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.

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.

MR-guided core biopsies of soft tissue tumours on an open 0.23 T imager.

PURPOSE: To assess the feasibility of MR-guided soft tissue core biopsies on an open 0.23 T magnet. MATERIAL AND METHODS: Twenty-nine consecutive patients with known or suspected benign or malignant soft tissue tumours underwent MR imaging. A one-slice dynamic enhancement sequence was used to obtain an enhancement curve of the tumour. MR-guided core biopsy of the tumour was performed in the same session. RESULTS: All biopsies could be performed on an open 0.23 T magnet. Standard MR images and dynamic enhancement curves were used in deciding biopsy route and target. The MR-guided core biopsy specimens were sufficient for histopathological diagnosis in 27 of 29 cases. CONCLUSION: Open magnet configuration allows easy access to the patient and near real-time imaging guidance of soft tissue tumours. Minimally invasive MR-guided core biopsies of soft tissue tumours are feasible and help to avoid open surgical biopsies.

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.

Decorin is produced by capillary endothelial cells in inflammation-associated angiogenesis.

Decorin is a small extracellular chondroitin/dermatan sulfate proteoglycan that has previously been shown to be involved in the angiogenesis-like behavior of endothelial cells (ECs) in vitro. There is also evidence that decorin plays a role in angiogenesis in vivo. In this study we sought to further explore the involvement of decorin in angiogenesis in vivo, especially in that associated with inflammation. We found by CD31 immunostaining of ECs that in giant cell arteritis there are capillary blood vessels not only in the adventitia as in uninvolved temporal artery wall, but also in the media and the external zone of the thickened intima. Localization of decorin by antiserum LF-30 in adjacent sections showed that in normal temporal artery wall decorin resides mainly in the media and the adventitia, whereas in inflamed temporal artery wall decorin is distributed throughout the vessel wall including the intima. Furthermore, the most intense reaction for decorin was evident in ECs of capillary neovessels within the media and the thickened intima of inflamed temporal artery wall. Decorin was also found in capillary ECs in certain pathological and physiological conditions in which the pivotal role of angiogenesis is more generally accepted. Pyogenic granulomas, granulation tissue of healing dermal wounds, and ovaries at different phases of follicle and corpus luteum formation all contained widely distributed CD31-positive capillaries. Decorin, on the other hand, was found in capillary ECs in pyogenic granulomas and granulation tissue, but not in those in the ovaries. The assessment of the degree of inflammation in the specimens with the presence of CD68-positive macrophages showed that the pyogenic granuloma, granulation tissue, and giant cell arteritis specimens were rich in macrophages around the decorin-positive capillaries. In contrast, the ovarian specimens were populated with fewer macrophages and even they were not located in close vicinity of capillaries negative for decorin. Our results confirm that decorin is involved in angiogenesis in vivo and, particularly, in conditions in which the inflammatory component is dominant.

Leiomyoma of the urinary bladder.

We report a case of rare benign bladder leiomyoma. The patient was a 42-year old man complaining impotence. Digital rectal examination revealed a palpable pelvic tumor. Transurethral ultrasonography, computed tomography (CT), and magnetic resonance image (MRI) examinations all showed a cystic tumor measuring 10 x 8 cm. On the basis of these findings leiomyoma was suspected already before operation. The tumor was removed by open resection of bladder, and the diagnosis was confirmed by histology and positive immunohistochemistry.

Leiomyoma of the urinary bladder.

We report a case of rare benign bladder leiomyoma. The patient was a 42-year old man complaining impotence. Digital rectal examination revealed a palpable pelvic tumor. Transurethral ultrasonography, computed tomography (CT), and magnetic resonance image (MRI) examinations all showed a cystic tumor measuring 10 x 8 cm. On the basis of these findings leiomyoma was suspected already before operation. The tumor was removed by open resection of bladder, and the diagnosis was confirmed by histology and positive immunohistochemistry.

Dynamic contrast-enhanced MR imaging and MR-guided bone biopsy on a 0.23 T open imager.

OBJECTIVE: To assess the feasibility of MR (magnetic resonance)-guided bone biopsies. DESIGN AND PATIENTS: Thirty-six consecutive patients with known or suspected benign or malignant bone lesions underwent comprehensive MR imaging. A dynamic contrast-enhanced sequence followed by stationary T1-weighted sequences were obtained and MR-guided bone biopsy of the tumor at the site with fastest enhancement was performed using an open 0.23 T MR imager. RESULTS: All MR-guided bone biopsies samples were estimated to be sufficient by the pathologists. The biopsy specimens were diagnostic in 34 of 36 cases. CONCLUSION: MR-guided bone biopsies combined with dynamic contrast-enhanced imaging are feasible and safe for the diagnostic investigation of equivocal bone lesions.

