HER-2 and INT-2 amplification estimated by quantitative PCR in paraffin-embedded ovarian cancer tissue samples.

Competitive polymerase chain reaction (PCR) systems were developed for rapid and quantitative estimation of HER-2 (c-erbB-2) and INT-2 oncogene amplification in paraffin-embedded ovarian cancer tissue samples. The beta-globin gene was used as reference and DNA from paraffin-embedded placenta tissue as single copy control. Reliability of the PCR method could be demonstrated by comparing dot blot data with PCR data of identical tumour samples. The PCR method was used to determine HER-2 and INT-2 copy numbers in 196 ovarian cancer samples. HER-2 and INT-2 were found to be amplified in 40 and 19%, respectively. In 8% HER-2 copy numbers were greater than five, but no high INT-2 copies were noted. Kaplan-Meier estimates did not reveal significant association with overall survival. Indirect correlation between HER-2 and INT-2 amplification was observed. The present PCR system is a valuable method for prospective and retrospective studies.

Ki-1-positive large cell lymphoma. A clinicopathologic study of 41 cases.

We report the clinicopathologic findings of 41 patients with Ki-1 (CD30)-positive large cell lymphoma. The median age was 50 years; 13 patients were under 40 years of age. Ten patients presented with extranodal disease. Fifty-five percent of the patients presented with stage I or II disease, and bone marrow involvement was histologically documented in 30% and occurred exclusively in patients over 40 years of age. Two cytomorphologically distinct groups of Ki-1--positive large cell lymphomas could be separated. Group A lymphomas consisted of pleomorphic large cells, sometimes with wreathlike and embryo-like nuclei, whereas group B lymphomas displayed a rather monomorphic appearance. Clinically the two groups of lymphomas differed with respect to stage of disease, frequency of bone marrow involvement, and median survival. On paraffin sections, the Ki-1--related antibody Ber-H2 provided excellent staining results in all cases. Immunologic phenotyping disclosed a T cell type in the majority of cases, revealed marked loss of differentiation antigens, and frequent expression of HLA-DR and IL-2 receptor. The overall median survival was 13 months. Age below 40 years, limited stage of disease (I and II), and, although not statistically significant, lymphoma morphology were associated with longer survival. We conclude, that Ki-1--positive large cell lymphomas represent a morphologically and immunologically heterogeneous category of hematolymphoid neoplasms derived from dedifferentiated and activated lymphoid cells with marked age-dependent prognosis.

Peripheral T-cell lymphomas: a clinicopathologic study of 75 cases.

Seventy-five peripheral T-cell lymphomas (PTLs) were classified according to the recently proposed "Updated Kiel Classification of Non-Hodgkin's Lymphomas" (mycosis fungoides and Sezary's syndrome excluded). Thirty-seven PTLs belonged to the low-grade category (T-cell chronic lymphocytic leukemia [T-CLL], 3; lymphoepithelioid, 4; angioimmunoblastic, 22; T-zone, 6; pleomorphic small cell, 2) and 38 belonged to the high-grade category (pleomorphic medium and large cell, 24; immunoblastic, 1; large-cell anaplastic Ki-1-positive, 13). Loss of pan-T antigens occurred exclusively in high-grade PTLs; on paraffin sections UCHL 1 was slightly more sensitive than MT 1. Sixty patients presented with lymphadenopathy and 15 patients (20%) presented with extranodal disease most frequently affecting the skin and upper aerodigestive tract. B-cell lymphoma symptoms were found in 43 cases (57%) and bone marrow involvement (T-CLL excluded) was found in 12 cases (17%). Staging (T-CLL excluded) revealed stage I in 13%, stage II in 15%, and stages III and IV in 72% of the cases. Among the intensively treated patients, 37% achieved complete remission and 15 are still in complete remission after 4 to 79 months (median: 24 months). The overall median survival (MS) rate was 23 months. Peripheral T-cell lymphoma of pleomorphic medium and large-cell type was the most aggressive lymphoma (MS: 8 months). B-cell lymphoma symptoms, bone marrow involvement, and Ki-67 positivity 60% or greater significantly shortened survival times, whereas age (under 60 versus over 60 years), stage (I and II versus III and IV), and grade had no significant influence. Ki-67 reactivity was found to be a prognostic factor which allows prediction of probable poor outcome, especially in cases with limited stage of disease.

