Nuclear morphology in breast lesions: refining its assessment to improve diagnostic concordance.

AIMS: Although evaluation of nuclear morphology is important for the diagnosis and categorisation of breast lesions, the criteria used to assess nuclear atypia rely upon the subjective evaluation of several features that may result in inter- and intraobserver variation. This study aims to refine the definitions of cytonuclear features in various breast lesions. METHODS AND RESULTS: ImageJ was used to assess the nuclear morphological features including nuclear diameter, axis length, perimeter, area, circularity and roundness in 160 breast lesions comprising ductal carcinoma in situ (DCIS), invasive breast carcinoma of no special type (IBC-NST), tubular carcinoma, usual ductal hyperplasia (UDH), columnar cell change (CCC) and flat epithelial atypia (FEA). Reference cells included normal epithelial cells, red blood cells (RBCs) and lymphocytes. Reference cells showed size differences not only between normal epithelial cells and RBCs but also between RBCs in varied-sized blood vessels. Nottingham grade nuclear pleomorphism scores 1 and 3 cut-offs in IBC-NST, compared to normal epithelial cells, were < ×1.2 and > ×1.4 that of mean maximum Feret's diameter and < ×1.6 and > ×2.4 that of mean nuclear area, respectively. Nuclear morphometrics were significantly different in low-grade IBC-NST versus tubular carcinoma, low-grade DCIS versus UDH and CCC versus FEA. No differences in the nuclear features between grade-matched DCIS and IBC-NST were identified. CONCLUSION: This study provides a guide for the assessment of nuclear atypia in breast lesions, refines the comparison with reference cells and highlights the potential diagnostic value of image analysis tools in the era of digital pathology.

Comparison of the Diagnostic Accuracy of Magnetic Resonance Imaging with Sonography in the Prediction of Breast Cancer Tumor Size: A Concordance Analysis with Histopathologically Determined Tumor Size.

BACKGROUND: In order to effectively treat patients with breast cancer, it is important to know the precise tumor size. We compared the rates of concordance of magnetic resonance imaging (MRI)-derived and sonography-derived breast cancer tumor size with histopathologically determined tumor size. METHODS: Accuracy of MRI and sonography in establishing tumor size was evaluated by comparing preoperative images with postoperative pathologic findings. The accuracy of MRI and sonography was graded as concordance, underestimation, or overestimation and was compared in different subgroups. RESULTS: A total of 682 patients comprised the study cohort. Mean tumor size was 3.64 ± 1.8 cm via MRI, 2.12 ± 1.0 cm via sonography, and 2.78 ± 1.7 cm via pathologic examination. The difference between breast sonography and MRI to pathologic tumor field size was -0.68 ± 1.4, and 0.85 ± 1.25 cm, respectively (P < 0.001). Sonography had a concordance rate of 54.3 %, an overestimated rate of 9.8 %, and an underestimated rate of 35.9 %. For MRI, the concordance rate was 44.1 %, the overestimated rate was 52.5 %, and the underestimated rate was 3.4 %. In subgroup analysis, breast MRI had a higher concordance rate in patients with T3 (>5 cm) lesions. When the results of MRI and sonography were considered together, the concordance rate increased from 54.3 to 62.2 %. CONCLUSION: MRI tends to overestimate the actual tumor size, while sonography frequently underestimates it. Combined sonography and MRI increases the accuracy of tumor size prediction.

La telorrea como manifestación del carcinoma intraductal de mama / Nipple discharge as a manifestation of ductal carcinoma in situ

Antecedentes. La secreción por pezón es un método diagnóstico inocuo y accesible que, en la práctica, se considera de baja sensibilidad y especificidad diagnóstica. Caso. Mujer de 45 años con telorragia derecha y escozor que como único dato en pruebas radiológicas presenta dilatación de 1mm. de un conducto retroareolar. En una de las exploraciones, se consigue muestra de secreción a la expresión que muestra un fondo con hematíes y algunos agregados de células epiteliales, con ligera superposición celular, contorno abollonado y otras veces liso. Las células muestran aumento de relación N/C, hipercromasia, contorno nuclear irregular e imágenes de canibalismo. Con la sospecha de lesión papilar se realiza biopsia excisional que evidencia un carcinoma intraductal de grado medio con necrosis de tipo comedo. Conclusión. Aunque el carcinoma ductal de mama, ya sea intraductal o infiltrante se expresa sólo raramente con telorrea, ante grupos epiteliales con atipia citológica no hay que olvidar esta posibilidad además de lesiones papilares (AU)

