Cysteine proteinases in chondrosarcomas.

The aim of the present study was to define the role of cathepsins B, H, K, L and S in the pathogenesis of human chondrosarcomas. For this purpose 40 tumour samples obtained from 12 patients with the diagnosis of conventional chondrosarcoma were systematically investigated for the expression of cathepsin mRNAs by Northern hybridisation, and for immunohistochemical localisation of the proteins. Northern analysis demonstrated the highest levels of cathepsins B and L in a recurring grade 1 chondrosarcoma, and in a grade 3 chondrosarcoma and in fibrous histiocytomas. Increased expression of cathepsin K mRNA was seen in seven chondrosarcomas, as well as in control tumours; fibrous histiocytomas, osteosarcomas, enchondromas and a giant cell tumour of bone. Cathepsin L was immunolocalised within the large chondrocytes, while cathepsin K was predominantly localised in large multinucleated osteoclastic cells and in some hypertrophic chondrocytes. These results suggest that chondrosarcoma can be included in the growing list of tumours, where cathepsins may well be involved in tumour progression. The simultaneous upregulation of cathepsins B and L, together with matrix metalloproteinase-13, and the association of cathepsin K with negative prognostic parameters suggests that an aggressive biological behaviour of chondrosarcoma may be related to the synthesis of cysteine proteinases and activation of other proteolytic enzymes. If this turns out to be the case, cathepsin inhibitors could provide the much needed adjuvant therapy for chondrosarcomas.

Expression of inhibin alpha, beta A and beta B messenger ribonucleic acids in the normal human ovary and in polycystic ovarian syndrome.

We studied the cellular distribution of inhibin alpha, beta A and beta B mRNAs in the normal human ovary and in polycystic ovarian syndrome (PCOS) by in situ hybridization. Our results show that human granulosa cells express inhibin alpha, beta A and beta B subunit mRNAs, and theca cells express inhibin alpha and beta A subunit mRNAs. The co-localization of alpha and beta A mRNAs in theca cells supports the hypothesis that inhibin also has an autocrine function in these cells. We did not detect any inhibin subunit mRNA in the granulosa cells of atretic follicles, while theca cells also expressed alpha subunit mRNA in those follicles. The present findings suggest that the expression of inhibin subunits is regulated differently in human follicular granulosa and theca cells. It has been speculated that inhibin may be involved in the development of PCOS. Our results show that the cellular localization of inhibin subunit mRNAs is not disturbed in PCOS ovaries.

Primary signet-ring cell carcinoma of the ureter. A case report.

We describe here a 65-year-old woman with a right-sided ureteral tumour that was treated by nephroureterectomy. The tumour appeared to be a signet-ring cell carcinoma obstructing the lumen and infiltrating the whole thickness of the ureteral wall. No other primary site could be found, and the tumour is believed to represent the first reported case of primary signet-ring cell carcinoma of the ureter.

Immunohistochemical characterization of an amphicrine mucinous islet-cell carcinoma of the pancreas. Case report.

Immunohistochemical characteristics of a mucinous islet-cell carcinoma of the pancreas are described. The tumour presented with jaundice in a 59-year-old male. It consisted of polygonal atypical cells forming a reticular pattern, and invaded the common bile duct. In DNA flow cytometry, the tumour cells showed a clear-cut aneuploid peak. Intercellular mucin was abundant. A panel of antisera and monoclonal markers was applied in the immunohistochemical analysis. In addition to general epithelial and endocrine markers, the tumour cells showed a focal positive immunoreaction with anti-glucagon, anti-insulin, anti-vasoactive intestinal polypeptide, anti-pancreatic secretory trypsin inhibitor and anti-phospholipase A2 antigen. At the ultrastructural level, mucous and neuroendocrine granules were demonstrated in the same tumour cells.

Ossifying fibromyxoid tumour of soft parts: report of four cases including one mediastinal and one infantile.

