Electrochemotherapy in treatment of tumours.

AIM: Electrochemotherapy is a local drug delivery approach aimed at treatment with palliative intent of cutaneous and subcutaneous tumour nodules of different histologies. Electrochemotherapy, via cell membrane permeabilising electric pulses, potentiates the cytotoxicity of non-permeant or poorly permeant anticancer drugs with high intrinsic cytotoxicity, such as bleomycin or cisplatin, at the site of electric pulse application. METHODS: An overview of preclinical and clinical studies is presented, and the treatment procedure is further critically evaluated. RESULTS: In clinical studies electrochemotherapy has proved to be a highly efficient and safe approach for treating cutaneous and subcutaneous tumour nodules. The treatment response for various tumours (predominantly melanoma) was approximately 75% complete and 10% partial response of the treated nodules. CONCLUSIONS: Electrochemotherapy is a new, clinically acknowledged method for the treatment of cutaneous and subcutaneous tumours. Its advantages are high effectiveness on tumours with different histologies, simple application, minimal side effects and the possibility of effective repetitive treatment.

Genetic susceptibility to environmental carcinogenesis in Slovenian melanoma patients.

The skin acts as the first defence barrier against external environmental pollutants, including chemicals and UV radiation. Cytochrome P450 CYP1A1 and glutathione S-transferases (GSTs) found in melanocytes and skin basal layers were shown to participate both in the metabolism of xenobiotics and in detoxification of reactive oxygen species (ROS). In our study we analysed the distribution of single and combined CYP1A1, GSTM1, GSTT1 and GSTP1 genotypes contributing to inter-individual differences in metabolism of xenobiotics and ROS in 125 Slovenian healthy individuals and in 140 patients with sporadic malignant melanoma. Our results showed no statistically significant differences between melanoma patients and healthy controls in the frequency of polymorphic CYP1A1 and GST genotypes. The risk of developing melanoma was not significantly increased in individuals homo- or heterozygous for the CYP1A1*2A allele combined with GSTM1*0 genotype (OR: 1.86; 95% CI: 0.36-7.71), but increased slightly in carriers of CYP1A1*2A combined with both GSTM1*0 and GSTT1*0 genotypes (OR: 3.42; 95% CI: 0.36-29.6). Our results indicate that factors other than the polymorphic genes coding xenobiotic metabolising enzymes play a major role in protection against environmental carcinogenesis in human skin.

The circulating auto-antibodies to p53 protein in the follow-up of lymphoma patients.

Mutant p53 proteins may be targets of the host immune system - consequently a certain proportion of cancer patients (the percentage varies according to the type of cancer) with tumors that carry p53 missense mutations develop circulating p53 antibodies. The present study was aimed at defining the occurrence of circulating antibodies to p53 protein in patients with various types of non-Hodgkin's lymphomas (NHL). Altogether, the sera of 108 cases with various histological types of NHL and of 20 healthy controls were assessed for the presence of antibodies to p53 protein with an ELISA method. In 73 cases of NHL, also the immunohistochemical staining for p53 antigen was performed to make a rough estimation of the frequency of mutational events. The development of autoantibodies to p53 protein was observed in approximately 7% of NHL patients (predominantly in the more aggressive variants of the disease, but also in one case of small lymphocytic lymphoma). This proportion represents roughly one third of the number of patients assessed (immunohistochemically) to carry a missense p53 mutation in their tumors. The autoantibodies to p53 protein can be used as a tumor marker (early appearance, high specificity) in the follow-up of a subset of NHL patients, but, unfortunately, this subset comprises only approximately 7% of NHL patients.

Electrochemotherapy with cisplatin: the systemic antitumour effectiveness of cisplatin can be potentiated locally by the application of electric pulses in the treatment of malignant melanoma skin metastases.

