Soft-tissue sarcoma of the thigh: surgical margin influences local recurrence but not survival in 152 patients.

Between 1979 and 1998, 152 patients with a soft-tissue sarcoma arising in the thigh were surgically treated in the Sarcoma Center in Aarhus, Denmark. We studied clinicopathologic factors prognostic for local recurrence and survival. 27 patients (18%) had a low-grade tumor, 26 (17%) an intermediate-grade and 99 (65%) a high-grade tumor. 27 patients (18%) were amputated and 125 (82%) had a local resection. 21 (14%) underwent a marginal resection, 82 (54%) a wide resection and 49 (32%) a compartmental resection. 32 patients were also given radiotherapy, 11 of these had a marginal resection. The 5-year local recurrence-free rate was 91%. Multivariate analysis selected marginal resection and histological high grade as unfavorable prognostic factors for local recurrence. The 5-year survival rate was 68%. High age and histological high grade were unfavorable prognostic factors for survival in a multivariate analysis. Surgical margin influenced local recurrence, but not the overall survival.

Hypoxia in human soft tissue sarcomas: adverse impact on survival and no association with p53 mutations.

Clinical and experimental studies have suggested that tumour hypoxia is associated with poor treatment outcome and that loss of apoptotic potential may play a role in malignant progression of neoplastic cells. The tumour suppressor gene p53 induces apoptosis under certain conditions and microenvironmental tumour hypoxia may select for mutant tumour cells with diminished apoptotic potential due to lack of p53 function. The aim of this study was to evaluate the prognostic relevance of oxygenation status for treatment outcome and to compare pre-treatment tumour oxygenation measurements were done in 31 of those by PCR using DNA extracted from paraffin-embaedded sections (n = 2) or frozen biopsies (n = 29). The overall median of the tumour median pO(2)was 19 mmHg (range 1-58 mmHg). Only 6 tumours had functional p53 mutations and no association was found between mutant p53 and tumour hypoxia. Five out of 6 STS with lower histopathological grade were well-oxygenated whereas high-grade STS were both hypoxic and well-oxygenated. At a median follow-up of 74 months, 16 patients were still alive among 28 available for survival analysis. When stratifying into hypoxic and well-oxygenated tumours patients with the most hypoxic tumours has a statistically poorer disease-specific and overall survival at 5 years. In conclusion hypoxia was an indicator for both a poorer disease specific and overall survival in human STS but hypoxic tumours were not characterized by mutations in the p53 gene.

[Prognostic factors in soft tissue sarcomas. Experiences from the Sarcoma Center in Aarhus]. / Prognostiske faktorer ved bløddelssarkomer. Erfaringer fra Sarkomcentret i Arhus.

In the present study, the outcome, patterns of local recurrence and survival, as well as prognostic factors, were evaluated in patients surgically treated for soft tissue sarcomas. Between January 1979 and July 1993, 316 consecutive patients were referred to the Sarcoma Centre in Aarhus with localised malignant soft tissue sarcoma of the extremities or trunk. There were 161 men (51%) and 155 women (49%), the median age was 56 years (1-94). Histologically 52 patients (16%) had a grade I, 60 patients (19%) a grade 2 and 204 patients (65%) a grade 3A or 3B tumour. The five-year local recurrence rate was 18% and the five-year survival rate was 75%. Multivariate analysis indicated the following variables as independent unfavourable factors for local recurrence: extracompartmental location, histological high-grade (i.e. histologically highly malignant) local excision, no adjuvant radiotherapy and intralesional/marginal excision. Independent unfavourable factors for survival were advanced age, extracompartmental location, histological high-grade, lower extremity location and large tumour size. Based on these variables, a prognostic model was made.

[Agressive fibromatosis]. / Aggressiv fibromatose.

Aggressive fibromatosis is a rare soft tissue tumour with a high tendency to local recurrence. The patients often present with a deeply sited, firm mass with or without pain. Genetic, physical and endocrine factors seem to be important in the development of the disease. Patients with Gardner's syndrome and familial adenomatous polyposis have a higher risk of developing aggressive fibromatosis and often develop tumours in the mesentery, retroperitoneally or in the abdominal wall. Infantile fibromatosis is a variant of aggressive fibromatosis. The standard treatment of aggressive fibromatosis is surgical resection with wide free margin, in some cases supplemented with radiotherapy. Chemotherapy and hormonal treatment of non-resectable tumours has been described without convincing results and more data from clinical studies are needed.

