Elevated expression of the CC chemokine regulated on activation, normal T cell expressed and secreted (RANTES) in advanced breast carcinoma.

Breast carcinoma is the most common malignant disease among women and the second most lethal one. In search for a better understanding of the role of cellular mediators in the progression of this disease, we investigated the potential involvement of the CC chemokine Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES) in breast carcinoma progression. To this end, RANTES expression was determined in breast tumor cell lines and in sections of breast carcinomas, followed by analysis of the incidence and intensity of its expression in different stages of the disease. Our study reveals that high and physiologically relevant levels of RANTES are constitutively produced by T47D and MCF-7 breast tumor cell lines. Analysis of RANTES expression in sections of breast carcinomas demonstrates a high incidence of RANTES expression in epithelial tumor cells; the chemokine was expressed in 74% of the sections. RANTES expression was rarely detected in normal duct epithelial cells or in epithelial cells that constitute benign breast lumps, which were located in proximity to tumor cells. High incidence and intensity of RANTES expression were detected in sections of most of the patients with stage II and stage III of the disease (expression was detected in 83 and 83.3%, respectively), whereas RANTES was expressed at a lower incidence and intensity in sections of patients with stage I of breast carcinoma (55% of the cases). Most importantly, the expression of RANTES was minimally detected in sections of patients diagnosed with benign breast disorders and of women that underwent reduction mammoplasty (15.4% of the cases). These results indicate that the expression of RANTES is directly correlated with a more advanced stage of disease, suggesting that RANTES may be involved in breast cancer progression. Moreover, it is possible that in patients diagnosed with benign breast disorders, RANTES expression may be indicative of an ongoing, but as yet undetectable, malignant process.

Interferon-related mental deterioration and behavioral changes in patients with renal cell carcinoma.

Five out of 38 patients (13%) with metastatic renal cell carcinoma had mental deterioration 3 weeks to 13 months after the start of treatment with recombinant interferon alpha-C. Metastatic spread to the brain, paraneoplastic effect of the tumor on the central nervous system and other causes of dementia were excluded. Computed tomography of the brain in these patients was normal and the width of the cerebral sulci and ventricles did not correlate with the severity of dementia. Specific patterns of atrophy were not seen. General deterioration, assessed by the change in Karnofsky performance status, was associated with dementia. The dementia may have been caused by a neurotoxic effect of interferon.

Serial serum MCA measurements in the follow-up of breast cancer patients.

Mucin-like carcinoma-associated antigen (MCA) was serially assayed in 58 women with histologically proven breast cancer after their treatment for primary disease. MCA sensitivity and specificity were 87.5% and 76.9%, respectively, and the positive predictive value 82.4%. 10 patients had elevated MCA and no evidence of disease (NED) during their follow-up, of whom 4 finally developed overt metastases. The 4 had a mean (S.D.) MCA value of 46.48 (18.26) U/ml during the lead time, versus 18.76 (2.69) U/ml in the other 6, who are still at high risk for developing overt metastases. Raised levels of MCA in patients with NED create a dilemma of "treat" versus "wait and see". Early treatment of patients with serially uprising MCA levels should be evaluated in a prospective randomised study to assess its ability to prevent or delay the development of overt metastatic disease and influence survival.

Fluorescence polarisation changes in lymphocyte cytoplasm as a diagnostic test for breast carcinoma.

Lymphocytic cytoplasm from individuals with malignant disease, and from those without, differ in such a way as to be diagnostic both of malignancy generally and of specific types of cancer. Mitogenic stimulation of lymphocytes by phytohaemagglutinin (PHA) and antigenic stimulation by encephalitogenic factor (EF) and certain specific tumour-associated antigens, provokes changes in the structure of the cytoplasmic matrix (SCM) which are detectable upon fluorescence polarisation. The degree of change is quantifiable both by calculating the polarisation ration (PR, polarisation before and after stimulation) and the relative ratio (RRSCM, the ratio between the polarisation obtained after exposure to EF [PEF] and to the polarisation measured after exposure to PHA [PPHA]). A new tumour-associated antigen specific for breast cancer, CaBr, was tested for its diagnostic efficacy in comparison with that of EF, by prospectively testing blood samples from 138 consecutive women with suspicious breast masses. The previously known discriminatory power (sensitivity 60.7% and specificity 90.7%) of the polarisation-derived RRSCM was reconfirmed. However, the RR'SCM (the new ratio using CaBr instead of EF), was significantly more sensitive (77.4%; P < 0.01) and specific (94.4%) than the RRSCM in detecting breast cancers. The polarisation changes in the cytoplasmic matrix after stimulation by CaBr alone suggest the best discriminatory power (sensitivity 90.5% and specificity 94.4%) between cancerous and non-cancerous patients.