MACOP-B treatment in patients with Ki-1-positive large-cell anaplastic lymphoma.

Nine adult patients with Ki-1-positive large-cell anaplastic lymphoma were treated with MACOP-B. Two suffered from relapsed disease and had previously received chemotherapy; a third patient had received a single dose of 100 mg/m2 cisplatin before initiation of MACOP-B. The stage of lymphoma was determined according to the Ann Arbor Conference criteria and was II in one, III in two and IV in six patients. All patients had constitutional symptoms. Five patients had achieved complete remission 4 weeks after termination of the protocol and there were two partial remissions. One patient died of massive pulmonary embolism during the 4th week of treatment; another patient, who had received MACOP-B as salvage therapy, died of progressive lymphoma 1 month after completion of the regimen. Maximal observed toxicities according to WHO were mucositis grade 3 (n = 3) and there were three cases with thromboembolic complications, including a fatal pulmonary embolism in a young patient. However, MACOP-B appears an effective, fairly well-tolerated and feasible therapy for patients with Ki-1-positive large-cell anaplastic lymphoma.

Classification of intestinal T-cell neoplasms and their differential diagnosis.

We studied 42 intestinal T-cell lymphomas (ITLs) to establish a simple and reproducible classification. The ITLs were classified into pleomorphic small cell (n = 3), monomorphic medium-sized cell (n = 10), pleomorphic medium and large cell (n = 17), immunoblastic (n = 1), and anaplastic large cell (n = 9) lymphomas. Two cases were unclassifiable. Overlapping histologic features were noted between some cases and also within the same tumor and between multiple lesions of the same patient. Histologic evidence of enteropathy was present in most cases. The ITLs expressed cytoplasmic CD3 and antigens associated with cytolytic function (TIA-1, granzyme B), but not CD4 and CD5. Six of the 10 monomorphic medium-sized cell lymphomas were CD56+ T-cell lymphomas. Because of the histologic heterogeneity in some ITLs, we suggest a classification system with 2 main categories: (1) small to medium-sized cell, comprising pleomorphic small and monomorphic medium-sized cell lymphomas; and (2) large cell, comprising the remaining subtypes. The differential diagnosis includes B-cell lymphomas, tumors of histiocytic origin, anaplastic carcinoma, and malignant melanoma.

[Lymphatic reaction presenting as immunocytoma within the framework of carcinomas]. / Lymphatische Reaktion unter dem Bild eines Immunozytoms im Rahmen von Karzinomen.

Case histories are presented of 2 patients in whom the occurrence of several signs indicated the presence of an immunocytoma. A 56 year-old man presented with marked splenomegaly and a lymphocytosis in peripheral blood picture. Lymphatic infiltrates were demonstrated in the bone marrow and liver. During the course of the disease the initial monoclonal gammopathy and the lymphocytosis disappeared. An extensive carcinoma of the stomach became clinically manifest one year later. In the second patient, a 74 year-old man, lymph node swelling, monoclonal gammopathy and bone-marrow cytology indicated lymphatic disease. There was no indication of carcinoma in spite of extensive investigations. The patients died three years later of a hypernephroma. Both patients demonstrated no sign of lymphatic disease at autopsy and, hence, a lymphatic reaction must be diagnosed retrospectively.

[Clinical value of rectal digital examination in early diagnosis of colorectal cancer (author's transl)]. / Effizienz der rektal-digitalen Untersuchung zur Früherkennung des Dickdarmkarzinoms.

