Janos Fruhling

Intravenous infusion of high doses of liposomes containing NSC 251635, a water-insoluble cytostatic agent. A pilot study with pharmacokinetic data.

In patients with resistant malignant tumors, we performed a pilot trial of intravenous infusion of a water-insoluble cytostatic agent, NSC 251635, entrapped in large volumes of liposomes made of egg yolk lecithin, cholesterol, and stearylamine (4:3:1). Forty liposome infusions were given to 14 patients in 38 courses. The volume of liposomes (20 mg of lipids/mL) varied from 205 to 1,000 mL or 124 to 617 mL/m2 of body surface, and amounts of NSC 251635 varied from 82 to 456 mg/m2. Three patients received repeated single courses. Liposomal therapy was very well tolerated. Side effects observed during some infusions were mild sedation, fever, chills, lumbar pain, urticarial rash, and bronchospasm. In all patients investigated, an important activation of the complement system was observed. No objective regression of the tumors was observed. The limiting factor in the phase I study was not toxicity but the volume of liposomes that could be prepared at once because of the long time required for its preparation. Pharmacokinetic data showed that maximal serum phospholipid and NSC 251635 concentrations were obtained at the end of the liposome infusion. The drugs peak was followed by a decreasing phase leading to a kind of plateau and a prolonged presence of the drug in the blood until 120 hours after its administration. Comparison of the pharmacokinetics of phospholipids and NSC 251635 suggests a rather rapid dissociation of the drug from the liposome.

Evaluation of squamous cell carcinoma antigen as a new marker for lung cancer

Carcinoembryonic antigen (CEA) is the only tumor marker of proven, although limited, value for the management of patients with non‐small cell lung cancer (NSCLC). The authors have prospectively assessed the potential value of a new tumor marker, squamous cell carcinoma antigen (SCC Ag), in a large series of patients with advanced lung cancer (LC). Squamous cell carcinoma antigen and CEA levels were measured in 382 healthy persons (N1 group), 90 patients with benign pulmonary diseases, and 291 patients with LC (129 with SCLC and 162 with NSCLC, including 96 with squamous LC). Carcinoembryonic antigen levels were higher in smokers than in nonsmokers, but smoking habits did not influence the serum concentrations of SCC Ag. Elevated values (above the 95th percentiles of N1, i.e., 7.5 ng/ml for CEA and 3.0 ng/ml for SCC Ag) were observed in 11.1% of patients with benign pulmonary diseases for both markers. Carcinoembryonic antigen was more sensitive than SCC Ag, even for squamous LC (56% versus 35% of elevated values, P < 0.01). The specificity toward squamous LC was better, however, for SCC Ag, for which levels were elevated in only 8.5% of SCLC and in 18% of other forms of NSCLC, compared with 49% and 55%, respectively, for CEA. Moreover, measurement of SCC Ag and CEA levels did not give redundant information: thus, in squamous LC the SCC Ag level was elevated in 32% of the patients with a normal CEA level, increasing from 57% to 71% the proportion of patients with at least one elevated marker. Lastly, elevation of CEA or SCC Ag levels was an adverse prognostic factor in squamous LC (P = 0.05 for CEA; P = 0.07 for SCC Ag). In conclusion, SCC Ag appears to be worthwhile of further investigation in squamous LC. The authors found that this new marker provided additional information on CEA and that it was more specific for squamous LC than CEA.

111Indium-oxine-labeled leukocytes in the diagnosis of localized infection in patients with neoplastic disease

One hundred twenty‐nine 111In‐oxine‐labeled leukocyte scintiscans have been performed in 117 patients with cancer in order to diagnose localized infectious disease. Of the 115 contributive scans, 40 were in patients with localizing signs, whereas in 75 fever of unknown origin constituted the indication for this examination. The overall specificity of the method was 95.4%, the overall sensitivity 86%, and the global accuracy 91.3%. In 10 cases with localizing signs, the 111In‐oxine granulocyte scintigram allowed exclusion of the diagnosis of infection, whereas in 17 instances without localizing signs, a focal infectious process was demonstrated. Heterologous donor leukocytes were used successfully in five instances. With the exception of accumulation of label at the site of an osteolytic metastasis in one case, no uptake was observed in primary or secondary tumors. It is concluded that 111In‐oxine‐labeled leukocytes constitute a valuable tool in the diagnosis and localization of infection in patients with malignant disease.