Adaptation of the cellscan technique for the SCM test in breast cancer.

The value of the SCM (Structuredness of Cytoplasmic Matrix) cancer test, a procedure based on the detection of differences in lymphocyte activation in the presence and absence of cancer, has remained controversial, with inconsistent results having been reported among investigators. The Cellscan, a high-precision static cytometer system, has been designed to perform the SCM test; the apparatus facilitates the polarisation measurements and can examine cells which have been separated by simpler procedures than were originally described. In this study, using methods and diagnostic criteria adapted for the Cellscan system in a hospital environment, the SCM test correctly classified over 90% (76/80) of patients with breast cancer and differentiated over 90% (72/73) of individuals without cancer.

Chemoimmunotherapy of advanced malignant melanoma: sequential administration of subcutaneous interleukin-2 and interferon-alpha after intravenous dacarbazine and carboplatin or intravenous dacarbazine, cisplatin, carmustine and tamoxifen.

Both chemotherapy and interleukin-2 and/or interferon-alpha produce objective responses in a proportion of advanced malignant melanoma patients. While duration of response to chemotherapy is short, i.e. usually below 4 months, immunotherapy has resulted in a small number of long-lasting remissions in patients with metastatic melanoma. In two consecutive phase II trials in a total of 67 patients, we assessed the potential synergism between both modalities, i.e. chemo- and immunotherapy. Treatment consisted of intravenous (i.v.) carboplatin (CBDCA, 400 mg/m2) and dacarbazine (DTIC, 750 mg/m2) given twice (i.v. bolus over 30 min) at 3-week intervals, or 4 cycles of DTIC (220 mg/m2 i.v. 3 days), cisplatin (DDP, 35 mg/m2 i.v. 3 days), carmustine (BCNU, 150 mg/m2 i.v. cycles 1 and 3) and tamoxifen (TAM, 20 mg oral/daily) at 3-week intervals. Chemotherapy was followed by immunotherapy with combined subcutaneous (s.c.) interleukin-2 (rIL-2) and SC interferon-alpha 2 (rIFN-alpha). Among 40 patients who received a full cycle of chemotherapy with CBDCA/DTIC and sequential immunotherapy, there were 3 (7.5%) complete remissions (CRs) with a median duration of 19 months (range 13-26+). Partial remissions (PRs) were noted in 11 (27.5%) patients with a median response duration of 8 (range 5-14) months. Among 27 patients who received DTIC/DDP/BCNU/TAM and rIL-2/rIFN-alpha, there were 3 (11%) complete remissions and 12 (44.5%) partial remissions. Duration of complete and partial remissions ranged from 9+ to 13+ (median, 11+), and 5 to 15+ (median, 7+) months, respectively. Chemotherapy produced mostly moderate toxicity. Thrombocytopenia was common with the nadir after a median time of 18 days following start of CBDCA/DTIC and DTIC/DDP/BCNU, respectively. 10 patients required transfusion of thrombocytes. Nausea and vomiting due to chemotherapy were well tolerated using concomitant ondansetrone (8 mg i.v.). Immunotherapy was self-administered at home with mild to moderate side effects; malaise, fever, chills, nausea/vomiting, diarrhoea, anorexia and arthralgias were most frequent, but were spontaneously reversible after ending rIL-2/IFN-alpha. A mean 87 and 88% of the projected doses of rIL-2 and rIFN-alpha were administered on either protocol. There were no life-threatening complications and no treatment-related deaths. The sequential combination of chemotherapy and rIL-2 plus rIFN-alpha had at least additive therapeutic activity against metastatic malignant melanoma. The schedules produced long-lasting remissions and were tolerated well overall. These trials substantiate a potential role for low to intermediate dose immunotherapy in maintaining and consolidating therapeutic effects of chemotherapy in metastatic melanoma.

Adjuvant therapy for Dukes C adenocarcinoma of colon.

Forty-two patients with adenocarcinoma of the colon, who received surgery between 1975 and 1978 and were to found to have pericolonic fat infiltration and lymph node metastases, were analyzed for disease free period and overall survival. Twenty-one patients had received post-operative X ray therapy, and post-X ray therapy intravenous 5-Fluorouracil adjuvant therapy. Twenty-one patients, matched by age, sex, ethnic origin and site of disease were untreated. The 5 year survival rate for the treated group was 65% compared with 36% for the control group (P greater than 0.2). At 5 years 55% of the treated group were disease free but only 12% of the control group remained disease free (P = 0.04). The significance of this work needs to be established by a randomized and prospectively controlled clinical trial.