Introduction. Nipple discharge is an innocuous and accessible diagnostic method that, in practice, is considered to have low sensitivity and specificity. Case. A 45-year-old woman presented with bloody right nipple discharge and burning sensation. Imaging tests revealed a 1-mm dilatation as the only finding of interest. In one of the examinations, a sample of the secretion was obtained that showed a bloody background and some clusters of epithelial cells, with slight cellular overlapping, and contour that was smooth in parts and dented in others. The cells showed an increase in the nucleus/cytoplasm ratio, hyperchromatic nuclei, irregular nuclear membrane and images of cannibalism. Because a papillary lesion was suspected, excisional biopsy was performed, revealing intermediate-grade ductal carcinoma in situ with comedo-like necrosis. Conclusion. Though the ductal carcinoma of the breast, whether in situ or invasive, is rare expresses nipple discharge, if there are epithelial clusters with cytologic atypia we have not forget this possibility besides papillary lesions (AU)

Correlation of ultrasound findings with histology, tumor grade, and biological markers in breast cancer.

BACKGROUND AND PURPOSE: Ultrasound has been used successfully to differentiate benign and malignant breast lesions. The aim of this study was to investigate the correlation between ultrasound and prognostic indicators in breast cancer such as histological type, tumor grade, and biological markers. MATERIALS AND METHODS: Ultrasound findings (shape, margin, orientation, boundary, echo pattern, posterior acoustic feature, and presence of calcifications) of 458 breast cancers were analyzed and correlated with the tumor type, tumor grade, and biological markers by univariate and multivariate logistic regression analyses. The biological markers were estrogen receptor, progesterone receptor, and HER-2/neu. RESULTS: Invasive cancers displayed more frequently an irregular shape, a not parallel orientation, and a hypoechoic or complex echo pattern than carcinoma in situ cases (p < 0.05). Poorly differentiated invasive cancers had more frequently not circumscribed margins, an abrupt boundary, and a hypoechoic or complex echo pattern than moderately/well differentiated cancers (p < 0.05). Estrogen or progesterone receptor negative cancers more often displayed a hypoechoic or complex echo pattern and HER-2/neu positive cancers had more calcifications (p < 0.05). CONCLUSION: Ultrasound pattern is correlated with tumor type, tumor grade, and biological markers in breast cancers and it may be useful for prediction of prognosis.

Nuclear morphometry in columnar cell lesions of the breast: is it useful?

AIMS: To evaluate the nuclear morphometric features of breast columnar cell lesions (CCLs) observed on mammotome core biopsies, to determine if there are significant measurable differences between those with atypia and those without. Correlation with follow-up open excision specimens was made. METHODS: Mammotome core biopsies performed on patients that contained CCLs were derived from the departmental case files. Histological material was reviewed and foci of CCLs demarcated for nuclear morphometric assessment, which was accomplished using an imaging system. Nuclear parameters studied were nuclear area and perimeter, circularity factor and feret's diameter. Statistical analysis used the GraphPad Prism software, with p<0.05 indicating significance. RESULTS: On examination of core biopsies of 40 patients with CCLs, 8 lesions were benign, 4 showed atypical lobular hyperplasia, 8 showed CCLs with nuclear atypia, 19 disclosed atypical ductal hyperplasia (ADH) and 1 showed ductal carcinoma in situ (DCIS). The nuclear area, perimeter and feret's diameter of CCLs with atypia were significantly greater than those without (p = 0.04, 0.03 and 0.019, respectively), whereas no difference was observed in the circularity factor. Follow-up open excision biopsy specimens in 24 patients showed upgrading to DCIS in 40% of cases diagnosed initially with ADH on core biopsy compared with 20% of CCLs with atypia. CONCLUSIONS: Nuclear morphometry in CCLs confirms nuclear size as the key parameter in the assessment of nuclear atypia. Whether it can be potentially used as an adjunctive tool depends on the establishment of appropriate cut-offs.