A prospective study was carried out on a series of 1500 patients of a gastro-enterological and surgical unit in order to evaluate critically the accuracy of rectal digital examination as a single tool in the early diagnosis of colorectal tumours. The patients in this study were explored independently by two investigators, first by rectal digital palpation and afterwards by means of the proctosigmoidoscope. Endoscopy confirmed the presence of 239 tumours, 96 of these being diagnosed histologically as carcinoma. Comparative analysis of the results of physical and endoscopic examination demonstrated that digital palpation provided a correct diagnosis in 58 out of 239 tumours, equivalent to an accuracy of 24.3% for tumours within the reach of the rectoscope, and 10% for all colorectal tumours. Hence, rectal examination should not be used as single diagnostic tool in screening for large bowel cancer. A multiphasic screening system, including investigations for occult blood, proctosigmoidoscopy, endoscopic polypectomy and determination of colon-embryonic-antigen levels is discussed to obtain better results in general health screening, early diagnosis and avoidance of incurable colorectal carcinomas and the early detection of recurrence.

[Immunologic phenotypes of germinal center cell tumors]. / Immunologische Phänotypen von Keimzentrumstumoren.

A panel of monoclonal antibodies applied to frozen sections of non-Hodgkin's lymphomas was used to establish clear-cut differences among the different entities of malignant lymphomas of germinal centre cell origin. 51 cases (18 centrocytic, 25 centroblastic-centrocytic and 8 centroblastic lymphomas) were included in this study. A clear-cut difference in the expression of the T65 antigen (Leu 1+) and the common acute lymphoblastic leukaemia antigen (CALLA) was found. Thus, centrocytic lymphomas predominantly expressed Leu 1, but not CALLA, whereas centroblastic-centrocytic lymphomas were always positive for CALLA, but not for the T65 antigen. Centroblastic lymphomas are virtually never positive with respect to either antibody. These findings suggest that, perhaps, two different phenotypes of centrocyte exist in centrocytic and centroblastic-centrocytic lymphomas.

[67-Galliumscintigraphy as an auxiliary method for differentiation of mass lesions of the liver (author's transl)]. / Die 67-Galliumszintigraphie als Hilfsmittel zur Abklärung von Parenchymdefekten der Leber

Liver scintigraphy with 67-Ga citrate and alphafetoprotein (afp) determinations in the serum were carried out in 84 patients with liver mass lesions in the preceding sulphur colloid scans. Among these patients 51 cases were histologically verfied and 33 patients were regarded as clinically-proven cases. Scanning was carried out 72 hours after the intravenous injection of 3 mC 67-Ga-citrate. Corresponding to the intensity of 67-Ga uptake within the former liver lesions 3 groups of 67-Ga scans were differentiated: Ga 0 (the lesion showed no Ga uptake), Ga plus (the Ga uptake within the lesion was equal to that of the surrounding liver tissue) and Ga plus plus (the Ga uptake within the former lesion exceeded the physiological Ga uptake in the normal liver tissue). The number of cases, results of Ga scintigraphy and afp examinations as well as histological, clinical and nuclear medical diagnosis were correlated. It was shown that Ga plus plus cases were strongly suspect of hepatoma, whereas in Ga 0 cases a diagnosis of hepatoma could be excluded. In patients with Ga plus further investigations have to be performed (repeated afp examinations, angiography of the coeliac artery), because cirrhotic regeneration nodules, metastases and necrotic hepatomata were all found within this group. According to our experience liver scanning with 67Ga represents a useful auxiliary examination in liver diagnosis. Ga citrate scintigraphy of the liver is indicated in all cases with mass lesions detected by the routine sulphur colloid scan and in all patients in whom there is clinical suspicion of hepatoma, inorder to differentiate the origin of the lesions. In 2 cases of hepatoma marked Ga uptake was observed at a time when the afp was still negative.

