Thymidylate synthase expression in patients with colorectal carcinoma using a polyclonal thymidylate synthase antibody in comparison to the TS 106 monoclonal antibody.

Colorectal cancer is one of the most common human cancers, for which 5-fluorouracil (5FU) is usually part of the treatment. Thymidylate synthase (TS), the target enzyme for 5FU, can be predictive for the outcome of 5FU-based therapy. TS levels in tumor samples can be determined with radiochemical enzyme assays, RT-PCR, and immunohistochemical staining. We validated TS immunohistochemistry with a polyclonal rabbit anti-human TS antibody using the avidin-biotin method. This antibody can be used on paraffin-embedded, formalin-fixed material using an antigen retrieval method with citrate buffer and microwave treatment. The antibody shows a granular cytosolic staining pattern. The reproducibility in cross-sections from colorectal tumors from 50 patients was 90% and the interobserver variability was acceptable with a kappa of 0.45. On Western blotting it detects purified TS at 36 kD, while in 5FU-treated cells the ternary complex between FdUMP, TS, and 5, 10-methylene-tetrahydrofolate is clearly visible at 38 kD, with no other interfering bands. In a separate set of tumors, immunostaining was compared with enzyme levels; Western blots correlated with enzyme levels. Because both this polyclonal antibody and the monoclonal antibody TS-106 are being used for large-scale studies, we also determined whether they could be used interchangeably. No differences were observed. This polyclonal antibody is specific and gives reproducible results. A study on a larger scale is ongoing to determine the role of TS as a predictive parameter in patients with colorectal cancer treated either with postoperative adjuvant 5FU/levamisole or with surgery only.

Correlative 201Tl SPECT, MRI and ex vivo 201Tl uptake in detecting and characterizing cervical lymphadenopathy in head and neck squamous cell carcinoma.

UNLABELLED: The value of SPECT with 201Tl chloride, in combination with MRI (particularly short inversion-time inversion recovery [STIR] sequences that suppress fat signals) to detect and characterize cervical lymphadenopathies (nodes > or = 1 cm), and ex vivo lymph node 201Tl uptake were studied in patients with squamous cell carcinoma of the head and neck. METHODS: Preoperative SPECT and MRI, displayed in similar planes, were compared with the histologic findings in 15 neck dissection specimens from 12 patients with squamous cell carcinoma of the head and neck (9 with unilateral and 3 with bilateral neck dissection). Results were evaluated topographically with regard to the lymph node compartments (levels) of the neck. In addition, in 8 of these patients, the 201Tl activity of dissected lymph nodes of 10 neck sides was measured immediately after surgery in a gamma counter and expressed as percentage of the injected dose per gram tissue (%ID/g). RESULTS: Sixty-two lymph node levels were evaluated histologically. The high sensitivity of MRI (92% versus 71% for 201Tl SPECT), which correctly detected lymph node involvement in 22 of 24 levels, and the high specificity of 201Tl SPECT (92% versus 71% for MRI), which correctly characterized as negative 35 of 38 lymph node levels without metastasis on histology, led to a combined 201Tl SPECT/MRI accuracy of 92%. 201Tl SPECT was particularly effective in excluding involvement in 9 tumor-free neck levels with pathologically enlarged lymph nodes on MRI but failed to confirm involvement in 5 other tumor-positive levels. Mean 201Tl uptake in 53 lymph nodes with confirmed histologic involvement was significantly higher than uptake in 145 tumor-free lymph nodes (0.0043+/-0.0022 %ID/g versus 0.0023+/-0.0014 %ID/g, P = 0.0001), muscle and fat tissue but clearly lower than salivary gland uptake. CONCLUSION: Although 201Tl SPECT is not sensitive enough to be used as an independent imaging modality for staging of the neck, its correlative application with MRI appears to be an accurate method for the assessment of regional spread in head and neck squamous cell carcinoma. The ability of 201Tl SPECT to characterize neck lymphadenopathies detected by MRI appears to be based on the difference in 201Tl concentration found in lymph nodes with and without tumor involvement.

Metastasizing pleomorphic adenoma: a report of three cases.

Three cases of metastasizing pleomorphic adenoma are described. All three had a history of incomplete surgery of the primary tumour and several surgical procedures for local recurrences before systemic metastases became apparent. The histology of the primary tumour, recurrences and metastases were typical of pleomorphic adenoma. In view of the lack of information in the literature, it is concluded that inadequate surgical procedures, such as incomplete surgery or enucleation, leading to multiple recurrences, appear to be a prerequisite for the development of systemic disease.