Objective assessment of photoreceptor displacement and metamorphopsia: a study of macular holes.

BACKGROUND: We have developed a binocular perimetry technique for the quantitative assessment of retinal photoreceptor displacement and metamorphopsia. OBJECTIVE: To study the direction and amplitude of retinal photoreceptor displacement in eyes with idiopathic macular holes using our binocular perimetry technique. SUBJECTS: Five healthy control subjects and 10 patients with unilateral stage 3 to 4 macular holes in one eye and a healthy fellow eye. METHOD: Kinetic perimetry using red and green filter glasses, black binocular fixation targets, red and green selective monocular stimuli (Goldmann III-4-e), and fundus image superimposition of perimetry data. RESULTS: We found no discrepancy between the 2 visual fields in any healthy subjects. In patients with a unilateral macular hole, the central scotoma invariably extended beyond the rim of the hole. In 8 patients, each point on the rim of the scotoma had a perceptually corresponding location in the visual field of the fellow eye that was closer to the center of the visual field. In the 2 patients with the longest duration of symptoms (>2 years), no such discrepancy was found. CONCLUSIONS: Differential perimetry enables the objective study of retinal photoreceptor displacement and metamorphopsia. We found objective evidence for radial centrifugal photoreceptor displacement in most patients with idiopathic macular holes.

Proliferative activity (MIB-1 index) is an independent prognostic parameter in patients with high-grade soft tissue sarcomas of subtypes other than malignant fibrous histiocytomas: a retrospective immunohistological study including 216 soft tissue sarcomas.

AIMS: To evaluate the prognostic value of tumour proliferative activity, p53 accumulation and bcl-2 expression in a retrospective series of 216 patients with soft tissue sarcomas (STS). METHODS AND RESULTS: The immunohistochemical analyses were performed on formalin-fixed, paraffin-embedded tissue. The proliferative activity was assessed by use of the monoclonal antibody MIB-1 and evaluated in multiple, random systematic sampled fields of vision. The percentage of proliferating cells (the MIB-1 index) ranged between 1% and 85% (median 12%). A significant increase in mean MIB-1 index was seen with increasing histological malignancy grade. Variation in the incidence of p53 accumulation and bcl-2 positivity among different histological subtypes was observed. p53 accumulation was frequent in synovial sarcomas and leiomyo- and rhabdomyosarcomas, whereas bcl-2 preferentially was expressed in synovial sarcomas. Univariate analysis identified patient age, tumour size, histological grade of malignancy, MIB-1 index and p53 accumulation as significant prognostic parameters. Multivariate Cox analysis, including tests for interaction terms between histological subtypes and MIB-1 index, showed independent prognostic effect of MIB-1 index and tumour size in patients with high-grade tumours of other subtypes than malignant fibrous histiocytoma (MFH). CONCLUSIONS: Histopathological malignancy grading is the most important single prognostic factor for overall survival in STS, but estimation of MIB-1 index is useful for identifying the least favourable subgroup of high grade STS of other subtypes than MFH, for whom adjuvant therapy may be indicated.

Prognostic factors in soft tissue sarcomas: the Aarhus experience.

In the present study, the outcome, patterns of local recurrence and survival, as well as prognostic factors, were evaluated in patients surgically treated for soft tissue sarcomas. Between January 1979 and July 1993, 316 consecutive patients were referred to the Sarcoma Centre in Aarhus with localised malignant soft tissue sarcoma of the extremities or trunk. If possible, the patients were treated with a limb-sparing resection, primarily by use of a wide excision. 50 patients received adjuvant radiotherapy. There were 161 men (51%) and 155 women (49%) median age 56 years (range 1-94 years). 94 patients (30%) had tumours in the trunk, including shoulder and buttock lesions, 163 (52%) in the lower extremity and 59 (19%) in the upper extremity. 52 patients (16%) had grade 1 tumour, 60 (19%) grade 2 and 204 (65%) grades 3A-3B. The 5-year local recurrence rate was 18% and the 5-year survival rate was 75%. Multivariate analysis indicated the following variables as independent unfavourable factors for local recurrence: extracompartmental location, histological high grade, local excision, no adjuvant radiotherapy and intralesional/marginal excision. Independent unfavourable factors for survival were advanced age, extracompartmental location, histological high grade, lower extremity location and large tumour size. If the variable local recurrence was included in the analysis, it was found to have a very strong influence on survival. Based on these variables, a prognostic model was developed.

