c-erbB-2 protein expression in node negative breast cancer.

We investigated the expression of c-erB-2 protein in two matched groups of breast cancer patients, one with and one without relapse. 37 patients with relapse were compared with 42 patients without recurrence for time of observation, adjuvant treatment, age, menopausal status and estrogen receptor content. Paraffin-embedded sections were stained with the polyclonal antibody 21N, raised against a synthetic peptide from the predicted sequence of the c-erbB-2 protein. The staining of c-erbB-2 was measured on a scale of 0 to 3+. C-erbB-2 staining was negative in 16 (38%) patients in the relapse-free group, and in 8 (22%) of the patients with metastases. Neither disease-free survival (DFS) nor overall survival (OS) were dependent upon the extent of c-erbB-2 expression. An analysis by estrogen receptor (ER) status (i.e. positive or negative) and by c-erbB-2 expression (i.e. positive or negative) revealed that patients with ER-positive primaries and negative c-erbB-2 have the longest disease-free survival (DFS) and overall survival (OS). We conclude that c-erbB-2 expression might be clinically useful only if other prognostic variables (e.g. estrogen receptor content in the tumor) are also considered.

Correlation of c-erbB-2 gene amplification and protein expression in human breast carcinoma with nodal status and nuclear grading.

Fifty-one primary human breast tumors were analyzed for amplification of the c-erbB-2 protooncogene. Thirteen (25%) of the DNA samples contained multiple gene copies. Paraffin-embedded tumor sections, available from 47 of the cases, were stained with a c-erbB-2 specific antiserum. Eighty-three % (10 of 12) of the tumors containing amplified c-erbB-2 gene copies stained positively with the c-erbB-2 specific antiserum (P = 0.03). Thirteen tumors containing single copy c-erbB-2 sequences also stained positively with the antiserum. This suggests that mechanisms other than gene amplification may lead to elevated levels of c-erbB-2 protein. Finally, there was a statistically significant correlation between c-erbB-2 protein expression and parameters used in breast cancer prognosis. Positive staining was associated with positive nodal status of the patient (P = 0.02) and with tumors showing a poor nuclear grade (P = 0.02). This is the first study showing that a determination of the level of c-erbB-2 protein in paraffin-embedded tumor sections may have prognostic value for the course of human breast cancer.

Sensitive detection and typing of human papillomavirus DNA in gynecological cell scrapings by slot-blot hybridization.

A rapid and sensitive method for detecting and typing human papillomaviruses (HPVs) in cell scrapings is presented. DNA from scrapings is extracted and bound to nitrocellulose filters (Slot-Blot). By DNA-DNA hybridization with specific 32P-labelled HPV-probes (types 6/11 or 16/18) the patient's DNA is then analyzed for the presence of, and for the type of, HPV DNA sequences. A parallel hybridization with a human repetitive element (Alu sequence) allows quantitation of the different hybridization results. Experiments with HeLa cell DNA show that as little as 10(4) HPV sequences can be detected and typed specifically with this test. Evaluation of this test is completed within 6 to 7 days after cell collection. This Slot-Blot method was used to analyse 1330 specimens taken at the Bernese Dysplasia Outpatient Clinic. The results reveal a very high percentage (90%) of HPV-positive cases in the patient group examined.

[Intramural pregnancy with vague symptoms]. / Symptomarme intramurale Gravidität.

The clinical course of an intramural pregnancy in a 28-year-old nullipara is reported. Clinical and sonographic symptoms were not specific. The diagnosis was suspected on laparoscopy. At laparotomy, evidence of incipient rupture of the uterus was found, but no intraperitoneal hemorrhage. The gestational sac was removed without opening the uterine cavum, and the localization within the myometrium was confirmed histologically. Intramural pregnancy is a rare event whose pathogenesis is not clear. Surgical treatment with conservation of the uterus is possible in some cases.

[Human papillomavirus infections of the cervix uteri. I. Pathobiology, epidemiology and relation to cancer pathogenesis]. / Humane Papillomavirus-Infektionen der Cervix uteri. I. Pathobiologie, Epidemiologie und Beziehungen zur Krebspathogenese.