Thallium-201 SPECT in the diagnosis of head and neck cancer.

UNLABELLED: The accuracy of SPECT with 201Tl-chloride for the diagnosis of primary tumors, lymph node metastases and recurrences in head and neck cancer was evaluated for clinical applicability. METHODS: SPECT images, obtained 60 min after administration of 150 MBq 201Tl-chloride, were compared with clinical, CT and/or MRI and histology results. In addition, whole-body images were obtained to detect distant metastases. RESULTS: In 79 patients studied for primary tumors (principally larynix, hypopharynx, oropharynx, nasopharynx and oral cavity), 201Tl SPECT correctly identified 69 of 73 (95% versus 88% for CT/MRI) histologically confirmed malignancies including 63 squamous-cell carcinomas. The method localized four occult naso- and oropharynx carcinomas not seen on CT/MRI and was correctly negative in two patients without tumor and in three of four patients with no confirmed primary tumor in the head and neck. With respect to regional spread, only patients who had cervical lymph node dissection were evaluated, and the findings were recorded per side of the neck. Thallium-201 SPECT correctly identified metastases in 31 of 36 neck dissections with proven lymph node involvement (86%), was correctly negative in nine and false-positive in one. Although the sensitivity of CT/MRI was clearly higher (97%), considerably more false-positive cases affected its accuracy (81% versus 87% for SPECT). In 30 patients investigated for recurrences, 201Tl SPECT correctly identified 27 of 29 microscopically confirmed tumor sites (93%) and was correctly negative in seven. Sensitivity of CT/MRI was lower (76%), and a greater number of false-positives (seven versus three for SPECT) further decreased its accuracy (64% versus 87% for SPECT). Distant metastases were detected in five patients. CONCLUSION: Thallium-201 SPECT appears to be an accurate method for the diagnosis of head and neck cancer. The method is particularly useful for detection of occult head and neck tumors and for assessing recurrences. It also may be of complementary value in the staging of primary tumors, in the differentiation of metastatic from reactive lymph nodes in the neck and, on the basis of whole-body scanning, for screening of distant metastases.

The role of radiotherapy in the local management of dermatofibrosarcoma protuberans. Soft Tissue Tumours Working Group.

Dermatofibrosarcoma protuberans (DFSP), a fibrohistiocytic tumour of intermediate malignancy, has a strong tendency to recur locally. Wide local excision is the recommended treatment modality. A retrospective analysis was performed on 38 consecutive DFSP patients presenting to The Netherlands Cancer Institute, to define the role of radiotherapy. Of the 21 patients treated surgically (all with negative resection margins) seven recurred, a local control probability of 67%. Combined modality treatment was given to 17 patients. Prior to radiotherapy, these patients experienced 33 occurrences of DFSP, but after irradiation only three recurrences were seen, a local control probability of 82%. These results are in keeping with the recent literature where increasing value is being given to both adjuvant and curative radiotherapy in the local management of DFSP. We recommend radiotherapy in DFSP patients where repeated surgery may cause mutilation or functional impairment.

Overexpression of cyclin D1 indicates a poor prognosis in squamous cell carcinoma of the head and neck.

OBJECTIVES: To evaluate the overexpression of cyclin D1 and p53 as a prognostic marker of squamous cell carcinoma of the head and neck and to investigate whether deregulation of these genes is associated with an unfavorable course of disease. DESIGN: Retrospective study. MATERIALS AND METHODS: Formalin-fixed, paraffin-embedded tumor materials that were obtained from a well-characterized series of 115 patients with resectable head and neck cancer at The Netherlands Cancer Institute, Amsterdam, were analyzed by immunohistochemical methods using antiserum samples that were directed against 2 proteins (ie, cyclin D1 and p53), which are crucial in the regulation of the G1 phase of the cell cycle. RESULTS: Overexpression of cyclin D1 protein was found in 49% of the patients with squamous cell carcinoma of the head and neck. This overexpression was not associated with known prognostic factors (eg, the T and N stages). Tumors recurred more frequently and in a shorter period in patients whose primary tumors showed an overexpression of cyclin D1 protein. This difference (P = .05) was statistically significant in a stepwise proportional hazard regression analysis. However, since a discrepancy in staining results was observed between the biopsy and resection materials that were taken from the same patient, this result may not have been applicable in the evaluation of biopsy specimens only. This discrepancy is most likely owing to tissue heterogeneity. The overexpression of p53 that was found in 42% of the patients was of no prognostic significance. CONCLUSIONS: These data provide evidence that overexpression of cyclin D1 protein in resection material of squamous cell carcinomas of the head and neck is indicative of a poor prognosis, independently of other known prognostic factors. Whether overexpression of cyclin D1 may therefore be used to select patients for more intensive treatment should be examined in the context of a clinical trial.