The application of electric pulses to skin tumour nodules enhances the antitumour effectiveness of cisplatin. This treatment approach, known as electrochemotherapy, was employed in the treatment of skin metastases and lymph node metastases in malignant melanoma patients. Electric pulses were applied to tumour nodules in order to potentiate locally the antitumour effectiveness of systemic cisplatin-based chemoimmunotherapy. The study included nine malignant melanoma patients with skin metastases and metastases in lymph nodes not amenable to surgery, undergoing cisplatin-based chemoimmunotherapy. The antitumour effectiveness of the chemoimmunotherapy was compared with the antitumour effectiveness of electrochemotherapy, i.e. application of electric pulses to tumour nodules together with cisplatin-based chemoimmunotherapy. Application of electric pulses to the 27 skin tumour nodules potentiated locally the antitumour effectiveness of cisplatin. Four weeks after the treatment, 48% of the tumour nodules had an objective response (OR), compared with 22% of the 18 tumour nodules treated with cisplatin-based chemoimmunotherapy alone. Furthermore, the median time to progression was longer in the electrochemotherapy-treated nodules (21 weeks) than in the chemoimmunotherapy-treated nodules (4 weeks). This study shows that application of electric pulses to malignant melanoma tumour nodules can potentiate the antitumour effectiveness of cisplatin in patients undergoing systemic cisplatin-based chemoimmunotherapy. Therefore, electrochemotherapy may be used as an adjunct to systemic ongoing cisplatin treatment, predominantly in patients in whom antitumour effectiveness needs to be potentiated locally.

Cathepsin D expression in early cutaneous malignant melanoma.

BACKGROUND: The aspartic proteinase cathepsin D is believed to be associated with proteolytic processes leading to the invasion and seeding of tumor cells. An association between cathepsin D tissue concentration and aggressiveness of tumors has been detected in different cancer types, as well as in metastatic melanoma. METHODS: The concentration of cathepsin D was measured immunoradiometrically (ELSA-CATH-D kit, CIS Bio International) in the cytosols of 51 primary cutaneous melanomas (with Breslow index < 4 mm) to estimate the tissue concentrations of cathepsin D in early cutaneous melanoma. RESULTS: A significantly elevated concentration of cathepsin D was measured in the tumor cytosols as compared to adjacent normal tissue (44.2 vs. 14.7 pmol/mg of total protein, P < 0.001). CONCLUSIONS: Our results indicate that cathepsin D is expressed at high levels by melanoma cells. The extremely high expression of cathepsin D in two of our patients, with later progression of the disease over a 42-month follow-up period, suggests a possible correlation between the cathepsin D tissue concentration and the prognosis of primary cutaneous malignant melanoma.

Serum values of tumour necrosis factor-alpha and of soluble tumour necrosis factor-R55 in melanoma patients.

Tumour necrosis factor-alpha (TNFalpha) is a cytokine with a variety of biological activities, including an effect on tumour growth. The soluble TNF receptor TNF-R55 (sTNF-R55) inhibits TNFalpha functioning. Serum values of TNFalpha and TNF-R55 have been observed in patients with different cancers. To determine the serum values of TNFalpha and its soluble receptors and to investigate the prognostic value of sTNF-R55, we studied the sera of 68 patients with metastatic melanoma, 109 patients with no recurrent disease after surgical removal of melanoma, and 69 healthy controls. At least four different monoclonal antibodies against human TNFalpha and human TNF-R55, respectively, were prepared to obtain sensitive and reliable sandwich enzyme-linked immunosorbent assays. We found that in patients with metastatic melanoma the serum values of sTNF-R55 were significantly higher (2.41 ng/ml; range 0.02 23.0 ng/ml; P < 0.05) than in the melanoma patients without recurrence (0.54 ng/ml; range 0.02-6.25 ng/ml) and healthy controls (0.5 ng/ml; range 0.02 5.0 ng/ml). The sTNF-R55 concentrations increased as the disease progressed. Patients with metastatic melanoma also had significantly higher concentrations of TNFalpha (3.34 ng/ml; range 0.03-30.0 ng/ml; P<0.05) than patients without recurrence (1.24 ng/ml; range 0.02 23.0 ng/ml). Patients with metastatic melanoma, a high sTNF-R55 concentration, a low TNFalpha concentration and a low TNF:sTNF-R55 ratio had the worst prognosis. Low values of sTNF-R55 (<0.6 ng/ml) were associated with favourable response to chemotherapy (P = 0.007). According to our findings, patients with metastatic melanoma have higher values of sTNF-R55 than the controls and melanoma patients without recurrence. sTNF-R55 values higher than 0.6 ng/ml and a TNF:sTNF-R55 ratio lower than 1.5 are unfavourable prognostic factors for response to chemotherapy.