Cystic hypersecretory carcinoma: rare and poorly recognized variant of intraductal carcinoma of the breast. Report of five cases.

AIMS: To report five cases of a rare variant of intraductal carcinoma of the breast, so-called cystic hypersecretory carcinoma. The clinical and pathological characteristics of the lesion are described, along with a review of the literature. METHODS AND RESULTS: The patients were females aged between 53 and 78 years (average 66.8 years). The size of the lesions ranged between 70 and 80 mm in largest dimension. In two cases, the development of high-grade invasive ductal carcinoma was observed; in one additional case there was recurrence of high-grade in-situ carcinoma after 3 years. This emphasizes the importance of correct diagnosis of this potentially aggressive lesion. Strong over-expression of HER-2/neu protein was observed in three cases, including the two with an invasive component. Protein p53 was variably positive in all cases. Steroid receptor immunohistochemistry yielded variable results with only one case being positive for both oestrogen and progesterone receptors. Interestingly, in most cases (4/5) staining for androgen receptors was observed. CONCLUSIONS: Cystic hypersecretory ductal carcinoma of the breast is a rare distinctive variant of ductal carcinoma in situ. It has the potential for invasive growth and the development of metastases.

Ductal carcinoma in situ with spindle cells: a potential diagnostic pitfall in the evaluation of breast lesions.

AIMS: To evaluate the morphological features of 11 cases of breast ductal carcinoma in situ (DCIS) with spindle cells and to propose an approach to distinguish it from benign mimics. The association with neuroendocrine differentiation was also investigated. METHODS: Cases of breast DCIS with a spindle cell component diagnosed in the Department of Pathology, Singapore General Hospital, between June 1996 and January 2003, were included in the study. The histological characteristics were documented, and immunohistochemistry for neuroendocrine markers, hormone receptors, cerbB2, smooth muscle actin (SMA) and high-molecular-weight (HMW) cytokeratins, was carried out. Electron microscopy was carried out on reprocessed paraffin-embedded material in three cases. RESULTS: Of 11 women diagnosed with DCIS with spindle cells, four presented with nipple discharge, six with a breast lump, while one was discovered to have a screen detected density. The tumour size ranged from 3 to 41 mm. The proportion of spindle cells varied from 10% to 80% of the in-situ tumour cell population. Nuclear grade was low in seven cases and intermediate in four. Necrosis was observed in two cases. Architectural pattern was papillary in six cases, and mixed in the rest. Microinvasion was present in two cases, with possible microinvasion in another two. Immunohistochemistry for neuroendocrine markers synaptophysin and chromogranin showed positive reactivity for at least one marker in all but three cases; one of these latter cases demonstrated ultrastructural neurosecretory granules. Oestrogen and progesterone receptors were expressed in 10 and nine cases, respectively, while cerbB2 was positive in only one case. HMW cytokeratin immunoprofile revealed a general lack of immunostaining within the abnormal cell population; likewise, no positivity for SMA of the cellular proliferation was detected. CONCLUSIONS: Almost all DCIS lesions with spindle cells disclose neuroendocrine differentiation. Although the distinction from benign florid usual hyperplasia may pose a diagnostic histological problem, the presence of diffuse neuroendocrine expression, in conjunction with the pattern of HMW keratin profile on immunohistochemistry, supports an in-situ neoplastic process. The absence of SMA immunostaining, in conjunction with negative reactivity for cytokeratins 5/6 and 14, makes the possibility of a myoepithelial proliferation unlikely.

Aggressive giant fibroepithelial lesion with unusual vascular stroma--a case report.