Internal mammary lymphoscintigraphy: Current status in the treatment of breast cancer

In the management of breast carcinomas, the internal mammary lymphoscintigraphy represents a simple, nontraumatic and reproducible technique to visualize and investigate the internal mammary nodes. As suggested by the total absence of visualization of nodes on the operated side in all or in the two upper intercostal spaces, internal mammary chain invasion concerns 30% of the population. This frequency increases with the clinical staging of the tumor, its size, and the extent of the disease in the axilla. It is higher for tumors of inner rather than of outer quadrants. When internal mammary lymphoscintigrams are compared in frontal view to the limits of the usual irradiation fields, parasternal nodes outside these limits or in borderline position are observed in 15 to 34% of the population according to the X-ray technique used. Furthermore, 40% of the tangential irradiation fields does not give an adequate irradiation dose to the internal mammary nodes. In one case out of four, IMLSc allows the reduction of useless heart- and lung-irradiated volumes. Internal mammary node invasion as demonstrated by lymphoscintigraphy has been demonstrated to have a prognostic value as the anatomopathological axillary node status, concerning both survival and disease evolution rates. IMLSc, when compared to the other possible investigation techniques of these nodes (X-ray computed tomography, echography or surgery, etc.), represents at the present time the investigation method of choice with the widest implications (diagnostic, therapeutic, and prognostic).

Observations cytogenétiques dans la maladie de Vaquez.

A cytological study has been made of the chromosomes of bone marrow cells obtained from 9 patients suffering from polycythemia vera and from a patient with essential thrombocythemia. The karyotypes were normal in 7 cases, as found in previous studies of these diseases. In 2 cases, however, there was clinical evidence of leukemic transformation and cytogenetic examination confirmed the diagnosis of chronic myeloid leukemia with Ph1 in the first case, and of acute leukemia with an aneuploid line of 44 chromosomes in the second case. The remaining case exhibited a modal chromosome count of 46 with chromosome lesions resembling those observed after irradiation in 15% of the cells and the Philadelphia chromosome in 10%. Further examination of this last case might enable one to determine the aetiological role of the Philadelphia chromosome in the induction of chronic myeloid leukemia.

Postoperative axillary lymphoscintigraphy in the management of breast cancer

Postoperative axillary lymphoscintigrams have been performed according to a technique of two-staged injection in 313 patients who had undergone a radical mastectomy with axillary dissection for breast cancer. Total absence of visualization of residual nodes suggests that dissection could be said to be complete in only 35.8% of the investigated patients, or because of metastatic disease causing blockage. The visualized residual nodes in patients represent a risk of developing nodal relapse, especially with positive pathological demonstration of axillary node invasion and without nodal irradiation. On the other hand, when the investigation showed the interruption of the lymphatic pathways of the arm, patients developed upper limb edemas more frequently. It is concluded that this simple and non traumatic technique is of value in order to assess the immediate postoperative axillary nodal status, and that it may influence the treatment of the patients.

Labelled polycyanoacrylate nanoparticles for human In Vivo use

Isobutyl and isohexyl cyanoacrylate nanoparticles are used as drug carriers, particularly for some anti-cancer drugs. Body distribution as well as pharmacokinetics have been well studied in animal and partially in man. Labelling of the monomer itself or of the carried drug with beta-emitters allowed such studies. In man, however, organ distribution and uptake could easily be done and followed by means of scintigraphy (imaging) techniques if one could achieve nanoparticle labelling with gamma-emitting isotopes. We have developed labelling methods able to supply such carriers using gamma-emitters like radioactive iodine (125I or 131I), indium or technetium. We used DTPA as a spacer in order to fix the last two isotopes. This would mean that any other gamma-emitting cation can theoretically be tried pending on its ability to be chelated by DTPA. The preparations were obtained with high labelling yields, usually > 80% and were relatively stable in human plasma over the whole period of investigation. 111In and 99mTc labelled forms have been administered to rabbit and then to man with 60-75% accumulation in the reticulo-endothelial system.