Four cases of ossifying fibromyxoid tumour of soft parts are described. One of them was in the mediastinum, a hitherto unreported location of this rare neoplasm. Another was removed from the subcutaneous tissue of the head of a two-year-old girl, the youngest patient so far described. A peculiar feature of this tumour was haphazard spindle cell groups showing smooth muscle differentiation. One tumour was remarkably proliferative with 20 mitotic figures per 10 high power fields and 50% of cells positive for Ki-67 antigen. Immunohistochemical analysis revealed that all the tumours were diffusely positive for vimentin, and focally for S-100-protein. In addition to this the infantile tumour expressed focal alpha-smooth muscle actin, desmin and glial fibrillary acidic protein, while the mediastinal tumour expressed only alpha-smooth muscle actin and the highly proliferative one expressed none of these antigens. Background cells, including histiocytes, lymphocytes and mast cells were numerous. DNA cytometry analysis using both static and flow methods showed that the mediastinal tumour contained two cell clones, while the others were diploid. The proliferative fraction of cells (S plus G2 phases) was prominent in the proliferative and infantile tumours.

Expression of group II phospholipase A2 in Paneth cells of an adenoma of the rectum. A case report.

A case of tubulovillous adenoma in the rectum of a 51-year-old man is presented. The tumour contained numerous Paneth cells which formed well-developed glands in the basal areas. Group II phospholipase A2 and lysozyme were found in the tumour cells by immunohistochemistry. mRNA of group II phospholipase A2 was localized in the tumour cells by in situ hybridization. It was concluded that a considerable part of this rare type of tumour consisted of Paneth cells which were capable of synthesizing group II phospholipase A2.

Acinar cell carcinoma of the pancreas. Report of three cases.

Pancreatic acinar cell carcinoma is a rare neoplasm (comprising about 1% of pancreatic tumours). We studied three cases (61-year-old female; 42-year-old male; 57-year-old male), whose survival after diagnosis ranged from 1 year 2 months to 6 years 8 months. There were widespread metastases in each case. The tumours had acinar, trabecular and solid growth patterns. By immunohistochemistry, pancreatic acinar cell markers including carboxyl ester lipase, pancreatic secretory trypsin inhibitor and pancreatic phospholipase A2 (group I PLA2) gave a strong positive reaction in all three cases. By electron microscopy, zymogen granules were seen in the cytoplasm of the tumour cells. Immunostaining for prostate-specific antigen was positive in all three cases. Above-normal concentrations of pancreatic PLA2 were measured in the serum of one patient and the values decreased during chemotherapy concomitantly with the reduction in the size of the tumour mass. In conclusion, immunohistochemical demonstration of the secretory products of acinar cells including the new marker pancreatic PLA2 is useful in the differential diagnosis of pancreatic acinar cell carcinoma. Determination of the concentration of pancreatic group I PLA2 in serum may be helpful in the evaluation of therapy.

Borrelial lymphocytoma--a historical case.

We here describe a patient with a tick bite in the areola mammae in 1953 followed by erythema migrans. Twenty years later, after another tick bite in the axillary skin, also followed by erythema migrans, a large lymphatic infiltrate developed in the mammary skin, when the margin of the erythema reached the areola. The infiltrate resolved within a year without any therapy. Borrelial DNA was detected by polymerase chain reaction in the paraffin blocks of the lymphatic skin infiltrate. The patient died 9 years later of generalized lymphoma. A similar monoclonal immunoglobulin heavy chain gene rearrangement was detected both in the mammary skin lesion and in the lymphoma specimen.

Expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human chondrosarcomas.