The stroma of fibroadenoma and phyllodes tumor usually consists of fibroblastic proliferation. Rarely the stroma contains bundles of smooth muscle. Pseudoangiomatous hyperplasia of the mammary stroma has been described in fibroadenomas. However, true benign vascular stroma has not been reported. We report a case of a 34-year-old Chinese woman who presented with a large mass occupying the entire left breast. Left mastectomy was performed and showed a large, well-circumscribed, lobulated, rubbery-firm tumor measuring 13 x 10 x 6 cm. Microscopic examination revealed a fibroepithelial tumor formed by an organoid pattern of ductal structures with a very striking stromal appearance composed of extensive vascular proliferation and that demonstrated strong immunoreactivity for CD31, CD34, and Factor VIII. Ultrastructural examination revealed intercellular junctions, basal lamina, pinocytotic vesicles, and Weibel-Palade bodies in the cells lining the vascular spaces, confirming their endothelial nature. These findings rule out the diagnosis of pseudoangiomatous hyperplasia. The patient developed local recurrence a year later, and the resection showed malignant phyllodes tumor with ductal carcinoma in situ. The extensive vascular stroma noted in the primary tumor may have played a role in the malignant transformation of the epithelial and stromal components in this tumor.

Carcinoma ductal micropapilar hipersecretor. Estudio inmunohistoquímico y ultraestructural / An atypical case of hypersecretory micropapillary ductal carcinoma. An immunohistochemical and ultrastructural study

El caso descrito corresponde a una variante inusual de carcinoma ductal de mama llamada Carcinoma Ductal Hipersecretor Quístico. Esta lesión esta caracterizada por la formación de ductos quísticos con mínima atipia celular, lo cual plantea el diagnostico diferencial con la contrapartida benigna llamada Hiperplasia Hipersecretora Quística. A la paciente se le detecta, en una revisión periodica mamográfica, multiples microcalcificaciones en el cuadrante superointerno de mama izquierda. Tras biopsia extemporánea se practica cuadrantectomía con linfadenectomía axilar. La paciente esta libre de enfermedad dos años después del tratamiento. De este caso se realizaron estudios ultraestructurales e inmunohistoquímicos complementarios (AU)

Ultrastructure of the periductal area of comedo carcinoma in situ of the breast.

We have previously shown that the different biological natures of comedo ductal carcinoma in situ (DCIS) and non-comedo DCIS may, in part, be explained by the different expression patterns of tenascin, a large extracellular matrix protein, as observed by immunohistochemical studies. In the present study, we compared 8 cases of comedo DCIS with 5 cases of non-comedo DCIS by ultrastructural analysis, focusing on the myoepithelium, basal lamina, and tenascin-positive extracellular periductal stromal matrix. Our observations show that the comedo type DCIS frequently has an altered basal lamina, a looser and more disorganized collagenous matrix, and a general increase in stromal cellularity, including fibroblasts, lymphocytes, histiocytes and small blood vessels. In addition, in comedo DCIS, the lateral intercellular spaces between large myoepithelial cells that border the basal lamina are often expanded, compared to those of non-comedo DCIS. These results identify structural characteristics of comedo DCIS that may play a role in its greater preinvasive potential. They may also provide a structural basis for the different strategies that are needed for for clinical management of comedo DCIS, compared to non-comedo DCIS.

Mammary "comedo"-DCIS: apoptosis, oncosis, and necrosis: an electron microscopic examination of 8 cases.

The terms apoptosis and necrosis are commonly used to imply two distinct types of cell death. Apoptosis reflects a genetically mediated. ATP-dependent form of cell death. A passive form of cell death (oncosis) also occurs, often in response to some form of injury. Both pathways can lead to necrosis (postmortem autolytic cell changes). The nature of intraluminal necrosis in mammary ductal carcinoma in situ (DCIS) was evaluated using ultrastructural analysis on paraffin-embedded material of 8 cases with "comedo"-DCIS. In each case, intraepithelial proliferation zones and intraluminal zones (peripheral and central luminal zones) were examined. All cases with "comedo"-DCIS revealed abundant apoptosis, characterized by apoptotic cells showing chromatin condensation and margination with sharply circumscribed, uniformly dense crescents, as well as cytoplasmic condensation. Numerous membrane-bound apoptotic bodies with condensed cytoplasm (with or without nuclear fragments) were also observed. The central luminal zones of "comedo"-DCIS, however, revealed necrotic debris characterized by severe degradative changes, largely devoid of recognizable cell structures. In addition, two cases displayed features of oncosis, characterized by nuclear and cytoplasmic swelling, vacuolization of cytoplasm, and mitochondrial swelling with occasional dense bodies. The results indicate that necrosis (postmortem, secondary degradative cell changes) in "comedo"-DCIS is the end result of either apoptosis (programmed cell death) alone or a combination of apoptosis and oncosis (passive or "accidental" cell death).