Sucrose has no beneficial effects on wound healing in rats.

OBJECTIVE: To evaluate the effects of sucrose treatment on the formation of granulation tissue in a standard wound model. DESIGN: Animal study. SETTING: University hospital, Finland. ANIMALS: 32 male Sprague-Dawley rats divided into 4 groups. INTERVENTIONS: Implantation of viscose cellulose sponge subcutaneously, and daily injection of three concentrations of sucrose (0.01, 0.1 or 1 M) or vehicle for 7 days. MAIN OUTCOME MEASURES: The amount of granulation tissue measured by chemical analysis and histology. The amount and distribution of types I and III collagen assayed by immunofluorescence. RESULTS: None of the three concentrations altered the amounts of DNA, RNA, hydroxyproline, nitrogen, hexosamines, and uronic acids in granulation tissue. Neither improvement nor deterioration was seen in the growth of granulation tissue in histological specimens. The amount and distribution of types I and III collagen was similar in controls and sucrose-treated rats. Type III collagen was most abundant near newly-formed vessels. Neither sucrose nor fructose was found in wound fluid while the concentration of glucose was significantly lower in all test groups than in controls. CONCLUSIONS: Sucrose solution had neither beneficial nor deleterious effects on the amount of developing granulation tissue in an experimental wound model. The amount and distribution of types I and III collagens were also not altered by sucrose treatment.

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.

Effects of hexose sugars: glucose, fructose, galactose and mannose on wound healing in the rat.

The effects of four hexose sugars (D-glucose, D-fructose, D-galactose, D-mannose) on the developing granulation tissue in rats were examined. Cylindrical hollow sponge implants were used as an inductive matrix for the growth of granulation tissue. In the test group, the implants were injected with 0.1 ml of solution containing the different hexoses in 0.01, 0.1 and 1 M concentrations daily for 7 days while the implants of the control groups were injected with 0.1 ml of phosphate-buffered saline solution only. Analyses of granulation tissue and wound fluid in the sponge implants were carried out 7 days after implantation. The results demonstrated that galactose caused a significant increase in the accumulation of granulation tissue as estimated by histological analyses, but no significant differences were observed in various chemical analyses. In striking contrast, statistically significant decreases were observed in the number of leukocytes in wound fluid, in the amount of DNA, RNA, collagen hydroxyproline, nitrogen, hexosamines and uronic acids in sponges treated with 0.1 or 1 M mannose, reflecting decreased granulation tissue formation. This effect was also observed in histological analyses of the specimens. There were no major changes in sponges treated with glucose or fructose. In summary, the findings of the present study demonstrate that galactose may enhance wound healing and mannose treatment inhibits the inflammatory reaction in wound healing and decreases granulation tissue formation in an experimental wound model.

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 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.

Glomus tumor of the trachea.

We report a case of a glomus tumor in the trachea which was an incidental finding in a 66-year-old man. The histological picture and immunohistochemical profile were typical for this tumor. The glomus tumor is an exceedingly rare mass lesion in the trachea, but it is useful to keep it among differential diagnostic alternatives when a tracheal tumor is seen on radiographs or endoscopy.

Immune responses and clinical outcome of massive human osteoarticular allografts.

Cell mediated immune responses as measured by lymphocyte proliferation induced by the mitogens phytohemagglutinin and concanavalin A and antigen extracts of donor derived bone were studied within 2 years after wide resection of bone tumors in 18 patients receiving fresh frozen massive osteoarticular allografts. No uniform changes were seen in mitogen induced responses in 18 patients. However, five of nine patients tested with antigen extracts of donor derived bone showed elevated immune responses, one moderate and four weak. The incorporation of the allograft (evaluated by repeated radiographs; specific isotope techniques; clinical outcome assessed by the functional rating scores of Mankin-Waber and the Musculoskeletal Tumor Society; and histologic biopsy findings on new bone formation) did not differ in these patients from those in patients without any response to donor derived tissue. During a long term followup (mean, 11 years; range, 2-20 years), degenerative joint and sclerotic density bone changes developed after 2 to 4 years without correlation to immune responses. Histologic specimens showed no signs of immunologic reaction, and no clinical rejection episodes were recorded. A slightly variable immune response to allograft bone seems to occur, but its clinical significance for outcome of the grafts remains to be determined. The low immune responses might reflect a low antigen release rate through an indirect pathway or immunologic tolerance to antigens or proteins shed from massive allografts that are nonliving scaffold implants during the creeping substitution process, corresponding to the low immune response and the slow histologic repair.