In B-cell chronic lymphocytic leukaemia chromosome 17 abnormalities and not trisomy 12 are the single most important cytogenetic abnormalities for the prognosis: a cytogenetic and immunophenotypic study of 480 unselected newly diagnosed patients.

Of 560 consecutive, newly diagnosed untreated patients with B CLL submitted for chromosome study, G-banded karyotypes could be obtained in 480 cases (86%). Of these, 345 (72%) had normal karyotypes and 135 (28%) had clonal chromosome abnormalities: trisomy 12 (+12) was found in 40 cases, 20 as +12 alone (+12single), 20 as +12 with additional abnormalities (+12complex). Other frequent findings included abnormalities of 14q, chromosome 17, 13q and 6q. The immunophenotype was typical for CLL in 358 patients (CD5+, Slg(weak), mainly FMC7-) and atypical for CLL in 122 patients (25%) (CD5-, or Slg(strong) or FMC7+). Chromosome abnormalities were found significantly more often in patients with atypical (48%) than in patients with typical CLL phenotype (22%) (P < 0.00005). Also +12complex, 14q+, del6q, and abnormalities of chromosome 17 were significantly more frequent in patients with atypical CLL phenotype, whereas +12single was found equally often in patients with typical and atypical CLL phenotype. The cytomorphology of most of the +12 patients was that of classical CLL irrespective of phenotype. In univariate survival analysis the following cytogenetic findings were significantly correlated to a poor prognosis: chromosome 17 abnormalities, 14q+, an abnormal karyotype, +12complex, more than one cytogenetic event, and the relative number of abnormal mitoses. In multivariate survival analysis chromosome 17 abnormalities were the only cytogenetic findings with independent prognostic value irrespective of immunophenotype. We conclude that in patients with typical CLL immunophenotype, chromosome abnormalities are somewhat less frequent at the time of diagnosis than hitherto believed. +12single is compatible with classical CLL, and has no prognostic influence whereas chromosome 17 abnormalities signify a poor prognosis. In patients with an atypical CLL immunophenotype, chromosome abnormalities including +12complex, 14q+, del 6q and chromosome 17 are found in about 50% of the patients, and in particular chromosome 17 abnormalities suggest a poor prognosis.

The bone-marrow infiltration pattern in B-cell chronic lymphocytic leukemia is not an important prognostic factor. Danish CLL Study Group.

In a multicentre study of 635 consecutive newly diagnosed patients with B-CLL, the histological bone marrow (BM) specimens were reviewed independently by each of 3 pathologists and found evaluable for BM infiltration pattern in 575 patients, 404 of whom had a CD5+, mainly FMC7-, faint surface-membrane immunoglobulin (SIg) fluorescence-intensity ppenotype. In these 404 patients the following BM infiltration patterns were found: mixed nodular-interstitial (30%), moderate interstitial (44%), heavy interstitial (20%) and diffuse packed (6%). In univariate survival analysis, significant differences were found according to BM pattern (p < 0.05), the presence of nodules being a favorable prognostic sign. In multivariate survival analysis in a model including age, clinical stage, BM pattern, BM lymphocytosis, WBC and sex, only age and stage but not BM pattern or BM lymphocytosis had independent prognostic significance. In stage A, progression-free survival was significantly longer in patients with nodular than in patients with non-nodular bone-marrow pattern. The overall survival of these patients, however, did not differ, possibly owing to the prompt and prolonged treatment given to most patients at the time of progression to stage B or C. We conclude that in CD5+, SIg(faint), mainly FMC7-B-CLL, bone-marrow histology may predict unstable disease in early clinical stage but is not important for treatment decisions, when these are based on clinical stage.

Intradermal and subcutaneous leiomyosarcoma: a clinicopathological and immunohistochemical study of 41 cases.