Immunohistochemical localization of metallothionein in human breast cancer in comparison with cathepsin D, stromelysin-1, CD44, extracellular matrix components, P53, Rb, C-erbB-2, EGFR, steroid receptor content and proliferation.

Metallothionein (MT) is a low molecular weight, cysteine-rich, zinc-binding protein that may have a function in cellular repair processes, growth and differentiation. Using a monoclonal antibody (E9) to metallothionein, we investigated the immunohistochemical expression of MT in routinely fixed and paraffin-embedded tissue from 98 cases of female breast carcinomas. The MT expression was studied in comparison with the expression of the basement membrane (BM) antigens (type IV collagen, laminin), fibronectin, cathepsin D, adhesion molecule CD44, p53 protein, the pRb, c-erbB-2 oncoprotein, EGFR, stromelysin-1, proliferation indices (Ki-67, PCNA), steroid receptor content as well as with other conventional clinicopathological parameters of breast cancer. Strong MT expression was observed in the majority of tumour cells in 18.4% of tumours, focal MT positivity in 13.3% and almost complete lack of MT expression in 68.4% of cases (mean value 33.36 +/- 26.36). The MT expression in carcinoma cells was strongly associated with the DCIS component of the tumour (p < 0.0001). High values of MT were correlated with low steroid receptor status (p = 0.08 for ER receptor and p = 0.019 for PgR receptor content). MT positive cases were correlated with stromelysin-1 expression (p = 0.059) and cathepsin D (p = 0.058). These findings suggest that MT expression is characteristic of the early phase of breast carcinogenesis, possibly regulated by hormones, and could be a new potential prognostic marker in breast cancer.

Immunohistochemical localization of beta-1,4-galactosyltransferase in human pancreatic tissues.

beta-1,4-Galactosyltransferase (GalTase) is the glycosyltransferase in the Golgi apparatus that transfers galactose from UDP-galactose to terminal N-acetylglucosamine residues in glycoconjugates with formation of a beta-1,4 linkage. Neoplasms undergo various changes in the carbohydrate moieties of their glycoconjugates. This process also indicates the possibility of changes in glycosyltransferases themselves. Therefore, we compared the binding pattern of a monoclonal antibody (MAb8628) against GalTase in both normal and neoplastic exocrine pancreatic tissues. Ten normal and 11 neoplastic human exocrine pancreatic tissues obtained from surgery were used. Frozen sections were incubated with this antibody. Supranuclear regions and terminal bars of normal duct cells and acinar cells revealed positive staining for GalTase at the light microscopic level. Centroacinar cells revealed positive staining in their perinuclear region. Neoplastic cells were also stained in their supranuclear regions and terminal bars. Supranuclear regions were well developed in neoplastic cells and intensely stained compared with those in normal cells. The supranuclear regions and the terminal bars corresponded to the trans cisternae of the Golgi apparatus and the junctional complex (i.e., tight junction and adherens junction), respectively, seen at the electron microscopic level. Pancreatic neoplastic changes thus led to an increase in the expression of GalTase in the Golgi apparatus, the increase of which may have an important effect on the intercellular adhesion and communication among pancreatic epithelial cells. Measurement of this enzyme is useful for diagnosis of exocrine pancreatic neoplastic changes from normal tissues.

Intraluminal crystalloids in breast carcinoma. Immunohistochemical, ultrastructural, and energy-dispersive x-ray element analysis in four cases.

