Spontaneous regressive epitheliomas in the Japanese newt, Cynops pyrrhogaster.

Spontaneous tumors in urodele amphibians have been considered uncommon, and this resistance has sometimes been associated with the natural regenerative capacity of tissues in such species. This report describes spontaneous, nonpigmented, benign epitheliomas which were found in 44 of 1586 (2.8%) adult newts, Cynops pyrrhogaster, captured in central Japan. Both sexes were affected equally, usually with single tumors occurring at any epidermal site. Under laboratory conditions, these large tumors rapidly regressed or disappeared. Lesions were histologically noninvasive, hyperplastic epidermal reactions accompanied by loss of basal, subdermal melanocytes. Ultrastructurally, enlargement of intercellular spaces between tumor cells, increased pigment granules and membrane-bound cytoplasmic aggregates within the spaces, swollen rough endoplasmic reticula, degenerating pigment granules, and altered corneal cells were noted. Virus-like particles were observed in one tumor cell. Prelminary attempts failed to demonstrate transmissibility of the tumor, and no new cases arose under laboratory conditions. The cause of these tumors in the Japanese newt remains unknown, and it is suggested that if a viral agent is involved then other environmental cofactors (diet, temperature, or water constituents) are required for its expression.


Effects of hyperthermia (41.5 degrees) on Chinese hamster ovary cells analyzed in motisis.

The mitotic cells from an asynchronous population of Chinese hamster ovary cells exposed to 41.5 degrees for 7 hr were examined by light and electron microscopy to determine if there were any morphological abnormalities related to cell death or lengthening of metaphase induced by hyperthermia. All components of the mitotic apparatus were formed during exposure to heat, and the mitotic apparatus was functional as demonstrated by eventual cell division. However, heat caused the nuclear envelope to reform precociously around the chromosomes except in the region of the kinetochores, and the nuclear envelope remained associated with the chromatids during segregation. The precocious reformation of the nuclear envelope may be responsible for the lengthening of metaphase. Cells undergoing mitosis during the heat treatment possessed large evaginations of the plasma membrane, and the ubiquitous cortical microfilaments were absent in the region of these evaginations. Possibly related to the membrane damage were osmotic changes resulting in swollen mitochondria observed in heated cells entering mitosis. Since hyperthermic damage to the plasma membrane-microfilament complex was not observed in interphase cells or in cells completing division but was morphologically expressed during mitosis, the thermal lability of the plasma membrane must increase as the cells enter mitosis.


Glycosyltransferase and glycosidase activities in ovarian cancer patients.

In order to elucidate the mechanism of appearance of abnormal glycoproteins in cancer, activities of glycoprotein glycosyltransferases and glycosidases were determined in the homogenates prepared from normal ovaries and ovarian epithelial adenocarcinoma. Significantly high activities (more than normal mean + 2 S.D.) of these enzymes were found as follows: galactosyltransferase and sialytransferase in 100%; fucosyltransferase 1 (exogenous acceptor, fetuin minus sialic acid and galactose) in 86%; fucosyltransferase 2 (fetuin minus sialic acid, acceptor) in 45%; N-acetylglucosaminyltransferase 1 (ovalbumin acceptor) in 53%; N-acetylglucosaminyltransferase 2 (ribonuclease A as acceptor) in 10% of the samples analyzed. Among the glycosidases, substantially elevated activities above normal controls were found as follows: N-acetyl-beta-D-glucosaminidase in 85%; N-acetyl-beta-D-galactosaminidase in 63%; N-acetyl-beta-D-galactosidase in 50%, and those of alpha-L-fucosidase in 35% of the tumors. In serum of these cancer patients, only levels of galactosyltransferase were consistently elevated compared to controls. Increases in serum levels for other transferases were as follows: fucosyl-1, 10%; fucosyl-2, 60%; sialyl-, 20%; N-acetylglucosaminyl-1, 90%; N-acetylglycosaminyl-2, in 80% of the serum samples from ovarian carcinoma patients. Galactosyltransferase thus appears to be an excellent marker for ovarian carcinoma.


Gluconeogenesis from alanine in patients with progressive malignant disease.