The aim of the present study was to characterise the ability of malignant chondrosarcomas to invade normal bone by analysing their production of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). For this purpose 12 chondrosarcomas were investigated for the expression of mRNAs for several MMPs and all 4 TIMPs by Northern hybridisation, and for immunohistochemical localisation of the proteins. A characteristic finding of these analyses was increased expression of MMP-13, MMP-14 and TIMP-2 mRNAs in chondrosarcomas when compared with nonmalignant control samples. Individual chondrosarcomas also exhibited elevated levels of MMP-1, MMP-7 and MMP-9 mRNAs. The results of Northern hybridisations were supported by immunohistochemical stainings of the corresponding tumour areas for MMP-2, MMP-14 and TIMP-2, further suggesting that these may have prognostic value for determining whether individual chondrosarcomas are locally aggressive or have a probability of recurrence. Another finding of the present study was a marked heterogeneity in histologic appearance and gene expression of the chondrosarcomas, emphasising the importance of analysing several areas of these tumours to get representative results. These findings suggest that analysis of MMPs could be a useful diagnostic indicator in patients with cartilaginous tumours and could help in differentiating between a low-grade malignant chondrosarcoma and a benign growing enchondroma.

Proton relaxation times in arterial wall and atheromatous lesions in man.

The proton relaxation times of autopsy samples of arterial intima-media were measured with an NMR spectrometer and results correlated to the microscopically estimated lipid content of the vascular wall. The normal arterial intima-media contained two T1 relaxation components. The short T1 component (T1s) was 90 +/- 13 ms and its relative weight was 11%. The long T1 component (T1l) was 523 +/- 89 ms and relative weight 88%, respectively. The average T2 was 99 +/- 18 ms. In diseased vessels, a positive correlation was found between the lipid content of the vessel wall and the relaxation rate of the fast component. T1s of the intima-media was significantly shorter (P less than .01) in severe atheromatosis compared with vessels without fat deposition. The results suggest that atheromatous lesions should be best highlighted in spin-echo images by using short TR and TE to suppress the influence of T1l and to avoid (noncontrast contributing) T2 decay of the signal intensity.

Alveolar soft part sarcoma. Immunohistochemical evidence for muscle cell differentiation.

Seven cases of alveolar soft part sarcomas (ASPSs) were studied immunohistochemically for the presence of a number of differentiation markers in an attempt to define the cellular nature of this tumor. Desmin-positive tumor cells were found in three and muscle actin-positive cells in four cases when studied in formaldehyde-fixed and paraffin-embedded material. In one case studied in frozen sections, focal desmin positivity but no other intermediate filaments were found. Immunostaining for synaptophysin, a general neuroendocrine marker, was negative in all cases. All tumors were positive with a monoclonal antibody NK1C3, which consistently stains melanomas, and three cases showed significant numbers of S-100 protein-positive tumor cells, but immunostaining with HMB-45, a melanoma-specific monoclonal antibody, was negative in all cases. However, several rhabdomyosarcomas studied for NK1C3 and S-100 protein for comparison were also at least focally positive. Electron microscopic examination, performed in three cases, showed uniform paucity of all kinds of filaments in the tumor cells of ASPS, and it specifically failed to reveal any signs of smooth or striated muscle cell differentiation. Thus, the results of the present study do not unequivocally define the nature of ASPS but speak against its paraganglionic character and present evidence for muscle cell differentiation.

Pure and mixed mucinous breast carcinomas: DNA stemline and prognosis.

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.

Clinical correlations of genetic changes by comparative genomic hybridization in Ewing sarcoma and related tumors.

Our previous comparative genomic hybridization (CGH) study of Ewing sarcoma and related tumors showed that DNA sequence copy number increases of 1q21-q22 and of chromosomes 8 and 12 were associated with trends toward poor survival (Armengol et al., Br J Cancer 1997, 75, 1403-1409). These trends were not statistically significant. In the present study, we analyzed 28 primary Ewing sarcomas and related tumors by CGH to study whether these (or other) changes have prognostic value in these tumors. Twenty-one tumors (75%) had changes with a mean of 1.9 changes per tumor. The most frequent aberration was gain of chromosome 8 in 10 tumors (36%). Five tumors (18%) had copy number increases at 1q21-22 and 5 had gain of 7q. Copy number increase of 6p21.1-pter, gain of chromosome 12, and loss of 16q were seen in 11%. Copy number increases of 1q21-q22 and of chromosomes 8 and 12 were associated with trends toward worse outcome, but the differences did not reach statistical significance. A novel finding is the association of copy number increase at 6p with worse distant disease-free (P = 0.04) and overall survival (P = 0.004). To confirm this finding and to see whether copy number increases of 1q21-q22 and of chromosomes 8 and 12 have definite prognostic value, a larger number of cases needs to be studied.