Infection of the uterine cervix by human papilloma viruses (HPV) is a ubiquitous yet only recently recognized lesion. The morphological findings correlate with early and late gene expression. The pathological events presumably depend upon the HPV type involved and upon environmental and host factors. The recognized risk factors are those described for epithelial dysplasias and carcinomas. The prevalence is unclear: it varies among different population groups and depends upon the investigation methods employed. The natural history of the infection is unknown, although relations between cervical HPV infection and carcinogenesis are evident. Research is handicapped by the lack of suitable animal or in vitro models.

[Human papillomavirus infections of the cervix uteri. II. Diagnosis, terminology, clinical significance of the diagnosis]. / Humane Papillomavirus-Infektionen der Cervix uteri. II. Diagnostik, Terminologie, klinische Bedeutung der Diagnose.

Cervical HPV infection may be diagnosed by colposcopy, cytology, histology and DNA hybridization. Each of these techniques alone may result in the detection of no more than 70% of the cases which are found by combining all methods. Typical cytological and histological findings are illustrated. A morphological continuum exists between HPV associated epithelial atypias (VAE), dysplasias and carcinomata in situ. The higher the grade of dysplasia, the lower the rate of cytological VAE. HPV types 16/18 are more frequently found in severe rather than in mild dysplasias. The use of hybridization techniques in mass screening programs appears premature, since the natural history of the infection is unknown. Microscopic grading of epithelial dysplasia remains the basis for patient care.

Sonographic patterns of ovarian tumors: prediction of malignancy.

Four hundred four women with suspected pelvic masses or histories suggesting ovarian cancer (eg, low abdominal pain) entered a prospective study designed to assess the accuracy of sonography in confirming or excluding the presence of ovarian cancer. Three hundred twelve of these patients were operated on and evaluated within three weeks after sonography. The predictive value of the sonographic evidence of malignancy was 73% (38 of 52 patients), whereas benign tumors were predicted correctly in 95.6% (177 of 185). Sonographic reassessment of masses with patterns suggesting benign disease may be an alternative to immediate surgical exploration in a selected population (ie, those with poor surgical risk). The sonographic detection of ovarian malignancy requires further improvement; as a diagnostic tool it continues to present a challenge.

Activities in leiomyomas and myometrium of enzymes involved in energy metabolism.

The activities of six enzymes involved in energy metabolism were measured in leiomyoma specimens and in the adjacent normal myometrium from the uterus of 17 patients. In leiomyomas the specific activities of hexokinase, lactate dehydrogenase and hydroxyacyl-CoA dehydrogenase (HAD) were higher than in myometrium. The soluble protein content was lower in leiomyomas. Therefore, most of the differences of specific activities were not found when the enzyme activity was expressed per gram wet weight of tissue, except for HAD activity, which was still higher in leiomyomas (p less than 0.05). This result is compatible with increased fatty acid utilization by leiomyomas, and with the increased growth potential of such tumors.

Prognostic significance of tumor grade in clinical trials of adjuvant therapy for breast cancer with axillary lymph node metastasis.

The prognostic significance of histologic tumor grade has been evaluated in 1537 women entered into the Ludwig Trials I-IV of adjuvant therapy for node-positive breast cancer. Tumor grade was determined on histologic review of primary tumor sections by two central review pathologists using a modification of the Bloom and Richardson grading system. The 5-year overall survival rates (+/- SE) were: Grade 1, 86% +/- 2; Grade 2, 70% +/- 2; and Grade 3, 57% +/- 2 (P less than 0.0001). This survival difference was seen in both premenopausal (P less than 0.0001) and postmenopausal (P less than 0.0001) women. Significant differences in disease-free survival (DFS) by tumor grade were also observed (P less than 0.0001). The tumor grade determined by the 75 contributing local clinic pathologists was also highly significant for predicting DFS and overall survival. Tumor grade remained a statistically significant prognostic factor for DFS (P less than 0.0001) and overall survival (P less than 0.0001) in multivariate analyses controlling for nodal status, tumor size, estrogen receptor status, menopausal status, age, peritumoral vessel invasion, and treatment assigned. In postmenopausal patients for whom adjuvant treatment was compared with no adjuvant therapy, the prognostic significance of tumor grade was modified by the effect of treatment. The presence of vessel invasion by primary tumor cells was a stronger predictor of early recurrence than was increasing tumor grade in postmenopausal patients who received no adjuvant therapy. The higher failure rates for patients with high-grade tumors was due to a larger number of failures in regional and visceral sites. Tumor grade can be determined by any pathologist and allows for selection of a subpopulation of breast cancer patients at high risk for early mortality.