Superficial leiomyosarcomas are rare tumours. The lesions confined to the dermis, contrary to those involving the subcutis, have been reported to carry a favourable prognosis. A retrospective study of 41 consecutive cases of surgically treated intradermal and subcutaneous leiomyosarcomas was undertaken in order to determine the prognostic factors that may influence the survival of these patients. Seven tumours were predominantly intradermal and 34 involved the subcutaneous tissue. Fifty-four percent of the tumours were located in the lower extremities. All cases stained positively for smooth muscle antigen and 66% for desmin. The tumours were classified with regard to tumour grade I (low grade, 3%), II (intermediate, 12%), IIIA (high grade, 46%) and IIIB (high grade, 39%). In all patients, follow-up information was available. Mean follow-up time was 5 years. The patients with intradermal tumours were all alive without signs of recurrence, whereas 14 of those with leiomyosarcomas involving the subcutis have died with pulmonary metastases. Our study confirms that "pure" intradermal leiomyosarcomas independent of tumour grade behave in a benign fashion, probably due to small tumour size. Tumour size > or = 5 cm, deep localization with fascia involvement, and high malignancy grade (IIIB) were found to deteriorate survival based on a univariate analysis. However, in a multivariate analysis only tumour size was found to be an independent prognostic factor.

Fibrous dysplasia of the spine, costae and hemipelvis with sarcomatous transformation.

We describe a patient with polyostotic fibrous dysplasia and secondary malignant fibrous histiocytoma in a spinal lesion.

The relationship between tumor oxygenation and cell proliferation in human soft tissue sarcomas.

PURPOSE: In malignant tumors the oxygenation status and tumor cell proliferation are known to influence local tumor control after radiotherapy. However, the relationship between oxygenation status and tumor cell kinetics in human tumors has not yet been described. Newly developed clinically applicable techniques such as oxygen electrode measurements and assessment of tumor cell proliferation rates have been suggested as promising predictive assays. The purpose of the present study was to characterize tumor oxygenation status in soft tissue sarcomas and to compare this with tumor cell kinetics and clinical parameters. METHODS AND MATERIALS: Pretreatment tumor oxygenation status was measured by polarographic oxygen needle electrodes and evaluated as the median pO2 and the percentage of pO2 values < or = 5 mmHg and < or = 2.5 mmHg in 22 patients with primary soft tissue sarcomas. All tumors were characterized by histology, grade of malignancy, the level of microscopic necrosis, the level of effective hemoglobin, and magnetic resonance imaging estimation of tumor volume. The tumor cell potential doubling time and labeling index were measured by flow cytometric and immunohistochemical analysis of tumor biopsy specimens after in vivo incorporation of iododeoxyuridine. RESULTS: There was a significant correlation between the median pO2 and the tumor cell potential doubling time (p = 0.041), whereas no correlation was found between the level of hypoxia expressed by the percentage of pO2 values < or = 2.5 and < or = 5 mmHg, respectively, and tumor cell potential doubling time. Furthermore, no correlation was found between either of the three tumor oxygenation parameters and labeling index. The material represented large intertumor heterogeneity in oxygenation status, cell kinetics, and tumor volume, and no correlation was found between oxygenation status and either volume, histopathology, grade of malignancy, or effective hemoglobin. CONCLUSION: This report is the first to suggest a correlation between tumor oxygenation and tumor cell doubling time, as the fastest proliferating tumor cells were found in the poorest oxygenated soft tissue sarcomas. More data are needed to clarify if this relation is really a true biological phenomenon. Furthermore, tumor oxygenation status of soft tissue sarcomas was heterogeneous and independent of clinical and histopathological parameters.

MIB-1 expression and iododeoxyuridine labelling in soft tissue sarcomas: an immunohistochemical study including correlations with p53, bcl-2 and histological characteristics.