Electrochemotherapy with cisplatin: clinical experience in malignant melanoma patients.

Electrochemotherapy consists of chemotherapy followed by local application of electric pulses to the tumor to increase drug delivery into the cells. The aim of this Phase II clinical study was to evaluate the antitumor effectiveness of electrochemotherapy using intratumoral cisplatin administration on cutaneous tumor nodules in malignant melanoma patients. In 10 patients, 133 tumor nodules of different sizes were treated: (a) 82 tumor nodules were treated with electrochemotherapy; (b) 27 tumor nodules were treated with cisplatin; (c) 2 tumor nodules were treated with electric pulses; and (d) 22 tumor nodules were untreated. Four weeks after therapy, 78% objective responses were obtained in the electrochemotherapy group, and 38% objective responses were obtained in the cisplatin group. Exposure of tumor nodules to electric pulses without cisplatin treatment had no effect on tumor growth. Electrochemotherapy was well tolerated by all patients, and a good cosmetic effect was obtained, with only minimal scarring and a slight depigmentation of the skin. At 124 weeks of follow-up, a 77% control rate of the tumor nodules treated by electrochemotherapy was observed, compared to 19% for those that were treated with cisplatin only (P < 0.0001). Our results clearly demonstrate that electrochemotherapy with cisplatin is a highly effective approach for treatment of cutaneous malignant melanoma nodules. The advantages of this therapy include its simplicity, the short duration of treatment sessions, low cisplatin doses, and insignificant side effects, as well as the fact that it can be done on an outpatient basis.

Genetic polymorphism of xenobiotic metabolising enzymes in Slovenian lung cancer patients.

Most carcinogenic substances require metabolic activation in order to become ultimate carcinogens. Genetic polymorphism of xenobiotic metabolising enzymes cytochromes P450 may therefore influence human cancer susceptibility. The aim of our study was to investigate if CYP1A1 gene polymorphism contributes to lung cancer susceptibility in Slovenian patients. Two polymorphic sites in CYP1A1 gene were analysed in DNA samples from 100 healthy controls and 199 lung cancer patients using genotyping approach. Our results indicate that CYP1A1 may be one of the factors determining susceptibility to squamous cell carcinoma of lung in Slovenian population. However the frequency of CYP1A1 polymorphisms is too low to be a potentially useful marker of increased lung cancer risk.

Electrochemotherapy with cisplatin: potentiation of local cisplatin antitumour effectiveness by application of electric pulses in cancer patients.

This study was aimed at assessing the response to electrochemotherapy with cisplatin of cutaneous tumour nodules in patients with malignant melanoma, squamous cell carcinoma and basal cell carcinoma. In 4 patients, 30 tumour nodules of different sizes were treated; five without treatment, one with electric pulses, five with cisplatin injected intratumorally and 19 with electrochemotherapy, i.e. intratumoral administration of cisplatin followed by delivery of electric pulses to the tumour nodule. After 4 weeks, a complete response (CR) in all 19 electrochemotherapy treated nodules was obtained. All electrochemotherapy treated nodules remained in CR (range 7-11 months), regardless of histological type, except for the metastasis of a squamous cell carcinoma that progressed after 9 months. CR was also obtained in two of five tumour nodules treated with cisplatin intratumorally, but the other three nodules progressed within 3-7 months. Exposure of the tumour nodule to electric pulses without cisplatin treatment had no effect on tumour growth. Electrochemotherapy was well tolerated by all patients and a good cosmetic effect was obtained, with only minimal scarring and a slight depigmentation of the skin. Electrochemotherapy with cisplatin has proved to be effective in patients with cutaneous tumour nodules. Furthermore, electrochemotherapy is easy to perform and can be carried out on an out-patient basis.