[Intratumoral regional differences in hormone receptor status of breast cancer]. / Intratumorale regionäre Unterschiede im Hormonrezeptor-Status des Mammakarzinoms.

Biopsy specimens of tissue were taken from 32 invasive carcinomas of the breast, in each case from the centre of the tumour, the tumour margin and the surrounding mammary gland tissue. After preparation of a frozen section for tissue identification, the concentration of oestrogen and progesteron receptors (ER and PR, respectively) was determined in each specimen by means of the dextrane-coated charcoal method. On the basis of these measured values, the receptor status of each specimen was classified either as positive (R+), borderline value (BL) or as negative (R-). Various morphological parameters were compared with the measured data. 9 (28%) of the 32 carcinomas demonstrated such high intratumoural differences in receptor concentration that the receptor status was classified quite differently in each of them. Formation of tubuli of the invasive ductal carcinomas was more marked with the R+ tumours (p = 0.005) than with the R- tumours. The quantitative ER content did not correlate with the abundance of the tissue specimen (r = 0.182). The regional differences in receptor status could not be explained, neither by the abundance in cells nor by morphological peculiarities of the tumour. - It follows from the results of this study that intratumoural regional variations in ER and PR concentrations can occur in carcinoma of the breast, and that such variations can have clinical significance.

Receptor heterogeneity of human breast cancer as measured by multiple intratumoral assays of estrogen and progesterone receptor.

Multiple intratumoral tissue samples from the primary mass of 30 consecutive invasive breast cancer patients were assayed for estrogen receptor (ER) and progesterone receptor (PR) by the dextran-coated charcoal method following frozen section histopathological examination. Steroid receptor status of each sample was classified as positive (R+) or negative (R-), based only upon quantitative guide lines from the ER/PR results. Four out of 32 (12.5%) of the invasive cancers had an intratumoral sample classified as R+ and one sample as R-. R+ invasive ductal carcinomas has a highly significant degree of tubule formation (P = 0.005) when compared with R- invasive ductal cancers. While the quantitative ER content (r = 0.18) and the degree of quantitative variation in ER content (P = 0.04) did not correlate with the tumor cellularity of the individual samples, tumor cellularity (P = 0.005) and ER content (P = 0.005) were lower in the samples from the tumor border than from the central tumor samples. Variations in ER and PR content may be found on a regional basis within a breast tumor mass resulting from heterogeneity of tumor subpopulations and/or differences in tumor cellularity.

Pathologic data of prognostic significance for remission induction in advanced ovarian carcinoma.

Sixty-eight patients with "advanced ovarian carcinoma" were entered into an ongoing phase-II trial for remission induction with cis-platinum (DDP) 80 mg/m2 i.v. on day 1 followed by forced saline diuresis, melphalan (L-PAM) 12 mg/m2 i.v. on day 2 and hexamethylmelamine (HMM) 130 mg/m2 p.o. X 14 days from days 8-21 in six monthly cycles following operative resection and/or staging. Fifty-one patients were evaluable for response, ten had not completed six courses and could not be assessed, two patients died early (one probably of toxicity), and five patients refused treatment and follow-up. Thirty-Two patients had serous, endometrioid or undifferentiated carcinomas of the ovary. Of these, 11 (35%) achieved a pathologically proven complete remission (CR), five (16%) were NED after second-look (residual disease in ovary or removed omentum with all other biopsies and cytology washings negative), eight (32%) achieved a partial remission (PR), and three (12%) had progressive disease. None of the seven patients with clear-cell carcinoma and none of the three patients with Mullerian tumor of the ovary responded. Six of nine patients with tumors of uncertain origin or proven metastasis to ovary did not respond to treatment. These preliminary results indicate that advanced ovarian carcinomas form a heterogeneous group of recognizable neoplastic diseases with striking variation in response to treatment.

