Martin Tötsch

Relationship between quantity of silver stained nucleolar organizer regions associated proteins (Ag-NORs) and population doubling time in ten breast cancer cell lines.

Using a one-step silver staining technique ten human breast cancer cell lines were investigated, determining by means of automated image analysis the mean area of Ag-NORs per nucleus, the total number of Ag-NORs, and the so-called scattered and clustered Ag-NORs. These parameters were statistically correlated with the population doubling time (PDT) of the various carcinoma cell lines of this series. Our results show that the mean area of Ag-NORs per nucleus (p less than 0.001) is highly significantly correlated to the PDT, whereas all other parameters were less or not significant. It is concluded that the assessment of the area of Ag-NORs by automated image analysis is a simple and reliable method for the determination of cell duplication rates.

Diagnostic value of different PCR assays for the detection of mycobacterial DNA in granulomatous lymphadenopathy.

Diagnosis of mycobacterial infection is made by assessment of characeristic histological features, staining of acid‐fast bacilli, or agar culture. Recent advances in molecular biology have provided alternative approaches for the detection of mycobacteria, but only limited data are available dealing with the comparative evaluation of these methods. In order to determine the diagnostic applicability of polymerase chain reaction (PCR)‐based assays, 20 formalin‐fixed and paraffin‐embedded lymph nodes with bacille Calmette–Guérin (BCG) lymphadenitis were investigated which in Löwenstein Jensen agar culture were either positive or negative (ten cases each); ten lymph nodes with non‐specific lymphadenitis served as negative controls. Ziehl–Neelsen staining as well as three different PCR assays (including nested PCR), amplifying a specific sequence of theMycobacterium tuberculosis complex or sequences of the 65u2009kD antigen gene, were performed. Positive culture was only obtained from lymph nodes which had been surgically removed within 20 weeks after vaccination (P<0·001). In contrast to microscopic examination, which yielded no more information than agar culture, PCR detection of mycobacterial DNA was unrelated to culture findings. Combined use of different assays, as well as DNA extraction from at least three paraffin sections from each specimen, resulted in the detection of mycobacterial DNA in all lymph nodes with amplifiable DNA (18 out of 20 cases). Controls remained consistently negative. Thus, the combined use of different PCR assays is proposed as a rapid and sensitive technique for the detection of mycobacterial DNA in formalin‐fixed and paraffin‐embedded tissue.

Fine needle aspiration cytodiagnosis of anaplastic carcinoma and malignant haemangioendothelioma of the thyroid in an endemic goitre area

Between 1970 and 1987,20 028 fine needle aspirates (FNA) of the thyroid have been examined in the Department of Pathology of the University of Innsbruck, Austria. During this period 92 cases of anaplastic carcinoma and 16 cases of malignant haemangioendothelioma (MHE) of the thyroid were diagnosed. Forty‐three out of these 108 highly malignant tumours of the thyroid underwent FNA pre‐operatively (39.1%). Thirty‐seven FNA contained numerous cells of a highly malignant tumour. Five specimens (11.8%) contained only necrotic material and inflammatory cells. In one case of an anaplastic carcinoma no malignant cells could be demonstrated in FNA. We conclude that pre‐operative FNA of highly malignant thyroid tumours may contribute substantially to subsequent clinical management.

Fine needle aspiration cytodiagnosis of subacute (de Quervain's) thyroiditis in an endemic goitre area.

Between 1977 and 1989 252 fine needle aspirates (FNAs) of the thyroid from patients with a clinical suspicion of subacute granulomatous (de Quervains) thyroiditis were examined in the Department of Pathology of the University of Innsbruck, Austria. In the same period 31 cases with preoperative FNA were diagnosed histologically as subacute thyroiditis. Only in three of these cases were the cytological features of de Quervains thyroiditis found in the preoperative FNA. However, in 13 of these 31 cases a cytological suspicion of malignancy was obtained. Subsequent histological examination revealed an acute phase inflammation of de Quervains thyroiditis in most of these cases. We conclude that an accurate FNA diagnosis of de Quervains thyroiditis, particularly in the acute stage, may cause difficulties due to a lack of typical features and the appearance of atypical thyroid follicular cells. For the cytopathologist, accurate clinical information relating to the possibility of de Quervains thyroiditis is essential if unnecessary surgery is to be avoided.

Secretoneurin in bronchopulmonary carcinoids—immunohistochemical comparison with chromogranins A and B and secretogranin II

Ninety‐nine classical and 11 atypical bronchopulmonary carcinoids were investigated immunohistochemically with an antibody against secretoneurin, a peptide proteolytically processed from secretogranin II (chromogranin C), as well as antibodies against chromogranin A and B and secretogranin II. Secretoneurin was immunolocalized in 86 tumours (78 classical and eight atypical carcinoids); secretogranin II was found in the same tumours in a similar distribution, whereas chromogranin A was present in all 100 and chromogranin B in 106 tumours investigated. Bronchopulmonary carcinoids are usually not associated with clinically or biochemically distinct syndromes. Although we found bronchial carcinoids with different immunohistochemical chromogranins/secretogranin patterns, no correlation with the biological behaviour of these tumours could be demonstrated.

Proliferative (MIB1, mdm2) Versus Anti-Proliferative (p53) Markers in Head and Neck Cancer. An Immunohistochemical Study.