The production of human monoclonal anti-MMTV antibodies by in vitro immunization with anti-idiotypic antibodies.

We have previously reported the production of a series of human monoclonal antibodies reacting with mouse mammary tumor viral antigens as well as the human counterpart HuMTV antigen. Against two of these antibodies (B11 and 4.6/6), anti-idiotypic antibodies were generated, which appeared to be the internal image of viral antigen. Vaccination with the anti-idiotypic antibodies induced in mice humoral and cellular immunity against both MMTV and HuMTV. The current study describes a novel method to produce human anti-MMTV antibodies derived monoclonal antibodies. Normal peripheral blood lymphocytes (PBLs) were immunized in vitro in the presence of IL-2, with rabbit anti-B11 antibodies bound to silica beads. Following in vitro immunization, the lymphocytes were fused with the UC-792-6 human lymphoblastoid cell line. Four human hybridomas were found to secrete human monoclonal antibodies which bound to MMTV and HuMTV. Three of the secreted antibodies were of the IgG class and one of the IgM class. The specificity of binding was confirmed by direct and competitive ELISA assays and by radioimmunoprecipitation. Our study demonstrates the ability to generate human monoclonal anti-viral antibodies by in vitro immunization, employing "internal image" anti-idiotypic antibodies. The method can be used to generate human antibodies, especially against pathogenic agents or ill-defined antigens.

Combined postoperative radiotherapy and pcv-chemotherapy (ccnu, procarbazine and vincristine) in managing anaplastic astrocytoma - a report of a long-term follow-up.

Postoperative radiotherapy together with CCNU, procarbazine and vincristine (PCV-chemotherapy regimen) were analysed in patients with histologically confirmed anaplastic astrocytoma. This report evaluates the long-term survival correlated with important prognostic variables. Patients who received more than 6 courses of the PCV-chemotherapy had significantly longer survival (P=0.03). Age and performance status, identified as important prognostic factors in previous studies, are confirmed in this analysis.

Postoperative 5-fluorouracil plus levamisole combined with radiation therapy for high risk rectal cancer patients.

Adjuvant chemotherapy in rectal cancer patients is aimed at decreasing the relapse rate of the disease, increasing the disease-free and the overall survival of the patients. The combinations of radiation therapy and several drugs have been tested, but the efficacy of 5-fluorouracil plus levamisole in rectal cancer is not determined yet. Sixty-two consecutive Dukes' B2 or C rectal cancer patients were enrolled into a prospective phase II study. Within 4 to 6 weeks following en-block resection of the tumor and lymphatics, adjuvant chemotherapy had to be started. Combination of 5-FU 375 mg/m(2)/day was given intravenously over 15-20 min for 5 consecutive days, every 4 weeks for 1 year. Levamisole 50 mg t.i.d. was administered orally during the first 3 days of each course of chemotherapy. Radiation therapy included 50 Gy, given by a linear accelerator 8 MV, to a pelvic BOX field. Dose per fraction was 1.8-2.0 Gy daily, administered for 5 consecutive days a week. Follow-up time of the whole group ranges from 7 to 53 months (median 18). Failure rate was 32.3%. The most common site for first relapse was the liver followed by recurrence in the pelvis or the surgical bed. Age, sex, tumor grade and number of involved lymph nodes were not associated with rate nor with site of relapse. Fifty-five percent of the relapses were observed in patients during the year of the adjuvant treatment. Toxicity included diarrhea (33.9%), nausea and vomiting (19.4%), weakness and mucositis. The disease-free survival rate for a median follow-up of 18 months in our study is 75%. The 3.5 year disease-free survival is 67% for the whole group. Our phase II study results point to the feasibility and acceptable toxicity of post-operative combination of 5-FU plus low-dose levamisole and radiation therapy in rectal cancer patients.

Treatment of disease-negative but mucin-like carcinoma-associated antigen-positive breast cancer patients with tamoxifen: preliminary results of a prospective controlled randomized trial.

Increasing levels of tumor markers such as carcinoembryonic antigen, mucin-like carcinoma-associated antigen (MCA), CA 15.3, and monoclonal antibody H23 in breast cancer patients following the treatment of the primary disease and adjuvant radiation and chemotherapy reflect subclinical development of metastatic disease. Overt metastatic disease is usually incurable and prolongation of life at this stage is impossible, and the treatment is only palliative. The efficacy of tamoxifen, a least-toxic agent, in the treatment of early and minimal metastatic disease detected only by increasing serum levels of MCA was studied prospectively in a randomized study. Our preliminary, albeit encouraging, results showed that the rate of relapse within a median follow-up period of 11 months was 24.1% in the control arm as compared with 0% in the tamoxifen arm (Fisher's exact test, P = 0.012). None of the patients with a relapse had positive progesterone receptors (PR). We may carefully conclude that early treatment may be warranted in young patients with negative PR and continuously increasing serum levels of the marker.