Immunohistochemical localization of galactosyltransferase in the normal human ovary and fallopian tube.

Galactosyltransferase (UDP-galactose: 2-acetamido-2-deoxy beta-D-glucopyranose beta-(1-4) transferase) in human tissue specimens from ovaries and the corresponding fallopian tubes was localized immunohistochemically for light microscopy. An affinity-purified rabbit anti-human milk galactosyltransferase antibody was used. Intracellular galactosyltransferase was found to be localized to the juxtanuclear (Golgi) region of the secretory cells of the fallopian-tube epithelium and to the ovarian stromal cells involved in steroid-hormone production. Cell-surface galactosyltransferase was localized to ciliated cells of the fallopian-tube epithelium. During the follicular phase of the menstrual cycle, galactosyltransferase was found only in the Golgi regions of theca interna cells of the ovarian graafian follicle, and in the fallopian tube was found predominantly on the cilia of epithelial cells. During the luteal phase, galactosyltransferase was abundant in the Golgi regions of granulosa lutein cells of the corpus luteum, and was predominant in the secretory cells of the tubal epithelium. Galactosyltransferase was not detected on the mesothelial ovarian surface. The results demonstrate that the cellular distribution and location of galactosyltransferase correlates with phenotypic differentiation and varies during the human female hormonal cycle.

[Ultrasonic diagnosis of ovarian carcinoma]. / Ultraschalldiagnose des Ovarialkarzinoms.

Gray-scale ultrasonography was performed prior to surgery in 68 patients with tumors of the ovary. Thirty-four of the 38 benign tumors were considered benign by sonographic criteria. Three of 4 borderline epithelial tumors were considered malignant sonographically, as were 22 of the 26 cancers of the ovary (sensitivity 84%). Nineteen of them were malignant epithelial neoplasms and most were classified as FIGO stages III and IV (76%). In 6 of the 7 patients with stage I and II cancers the latter were considered malignant by sonographic criteria. Twenty-three of the 26 cancers were classified as having a complex structure using the classic ultrasonographic criteria. Histopathological evaluation did not correlate with the ultrasonic findings: sonography could not predict the correct stage and a particular histological type. Nevertheless, ultrasonography is a valuable diagnostic tool as a preoperative procedure in patients with a suspected ovarian neoplasm.

Cellular localization of keratin in proliferative epithelial processes and neoplasms of the human ovary.

Cytokeratins are one of the intermediate cytoplasmic filaments which contribute to the cytoskeleton. Keratins have recently been demonstrated in normal and neoplastic tissues as well as in human cell lines. It has been suggested that the cellular location of keratin may reflect tissue-specific or epithelial type differentiation. Twenty-three examples of human ovaries containing the full spectrum of epithelial proliferations from inflammatory reactive processes to malignant neoplasia were studied for the cellular distribution of cytokeratin using antisera to human keratin. Nineteen cases contained immunoreactive keratin which was limited to the epithelial cells: 2/2 inflammatory, 8/10 benign tumors, 5/7 borderline tumors, 4/4 carcinomas. There was marked regional heterogeneity in keratin expression such that adjacent morphologically-identical cells could be functionally distinguished by the immunoreactive staining. The predominant cellular localization of keratin varied between histological tumor types in the benign neoplasms: serous = apical, subciliary; endometrioid = apical; mucinous = basal. This pattern was lost in the cytological progression to borderline and malignant tumors. In borderline tumors, the most intense reactivity was noted in areas of cellular atypia and proliferation. In borderline and malignant tumors, keratin was usually present in basal cytoplasmic regions contiguous with stroma.

Humoral antibodies against spermatopoietic cells associated with mediastinal germinoma.

A case of mediastinal germinoma in a 21-yr-old male is reported. The neoplasm exhibited a morphology similar to that of a testicular seminoma and was heavily infiltrated by lymphocytes and plasma cells. Germinal centres were also present in lymph follicles. Humoral antibodies (IgG), specific for spermatopoietic tissue, were present in the patient's serum. Antibody titres declined after surgical removal of the germinoma over a period of 2 yr, and antibody is no longer detectable at the present time. The implications of this observation are discussed with regard to defence mechanisms of the tumour host, and to the histogenesis of these rare neoplasms.