[Immunohistochemical studies on the association between hepatitis B surface antigen and primary hepatocellular carcinoma (author's transl)]. / Immunhistochemische Untersuchungen zur Assoziation des Hepatitis-B-Oberflächenantigens mit dem primären Leberzellkarzinom in Biopsie- und Autopsiematerial.

In many geographical areas especially in African and South-east-Asian countries hepatitis B virus infection is considered to be a major etiological factor in the development of primary hepatocellular carcinoma. 107 autopsy- and 15 biopsy-specimens were studied by means of immunohistochemistry [peroxidase-antiperoxidase-(PAP-)method] to demonstrate the association between primary hepatocellular carcinoma and hepatitis B surface antigen (HBsAg). HBsAg was found in 8 of 107 tumour specimens (7.4%) and liver cirrhosis in 102 of the 107 autopsy specimens with hepatocellular carcinoma (95%). 10 of the 15 biopsy-specimens showed neoplastic and non-neoplastic liver tissue, and in 2 of these 10 cases HBsAg was found. Liver cirrhosis could be seen in 9 of those 10 specimens. HBsAg was also studied in 90 cases with liver cirrhosis and was found to be positive in 2 of them (2,2%). HBsAg associated with primary hepatocellular carcinoma was only found in non-neoplastic liver cells of cirrhotic livers. Our studies indicate that in our geographical area the association of HBsAg with primary hepatocellular carcinoma is much less conspicuous than in Asian, African and even Southern European communities.

[The high autopsy rate in Vienna]. / Zur hohen Autopsierate in Wien.

In 1983 34% of all persons who died in the Republic of Austria were autopsied. We examined the frequency of postmortem examinations in the capital city, Vienna, between 1983 and 1987 in view of this high overall autopsy rate. The annual autopsy rate in all 9 pathology institutes and the department of legal medicine of the University of Vienna was on average 51 to 53%. There was a slight statistically verifiable decline in 1987. The reason for this high necropsy rate is that Austrian law permits the autopsies without the consent of next of kin if it appears indicated for medical, scientific or educational reasons; further more the fact, that the chairman of the department is at the same time the coroner for the hospital. A high autopsy rate is necessary to maintain a high standard of diagnostic accuracy even in modern medicine. The diagnoses were inaccurate or incomplete in 15% of all cases. The high autopsy rate is the basis for extremely accurate mortality and morbidity statistics and provides the basis for long range public health planning. Medical students develop an insight in disease patterns, which cannot be gained in any other way. The pathology departments of Vienna introduce a new city-wide computer-supported diagnostic file system, which will provide a baseline for comparative scientific studies.

[Lymphomas of the gastrointestinal tract]. / Lymphome des Gastrointestinaltrakts.

Recent progress in polychemotherapy employed in the treatment of gastrointestinal lymphomas has markedly improved prognosis. Between 1980 and 1986, 22 patients with malignant lymphomas of the gastrointestinal tract were treated at the Department of Surgery at the Hanusch Medical Center, in cooperation with the Department of Hematology and Oncology. 19 patients underwent surgery. The average observation time was 29 months. The rate of survival after 36 months was 64% in the entire patient population. Palliative-surgical interventions combined with chemotherapy equally produced satisfactory results.

[Epidemiology of primary carcinoma of the liver in Austria (author's transl)]. / Epidemiologie des primären Leberkarzinoms in Osterreich.

29 348 in patients were treated from 1945-1975 on a gastroenterological ward; 1 892 (6.47%) of these patients had cirrhosis and 183 (0.63%) carcinoma of the liver. In the groups of patients who died, resp. came to autopsy percentages were considerably higher: 17% resp. 25.6% had cirrhosis, 4.9% resp. 13% had primary carcinoma of the liver. Analysis of these data shows, that the incidence of both diseases is increasing. This seems to be due in the first line to increasing alcoholism, and only to a lesser degree, at least in Middle Europe, to viral hepatitis. Taking into account other data published in Austria concerning absolute and relative incidence of primary carcinoma of the liver it can be stated, that this malignancy ranges among the 10 most frequent tumor diseases already.