Pharmacokinetics of recombinant interferon alpha-C.

Recombinant interferon alpha-C (rIFN alpha-C, Interpharm), is a new type of alpha-interferon that has a specific activity of 1-2 x 10(9) units/mg protein. The pharmacokinetics of rIFN alpha-C were studied in 11 patients with metastatic renal-cell carcinoma. A total of 10 million units IFN alpha-C were injected intramuscularly and the serum level of IFN was evaluated up to 72 h post-administration. Measurable IFN concentrations appeared in the serum as early as 0.5 h, and levels peaked at 4-6 h (Cmax = 53.2 +/- 4.6 units/ml). Relatively high levels persisted for 24 h and declined thereafter with an apparent half-life of 3-4 h. The mean area under the serum-concentration curve (AUC) was 1,259 +/- 145 units h ml-1, indicating good bioavailability of the preparation from the intramuscular injection.

Interferon-related cortical blindness.

The term cortical blindness indicates loss of sight due to bilateral lesions in the occipital lobes. It is a rare but severe side effect produced by chemotherapeutic agents. Cortical blindness was diagnosed in a 75-year-old man who had been treated with alpha-interferon for metastatic renal-cell carcinoma. The absence of focal neurological signs and of abnormal findings as determined by two repeated computed tomography (CT) scans of the brain, which excluded structural damage to the occipital lobes, suggest that metabolic or toxic reactions may have caused the cortical blindness diagnosed in our patient. The temporal relationship between the treatment with alpha-interferon and the development of cortical blindness indicates that this substance may have been the causative agent for this phenomenon.

Breast-cancer associated brachial plexopathy - still a diagnostic and treatment challenge.

Brachial plexopathy (BP) in breast cancer patients is a rare event, attributed mainly to radiation damage or tumor infiltration of the plexus. Differentiation between these etiologies is a diagnostic challenge. We have studied retrospectively eight female patients with breast cancer who developed a clinical syndrome of brachial plexopathy following the treatment of the primary disease, out of more than 900 during the last 10 years. None of the available ancillary tests such as plain films, CT or MRI studies, EMG or tumor markers, provided reliable data regarding the cause of the plexopathy. Biopsy, on the other hand, was not always feasible. In our series, all the patients who developed BP did not have any blood-borne metastases before developing the syndrome. In 3 of the patients BP was the first sign of recurrence. In the other 5, only local or locoregional relapse preceded. In 7 of the 8 patients the left side was affected. Treatment should be tailored in each case according to course of the disease. The optimal treatment has not yet been defined.

Radiofrequency and testicular cancer (review).

Physical agents such as ultraviolet or ionizing radiation, heat, and repetitive trauma have been related to the causation of cancer in humans. Much less clear is the association between exposure to radiofrequency, i.e. radar and microwave radiation, emitted from television screens, antennas and detection equipment, to the development of cancer. Sporadic case reports and small series suggest that this type of radiation might lead to cancer or contribute to its evolution. The association between radiofrequency and testicular damage and cancer is unproved, but clinical and experimental data are suggestive of such possibility. We have recently encountered a case of secondary severe oligospermia, followed by seminoma, most probably induced by exposure to radar and microwave radiation, in an 18-year-old man. The population handling or repairing dangerous radar facilities is too small to be evaluated by epidemiological surveys. Even it were so, this material is handled in secrecy, either military or industrial. Arousing the alertness of the medical team in those facilities and bringing better protection to the employees is the aim of our report.

Adjuvant levamisole and fluorouracil in high risk colorectal cancer patients.

Adjuvant chemotherapy in colorectal cancer patients is aimed at decreasing the relapse rate of the disease and increasing the disease-free and the overall survival of the patients. In a prospective study we evaluated the efficacy of 5-FU plus levamisole as an aduvant therapy for 153 patients with Dukes' B-2 or C colon or rectal cancer following a curative-intended surgery. Adjuvant chemotherapy was started within 4 to 6 weeks following the operation. Combination of 5-FU 375 mg/m(2)/day was given intravenously over 15-20 min for 5 consecutive days, every month for 1 year. Levamisole 50 mg t.i.d. was administered orally during the first 3 days of each course of chemotherapy. Rectal cancer patients were also irradiated to the tumor bed and pelvic lymphatics. The dose intensities (DI) of 5-FU and levamisole in our study were 432.6 mg/m(2)/w and 103.8 mg/m(2)/w, respectively. Failure analysis in Dukes' B and C patients showed that the rectum accounted for 47.5% of the relapses, of which only 3 cases were in the vicinity of the resected area. Almost half of the failures were observed within the year of adjuvant treatment. The liver was the most common site for first relapse (50%). The 3-year disease-free survival of Dukes' B-2 patients group was 84%, compared with 64% in Dukes' C. The main toxic manifestations were diarrhea, nausea and vomiting, weakness and mucositis. No dose reduction was needed. Our protocol, using lower DI of levamisole yielded similar results with a lower rate of toxicity than other common protocols.