We investigated the relationship between immunohistochemical estimates of proliferative activity and expression of bcl-2 protein and mutant p53 protein in 23 cases of soft tissue sarcoma. Furthermore, the reproducibility of estimates of proliferative activity was analysed and correlations between the variables and with mitotic score were investigated. Proliferative activity was assessed by use of monoclonal antibody MIB-1 and staining for iododeoxyuridine (IdUrd), and evaluated in multiple, random, systematically sampled fields of vision. MIB-1 indices were higher than those of IdUrd but for each case the two values were positively correlated (r = 0.78). The MIB-1 index correlated positively with mitotic score (2P < 0.001) and malignancy grade (2P = 0.001). The intraobserver reproducibility of the MIB-1 and IdUrd indices were excellent (r = 0.98 and r = 0.90, respectively). p53 expression was detected in 43% and strong bcl-2 expression was present in 57% of the studied cases. Expression of p53 and bcl-2 were not significantly correlated to proliferative activity or the histological features. We conclude, that the MIB-1 index is a reliable and reproducible estimate of proliferative activity and might improve the accuracy of conventional malignancy grading of soft tissue sarcomas. Furthermore, the results indicate that neither mutant p53 protein nor bcl-2 oncogene alone are sufficient to induce increased proliferation in these sarcomas.

[Can magnetic resonance imaging differentiate between benign and malignant conditions in patients with skeletal or soft tissue tumors?]. / Kan magnetisk resonans-undersøgelse adskille benigne tilstande fra maligne hos patienter med skelet- eller bløddelstumorer?

The records of 437 patients referred preoperatively for magnetic resonance imaging (MRI) due to clinical or radiological suspicion of a bone or soft-tissue malignancy were examined retrospectively in order to evaluate how good MRI is at distinguishing malignant from benign lesions. The MR-examination tends to overestimate malignancy. In all cases, the preoperative MR-examination correctly identified all malignant tumours as malignant, but also estimated a number of benign lesions as malignant. Sensitivity, specificity and predictive value was 100%, 71.2% and 76.4% respectively. In case of bursa/ganglion and pseudotumour, MR was able to correctly identify these lesions as benign. MR could not differentiate between the different types of sarcomas.

Genital anomalies and risk for testicular cancer in Danish men.

In a cohort of Danish boys characterized by (1) being born between 1941 and 1957, (2) having attended schools in a defined area of Denmark, and (3) having a school health record available, 183 were registered in the Danish Cancer Registry with testicular cancer diagnosed before January 1, 1985. We selected 366 age- and sex-matched controls from the same cohort. Using information recorded by school physicians, we performed logistic regression analyses to estimate the relative risks (RR) associated with various genital anomalies. We found the risk for testicular cancer to be raised for men with a history of cryptorchidism [RR = 5.2; 95% confidence interval (CI) = 2.1-13.0], inguinal hernia (RR = 1.8; 95% CI = 0.9-3.7), hypospadias (RR = 4.2; 95% CI = 0.4-42.7), and hydrocele (RR = 2.4; 95% CI = 0.6-9.0). We observed no decrease in the risk associated with cryptorchidism after correction of the maldescent in early childhood. The RR of testicular cancer in the contralateral, normally descended testis in unilateral cryptorchid men was increased to 3.6. The results add to the growing evidence for a common causal factor for both testicular cancer and cryptorchidism and support the findings from other studies of associations between other genital anomalies involving the closure of the processus vaginalis and the risk of testicular cancer.

Penetrating keratoplasty and transscleral fixation of posterior chamber lens.

We reviewed the charts of 21 patients who underwent penetrating keratoplasty and transscleral fixation of a posterior chamber lens. One lens was sutured in an aphakic eye and 20 lenses were sutured after removal of an anterior chamber lens. Postoperative follow-up averaged 13 months (2-39 months). Visual acuity improved in 20 patients (95%) and remained the same in 1 patient (5%). Postoperative visual acuity was less than 0.1 in 5 patients (23.8%), 0.1 to 0.33 in 14 patients (66.7%) and better than 0.33 in 2 patients (9.5%). Twelve patients (57.1%) expressed a substantial reduction in ocular pain, 7 patients (33.3%) had no pain either before or after the operation, 2 patients (9.5%) expressed no reduction in pain. No cases of endophthalmitis, choroidal hemorrhage or retinal detachment were found. In one case, the sutured lens was dislocated without disturbing vision. Intraocular pressure increased in 3 of 9 patients with preoperative glaucoma. New-onset glaucoma developed in 1 patient. We find transscleral fixation of a posterior chamber lens to be an acceptable procedure in penetrating keratoplasty with IOL implantation where capsular support is inadequate for conventional implantation of a posterior chamber lens.