We have studied by tracer technique the conversion of the carbon skeleton of alanine to glucose in patients with progressive malignant disease. These data have been compared to similar studies done in patients with chronic undernutrition from other causes. The results show increased conversion of alanine to glucose in the overnight fasting state as compared to the control group. Whereas the percentage increases are comparable to those found with pyruvate-glucose cycling in such subjects, the total amount of carbon conversion is considerably less (alanine carbon, 5.6 mmol/hr, versus pyruvate carbon, 39 mmol/hr). Exogenous glucose resulted in good suppression of alanine-to-glucose conversion as it does in normal subjects. It did, however, result in increased glucose-to-alanine conversion, increased alanine levels, and increased flux of alanine from the circulation. Although these latter data may not have specificity for the patient with advanced cancer, a strong dependence for carbohydrate and protein metabolism is suggested. We conclude that uncontrolled gluconeogenesis from alanine is probably not significant in terms of energy expenditure in the patient with uncontrolled cancer.


Leukocyte migration inhibition in vitro in bladder carcinoma.

The leukocyte migration inhibition response in vitro of peripheral blood leukocytes from patients with transitional cell carcinoma of the urinary bladder, other genitourinary diseases, and normal controls was studied. The agarose droplet migration inhibition method was used to determine the cell-mediated immune reactivity of bladder tumor patients to antigens derived from allogeneic bladder tumor cell lines by hypertonic potassium chloride extraction. Sixty-nine bladder tumor patients were tested 131 times and included 68 positive and 63 negative results. A positive assay was shown to correlate strongly with the presence of tumor at the time of testing (p less than 0.001). Five of 24 patients with other genitourinary diseases had positive inhibition assays, but none of the 26 normal controls were positive.  These results suggest that this in vitro assay method may prove valuable in monitoring bladder tumor patients for completeness of initial tumor resection and for clinical recurrence as well as for further studies of the tumor-specific immune response in this disease.


Transfer of tumor cells between cell aggregates as a model for adhesive changes in metastasis.

A model for cell detachment which may influence metastasis in vivo is described involving the transfer of hamster melanoma cells from aggregates of those tumor cells to hamster fibroblast aggregates. The aggregates were made by the spontaneous association of cultured cells. Tumor and fibroblast aggregates were then incubated together but separated by a nylon net that allowed only single cells to pass. Melanoma cells rapidly separated from the tumor aggregates, crossed the net, and attached to the fibroblast aggregates, but fibroblast cells did not. This model of metastasis reflects the postulated role of cell-to-cell adhesion in metastasis and will allow further study of the role of cell attachments in metastasis.


Alterations in platelet surface sialytransferase activity and platelet aggregation in a group of cancer patients with a high incidence of thrombosis.

Platelet aggregation, platelet surface sialic acid, and platelet surface sialytransferase activity were studied in a group of 12 cancer patients with a high incidence of thrombosis. These patients demonstrated accelerated coagulation, increased Factor VIII antigen and restocetin cofactor, and enhanced adenosine 5'-diphosphate-induced platelet aggregation. Platelet exogenous sialytransferase activity was increased in cancer patients (117.6 +/- 14 pmol/10(9) platelets) as compared to controls (59.0 +/- 4.3 pmol/10(9) platelets, p less than 0.01). Platelet exogenous sialytransferase activity and platelet aggregation were inhibited by aspirin. Thrombosis and bleeding have complicated the clinical course of half of these patients. This platelet membrane analysis provides additional data which may be related to current observations of increased levels of plasma sialytransferase activity and serum sialic acid in cancer patients.


Potentiation of anticancer agent cytotoxicity against sensitive and resistant AKR leukemia by amphotericin B1.

Amphotericin B was able to enhance the effects of actinomycin D (Act-D), adriamycin, and vincristine against AKR leukemia. AKR leukemia lines of increasing resistance to Act-D were obtained by passage in syngeneic mice treated with Act-D. The cells selected for Act-D resistance also eventually became cross-resistant to vincristine and Adriamycin, but resistance to these agents developed at a slower rate. Amphotericin B enhanced the effects of all these agents against the resistant cells, although the degree of enhancement varied among these antitumor agents and decreased as drug resistance increased in the late-passage leukemia lines.


Metabolism and excretion of benzo(a)pyrene 4,5-oxide by the isolated perfused rat liver.

