Preclinical studies on detection of circulating melanoma cells in patients: telomerase as a recognition marker of malignancy.

Preclinical studies based on a "simulation design", were performed with cultured melanoma cells prelabeled with 51Cr, added to normal blood and subjected to separation and recognition steps. Mononuclear cells (MNC) were isolated on ficollhypaque gradient, and melanoma cells were separated from lymphocytes using anti-CD45 immunomagnetic beads. Malignant cells were then recognized by measuring telomerase activity (TRAP and TRAP-ELISA assays). It was found that: (a)recovery of prelabeled cells present in MNC did not exceed 75%; (b) further recovery of prelabeled cells after separation from lymphocytes did not exceed 68%. Therefore, the overall recovery of prelabeled cells did not exceed 48%; (c) the entire procedure was able to reliably detect as few as 30 malignant cells added to normal blood, providing a telomerase signal significantly higher than that found in absence of melanoma cells. These results furnish the technical bases for developing a tumor detection assay in the blood of melanoma patients.

Adjuvant therapy of melanoma patients (stage II, III): a pilot immuno-toxicological study.

A pilot study was conducted to assess the tolerability and the effect on host immunity of a post-surgery adjuvant treatment of melanoma patients with an anti-angiogenic agent, Tamoxifen (TAM, 20 mg/die p.o., daily), combined with immunomodulating cytokines, i.e. recombinant interleukin-2 (IL-2, 4 MUI/m2 s.c., day 8,10,12) and alpha-2b-interferon (IFN, 3 MUI/m2 i.m., day 15,17,19), starting a new cycle on day 21, for a total of 12 cycles. Fifty patients (pts) entered into the study, 27 males and 23 females with a median age of 55 years (range 25-75), performance status (ECOG) 0 with melanoma stage IIA (12 patients), stage IIB (28 patients), stage III (10 patients). Preliminary in vitro studies showed that TAM does not interfere with up-regulation of natural immunity induced by IFN, IL-2, or IFN + IL-2 in normal peripheral blood mononuclear cells (MNC). The clinical study indicates that the protocol was well tolerated. Increase of NK and LAK activity of patient MNC was observed on day 15. The mean disease-free interval was 10 months and 40 pts were alive at 5 years of follow-up. Further investigations should be performed to test effectiveness of this protocol in a randomized study.

Lymphatic mapping and sentinel node identification in squamous cell carcinoma and melanoma of the head and neck.

AIM: The aim of our study was to evaluate the role of scintigraphy in lymphatic mapping and in the identification of the sentinel lymph node (SLN) in patients with head and neck cancer. METHODS: Between September 1999 and February 2001 we enrolled 22 consecutive patients with cancer in the head and neck region: five squamous cell carcinomas, one Merkel cell tumor of the cheek, and 16 malignant melanomas. Lymphoscintigraphy was performed three hours before surgery after injection of 30-50 MBq of 99mTc -Nanocoll in 0.3 mL; the dose was fractionated by injecting the radiotracer at two points around the lesion. Static acquisition (anterior and/or lateral views, 512 x 512 matrix, 5 mins pre-set time) was started immediately after the injections so as to visualize the pathways of lymphatic drainage. The skin projection of the SLN was marked with ink. Intraoperative SLN detection was performed with perilesional injection of patent blue. RESULTS: SLNs were found with lymphoscintigraphy in all patients. Thirty-three SLNs were identified: one occipital node, three nodes at the base of the tongue, 10 superficial lateral nodes (external jugular), five submandibular nodes, five submental nodes, three mastoid nodes and six supraclavicular nodes. Biopsy was performed in 21/22 patients. In 20/22 patients the first lymph nodes were visualized in the proximal cranial regions (retroauricular, jugular and submandibular) at five minutes post injection. The SLN positivity rate was 13.6% (three patients). All patients with tumor-positive SLNs were submitted to radical dissection. Poor concordance in the detection of sentinel nodes was observed with patent blue. CONCLUSIONS: The flow of nanocolloid in the lymph vessels of the head is rapid. In our experience immediate scintigraphic imaging was essential to visualize the pathways of lymphatic drainage and the first SLN. Radioguided SLN biopsy is therefore recommended within three hours. Injection of patent blue is inadvisable because of the poor concordance with lymphoscintigraphy and the risk of permanent tattooing of the face.

Surgical treatment of intestinal complications after radiotherapy and surgery for malignant lymphoma and carcinoma of the testis.

The aim of this paper is the analysis of the complications occurred after surgical and radiological procedures, for staging and treatment of malignant lymphomas and testicular carcinoma with an evaluation of the possible efficacy of their surgical treatment. At the Istituto Nazionale dei Tumori of Milan during the period 1970-1976, 718 patients with malignant lymphomas were staged with laparosplenectomy; 444 of them received radiotherapy on the abdominal area. Laparosplenectomy was not performed on other 123 cases that were irradiated as well on the abdominal area. Retroperitoneal lymphadenectomy or explorative laparotomy were carried out in 98 patients with carcinoma of the testis and 46 of them received adjuvant radiotherapy; other 29 were irradiated without previous surgical procedures. After surgery alone morbidity and mortality rate were 2.7% and 0.73% respectively. The mortality/morbidity ratio was 26%. The incidence of complications after radiotherapy alone was 3.3%. No deaths followed the treatment. Combined treatment had a morbidity rate of 5.5% and mortality rate of 2%. The mortality/morbidity ratio was 37%. Complications were treated mainly by surgery: out of 50 interventions, 22 were intestinal resections. New secondary complications were however frequent (more than 50% after intestinal resection) especially in patients originally treated with laparosplenectomy or retroperitoneal dissection plus radiotherapy.

Axillary lymph node metastases from occult carcinoma of the breast.

From the case records of the Istituto Nazionale Tumori of Milan from 1956 to 1976, 49 cases of metastases in axillary lymph nodes from an occult carcinoma of the breast were selected. Forty-four patients underwent removal of the suspicious breast. In 11 patients no tumor was found at the pathologic examination. The survival of these patients was no different from the survival of those in whom the tumor was found. The 5-year survival of the total series was 87%, confirming that this type of presentation of breast cancer has a prognosis that is better than that generally reported for stage II carcinoma of the breast.