Non-Hodgkin lymphoma following Hodgkin's disease. A case report.

A 40-year-old male developed 66 months after diagnosis of Hodgkin's disease a non-Hodgkin lymphoma of high grade malignancy. Initially he had been treated by irradiation because of Hodgkin's disease PS IIA. After 2 years of remission he had relapsed and had received MOPP and CCNU. After 2 years without any therapy he developed the secondary neoplasm. With increasing frequency of long term survivors late complications, either therapy related or caused by immunologic defects, are observed.

[Systemic mastocytosis]. / Systemische Mastozytose.

Systemic mastocytosis associated with urticaria pigmentosa seems to be strikingly more common than previously assumed. The diagnosis can be established by the investigation of bone marrow sections, whereas bone marrow smears are less reliable. Some mediators are produced by the enhanced number of mast cells; telemethyl imidazole acetic acid is the most suitable mediator to calculate the size of the mast cell pool. Investigations like this might offer an alternative to the examination of bone marrow sections in future.

Progress in the management of high risk non-Hodgkin's lymphomas. 10 years of experience of the 3rd Medical Department of Hanusch Hospital, Vienna.

An analysis of 173 cases of non-Hodgkin's lymphoma (NHL) admitted to our hospital from January 1973 to January 1983 is presented. Of the 173 cases, 124 patients suffered from NHL of high grade malignancy according to the Kiel classification (37 centroblastic lymphoma (CB), 30 immunoblastic lymphoma (IB), 43 lymphoblastic lymphoma (LB), 14 NHL high grade malignancy unclassifiable). In addition, 26 patients with secondary high grade malignant NHL were included in the analysis (14 secondary CB, 10 secondary IB, 2 secondary LB). Also investigated were 23 patients with anaplastic centrocytic lymphoma (CC) (20 primary CC, 3 secondary CC), an entity originally classified as low grade malignant lymphoma, but showing a poor outcome and need for aggressive therapy. Symptoms at presentation of all patients are described. Of the 173 patients, 71% had an advanced stage of the disease at the time of diagnosis (Ann Arbor stage III or IV). B-symptoms were observed in 81%. Extranodal involvement, (exceptive bone marrow involvement), determined by clinical examination was seen in 55%. Survival of patients changed significantly after replacing initial radiotherapy with aggressive chemotherapy (P less than 0.001). Improvement of survival statistics was due to the better outcome of patients with localized stages (Ann Arbor stages I and II) as compared to those with generalized disease (P less than 0.002). Prognostic factors influencing survival were elevation of lactic dehydrogenase (P less than 0.0001) and response to therapy (P less than 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

[Varied importance of endoscopic polypectomy as prevention of cancer in the stomach and colon]. / Die unterschiedliche Bedeutung der endoskopischen Polypektomie als Karzinomprophylaxe im Magen und Dickdarm.

15,300 endoscopies of the upper and lower gastrointestinal tract were done during the years 1974-1982. During this time 915 polyps were excised. 3.2% of the polyps excised from the stomach, as contrasted to 64.8% of the ones taken from the colorectal part of the gastrointestinal tract were classified as precancerous lesions. These results show, that endoscopic polypectomy is a very efficient procedure in regard to prophylaxis of colon carcinoma, however almost without significance in this regard for prophylaxis of gastric carcinoma.

[Preservation of ram seminal fluid in different diluents]. / Prouchvaniia vurkhu sukhranenieto na semenna technost ot kochove v razlichni razrediteli.