Osteoid and bone formation in a nasal mucosal melanoma and its metastasis.

AIMS: To present a literature review and a case history concerning bone and osteoid formation by a metastasizing (mucosal) melanoma. CASE DETAILS: Osteocartilaginous differentiation and production of osteocartilaginous structures in malignant melanoma have been described only in 12 previous cases (osteoid in 11. bone in four), all of which involved dermal melanomas. Five of these melanomas were recurrent and one was associated with neurofibromatosis. The case report concerns a 75-year-old man with a nasal mucosal melanoma which was treated surgically. One year later, the patient developed a local recurrence and a cervical lymph node metastasis. Both the recurrent tumour and the metastasis showed clear evidence of bone and osteoid formation. CONCLUSIONS: This case is the first report in the literature, clearly demonstrating bone and osteoid formation by a mucosal melanoma, not only at the primary site, but even more convincingly in a cervical lymph node metastasis.

Value of resection of pulmonary metastases in head and neck cancer patients.

BACKGROUND: Literature shows no data about a complete cohort of head and neck cancer patients who developed pulmonary metastases. In this study, we investigate factors related to survival, with emphasis on the role of a pulmonary metastasectomy. METHODS: A retrospective review of 138 patients who developed pulmonary metastases (5.5% of all head and neck cancer patients) in the period 1978 to 1994 is presented. In a stepwise regression analysis (Cox), factors were identified related to survival. Also investigated was whether the prognostic value of potential prognosticators differed between the group that underwent metastasectomy and the group that did not. RESULTS: One hundred thirty-eight patients had metastases originating from head and neck cancer. The 5-year survival rate for all these patients was 13%. Younger patients (P = .011), patients with a longer disease-free interval (DFI) (P = .011), patients with a longer disease-free interval (DFI) (P = .016), and patients with a nonsquamous cell carcinoma (P = .038) did better. No evidence of a relationship between survival and sex or survival and number of metastases was found. Twenty-one patients underwent surgical resection of their pulmonary metastases. In 18 patients the resection was complete. The 5-year survival rate for patients who underwent a metastasectomy was 59%, compared with 4% in the nonmetastasectomy group (P = .0033). CONCLUSION: Isolated pulmonary metastases from head and neck cancer are potentially curable by surgical resection. Preconditions for this approach are locoregional control of the primary lesion and technical resectability of the pulmonary metastases. Patients with a long interval between primary treatment and the diagnosis of pulmonary metastases may benefit more from resection therapy.

Survival of adult patients with head and neck soft tissue sarcomas.

A retrospective review and survival analysis is presented of 58 adult patients referred between 1963 and 1993 for treatment of a head and neck soft tissue sarcoma. The 5 year overall and disease-free survival was 60% and 52% respectively. Univariate analysis indicated that age, positive margins, bone invasion, distant metastases, single modality treatment, histological grading, sites below the deep fascia and size might be predictors of survival. On multivariate analysis, age, histological grade, distant metastases, site and referral for recurrent disease emerged as prognostic factors known before treatment. By including variables known after treatment, age remained the most important independent prognostic factor followed by the presence of free margins, histological grading and distant metastases at presentation.

Fibromatosis of the head and neck.

Fibromatosis is a locally infiltrative fibrous tissue proliferation with a tendency to recur locally. From a large series of head and neck patients treated between 1977 and 1994 in our institute, we retrieved the records of nine adult patients diagnosed with this disease. They serve as examples to demonstrate this rare entity in the head and neck. Five out of nine lesions were localized in level V (posterior triangle of the neck). The majority of patients were treated by surgery in combination with radiotherapy. None of the patients died of the disease.

Soft tissue sarcomas of the head and neck associated with surgical trauma.

Soft tissue sarcomas in the head and neck are rare. Aetiological factors relating to these tumours have not yet been identified. The association with von Recklinghausen's disease and with irradiation is however well recognized. In the literature it has been speculated that trauma may also play a role in the development of soft tissue sarcomas. In this article we present five patients with a history of surgical trauma at the site where a sarcoma later developed. Although we cannot prove a causal relationship, the relatively high incidence of possibly trauma-related soft tissue sarcomas in a series of 60 patients we have seen over a 30-year period, suggests that such a relationship could exist.