Effective treatment of cutaneous and subcutaneous malignant tumours by electrochemotherapy.

Electrochemotherapy (ECT) enhances the effectiveness of chemotherapeutic agents by administering the drug in combination with short intense electric pulses. ECT is effective because electric pulses permeabilize tumour cell membranes and allow non-permeant drugs, such as bleomycin, to enter the cells. The aim of this study was to demonstrate the anti-tumour effectiveness of ECT with bleomycin on cutaneous and subcutaneous tumours. This article summarizes results obtained in independent clinical trials performed by five cancer centres. A total of 291 cutaneous or subcutaneous tumours of basal cell carcinoma (32), malignant melanoma (142), adenocarcinoma (30) and head and neck squamous cell carcinoma (87) were treated in 50 patients. Short and intense electric pulses were applied to tumours percutaneously after intravenous or intratumour administration of bleomycin. The tumours were measured and the response to the treatment evaluated 30 days after the treatment. Objective responses were obtained in 233 (85.3%) of the 273 evaluable tumours that were treated with ECT. Clinical complete responses were achieved in 154 (56.4%) tumours, and partial responses were observed in 79 (28.9%) tumours. The application of electric pulses to the patients was safe and well tolerated. An instantaneous contraction of the underlying muscles was noticed. Minimal adverse side-effects were observed. ECT was shown to be an effective local treatment. ECT was effective regardless of the histological type of the tumour. Therefore, ECT offers an approach to the treatment of cutaneous and subcutaneous tumours in patients with minimal adverse side-effects and with a high response rate.

Immunochemotherapy with recombinant interferon-alpha 2b plus dacarbazine in the treatment of advanced malignant melanoma.

In order to estimate the therapeutic activity and tolerability of immunochemotherapy with recombinant interferon-alpha 2b (rIFN-alpha 2b) plus dacarbazine (DTIC), a study was carried out in 61 patients with cytologically and/or histologically confirmed metastatic malignant melanoma. The treatment regimen was as follows: rIFN-alpha 2b 2 x 10(6) IU intramuscularly on days 1 to 4, and DTIC 800 mg/m2 intravenously on day 5, repeated at 3-week intervals until the progression of the disease or, in the case of a complete response, for up to 6 months. The overall response rate was 28%-12% complete response (CR) and 16% partial response (PR). The median response duration was 10.9 months (CR 11.5 months, PR 9.3 months; P > 0.05). Responses occurred in soft tissue and lung metastases only. The median times to treatment failure for responding and non-responding patients were 10.9 and 3 months, respectively (P < 0.0001), and the median survival durations were 16.5 and 5.8 months, respectively (P < 0.0001). The stratification of the patients into a low-risk group (World Health Organization performance status [WHO PS] < or = 1 and soft tissue or lung metastases) and a high-risk group (WHO PS = 2 or disease localization other than skin, lymph nodes or lung) showed a significant advantage for the first group with respect to the response rate, median time to treatment failure and survival duration. A flu-like syndrome was recorded in 72% of patients, nausea and vomiting in 34%, haematological toxicity in 26%, hepatic toxicity in 5%, and neurotoxicity in 5%. In view of the results obtained in our study and those reported in the literature, IFN plus DTIC immunochemotherapy represents a reasonable treatment option, particularly for patients with soft tissue and lung metastases.

Clinical evaluation of potential usefulness of CEA, CA 15-3, and MCA in follow-up of breast cancer patients.

