectrum implies a simple repeating subunit structure closely related to that of the oligosaccharides.

Of 256 patients with carcinoma confined to the uterine corpus at the time of hysterectomy treated in the period 1959-1975 at Stanford University Hospital, 98 patients (38%) had neoplasms which demonstrated at least focal intracytoplasmic mucin. In 21 carcinomas (9%), intracytoplasmic mucin production was the dominant form of differentiation--a group which we designate primary mucinous carcinoma of the endometrium. Freedom from relapse and frequency of myometrial invasion were not statistically different for patients whose neoplasms contained intracytoplasmic mucin, regardless of the amount of mucin present, when compared with cases of nonmucin-containing carcinoma. Using histochemical methods, it was impossible reliably to distinguish between the intracytoplasmic mucin produced by carcinomas arising in endometrium and that produced by carcinomas primary in the endocervix. Differential biopsy and fractional curettage are stressed as useful tools in making this clinically important distinction. Since both benign mucinous metaplasia and mucinous carcinoma may arise in the endometrium, it is important to establish histopathologic criteria by which the malignant lesions may be recognized. The use of criteria illustrated in this paper (which include architectural complexity of proliferation, epithelial stratification, loss of epithelial polarity, and nuclear atypicality) resulted in the recognition of mucin producing proliferations which as a group manifest a 50% incidence of myometrial invasion.

Three to 5% of breast carcinomas are argyrophilic, including some which are mucinous and thus "composite", whereas there are no argyrophilic cells in normal breast nor in benign breast pathology. This raises the problem of the origin and type of these argyrophilic cells. We carried out a histologic and immunohistochemical study in 4 such cases of mucoid tumors containing at least 50% argyrophilic cells. Two of these tumors presenting node involvement were also studied immunohistochemically. The histologic study showed colloid and intragalactophoric proliferation areas in cell cases and some endocrine areas in 2 out of 4 cases. Argyrophilic cells were present in all of these areas. True mucoargyrophilic amphicrine cells were found primarily in colloid areas. None of these tumors were argentaffin. Immunohistochemical study was performed by the PAP method using antibody directed against VIP, ACTH, PP, somatostatin, bombesin, calcitonin, gastrin, prolactin and GH. Three out of four tumors were positive with VIP. Moreover one of them contained ACTH cells and a metastasis of this tumor contained bombesin cells. No tumor was positive with the other anti-sera tested. This study is related to the rare series in the literature which report secretion of ACTH, catecholamins, bombesin, gastrin, VIP, PP, somatostatin, prolactin, etc. The number of cases reported to date remains too low to show a significant prognostic difference between amphicrine tumors and other mammary carcinomas.

An improved immunohistochemical determination of the cytokeratin profiles of epithelia and their neoplasms is possible using monoclonal antibodies that will either identify all 19 cytokeratins (AE1/3) or delineate specific subsets (35 beta H11, 34 beta E12, 34 beta B4 and Cam 5.2). Ovarian common "epithelial" tumors (CET) contain cytokeratin filaments. To determine the nature and differences in the cytokeratin profiles of ovarian CET, eight benign Brenner tumors, four serous cystadenofibromas, 28 mucinous tumors, 27 serous tumors and six endometrioid, five clear cell and five undifferentiated carcinomas, as well as nine normal ovaries were immunostained with the above five antibodies. AE1/3 staining was predominant, while Cam 5.2 and 35 beta H11 displayed the most frequent staining thereafter. Statistically significant staining differences were found between a number of tumor groups using the antibodies 35 beta H11, 34 beta E12 and Cam 5.2. In this study, all ovarian CET, except the benign Brenner tumors, displayed a predominantly low molecular weight cytokeratin profile. The same profile in the normal surface epithelium lends credence to the belief that these tumors are derived from this epithelium. A significant staining difference between some of the tumor types using some of the antibodies suggests a possible ancillary, diagnostic role of cytokeratin profiling in situations where exact tumor typing is difficult.

Breast cancer tissue from 12 male patients was evaluated for the presence of estrogen receptors (ER), and at least 10 of 12 were found to be high in ER (ER+). A combination of previously published cases and our own experience indicates that 80% of patients (37/47) have ER+ tumors. This incidence of ER positivity in men is greater than that in women with breast cancer. Clinical responses to hormonal therapy have been observed in the ER+ patients but not in one ER--patient. Measurement of ER may be clinically important in men with breast cancer.

For the purpose of investigating cell the kinetics of human gastric carcinoma, the distribution of 3H-thymidine labeled cells and the labeling index were evaluated in biopsy specimens obtained from 52 patients with gastric cancer, using an in vitro labeling technique. Adenocarcinomas were scattered with labeled cells, while in carcinoid speciners labeled cells were confined to the periphery of tumor cell nests. Characteristically, small-sized cells were exclusively labeled with 3H-thymidine in signet-ring cell carcinomas but signet-ring cells bearing a large amount of mucin were not. Thus signet-ring cells could be regarded as cells in a differentiated condition which have lost their prolife-rating capability. The labeling index of gastric cancers ranged from 10 to 20% in most cases although it varied greatly from case to case (2.2-40.8%). The values of the labeling index were correlated with neither the degree of histological differentiation, macrascopic appearance nor the depth of mural invasion of carcinomas.

Estrogen and progesterone receptors were studied in 70 cases of human colorectal cancer by a cytochemical technique. 28.5% of the cases were estrogen-receptor positive and 42.8% progesterone-receptor positive. There was no difference between the sexes for estrogen receptors but the women had more tumours with progesterone receptors than men. The presence of receptors is unrelated to the differentiation of the tumor. More colon tumours were positive than those of the sigma and rectum. The concentration of cells with receptors in positive cancer cases tended to be low or medium-low.