Carcinoma arising in thyroglossal remnants.

Three patients with a papillary carcinoma arising in a thyroglossal duct cyst are presented and the literature is reviewed. This rare malignancy is seen mostly in women between the ages of 20 and 50 years. The distribution of carcinoma subtypes differs from that of thyroid carcinomas and thyroglossal duct carcinoma is recognized as a primary tumour. The diagnosis is seldom made pre-operatively though especially in older patients with midline swellings in the neck the diagnosis should be considered. Sistrunk's operation is the treatment of choice. In this operation the cyst, the middle part of the hyoid bone and the thyroglossal duct are removed in continuity.

Merkel cell carcinoma of the head and neck.

Merkel cell carcinoma is a rare cutaneous tumour that typically arises in the head and neck area of elderly patients. The tumour often follows an aggressive course with frequent local recurrences and (regional) metastases, especially when localized above the clavicles. Five patients with a Merkel cell carcinoma of the head and neck, treated in our institute since 1984, are presented, illustrating the need for radical initial treatment consisting of surgery and radiotherapy, and showing how rapidly progressive the disease can be. Four of the five patients were cured of disease by a combination of surgery and radiotherapy.

Intratubular germ cell neoplasia of the testis: identification by placental alkaline phosphatase immunostaining and argyrophilic nucleolar organizer region quantification.

We assessed the value of placental alkaline phosphatase (PLAP) immunostaining and argyrophilic nucleolar organizer region (AgNOR) quantification as techniques for the identification of intratubular germ cell neoplasia (ITGCN), and compared them with hematoxylin-eosin and periodic acid-Schiff staining. We examined 46 malignant testicular germ cell tumors for the presence of ITGCN; 43 had sufficient tubules available for assessment. We also examined 16 cryptorchid testes, 16 testicular biopsies from 10 subfertile men, and 12 normal adult intrascrotal testes. In tubules adjacent to invasive tumors, hematoxylin-eosin staining identified 30 cases (70%) of ITGCN, while PLAP and AgNOR staining identified 36 cases (84%). All the seminomas (18) and 22 of 28 nonseminomatous germ cell tumors were PLAP-positive and had high AgNOR counts. Intratubular germ cell neoplasia was not identified in the other groups examined; germ cells in these groups were PLAP-negative and had low AgNOR counts. Cells of ITGCN showed cytoplasmic block positivity with periodic acid-Schiff staining but this was not a consistent finding. We conclude that ITGCN is present adjacent to most invasive germ cell tumors, and is reliably identified by hematoxylin-eosin staining when fully developed. Periodic acid-Schiff staining was not helpful as normal spermatogonia were also positive. Staining with PLAP and AgNOR were useful diagnostic adjuncts, but results with PLAP were easier to interpret.

Are AgNORs useful in distinguishing follicular hyperplasia from follicular lymphoma?

Argyrophilic nucleolar organiser regions (AgNORs) in 28 follicular lymphomas and 30 lymph nodes showing reactive follicular hyperplasia were studied to see if they were helpful in distinguishing follicular lymphoma from follicular hyperplasia in paraffin wax tissue sections. Mean nuclear counts were greater in follicular hyperplasia (3.71 v 3.11). This difference was significant but counts overlapped so much that they were of no practical value in distinguishing between both conditions. Higher counts may reflect greater proliferative activity in follicular hyperplasia.

Breast carcinoma kinetics. Argyrophilic nucleolar organizer region counts correlate with Ki67 scores.

This study assessed the value of argyrophilic nucleolar organizer region (AgNOR) staining as a potential technique for the estimation of cell kinetics in conventional histology sections, in benign and malignant breast lesions. Using a silver staining technique and immunohistochemistry, the authors correlated the numbers of argyrophilic nucleolar organizer regions (AgNORs) and Ki67 scores in 70 breast carcinomas and 27 benign breast lesions. Epithelial cells in fibrocystic disease and fibroadenomas contained a mean of 2.65-6.8 small uniform AgNORs per cell, whereas malignant cells contained 4.6-26.9 frequently highly irregular AgNORs. In benign tissue, Ki67 scores ranged from 0 to 4%; in malignant tumors, Ki67 scores ranged from 3.0 to 98%. The correlation between AgNOR counts and Ki67 scores was highly significant (P less than 0.001). The authors concluded that AgNOR counts performed on routine formalin-fixed paraffin sections furnish significant kinetic information. Furthermore, the difference in AgNOR counts between benign and malignant tumors is such that they may be of diagnostic value.