The potential usefulness of MCA, CA 15-3 and CEA in monitoring of breast cancer patients was evaluated in 135 female patients with histologically confirmed breast cancer. The patients were classified into two groups as follows: group of patients with no evidence of disease, NED; and group of patients with progressive disease, PD. In total, 2106 measurements of CEA, CA 15-3, and MCA were performed using an enzyme immunoassay. Serum levels of all three markers in the NED group differed significantly from those of patients with PD. The observed differences in the sensitivity and specificity of CEA, CA 15-3, and MCA tests were not significant. The serum concentrations of a particular marker correlated well with the concentrations of the other two markers, except when CEA was correlated with MCA or CA 15-3 in NED group patients. The elevation of tumor markers preceded by some 7 months the clinical evidence of dissemination, and marker levels reflected at a high percentage the response to therapy in PD patients. Therefore, this clinical study confirmed that MCA, CA 15-3 and also CEA are suited to discriminate between disease and disease-free periods, and also validated the usefulness of markers for treatment response monitoring.

Prognostic value of cathepsins B, H, L, D and their endogenous inhibitors stefins A and B in head and neck carcinoma.

To estimate the prognostic value of cathepsins B, H, L, D and stefins A and B in head and neck carcinoma, their concentrations in cytosols of primary tumours and adjacent normal tissue were measured (cathepsins B, D stefins A, B in 45, cathepsin L in 24 and cathepsin H in 21 patients). Median concentrations of cathepsins B, L, and D were significantly higher in tumour than in the adjacent normal tissue (B and D: p < 0.0001; L: p = 0.004); cathepsin H concentration was higher in normal tissue (p = 0.001). Concentrations of either stefin did not differ significantly between normal and tumour tissue. Concentrations of cathepsins B, H, L, and D were higher in laryngeal than in non-laryngeal normal and tumour tissues. The difference was statistically significant for cathepsin B in tumour tissue (p = 0.045), and marginally significant in normal tissue (p = 0.07). Early tumours had lower concentrations of stefins A and B than locally advanced tumours (stefin A: p = 0.04; stefin B: p = 0.07). Disease-free and disease-specific survival rates were better in patients with concentrations of cathepsin L in tumour tissue below or equal to the cut-off values (p = 0.035; p = 0.05), whereas for cathepsin B the difference was established only for disease-free survival (p = 0.07). The opposite was true for stefin A (p = 0.0002; p = 0.002) and stefin B (p = 0.009; p = 0.003), and in disease-free survival also for cathepsin H (p = 0.055). The concentration of cathepsin D did not correlate with survival. Our data indicate that cathepsins B, H, L and stefins A and B might have prognostic value in head and neck carcinoma.

DTIC vs. IFN-alpha plus DTIC in the treatment of patients with metastatic malignant melanoma.

In our study we evaluated and compared the therapeutic success in 70 patients with cutaneous metastatic malignant melanoma (MM) treated at the Institute of Oncology in Ljubljana during the period 1985-1994. Twenty-nine patients received DTIC in a single 800 mg/m2 i.v. dose (Group 1) and 41 patients were receiving i.m. applications of IFN-alpha in 2 MU daily doses from days 1 to 4, completing the treatment with a DTIC application on day 5, given at the same dosage as in Group 1 (Group 2). The applications were repeated in three-week intervals until progression, or-in the case of a complete response-for up to 6 months. The rate and median duration of treatment response were higher in the group of patients treated by IFN-alpha plus DTIC (17% vs. 27%; 2.7 vs. 6.1 months), though the difference was not statistically significant. The survival of responders was either significantly higher (Group 2: p = 0.0007) or borderline-significantly higher (Group 1; p = 0.078) than that of non-responders. These patients also had significantly longer median survival (Group 1: 13.7 vs. 5.1 months, p = 0.019; Group 2: 19.3 vs. 4.9 months, p = 0.0003). The patients treated with IFN-alpha plus DTIC survived significantly better than those treated with DTIC alone (p = 0.043). There were no differences in the median duration of survival between both groups (6.6 vs. 6.7 months), and neither in the median duration of survival of responders (13.7 vs. 19.3 months) or non-responders (5.1 vs. 4.9 months) from both groups. The toxicity of combined therapy was higher than that of chemotherapy alone, though it was still moderate and acceptable. In view of our results, the addition of IFN-alpha to DTIC has shown an advantage over DTIC along.