Benzo(a)pyrene 4,5-oxide was metabolized in the isolated perfused rat liver by epoxide hydrase and glutathione S-transferases to the corresponding dihydrodiol and to thioether conjugates (derivatives of glutathione), respectively. Epoxide hydrase was more important relative to the glutathione S-transferases in the biotransformation of this oxide by the intact organ than was indicated by the results from earlier studies with subcellular fractions. The dihydrodiol was rapidly released into the circulation or conjugated with glucuronic acid; sulfuric acid esters were not found. All conjugated metabolites were rapidly excreted in the bile but some were also released into the circulation. The enzymatic systems responsible for the metabolism and excretion of benzo(a)pyrene 4,5-oxide remained viable in the isolated perfused liver for at least 60 min. The toxicological significance of the release of polycyclic aromatic hydrocarbon metabolites from the liver into the vascular circulation and the possible significance of UDP:glucuronyltransferase activity in preventing chemically induced carcinogenesis are discussed.


Induction of the DNA repair enzyme uracil-DNA glycosylase in stimulated human lymphocytes.

The capacity of human cells to modulate the synthesis of DNA repair enzymes has been investigated by measuring the induction of the uracil-DNA glycosylase during lymphocyte stimulation. Treatment of peripheral lymphocytes with phytohemagglutinin increased glycosylase activity 10-fold. Glycosylase stimulation was coordinate with the activation of DNA synthesis and DNA polymerase activity. Two chromatographically distinct species of the glycosylase have been resolved; only one species is induced during phytohemagglutinin stimulation. The effect of actinomycin D and cycloheximide on glycosylase induction was determined. Treatment with either inhibitor at 96 hr after phytohemagglutinin addition (maximal induction) decreased glycosylase activity after an appreciable lag period. This suggested that induction of the uracil-DNA glycosylase requires transcription and translation although the enzyme may be quite stable once induced.


Regional lymph node reactivity in explanted bladder cancer of mice as measured by flow cytometry.

The reactivity of lymphocytes in lymph nodes draining the site of a transplantable experimental bladder tumor (MBT2 in C3H/HeJ mice) has been measured in a multiparameter flow cytometry system. Acridine orange was used as a nucleic acid probe. This dye intercalates in helical DNA, emitting green (530 nm) fluorescence upon exposure to blue light; it stacks to single-stranded RNA, emitting red (640 nm) fluorescence. The relative magnitude of the increase of lymphocyte DNA and RNA has been evaluated simultaneously in tumor-draining nodes, in nondraining nodes of the same animal, and in untreated control animals. Stimulation of the regional node lymphocytes could be observed after 20 days but not after 10 days. It was uniformly high at 35 days. The transcriptive response (increased proportion of lymphocytes with high RNA) was more pronounced than the proliferative (increased proportion of lymphocytes with more than diploid DNA). The histological changes in the stimulated nodes resembled closely those described by others in human tumor-draining nodes. The described method has the advantage of being simple, rapid, and able to measure a representative part of the whole-cell population.


Effect of hepatocarcinogens on the adenine purine nucleotide cycle during the initiation phase of carcinogenesis.

Activities of the adenine purine nucleotide cycle enzymes, i.e., adenylosuccinate (SAMP) synthetase, SAMP lyase, and adenosine 5'-monophosphate deaminse, were determined in hepatic tissue of rats fed and/or given injections of 3'-methyl-4-dimethylaminoazobenzene, 4'-methyl-4-dimethylaminoazobenzene, thioacetamide, ethionine, or 2-acetylaminofluorene. SAMP lyase activity showed an early increase in all regimens containing hepatocarcinogens. Adenosine 5'-monophosphate deaminse showed increases with 3'-methyl-4-dimethylaminoazobenzene and thioacetamide but not with ethionine or 2-acetylaminofluorene. SAMP synthetase either was nonresponsive or else showed inhibition to the carcinogens. Increase in SAMP lyase activity was noted as early as 48 to 72 hr following i.p. injections of these carcinogens. The response of SAMP lyase was not duplicated by analogs of carcinogens such as 4'-methyl-4-dimethylaminoazobenzene or methionine. These data imply interaction of active carcinogens with SAMP lyase and to some extent adenosine 5'-monophosphate deaminse or to some mechanism responsible for their synthesis and/or release. This interaction may be a significant component of the initiation phase of carcinogenesis.


Resistance to Ehrlich ascites tumor in a strain of dystrophic mice.