Treatment results of osteogenic sarcoma. An evaluation of 36 patients treated during 30-year period in south-western Finland.

Histological re-evaluation revealed 36 osteogenic sarcoma (OS) patients for analysis in South-Western Finland treated between 1958 and 1987. In 21 cases (58%) the tumour was located in the knee region. The mean age at diagnosis was 28 years (range: 5-62 years) and the follow-up time of the patients was at least 5 years or until death. In 29 patients without metastases at the time of diagnosis, the extent of the primary tumour was T2 in 37% and T3 in 63% of the patients. There were no differences regarding the extent of the primary tumour, delay of the diagnosis, mean age of the patients, or duration of the symptoms while comparing the three decades of the study. Before the 1970s the treatment consisted of surgery with or without radiotherapy in most cases. Since the 1970s the combination of surgical treatment (amputation or wide excision) and adjuvant chemotherapy was the most common treatment modality. Since the late 1970s limb-salvage surgery has been applied in selected cases, and it seems to be justified. None of the patients treated before 1970 survived for 5 years. The 5- and 10-year survival of all 36 patients was 44.4% and 33.6%, respectively. In non-metastatic patients both the 5- and 10-year disease-free survival was 46.7%. A certain group exhibiting a good prognosis was found; the 10-year survival of the 10 patients with OS in extremities, treated with combined chemotherapy and surgery, was 70%. The median survival time was significantly longer for the patients with an intracompartmental tumour extent of T2 (112 months) compared with an extracompartmental extent of T3 (23 months), and for the patients with the primary tumour in the knee region (112 months) compared with other locations (18 months). The long-term survival of the OS patients has improved concomitantly with the multimodality of the treatment.

Clear cell sarcoma of tendons and aponeuroses: malignant melanoma of soft tissues? Report of four cases.

In a survey of 246 soft tissue sarcomas in the extremities and limb girdles reported to the Finnish Cancer Registry in 1960-1969, two clear cell sarcomas of tendons and aponeurones were diagnosed. We subsequently diagnosed two more examples. Three tumors were located in the foot and one in the wrist. The histology of the tumors was characteristic, and three of them were shown to contain brown pigment tinctorially indistinguishable from melanin. The pigment could be bleached with potassium permanganate. Ultrastructural studies performed in the most recent case revealed premelanosomes in the cytoplasm of tumor cells. Increasing evidence in the literature and results of the present study seem to suggest that clear cell sarcoma of tendons and aponeuroses represents a soft tissue malignant melanoma.

Immunohistochemical expression of p53 protein, mitotic index and nuclear morphometry in primary malignant melanoma of the skin.

The immunohistochemical expression of the p53 tumor suppressor protein and the nuclear morphometric parameters were studied in 80 primary skin melanomas. The mitotic index was counted in 64 tumors. In 95% of the tumors p53 positive nuclei were detected, but in only 31% of the cases was the proportion of positive nuclei 3% or higher. The mitotic indices and the mean nuclear areas were not significantly different in melanomas with different p53 expression levels. The p53 positive nuclei had a larger mean nuclear area than the p53 negative nuclei. p53 expression did not increase with melanoma thickness. In Cox's stepwise proportional hazards model, the ratio of mean nuclear long and short axis and melanoma thickness had independent prognostic value, while the level of p53 expression, the mitotic index, and the mean nuclear area were not significantly associated with survival. We conclude that the nuclei expressing p53 protein are larger than p53 negative nuclei. The level of immunohistochemical expression of p53 is low in primary skin melanoma, and it is not valuable as a general prognostic marker for this tumor. p53 expression is not associated with melanoma thickness, indicating that high p53 expression is not a late phenomenon in the progression of this tumor.