Human CYP2D6 gene polymorphism in Slovene cancer patients and healthy controls.

The polymorphic CYP2D6 gene encoding debrisoquine hydroxylase has attracted much interest for its possible role in human pulmonary carcinogenesis. The purpose of this work was to determine the frequency of poor metabolizers (PM) and extensive metabolizers (EM) of debrisoquine in Slovene population of healthy individuals (n = 107), lung cancer patients (200) and melanoma patients (121). Polymorphism of CYP2D6 gene was studied by genotyping based on PCR analysis of the intron 3 exon 4 junction containing G to A mutation and one base pair deletion in exon 5, which are responsible for approximately 95% of poor metabolizer phenotype in Caucasians. In the healthy Slovene population 62.5% of individuals were identified as extensive metabolizers, 31% as extensive-heterozygous metabolizers and 6.5% as poor metabolizers of debrisoquine. The frequency of EM individuals was 70.5% in lung cancer patients and 64% in melanoma patients, the frequency of extensive-heterozygous subjects was 27% in lung cancer patients and 31% in melanoma patients. The frequency of PM individuals in the lung cancer patients was 2.5% and in melanoma patients 5%. The decrease in PM genotype in the group of Slovene lung cancer patients is similar to the decrease published for some other ethnic groups. Our results support the hypothesis that polymorphic CYP2D6 gene probably plays some, though not a prevalent role in chemical carcinogenesis. Poor metabolizer individuals appear to be less susceptible to lung cancer than EM individuals.

Radiotherapy, combined with simultaneous chemotherapy with mitomycin C and bleomycin for inoperable head and neck cancer--preliminary report.

PURPOSE: Prospectively designed randomized clinical study was undertaken to assess the efficacy of simultaneous application of irradiation, Mitomycin C, and Bleomycin in treatment of patients with inoperable head and neck carcinoma. METHODS AND MATERIALS: Between March 1991 and October 1993, 49 patients with inoperable head and neck carcinoma were randomly assigned to receive either radiation therapy alone (group A) or radiotherapy combined with simultaneous application of Mitomycin C and Bleomycin (group B). Patients in both groups were irradiated five times weekly with 2 Gy to the total dose of 66-70 Gy. Chemotherapy regimen included intramuscular application of Bleomycin 5 units twice a week, with the planned dose being 70 units and Mitomycin C 15 mg/m2 applied intravenously after delivery of 9-10 Gy of irradiation. The application of Mitomycin C was planned to be repeated on last day of radiotherapy in the dose of 10 mg/m2. In attempt to enhance the effect of chemotherapeutic drugs, patients in group B received also Nicotinamide, Chlorpromazine, and Dicoumarol. RESULTS: The difference in complete response rate between both treatment groups (24% in group A and 63% in group B) was statistically significant (p = 0.015). The difference in response rate was much more pronounced in patients with oropharyngeal carcinoma only (18% in group A compared to 81% in group B; p = 0.0003), while for all other subgroups added together, there was observed no benefit of multidrug therapy. Median follow-up was 18 months. Disease-free survival of patients in group A (9%) was significantly lower then in group B (48%) (p = 0.001). The difference between both treatment groups was even greater in patients with oropharyngeal carcinoma only: disease-free survival of these patients in group B was 66%, while in group A, all recurred (p = 0.00001). CONCLUSION: From results of our prospective randomized study it seems that the group of patients that received multidrug treatment with Mytomycin C, Bleomycin, Nicotinamide, Chlorpromazine, and Dicoumarol as enhancers of radiotherapy fared better than patients treated by radiotherapy alone.

In vitro monocyte maturation in patients with malignant melanoma and colorectal cancer--clinical significance.