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.

P90 exceeding rate as a prognostic factor in primary malignant melanoma of the skin. A DNA image cytometric study.

OBJECTIVE: To study the prognostic value of DNA image cytometry in primary skin melanomas. STUDY DESIGN: DNA image cytometry was performed on 62 stage I, Clark level II-V, primary skin melanomas. The DNA histograms were classified into three categories (diploid, nondiploid and aneuploid) according to the percentages of cells with higher-than-diploid and higher-than-twice-the-diploid DNA content (the P90 and 2P90 exceeding rates [ERs]). The prognostic value of P90ER, 2P90ER, type of DNA histogram, melanoma thickness, Clark level, and patient age and sex were analyzed for disease-specific survival with Cox's stepwise proportional hazards model. RESULTS: Aneuploid DNA histograms were as common in thin as in thick melanomas. Melanoma thickness and P90ER had prognostic value in univariate analysis, but in the multivariate analysis only P90ER had independent and significant prognostic value. CONCLUSION: Aneuploidy is a common feature of malignant melanoma, and it is as common in thin as in thick melanomas. P90ER has more prognostic value than the type of DNA histogram. The prognostic value of P90ER as compared with melanoma thickness should be studied further.

Loss of expression of the p16INK4/CDKN2 gene in cutaneous malignant melanoma correlates with tumor cell proliferation and invasive stage.

The G1/S checkpoint of the cell cycle is regulated by p16, p53 and RB tumor suppressor genes. Loss of expression of the p16INK4 tumor suppressor protein, the product of the CDKN2 gene, has been associated with a wide variety of human malignancies. Mutations, loss of heterozygosity and deletions of the CDKN2 locus have been reported in sporadic and familial cutaneous malignant melanomas (CMM). To investigate the role of the alterations of p16 expression in melanoma, we evaluated by immunohistochemistry the p16 expression and cell proliferation in 79 primary CMM and 10 benign melanocytic nevi (BMN). Forty-six melanomas (58%) and all BMN were found to be p16 positive; 33 melanomas (42%) were considered p16 negative. The extent of invasion according to Clark was significantly higher in p16-negative tumors than in p16-positive tumors. Cell proliferation as expressed by the proportion of positive cells in Ki-67 immunostaining was found to be significantly higher in p16-negative tumors than in p16-positive tumors, although there was no significant difference in the mitotic index between p16-positive and p16-negative tumors. In p16-positive tumors, the number of Ki-67-positive cells correlated with the mitotic index; in p16-negative tumors, there was no correlation between these parameters. Our data suggest that loss of p16 expression is more common in advanced melanomas, and that G1/S checkpoint regulation is disrupted in p16-negative melanomas. Our results show that loss of p16 expression is a common event in primary melanomas, which further substantiates the role of p16 as a major tumor suppressor.

Primary malignant melanoma of the skin. Relationships of nuclear DNA content, nuclear morphometric variables, Clark level and tumor thickness.

OBJECTIVE: To investigate how nuclear morphometric variables, tumor thickness (measured according to Breslow), invasion depth (classified according to Clark), nuclear DNA content and type of DNA histogram are associated with each other in primary malignant melanomas of the skin. STUDY DESIGN: Image analysis DNA cytometry and nuclear morphometry were performed on 85 primary skin melanomas. The relationships of size, sphericity and DNA content of melanoma cell nuclei; melanoma thickness; and Clark level were analyzed in detail. The effect of melanin bleaching on DNA cytometry results was studied. RESULTS: Melanoma thickness correlated with nuclear size in aneuploid, but not diploid, melanomas. The prevalence of aneuploidy did not increase with tumor thickness. In aneuploid melanomas the proportion of cells with higher-than-diploid and higher-than-tetraploid DNA content increased with tumor size. CONCLUSION: Aneuploidy is as common in thin as in thick melanomas. Genetic instability in aneuploid melanomas correlates with melanoma thickness. This correlation in aneuploid melanomas partially explains the correlation between nuclear size and melanoma thickness. In diploid melanomas no correlation was observed between nuclear size and melanoma thickness. DNA cytometry is a valuable tool for studies on the background of phenotypic changes in skin melanomas.