Postoperative irradiation combined with chemotherapy for high-risk rectal cancer patients.

In a prospective open-study we evaluated the combination of radiation therapy and adjuvant 5-FU plus levamisole in controlling Modified Astler-Coller (MAC) B2 or C rectal cancer following a curative-intended surgery. Sixty-four consecutive rectal cancer patients were treated by adjuvant radiation therapy (Linac 8 MV, 50-50.4 Gy to an isocentric pelvis brick volume in 5 fractions per week each of 1.8-2 Gy), and 12 monthly courses of 5-fluorouracil (375 mg/m(2)/day for 5 consecutive days) plus levamisole (50 mg t.i.d. for 3 days). Within a median follow-up period of 36 months, 19 patients (29.6%) experienced relapses. The 3-year-DFS of MAC B2 patients was 82%, compared with 60% in MAC C. Early radiation treatment was not associated with a higher proportion of relapses, while late radiation therapy initiation was associated with a significantly higher proportion of relapses. Early radiation therapy, not later than following the 2nd to 4th course of chemotherapy, is associated with the more acceptable proportion of relapses.

Fluorescence polarization changes in the lymphocytic cytoplasm in the various stages of breast cancer.

Post-surgical pathological staging and ancillary tests determine the initial treatment of breast cancer. Because change in the structuredness of the cytoplasmic matrix (SCM) of peripheral lymphocytes (as assessed by measurement of fluorescein fluorescence polarization, FFP) has already emerged as diagnostic for breast cancer and an aid to staging in other cancers, testing was carried out in 113 pre-surgical patients to see whether such change could contribute to accurate staging of breast cancer. The FFP test was able to distinguish grouped stages of locoregional disease from metastatic disease but was unable to distinguish between any single locoregional stage and the metastatic stage. Technological improvements now underway may create a role for assessment of SCM changes in breast cancer staging.

Potential for anatomic localization in colorectal cancer using the SCM test.

We analysed a series of 81 colorectal cancer cases in which the SCM (structuredness of the cytoplasmic matrix) test had already been performed with a diagnostic sensitivity of 92% and a specificity of 92.6%, yielding positive and negative predictive values of 96.3% and 84.7% respectively. We subdivided this group of 81 patients by anatomic location of the malignancy. Although the resultant subgroups were admittedly small, we noted a tendency for the most prominent changes in observed and calculated polarization parameters to be associated with cecal cancers. This finding was of special interest because the cecum is the most inaccessible site for colonoscopy. Ongoing site-specific surveillance in SCM-tested cases of colorectal cancer is necessary to validate this result.

Tamoxifen for disease-negative but MCA-positive breast cancer patients.

Increased levels of mucin-like carcinoma-associated antigen (MCA) in breast cancer patients with no evidence of disease following the treatment of the primary disease created a dilemma of 'to treat' or 'wait and see'. One might assume that early treatment of clinically undetectable disease on the basis of an elevated serum level of a sensitive and reliable tumor marker, may improve the treatment results, and even prolong the patient's survival. 'Wait and see' on acceptance of the notion that even early metastatic disease, still manifested only by uprising MCA levels, is incurable, and treatment should be kept in reserve for palliation of symptomatic disease. Sixty-one breast cancer patients with increasing MCA levels but without evidence of metastatic disease were randomized for tamoxifen 20 mg b.i.d. or to follow-up till relapse. The results for a median follow-up period of one year were encouraging. The non-treated patients experienced a significantly higher relapse rate (24.1%) than the tamoxifen-treated subjects (0%; p=0.012). The results for a median follow-up of 5 years were disappointing. The overall relapse rate was 22.2%. The relapse rate among the control patients was 25.8% while in the treatment arm it was 17.4% (p=0.46). The event-free survival and the pattern of relapse were similar in both arms. Tamoxifen may therefore be reserved for overt metastases, and not wasted on asymptomatic subclinical disease. It seems that there is no yield in terms of event-free survival for MCA measurements in breast cancer patients during the 5-year follow-up period.