Ehrlich ascites tumor grows in almost any mouse host. We have found a strain of dystrophic mice to be strongly resistant to Ehrlich ascites tumor. Mice of the dystrophic strain survived injection of Ehrlich Ascites tumor cells that had just been passaged through either dystrophic mice or C57BL X DBA/2 F1, (hereafter known as BD2F1) mice. However, most of the dystrophic mice died when they were given injections of Ehrlich ascites tumor cells that had just been passaged through Swiss white mice. All Swiss white and BD2F1 mice died when they were inoculated with Ehrlich ascites tumor cells, regardless of the type of mouse through which the Ehrlich ascites tumor cells had just been passaged. In some dystrophic mice, the tumor grew to a palpabale size and then disappeared. This provided the opportunity to passage the tumor from one dystrophic mouse to another and thereby to demonstrate resistance in the dystrophic mice to tumor that had grown previously in dystrophic mice. There was no detectable difference in the rate of tumor growth in either Swiss white or BD2F1 mice.


Ozonation of mutagenic and carcinogenic alkylating agents, pesticides, aflatoxin B1, and benzidine in water.

The effect of ozonation on the mutagenicity of selected chemicals in water was determined. The use of the Salmonella-microsome assay for mutagensis allowed kinetic studies to be performed on the ozonation of all chemicals tested. The results indicate that the mutagenicity of certain pesticides, including captan and Dexon, was inactivated by short periods of ozonation. The mutagenicity of certain alkylating agents including bis(2-chloroethyl)amine and sodium azide was rapidly inactivated by ozonation while other alkylating agents such as beta-propiolactone, propanesultone, and N-methyl-N'-nitro-N-nitrosoguanidine were unaffected by treatment with ozone. The mutagenicity of aflatoxin B1 was rapidly inactivated by treatment with ozone. Three chemicals were shown to be converted to direct mutagens by ozone treatment. Under certain conditions, dimethylhydrazine could be converted to a mutagen that was stable for 3 weeks. A similar chemical, 2-hydroxyethylhydrazine, was converted to an unstable mutagen that was inactive after 24 hr at room temperature. When benzidine was treated with ozone, there was a transient increase in mutagenicity which was lost after longer treatment with ozone.


Tumor eradication and cell survival after localized hyperthermia induced by ultrasound.

This study was designed to test the effect of localized ultrasound-induced hyperthermia on experimental mouse tumors. Transducers operating at 5.17 MHz with relatively uniform output over an area 1 cm in diameter were used to heat EMT6 and KHJJ tumors in 407 BALB/cKa mice. Treatments were at 43, 43.5, 44, and 44.5 degrees. At each temperature level, treatments were applied for 15, 30, or 45 min. Temperature profiles measured in tumors treated by ultrasound hyperthermia indicated that heating was not completely uniform. In general, both rate of tumor eradication and growth delay increased as temperature and/or time of exposure increased. The EMT6 and KHJJ tumors had comparable rates of eradication for the same temperatures and times of exposure. Cell survival studies indicated that there was considerable variation in cell killing between individual EMT6 tumors exposed to the same hyperthermic dose. In addition, cell death appeared to be progressive over a period 2 to 48 hr after hyperthermic exposure. The mechanism of this delayed cell death is not known but may be important in eradicating the tumors. Ultrasound was a relatively safe and effective method of heating tumor volumes up to 44 degrees, and hyperthermia alone resulted in high rates of tumor eradication in the EMT6 and KHJJ systems.


Cardiovascular lesions and various tumors found in rats given T-2 toxin, a trichothecene metabolite of Fusarium.

White rats given intragastrically 3alpha-hydroxy-4beta,15-diacetoxy-8alpha-(3-methylbutyryloxy)-12,13-epoxy-tricholthec-9-en (T-2 toxin), a trichothecene metabolite of several Fusarium species, developed various acute and chronic, topical and systemic lesions. The rats that survived 12 to 27.5 months after the first of three to eight doses of T-2 toxin (0.2 to 4 mg/kg body weight) alone or in conjunction with nicotinamide given i.p. (200 to 250 mg/kg body weight) developed cardiovascular lesions of various degrees of severity and/or tumors, benign and malignant, of the digestive tract and of the brain. T-2 toxin is known occasionally to contaminate cereals and other agricultural products, harvested or stored under damp and cold conditions. T-2 toxin was responsible for an often fatal disease in humans, known in the U.S.S.R. as "alimentary toxic aleukia," and also for outbreaks of hemorrhagic mycotoxicoses in livestock in various countries. T-2 toxin and other Fusarium mycotoxins may be involved in the etiology of cardiovascular lesions and of certain tumors considered as "spontaneous" in animals and humans.