In vitro monocyte maturation was studied in 52 malignant melanoma patients, 15 patients with colorectal cancer and 44 healthy donors. Index of maturation (IM) was in malignant melanoma patients 6.3 +/- 5.1%, in colorectal cancer 12.7 +/- 9.6%, and in healthy donors 40.4 +/- 18.0%. The difference between mean values in malignant melanoma patients and patients with colorectal cancer was significant when compared with healthy donors (p less than 0.001). The values in malignant melanoma patients decreased in accordance with stage of the disease and the difference between Stage I and Stage IV was significant (8.2 +/- 4.0% vs. 2.8 +/- 2.0%, p less than 0.01). In patients with colorectal cancer significant difference was established between operated patients and inoperable cases (21.4 +/- 10.0 vs. 5.1 +/- 1.9%, p less than 0.01). Monocyte maturation was influenced by the success of treatment. In patients with complete response the values were significantly higher than in patients with progression (11.7 +/- 6.7% vs. 2.2 +/- 1.5%, p less than 0.001). Autologous serum seems to inhibit the maturation process in vitro in cancer patients, while it apparently has no influence in healthy donors. When a response to treatment was achieved the previously low values of IM increased, while at progression a decrease was noted. The correlation between in vitro monocyte maturation and clinical factors leads to conclusion that the in vitro maturation reflects the in vivo process and may prove useful as a marker of tumor load or spread and may be a sensitive monitor of the treatment effect.

Combined BCG and irradiation treatment of skin metastases originating from malignant melanoma.

Treatment with BCG (Bacillus Calmette-Guérin) followed by irradiation was attempted to improve response to therapy by cutaneous metastases from malignant skin melanomas. Both agents were applied in low doses, known to cause minimal side effects. Nineteen patients, divided into three groups, entered the clinical trial. The first group consisted of five patients with relatively large metastases that usually appeared as a residual disease in the surgically treated area. Five patients with numerous, large metastases were included in the third group. The treatment sequence consisted of applying BCG intralesionally in doses from 4 x 10(5) to 1.17 x 10(7) viable units. After a free interval, the affected area was irradiated with doses from 1500--2500 ret. Patients with numerous small metastases and those with a small number of larger metastases, i.e., patients of the first and second group, showed a complete response and in these cases regression affected all the noninjected nodules and was also effective when regression could not have been achieved by BCG alone.

Chemotherapy of disseminated malignant melanoma with imidazol carboxamide (DTIC) and cyclophosphamide. Preliminary results.

Forty three patients with disseminated malignant melanoma were treated by combination chemotherapy with DTIC (800 mg/m2) and subsequent peroral cyclophosphamide (100 mg/m2) for 14 days. Overall response was observed in 18/43 (42%), and complete response in 9/43 (21%) cases. Complete response was achieved mainly in patients with metastatic involvement of lungs. Mean survival from onset of treatment for responding patients was significantly longer than mean survival of nonresponders (15 versus 5 months, respectively). Serious complications were not noted.

Serum immunoglobulin levels and survival rates in bronchogenic carcinoma patients.

Serum immunoglobulin IgG, IgA and IgM levels were estimated in 112 patients with bronchogenic carcinoma, and the obtained values were as follows: IgG = 1762 +/- 595 mg/100 ml; IgA = 332 +/- 104 mg/100 ml; IgM = 157 +/- 76 mg/100 ml. Subsequently the patients were divided in a group of 74 patients with epidermoid carcinoma, and a group of 38 patients with small cell anaplastic carcinoma of the bronchus. In patients with epidermoid carcinoma, in cases with disseminated disease the mean survival was shorter (7.9 months) when compared to the mean survival of patients with localized disease (20.8 months). Similarly, a depression of lymphocyte counts was observed in cases with disseminated disease. In cases with IgA concentrations ranging from 300 to 410 mg/100 ml longer survival rates were observed. In patients with small cell anaplastic carcinoma variety, the survival rates in patients with localized disease were almost identical with those in patients with disseminated disease (11.4 months versus 9.9 months). The longest survival rates were observed in cases with IgG concentrations ranging from 1600 to 2100 mg/100 ml, and IgM concentrations ranging from 190 to 320 mg/100 ml of serum.