Formalin fixed and paraffin embedded samples from 36 squamous cell carcinomas of the larynx and the oral cavity (pT2N0M0, R0) surrounded by non-tumorous mucosa were studied immunohistochemically using a panel of four different anti-p53 antibodies (CM1, PAbl801, D07, PAb240), a monoclonal anti-mdm2 antibody and MIB1, following wet autoclave antigen retrieval. P53 immunoreactivity was detected in 11/14 laryngeal and in 9/22 oral carcinomas. All p53 positive oral, and all but one laryngeal tumors revealed mdm2 positivity as well, whereas in p53 negative tumors 4/12 and 1/3 mdm2 immunopositive cases were demonstrated, respectively. MIB1 labeling indices of the tumors ranged between 18% – 64% in p53 positive cases, and 10% – 53% in p53 negative ones. The difference was not statistically significant. Close spatial coexpression of p53, mdm2 and MIB1 immunoreactivity was observed at the invasive front of the carcinomas and in the basal and suprabasal layers of the non-tumorous epithelium in all p53 positive cases. However, the MIB1 expression was similarly increased at the invasive margins in carcinomas lacking immunohistochemically detectable p53 alterations. Our results strongly suggest that p53 overexpression does not necessarily correspond to increased rate of proliferation, but rather to mdm2 overexpression and is largely dependent on the anatomical site in case of small and localized squamous cell carcinomas of the head and neck region.

Possible relation of p53 and mdm-2 oncoprotein expression in thyroid carcinoma: A molecular-pathological and immunohistochemical study on paraffin-embedded tissue

Routinely processed tissues from a series of benign and malignant thyroid lesions were immunohistochemically investigated with antibodies against p53 and mdm-2. p53 was immunolocalized in <10% of nuclei in 2/80 nodular goiters, 2/60 follicular adenomas, 26/68 follicular carcinomas, 7/40 papillary carcinomas, 3/10 “insular” carcinomas, and 10/31 anaplastic carcinomas. More than 10% positively stained nuclei were found in 2 widely invasive follicular, 2 insular, and 15 anaplastic carcinomas. All p53-positive cases showed a concomitant immunohistochemical mdm-2 expression; an immunohistochemical colocalization on serial section was demonstrated in 12 anaplastic carcinomas. Screening by polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis of these 12 cases revealed no relevant mutations in the coding regions of exons 2–11 of the p53 gene. Additionally, 1 follicular adenoma, 6 follicular carcinomas (4 minimally and 2 widely invasive), 1 papillary, and 2 poorly differentiated insular carcinomas were mdm-2 positive without immunohistochemically detectable p53 expression. These results provide evidence that wild-type p53 expression in thyroid carcinomas may be associated with mdm-2 induced formation of stable complexes. However, the role of p53 mutations and p53 protein inactivation owing to other factors (e.g., mdm-2) in the progression of thyroid carcinomas is still poorly understood.

Relationship between quantity of silver stained nucleolar organizer region associated proteins (Ag-NORs) and growth rate suppression of breast cancer cell lines after interferon-gamma and 4-hydroxy-tamoxifen treatment

SummaryThe correlation between the silver-stained nucleolar organizer region associated proteins (Ag-NORs) and the growth rate suppression (GRS) of ten established breast cancer cell lines which were treated with 4-hydroxy-tamoxifen (OHT) and interferon-gamma (g-IFN), respectively, was investigated by means of automated image analysis. Previous studies have shown a statistically significant relationship between the Ag-NOR quantity and the population doubling time (PDT) of these cell clones. The results of the present study showed a highly significant correlation between the GRS and the Ag-NOR quantity in estrogen receptor (ER) positive tumour cell lines after OHT treatment (P< 0.001) whereas no strict correlation of these parameters could be demonstrated after g-IFN treatment in both ER positive and negative cell lines. Our results suggest a different behaviour of NOR-proteins in breast cancer cell lines if treated either with g-IFN or OHT, probably reflecting the different mechanism of cell suppression mediated by OHT and g-IFN. It is concluded that quantitative assessment of Ag-NORs is not as suitable for the determination of the GRS as it is for the determination of cell duplication rates obtained on untreated tumour cell lines.

Intestinale Angiodysplasie als Ursache schwerer Dünndarmblutungen — Bericht über vier Fälle

Angiodysplasia of the small intestine alone cannot be differentiated histologically from hereditary hemorrhagic telangiectasia (Morbus Rendu-Osler-Weber). Isolated localisation in the small intestine is rare and requires surgery in the event of massive bleeding. Four cases of bleeding from angiodysplasias of the small intestine in patients having undergone surgery at our institution over a ten-year period are presented. In three patients the particular intestine was resected. Two of these patients had an uneventful follow-up of even and ten years, one patient relapsed and was rehospitalized for surgical treatment. The fourth patient, who showed angiodysplasias scattered over the entire small intestine, had no further bleeding over a three-year period after transmural ligations of the lesions found by intraoperative endoscopical diaphany.SummaryAngiodysplasia of the small intestine alone cannot be differentiated histologically from hereditary hemorrhagic telangiectasia (Morbus Rendu-Osler-Weber). Isolated localisation in the small intestine is rare and requires surgery in the event of massive bleeding. Four cases of bleeding from angiodysplasias of the small intestine in patients having undergone surgery at our institution over a ten-year period are presented. In three patients the particular intestine was resected. Two of these patients had an uneventful follow-up of even and ten years, one patient relapsed and was rehospitalized for surgical treatment. The fourth patient, who showed angiodysplasias scattered over the entire small intestine, had no further bleeding over a three-year period after transmural ligations of the lesions found by intraoperative endoscopical diaphany.