Inhibition of differentiation of cultured mouse myeloid leukemia cells by nonsteroidal antiinflammatory agents and counteraction of the inhibition by prostaglandin E1.

Mouse myeloid leukemia cells (M1) were induced to differentiate into mature macrophages and granulocytes by glucocorticoids or a protein inducer in ascitic fluid from tumor-bearing rats. Addition of nonsteroidal antiinflammatory agents to M1 cells in suspension cultures inhibited the induction of differentiation by glucocorticoid (dexamethasone) or the protein inducer. The inhibition was unrelated to cytotoxicity and was reversible. The nonsteroidal antiinflammatory agent indomethacin inhibited dexamethasone-induced differentiation only when added before the time of commitment of the cells to differentiation. The indomethacin-mediated inhibition was counteracted by prostaglandins E1 or E2 but not by prostaglandins F1alpha or F2alpha. Prostaglandin E stimulated phagocytosis induced by a suboptimal concentration of dexamethasone, but prostaglandin F did not. Moreover, lysozyme activity, which is a typical biochemical marker of macrophages, was induced in M1 cells by prostaglandin E alone, as well as by inducers of differentiation. These results suggest that prostaglandin E may be important in the induction of differentiation of myelod leukemia cells.


Active efflux of daunorubicin and adriamycin in sensitive and resistant sublines of P388 leukemia.

Significantly decreased uptake and retention of daunorubicin (DAU) and Adriamycin (ADR) have been reported in sublines of P388 leukemia resistant to these anthracyclines. We studied the effects of inhibitors of oxidative phosphorylation on uptake and retention of DAU and ADR in order to characterize the transport process for these anthracyclines and to clarify further the alteration in resistant cells. In glucose-free medium, uptake of DAU and ADR was accelerated by metabolic inhibitors to a greater extent in resistant cells than in sensitive cells. Under these conditions, drug uptake was similar in sensitive and resistant cells. When glucose was added to the incubation medium in the presence of 2,4-dinitrophenol, efflux of DAU from both sensitive and resistant cells was observed. Net efflux of ADR was also observed with the resistant cells, whereas glucose markedly inhibited 2,4-dinitrophenol-stimulated uptake of ADR by the sensitive cells. Furthermore, in sensitive and resistant cells preloaded with ADR and DAU, efflux of the drugs was inhibited by the addition of 2,4-dinitrophenol. These results suggest that there is an active outward transport mechanism for anthracyclines in P388 leukemia cells and that enhanced activity of this efflux process renders cells highly resistant to the cytostatic and cytotoxic effects of ADR and DAU.


Adequacies and inadequacies in assessing murine toxicity data with antineoplastic agents.

Previous retrospective analyses have suggested a very positive correlation in toxic doses of antineoplastic agents between mice and humans. Additional toxicological information has now been accumulated and reveals a noticeable variability in the existing data base. Nevertheless, it is likely that mouse toxicological studies will become a principal determinant for estimating initial doses to be used in humans. Recognition of the factors responsible for differences in determinations of toxic dose levels in mice will enhance the proper utilization of this approach.


Drug sequence-dependent toxicity and small bowel mucosal injury in mice treated with low doses of 3-deazauridine and 1-beta-D-arabinofuranosylcytosine.

The toxicity to mice of combinations of 1-beta-D-arabinofuranosylcytosine and 3-deazauridine was investigated. The drugs were administered daily i.p. on Days 1 to 5, each drug at 10 mg/kg body weight; these dosages are small fractions of the dosages at which 10% of the treated animals died when either drug was administered alone on the foregoing schedule. This drug combination was severely toxic when 3-deazauridine was administered 2 to 8 hr prior to 1-beta-D-arabinofuranosylcytosine; most mice treated in this way died within 3 days of the last treatment. Histological examination showed that severe damage to the small bowel mucosa resulted from treatment with the drugs in the above, lethal sequence. In contrast, treatments with this drug combination at the same dosages were tolerated when the two agents were administered simultaneously or when 1-beta-D-arabinofuranosylcytosine preceded 3-deazauridine. Under the latter conditions, small bowel mucosal injury was much less severe. Female mice were more sensitive to the toxic treatment regimen than were male mice and were protected against the latter when either the 3-deazauridine or the 1-beta-D-arabinofuranosylcytosine component was preceded by treatment with nitrobenzylthioinosine (100 mg/kg), a potent inhibitor of nucleoside transport.