Flow cytometric analysis of the DNA profile of renal cell carcinoma.

DNA flow cytometry was applied to one hundred consecutive renal cell carcinomas from years 1974-1979. Single cell suspensions were prepared from paraffin-embedded tissue. From 96 evaluable tumours 34 were considered diploid and 62 aneuploid. Ploidy did not correlate statistically significantly with local growth, venous invasion, nodal or distant metastatic spread, or nuclear grade of renal cell carcinomas. Also the survival of the patients with diploid and aneuploid tumours was not significantly different, but, excluding patients with distant metastases at the time of extrafascial nephrectomy, 83% of the patients with diploid tumour survived for five years, while only 57% of the patients with aneuploid tumour were alive after the same time interval. The difference between survival curves is statistically almost significant (p less than 0.05). The cytologic nuclear grade predicted the prognosis more significantly (p less than 0.01), and the combination of these methods is recommended for the prognostication of renal cell carcinoma.

Cystic osteolysis induced by silicone rubber prosthesis.

Synovitic symptoms and radiologically documented cystic lesions in several carpal bones and distal radius developed two years after the implantation of silicone rubber scaphoid prosthesis for pseudoarthrosis. Open biopsy revealed granulomatous synovitis and osteitis and foreign material in carpal tissues. Ultrastructurally dense material was found in the cytoplasm of histiocytic, often multinucleated cells. Electron-probe analysis of the material showed a definite peak for silicon, proving the causal relationship of symptoms and signs to the silicone rubber prosthesis.

Intrapleural endodermal sinus tumor arising from the diaphragm.

We report a primary endodermal sinus tumor (EST) of the right hemidiaphragm, presenting as a right-sided hemothorax in an 18-month-old female. Four operations were performed, the last three being indicated solely by an elevated level of serum alpha-fetoprotein (AFP). Cyclic chemotherapy with varying drug combinations together with irradiation therapy were not effective in controlling the disease. To our knowledge, this is the first instance of a primary EST arising from the diaphragm.

Summer spores of birch rust fungus in Papanicolaou smears from healthy mass screening participants.

OBJECTIVE: To evaluate the origin and identification of mysterious particles in Papanicolaou smears from healthy, asymptomatic women participating in a local mass screening program. STUDY DESIGN: The material consisted of 16,000 cerricovaginal smears studied at the cytology laboratory of Pathology Laboratory of South-Western Finland Ltd. Unidentified particles were found in four apparently independent smears. All the slides were rescreened, but no further positive cases were found. RESULTS: Several swab samples were collected from the investigation room, but all were negative. Attempts at identification included numerous consultations. A telepathology consultation with the Cellular Division, Armed Forces Institute of Pathology, Washington, D.C., U.S.A., was conducted. Another teleconsultation with the reference laboratoryfor intestinal parasites in Iceland was conducted. Finally, the Aerobiology Unit, University of Turku, was consulted. The aerobiologist suggested that the particles were most probably summer spores of birch rust fungus (Melampsoridium betulinum). To confirm this we obtained a reference specimen of the fungus from the herbarium at the University of Turku. The morphology of the spore particles was identical to that in the cervicovaginal smears, proving that the smears were contaminated with birch rust fungus spores. CONCLUSION: Birch rust is ubiqutitous in Finland. Outside the hospital window there is a wide lawn behind which, at a distance of 70 m, there is a forest of birch trees. It is most likely that the rust spores flew in through the open window, to settle down on the surface of material that was used in sample taking. Airborne dust may cause misleading and surprising artefacts. Protection of specimens and instruments against dust should minimize the problem.