OBJECTIVE: Intraluminal crystalloids have been described in the prostate, salivary gland, and ovary, but have not yet been reported in the breast. We report four cases of breast carcinoma in which these crystalloids were found in ducts with intraductal carcinoma or atypical hyperplasia. The presence of intraluminal crystalloids may be a useful adjunct in making a diagnosis of carcinoma or may be a feature to look for as a marker for the presence of carcinoma. DESIGN: Four cases of breast carcinoma containing intraluminal crystalloids were identified among 6900 surgical breast specimens between January 1990 and June 1995 at M. D. Anderson Cancer Center, Houston, Tex. Those sections with crystalloids identified by hematoxylin-eosin stain were stained with periodic acid-Schiff, Alcian blue, and mucicarmine stains. Immunohistochemical and ultrastructural studies and energy-dispersive x-ray analysis were also performed on these sections. RESULTS: The intraluminal crystalloids were eosinophilic, varied in shape and size, and did not exhibit birefringence under polarized light. Immunohistochemically, the crystalloids were negative for keratin, muscle-specific actin, and kappa and lambda light chains, but the surfaces stained positively for epithelial membrane antigen. By electron microscopy, the crystalloids had no limiting membrane and were composed of an electron-dense material with no discernible periodicity. By energy-dispersive x-ray element analysis, the crystalloids had no mineral content; however, sulfur was found, indicating a protein content. CONCLUSIONS: The pathogenesis and constituents of these intraluminal crystalloids remain to be determined. Inasmuch as intraluminal crystalloids have not been found in normal ducts or acini, or in ductal hyperplasia without atypia, their presence may serve as a marker for breast carcinoma.

Endocrine ductal carcinoma in situ (E-DCIS) of the breast: a form of low-grade DCIS with distinctive clinicopathologic and biologic characteristics.

Endocrine ductal carcinoma in situ (E-DCIS), first characterized by Cross et al. in 1985, is an uncommon entity, and there is little information on its pathobiologic features and natural history in the literature. This report describes the largest series of 34 cases: 14 cases were pure in situ (group A), and 20 were accompanied by an invasive component (group B). All except three patients were over the age of 60 years, with the mean being 69.5 years for group A and 72.6 years for group B. Except for six patients in group A who had nipple discharge, all had a breast mass. On follow-up, one of five group A patients developed local recurrence 5 years after mastectomy, and two of seven group B patients developed another invasive primary in the contralateral breast. Histologically, E-DCIS showed expansile intraductal growths forming solid sheets and festoons traversed by delicate fibrovascular septa. Accumulation of basophilic mucin might be found within the growth and the fibrovascular septa. There were variable degrees of stromal sclerosis. In some cases, the solid intraductal cellular proliferations were focally punctuated by microglandular spaces and rosettes. Comedo necrosis was absent. Intraductal papillomas were found in the immediate vicinity of the tumors in 18 cases and invariably showed pagetoid involvement by E-DCIS. Pagetoid spread into the adjacent ducts and ductules was also a common feature (17 cases). The tumor cells were polygonal, oval, or spindly, often with eccentrically placed, bland-looking, ovoid nuclei and abundant eosinophilic granular cytoplasm. Intracellular mucin was commonly demonstrable. Immunostaining for myoepithelium using muscle-specific actin antibody confirmed the in situ nature of the E-DCIS component. The majority of tumor cells showed strong staining with the neuroendocrine markers chromogranin, synaptophysin, and neuron-specific enolase (monoclonal). Immunostaining also dramatically highlighted the pagetoid spread into the papillomas and ductules by outlining the tumor cells between the negatively stained residual ductal epithelium and myoepithelium. All cases were immunoreactive for estrogen and progesterone receptor, but not p53 and c-erbB2. The Ki-67 index was < 5%. Ultrastructural studies on four cases showed many dense-core neurosecretory granules and larger mucigen granules. In group B cases, the invasive component, which comprised 5-95% of the tumor, included colloid carcinoma, 12; "carcinoid" tumor, 3; mixed "carcinoid"/colloid carcinoma, 4; and small cell neuroendocrine carcinoma, 1. Neuroendocrine markers were also consistently demonstrable in the invasive component. In conclusion, E-DCIS is predominantly a disease of older women that is frequently accompanied by papillomas in the vicinity and may present as nipple discharge (an uncommon presentation in the usual forms of DCIS). It can mimic epitheliosis histologically, but the pagetoid spread is a helpful clue to its neoplastic nature. The bland nuclear morphology, lack of necrosis, and biologic marker profile suggest that E-DCIS is a form of low-grade DCIS despite its solid growth pattern. The invasive carcinomas associated with E-DCIS are also neuroendocrine programmed rather than the usual types of ductal carcinomas, suggesting that E-DCIS represents a biologically distinctive category of DCIS.