Hyperthermic treatment of human tumors heterotransplanted in nude mice.

A new model is presented for the study of the effects of supranormal temperatures on human tumors in vivo. Human tumors heterotransplanted serially in nude mice were heated in vivo by means of local radio-frequency heating. A lung carcinoma, a breast carcinoma, a colon carcinoma, and a malignant melanoma were studied. The tumors were transplanted s.c. in the inguinal area or under the kidney capsule of adult nude mice. The s.c. tumors were heated for 30 min. Temperatures of 43--44 degrees C were reached in the surrounding normal tissues, whereas in the center of the tumor temperatures of 46--49 degrees C were recorded. In 11 of 16 randomized pairs of mice, the growth of the tumor treated by hyperthermia was inhibited by 75% or more as compared with the growth of the untreated tumor control. No mortality and only temporary damage to skin and muscle were observed. The kidney tumors were also treated for 30 min, but it was possible to reach only 40 degrees C in the abdomen. Seventy-five % mortality was observed. Of seven randomized pairs evaluated, five exhibited a reduction of growth varying from 37 to 63%9 The model proposed appears to be a workable and promising one, especially for s.c. growing tumors.


Normal tissue and solid tumor effects of hyperthermia in animal models and clinical trials.

Localized hyperthermia therapy by high-energy radio-frequency waves was evaluated in malignant and adjacent normal tissue of 30 patients with 10 types of cancer. Hyperthermia was delivered to superficial and deep visceral cancers in awake patients who had refractory disease. Histological and clinical responses were recorded serially. Toxicity tests in dogs, sheep, and pigs showed that progressive necrosis of normal and cancer tissue occurred at temperatures above 45 degrees C (113 degrees F). However, as normal tissues approached this temperature, intrinsic heat dissipation occurred (possibly due to augmented blood flow) so that temperatures below 45 degrees C could be maintained, whereas most solid tumors did not have this adaptive capacity and could be heated to 50 degrees C (122 degrees F) with virtually no injury to normal organs, s.c. tissue, or skin. To date, 69 treatments have been administered to 36 tumors in the 30 patients. Selective heating was observed in both primary and metastatic tumors located in surface tissues and internal organs. Response appeared to be related to tumor size in that differential heating was possible more often in the larger lesions. In tumors successfully heated, moderate to marked necrosis occurred. Radio-frequency hyperthermia appears to be a safe and potentially useful form of therapy for selected cancer patients. While other cancer treatments are more effective for small tumors, hyperthermia may be uniquely beneficial against larger lesions.


Results of eleven years' experience with heated perfusion for melanoma of the extremities.

Eleven years have elapsed since we first added heat to regional perfusion for treatment of melanoma of the extremities. This report describes briefly our laboratory findings and our technique of hyperthermic perfusion and brings up to date the survival figures for the 165 patients (185 perfusions) which were originally reported in 1975 (Stehlin et al., Surg. Gynecol. Obstet., 122: 3--14, 1966). A dramatic increase in the survival rate is documented for those patients with recurrent melanoma confined to the extremities.


Clinical and biological studies of localized hyperthermia.

In 1975, we initiated a clinical study using multiple cutaneous lesions as the model system to determine whether hyperthermia alone or as an adjuvant to radiation evokes better tumor control than that achieved with radiation alone. The clinical observations of the heating patterns observed in tumor tissues and the adjacent normal tissues indicate that there was selective heating of superficial cancers in more than 80% of the cases. Using fractionated heat alone, there was a transitory tumor response in 10 of 19 patients. When combined with radiation and heat, the complete response achieved was 78% (42 of 54). Investigations in animal tumors indicate that the importance of the site of growth is related to local tumor control by heat alone. Other studies designed to investigate whether localized hyperthermia alone or with radiation increased tumor metastases were negative.


Thermal dosimetry and clinical requirements.