136 breast carcinomas were investigated immunohistochemically for alpha-lactalbumin (ALA), lactoferrin (Lfr), human milk fat globule membrane antigen (HMFG-2), transferrin receptor (TrfR), Ki-67 and epidermal growth factor receptor (EGFR). The relationships of pathologic features and steroid receptor status previously shown to be of prognostic significance and the immunohistochemical parameters were examined. It was found that estrogen receptor (ER) was inverse related to TrfR, Ki-67 and EGFR whereas progesterone receptor (PgR) was inverse correlated to Ki-67 solely. Tumor grading revealed significant correlations between TrfR, Ki-67 and HMFG-2 (inverse correlation). Tumor diameter showed correlation between Ki-67 solely. Moreover there were significant relationships between lymphoid infiltration and TrfR, Ki-67 and HMFG-2 (inverse correlation). The comparison of the immunohistochemical parameters showed correlations between Ki-67 and TrfR, Ki-67 and HMFG-2 (inverse correlation) as well as HMFG-2 and ALA. Therefore it is suggested that functional and tumor kinetic properties determined by HMFG-2, Ki-67 and TrfR may be an additional indicator with prognostic significance. On the other hand immunoreactivities of ALA in conjunction with HMFG-2 possibly indicates a subpopulation of breast carcinomas that may have to be investigated further. Moreover these results showed that lymphoid infiltration was correlated with Ki-67 reactivity as well as TrfR expression.

The distribution of intermediate - sized filaments in human colon mucosa as well as in adenomas and carcinomas of the colon was studied by means of both immunohistology and electron microscopy. The epithelial cells of the colonic mucosa are definitely labelled with antibodies against prekeratin (cytokeratin). Interwoven filaments of the prekeratin type are present in the basal compartments of the epithelial cells; they surround the nuclei and mucus droplets and form an apical skeletal disc. Pericryptal connective tissue is prekeratin negative and vimentin positive. Benign hyperplastic polyps have a high content of prekeratin. The potential precursors of colonic carcinoma, i.e., the tubular and villous adenomas, also show an increase in intermediate-sized filaments of the prekeratin type. Correspondingly, electron microscopy reveals elongated bundles of intermediate-sized filaments arising from the desmosomes of the lateral and basal cell membranes. The prekeratin content is particularly high in adenocarcinomas and highest in mucinous carcinomas. As expected, the stroma of all neoplasms studied is prekeratin-negative, but distinctly vimentin-positive. In one moderately differentiated adenocarcinoma there was evidence of "vimentin-positive" tumor cells. These changes may be caused by binding of cytokeratins with an unknown substance in vimentin antisera, as observed similarly by Moll et al. (1982) in a transitional cloacogenic carcinoma.

The mucin secreted by mucoid carcinomas of the breast has been studied in a series of 20 tumours by histochemical methods and peroxidase labelled peanut lectin. The histochemical techniques showed that the carcinomas could be divided into two groups depending on whether the mucin was composed predominantly of acid or neutral mucopolysaccharides. The specific binding of peanut lectin to beta-galactosyl groups, in combination with neuraminidase treatment, enabled further divisions to be made and so demonstrated the heterogenous nature of the mucin secreted by mucoid carcinomas and also the value of using lectins as specific probes in histological methods.

We reviewed 18 cases of mucinous adenocarcinoma of the endometrium seen at this hospital during 1969-1985. Tumor histology was studied in curettage and hysterectomy material; the Hospital Tumor Registry provided follow-up data. All women were postmenopausal, 47-89 years of age, and presented with vaginal bleeding; few had abdominal pain; four of the 18 were nulliparous. Eight patients had been receiving estrogen therapy. The disease was in Stage IA in 15 patients and Stage IB in three patients. All patients had total hysterectomy and salpingo-oophorectomy; six had preoperative radium implants, and four had postoperative radium plus progesterone treatment. Histologically the tumor was grade 1 in 13 (72%) women, only five being grade 2 or 3 (28%). All tumors were positive for mucicarmine and carcinoembryonic antigen (CEA). Only two cases showed adjacent benign endometrial mucinous metaplasia. Myometrial invasion varied from 5-60% and was absent in three cases. The degree of invasion did not appear to affect the rate of recurrence. The tumor recurred 54 weeks after treatment in one of 13 patients with grade 1, in none in grade 2 and in two of two patients with grade 3 within 10 months of diagnosis. In summary, mucinous endometrial carcinoma is a rare tumor (1% of endometrial carcinomas in our hospital) of postmenopausal women; it tends to be well differentiated and has a relatively good prognosis.

The correct classification of carcinoma of the lung is not only of therapeutic and prognostic importance but is also considered to have epidemiological and aetiological significance. Histological tests for mucin are essential in the classification of lung tumours but there is little information available about the influence of the method of detection used on the results of classification. Five established staining techniques were tested using paraffin blocks from surgical specimens of 81 human lung tumours diagnosed as adenocarcinoma, i.e. tumours of WHO Type III. Mowry's alcian blue-periodic acid-Schiff (AB-PAS) technique gave the highest proportion of positives (93%) slightly fewer (90%) being obtained by the PAS technique alone. Both these methods were influenced by the presence of cytoplasmic hyaline globules, structures which cannot be regarded as mucin. The stain recommended by the World Health Organization was also influenced by the presence of hyaline globules, was less frequently positive than the PAS techniques and was considered to have no special advantages. The aldehyde fuchsin-alcian blue sequence was positive in only 83% of cases but provided some information about the type of mucin present. Southgate's mucicarmine also detected mucin in only 83% of cases. It was concluded that the apparent incidence of adenocarcinomas may be influenced by staining methods used. Some standardization of technique is desirable and the AB-PAS combination appears to be the most satisfactory.