Derivation of ductlike cell lines from a transplantable acinar cell carcinoma of the rat pancreas.

Two cell lines were derived from a transplantable acinar cell carcinoma that had been established from a primary carcinoma of the pancreas in an azaserine-treated Lewis rat. The cultured tumor cells initially produced amylase, but production of exocrine enzymes ceased after 1-2 weeks in culture. The cultured cells were tumorigenic in Lewis rats, and one line produced solid tumors composed of ductlike structures surrounded by dense fibrous tissue. The second cell line produced partially solid and partially cystic tumors with a mixed phenotype of squamous, mucinous, and glandular areas when it grew in vivo following regrafting. Both cell lines lost structural and immunohistochemical acinar cell markers while acquiring duct cell markers during culture and regrafting. These studies provide strong support for the hypothesis that ductlike carcinomas can arise from neoplastic pancreatic acinar cells in rats.

Laminin production and fibronectin immunoreactivity in breast carcinomas.

Eighty-nine cases of primary breast carcinoma were evaluated immunohistochemically for basement membrane laminin and fibronectin (FN). 58 of the cases were also examined by transmission electron microscopy (TEM). Intraductal carcinomas were all positive for laminin and FN and by electron microscopy a continuous BM was found in all of them. Among invasive tumors, immunoreactivity for laminin was expressed in 31% of cases and FN in 66%, whereas TEM revealed BM in only 4 out of 52 (8%) studied cases. Tumors measuring 10 mm or less were more often laminin-positive and FN-negative than larger tumors. There was a tendency for lymph node negative tumors to express laminin more often, but there was no relation between lymph node status and FN. Laminin immunostaining proved to be very useful in identifying vascular invasion. The laminin and FN immunoreactivity showed no significant correlation with overall survival and disease-free survival (DFS) of the breast cancer patients. DFS for cases with laminin positive tumors was 50% and for laminin negative tumors 46.8%. For cases with tumors immunoreactive for FN, DFS was 50%, and for tumors lacking FN 43.5%.

Ductal carcinoma of breast: nuclear grade as a predictor of S-phase fraction.

Nuclear grade (NG) and S-phase fraction (SPF) are established independent prognostic variables for ductal breast carcinomas. Nuclear grade can be assigned by a pathologist in a simple fashion during histopathologic evaluation of the tumor, while SPF requires flow cytometric evaluation of tumor samples. This prospective study was undertaken to determine whether elevated SPF could be predicted from NG alone and how NG and SPF correlate with c-erbB-2 expression. Eighty-two breast carcinomas of ductal type were assigned an NG of low (grade 1 or grade 2) or high (grade 3). S-phase fraction was recorded initially from fresh-frozen tissue samples and was designated as either low SPF (below the value designated as the cutoff for elevated SPF) or high SPF (a value at or greater than the cutoff value). On fresh tissue the NG predicted the range of SPF (low or high) in 89% of cases. Four percent of the cases that did not correlate could definitely be attributed to sample error. The remaining 7% that did not correlate could have been due to sample error, specimen quality, or tumor heterogeneity, as demonstrated by reversal of SPF range as performed on paraffin blocks of tumor. Eighty-eight percent of the tumors positive for c-erbB-2 were NG 3 and 12% were NG 2. All c-erbB-2 tumors were aneuploid. This study demonstrates the importance of carefully assigning NGs on tissue and indicates the importance of reviewing flow cytometric data side by side with histopathologic parameters to detect discrepancies between these two modalities. Careful nuclear grading assignment can accurately predict the range of SPF.