Conjugation to preactivated proteins using divinylsulfone and iodoacetic acid.

Two methods for the preactivation of proteins and conjugation of peptides to proteins under mild conditions are presented. Preactivation of proteins with divinylsulfone (DVS) permits peptide conjugation through either amino, hydroxyl or sulphydryl groups depending on the coupling pH used, while preactivation with iodoacetic acid (IAA) N-hydroxy-succinimide ester permits selective conjugation through sulphydryl groups. In addition, the latter method allows quantitation of the conjugation ratio through determination of carboxymethyl cysteine after acid hydrolysis. The divinylsulfone activated proteins can be stored for extended periods of time at -20 degrees C until required for conjugation, while the iodoacetic acid activated protein can be stored for a few days at -20 degrees C. These conjugation methods were investigated with respect to obtaining peptide/protein conjugates for immunization purposes and for use as reagents in immunoassays. The DVS activated proteins permitted direct conjugation of luteinizing releasing hormone (LHRH) through its tyrosine side chain and allowed synthesis of well defined conjugates. The DVS derivatives of bovine serum albumin (BSA), reduced and carboxymethylated BSA and purified protein derivative (PPD) were compared with respect to their potential value as carriers for obtaining antibodies to LHRH (M(r) 1000) and epidermal growth factor (EGF, M(r) 5000). IAA-PPD was evaluated as a carrier for the conjugation of glutathione specifically through its cysteine side chain and for obtaining antibodies to glutathione. The antisera obtained were specific and of high titer, and the methods described here will thus allow the convenient synthesis of carrier conjugates with well defined characteristics.

Quantitation of immunogold with an interactive image analysis system. A new, practical approach to antibody-induced modulation, internalization and intracellular transport of surface antigens in viable hematopoietic cells.

This work aimed to evaluate the value of computerized quantitative analysis in ultrastructural studies on internalization and intracellular transport of leukocyte antigens traced with immunogold in viable lymphoid cells. The time of analysis and the results were compared with those obtained with a standard method based on counting gold particles on electron micrographs. A commercially available, personal computer-based, single-screen system was used for capturing and processing the ultrastructural images, counting gold particles and presenting data. The viable lymphoblasts of RAJI and NALM-6 cell lines were labeled with B4 (anti-CD19) murine monoclonal antibody followed by rabbit antimouse immunoglobulins coupled to 12.8-nm colloidal gold, cultured for six hours to allow internalization of CD19 antigen, collected after various periods of time and processed for electron microscopy. Each sample was analyzed by counting the gold particles on 30 randomly chosen, equatorial cell sections. Surface-bound particles and those located within certain intracellular compartments (that is, cytoplasmic vesicles, multivesicular bodies and lysosomes) were counted separately. No significant differences were found between the results obtained using electron micrographs and those yielded by on-screen analysis of digitized images. However, the time of analysis varied considerably--approximately eight and two hours, respectively. Also, using computerized analysis saved the cost of photographic processing. In conclusion, the system provided a reliable, rapid and inexpensive alternative for quantitation of immunogold-labeled leukocyte antigens at the ultrastructural level.

Changing indications for penetrating keratoplasty.

The indications for penetrating keratoplasty have changed over the past several years. One hundred and eighty cases of penetrating keratoplasty performed in the Department of Ophthalmology, Naestved Centralsygehus, Denmark, from January 1984 to December 1993, were analysed. Overall pseudophakic bullous keratopathy was the most common indication for penetrating keratoplasty (28.3%). This was followed by keratitis (13.9%), Fuchs' dystrophy (13.9%), regraft (11.1%), aphakic bullous keratopathy (10.0%) and keratoconus (6.7%). Pseudophakic bullous keratopathy was the most common indication for penetrating keratoplasty in 1989 and in each year from 1991 to 1993. Before 1989 the most common indications were keratitis (18.0%) and aphakic bullous keratopathy (14.8%). The emergence of pseudophakic bullous keratopathy, as the most common indication for penetrating keratoplasty, correlates well with the dramatic increase in the number of cataract extractions with intraocular lens implantation performed since the early 1980s. Especially semiflexible, closed-loop anterior chamber lenses used in our department in the early years of the period, have been the cause of subsequent corneal edema.