Studied was the conformation of the protein macromolecules with the dilution of ram semen to a various extent, using three yolk diluents -- gkh18, zhgts, and Zlatno rouno. Studied was also the effect of some yolk extenders used in this country's practice on some qualitative indices of ram semen under storing conditions. It was found that following dilution with the media cited above various conformational changes of the proteins take place with the semen of rams. The different rates of dilution had no effect on the primary motility of spermatozoa. Statistically significant was the difference in favour of the gkh18 diluent in so far as the heat resistance of spermatozoa was concerned. Most effective dilution (in terms of retaining the motility of spermatozoa at storing) proved 1 + 3, followed by 1 + 5 with the use of gkh18, and 1 + 1 with zhgts and Zlatno rouno. gkh18 was shown to produce a varying blocking effect on motility of spermatozoa. All results concerning the conformation of protein macromolecules, quality indices, and particularly the resistance to heat in evaluating the storing capacity of semen showed that the cumulation of more proteins of higher electrophoretic mobility in III group developed more favourable conditions in preserving the vitality of spermatozoa at storage.

[Comparative studies of the lipoprotein complexes in bull seminal fluid diluted with different media]. / Sravnitelni izsledvaniia vurkhu lipoproteinnite kompleksi v razredena s razlichni sredi semenna technost ot bik.

Electrophoresis on celogel and lipid staining with Fettrot 7 were employed to investigate the lipid complexes in the media Hagase Niva, Triladil, Leciphos, and GH22L (after Kichev). Studied were likewise the changes that these lipid complexes underwent when using them to dilute bull semen to a varying extent. It was found that the lipid complexes in these media changed their electrophoretic characteristic and percent distribution following dilution. At 1 + 1 dilution all media were shown to build up a new lipoid complex having a protein base and comprising 84 to 96 per cent of the lipid components in the samples. On the basis of Kichev's phenomenon of complex forming and building new biologic molecules the possibility is discussed of the practical use of these investigations in the cryoprotection of bull semen.

Immunohistochemical and biochemical measurement of estrogen and progesterone receptors in primary breast cancer. Correlation of histopathology and prognostic factors.

OBJECTIVE: The authors investigated correlations of estrogen-receptor and progesterone-receptor with conventional risk factors as well as histopathology in patients with primary breast cancer. SUMMARY BACKGROUND DATA: Immunohistochemically determined hormone receptors have gained importance as prognosticators in primary breast cancer, but their definitive role has not yet been evaluated. METHODS: Tumor samples from 299 patients were examined for estrogen and progesterone receptors by biochemical and immunohistochemical assay. Correlations with established risk factors (tumor size, lymph node status, menopausal status, grading including subfactors) and histopathology were analyzed. RESULTS: The estrogen receptor, determined by immunohistochemical method revealed positivity in 80.6% of patients; biochemical measurement yielded 76.2% positive results. The progesterone receptor measured by immunohistochemistry yielded 61.3% positivity versus 55.8% detected by biochemical analysis. Invasive lobular, tubular, and ductal invasive carcinoma with prominent stroma content ("scirrhous carcinoma") rather than ductal invasive carcinoma was more frequently estrogen-receptor positive with immunohistochemistry than with biochemical assay. For progesterone receptor, the same pattern of positivity was seen with immunohistochemical assay. With progesterone receptor determined biochemically, "scirrhous" and lobular carcinoma showed positive results in a lower proportion than invasive ductal and tubular carcinoma. Significant correlations were observed between the estrogen-receptor status, the histologic grade of malignancy, nuclear polymorphism, and the rate of mitosis with both methods (p < 0.001 respectively). Different correlations were found between tumor size, menopausal status and estrogen receptor status with both assays respectively. For the progesterone receptor status, immunohistochemistry yielded significant correlations with the histologic grade of malignancy, nuclear polymorphism, rate of mitosis (p < 0.001 respectively) as well as growth pattern (p < 0.01), while biochemical analysis revealed a correlation with nuclear polymorphism (p < 0.05). The correlation analysis of both components of the immunoreactive score revealed a more significant impact of percentage of positive cells than of staining intensity. CONCLUSIONS: Immunohistochemistry detected a closer correlation between prognostic factors and receptor data than biochemical analysis.