Increased serum alpha-melanocyte stimulating hormone (alpha-MSH) in human malignant melanoma

ALPHA-MELANOCYTE stimulating hormonc (alphaMSH) is a pituitary peptide responsible for increasing melanin synthesis by melanocytes in vertebrates. Despite thc important physiological role of this hormone, alpha-MSH determination in normal human plasma has only been reported by a fcw investigators. High alpha-MSH levels (> 80 pg/ml) have been described in 50% of pregnant women plasma at the term of the gestation [ll. Similar levels (> 60 pg/ml) have been found in healthy adults, while variations of these levels have been described according to psoralen and UVA exposure, depending on the skin phototype [2, 31. Ether researchers could not detcct alpha-MSH in human plasma 141. Since human malignant melanoma cells can express alpha-MSH receptors [5], alpha-MSH determination in patients suffering from this disease became of interest. We have developed a sensitive and specific radioimmunoassay to study alphaMSH concentration profiles in melanoma patients’ and healthy pcrsons’ sera. Technically, a modifica-. tion of thc Chloraminc-T2 iodination (“.‘I) method, which we have already described [6], was used together with an antiserum that gave crossreactions of less than 0.001% with pcptides closcl) related to alpha-MSH [7]. Our updated results show 3 times more detcctable serum alpha-MSH levels in 64 (34.4%) out of 186 melanoma patients (rcgardless of thc clinical evolution) as comparcd to 11.2% in 71 control

Bone marrow scintigraphy in breast cancer.

Bone marrow scintigrams (BMS) have been performed in 101 women with histologically proven breast cancer (36 stage IV among them at the time of the first investigation, follow-up examinations available in 41, disease evolution observed in 17). They have been compared to conventional bone scintigram (CBS) as well as to radiological, clinical, biological and follow-up data obtained in the same patients. Only three (7.4%) out of the 41 patients with radiologically or/and histologically proven skeletal metastasis had normal BMS and 33 had corresponding bone marrow scintigraphic defects (BMS ++). On the other hand, only three patients of the 32 with normal BMS had documented skeletal abnormalities. Abnormal bone marrow extension, observed in 61.4% of the patients (62/101), seems to represent a reaction of the reticulo-endothelial system to the disease dissemination - even microscopic - and, as such, to have prognostic implications. As BMS demonstrated skeletal invasion earlier or more precisely than CBS in ten cases, the technique appears of value and of interest in breast cancer management.

Tumor type and vascularity: Important variables in infusional brachytherapy with colloidal 32P

PURPOSE This study investigated the role of histologic tumor characteristics, in comparison with a normal tissue, and of tumor vascularization on the uptake and retention of colloidal 32P used in infusional brachytherapy of solid cancers. The cytotoxicity of colloidal 32P was also evaluated for two tumors of different radiosensitivity, a melanoma, and a squamous cell carcinoma. METHODS AND MATERIALS An in vitro analysis of colloidal 32P uptake was carried out on a human melanoma cell line, HBL, a human squamous cell carcinoma, SCC1, and normal fibroblasts, F-NBB. Tumor retention of colloidal 32P was studied in vivo for the HBL and the SCC1 tumors implanted subcutaneously in nude mice. Tumor vascular density was determined by microscopic study of Massons trichrome slides of HBL and SCC1 tumors of about 1 cm diameter. RESULTS In vitro studies showed that the time required for maximal cell uptake of colloidal 32P was only 10-20 min for the SCC1 and HBL tumors, while it took at least 60 min for the fibroblasts. After intratumoral injection of macroaggregated albumin (MAA), followed by 50 microCi of colloidal 32P, Bremsstrahlung imaging performed at 6 and 24 h showed that the activity remained in the HBL tumor while some of the radiocolloids leaked from the SCC1 tumor and was trapped in the reticuloendothelial system of the liver. Organ activity counting confirmed this finding: 32P activity was three to four times higher in the HBL than in the SCC1 tumor, whereas the activity in the liver, insignificant in the HBL mice (less than 0.1 microCi/g), was as high as 24 microCi/g in the SCC1 mice. This phenomenon may be explained by the difference in tumor vascular density, estimated for the HBL to be about four times less than that of the SCC1 tumor (5.7 vs. 21.4 blood vessels per mm2 for the HBL and the SCC1 tumors, respectively). CONCLUSION Intratumoral infusion of colloidal 32P may be a useful complement of radiation therapy in the treatment of nonresectable but accessible solid tumors. Tumor vascularization must be taken into account for a successful vascular blockade by MAA prior to the infusion of colloidal 32P.