Major problems in clinical hyperthermia include (a) inhomogeneity of heat distribution in designated tumor volumes due to tissue characteristics and differential blood flow, (b) limitations of heat delivery and control systems for adequate depth penetration and adjustments of temperatures, and (c) the lack of capability of accurate temperature measurements, especially in the area of noninvasive techniques for deep-seated tumors. Examples were given to illustrate the clinical requirements of hyperthermia of superficial, intermediate, and deep-seated tumors.


Regional blood flow in human tumors.

Blood flow in 97 human tumor nodules, of which 31 were lymphomas, 31 were anaplastic carcinomas, and 35 were differentiated cancers, was measured using the 133Xe clearance method. The lymphomas showed statistically higher blood flow [34.6 +/- 21 (S.D.) ml/min/100 g] than did the anaplastic carcinomas (15.4 +/- 11.4 ml/min/100 g; p less than 0.001) and the differentiated cancers (22.8 +/- 14.9 ml/min/100 g; p less than 0.05). The size of the tumors did not correlate with the blood flow. In the group of differentiated cancers, the blood flow in nodules in areas of earlier irradiation was lower (9.0 +/- 6.3 ml/min/100 g) than in nodules in intact regions (25.0 +/- 14.5 ml/min/100 g; p less than 0.05). Nodules in cicatricial areas after surgical operation had lower blood flow than did nodules in the intact areas, but the difference is not statistically significant. It is obvious that most human tumors have a considerably lower blood flow than what one would expect to find in the surrounding normal tissue.


Aspects of thermoregulatory physiology pertinent to hyperthermic treatment of cancer.

Local hyperthermia is relatively safe, while whole-body hyperthermia is potentially dangerous because the therapeutically effective elevation of body temperature is close to the tolerable limit to hyperthermia beyond which permanent damage may be caused. Here consideration is given to the most reliable index of body temperature during hyperthermia and the techniques for raising body temperature. The possibility of effecting local brain cooling and thereby increasing the margin of safety during whole-body hyperthermia is also considered.


Thermal dosimetry and temperature measurements.

A crucial ingredient of any hyperthermia procedure is the accurate measurement of achieved temperature. In this paper, we present some accuracy and resolution suggestions and address the problem of temperature measurements when electromagnetic energy is used as the mode of heating. In such cases, conventional metallic thermometers can cause severe errors due to reradiated fields, high internal healing, and electronic interference. To avoid these problems, several groups have developed either high-resistance lead thermometers or optical-fiber probes using a variety of sensors, including birefringent and semiconductor crystals. A new noninvasive approach uses ultrasound-computed tomography and relies upon the change in the speed of sound versus temperature to achieve maps of internal tissue temperature.


Recommendations for future research in hyperthermia at the molecular, cellular, and animal levels.

Recommendations are made for hyperthermia research to be carried out at the molecular, cellular, and animal levels. Effects of heat alone should be distinguished from effects of heat combined with radiation or chemotherapy. Factors to be considered include tumor cell environment, treatment sequencing, thermal tolerance, cell kinetics, any possible nonthermal effects, and therapeutic gain.


Increased survival of rats bearing Morris hepatoma 7800 after induction of hypothyroidism.

The survival of Buffalo rats brearing Morris Hepatoma 7800 was increased significantly (23 to 31%) after induction of hypothyroidism by propylthiouracil (PTU) (0.1% in Purina rat chow) or 131I. The concentration of PTU used was in the optimal range as 0.03% PTU was less effective than 0.1%, while 0.4% PTU appeared to be toxic. Exogenous thyroxine (8 microgram/kg body weight) reversed the effects of PTU and actually shortened survival. Because food consumption and body weights of hypothyroid rats were decreased, the survival of pair-fed controls was studied and found to be the same as in untreated controls. We conclude that the hypothyroid state increases the survival of rats bearing Morris Hepatoma 7800. We have not yet been able to define any anatomical or biochemical parameters which may be responsible for this effect.


Persistent binding of 3-methylcholanthrene to mouse lung DNA and its correlation with susceptibility to pulmonary neoplasia.

A/J, C3H/HeJ, DBA/2J, and C57BL/6J mice have different susceptibilities to polycyclic aromatic hydrocarbon-induced pulmonary neoplasia, whereas the livers from these animals are uniformly resistant to the carcinogenic actions of these substances. After i.v. injection of [3H]-3-methylcholanthrene, radiolabel was detected in the DNA of both lung and liver of all these strains. The DNA was digested to deoxyribonucleosides and chromatographed on Sephadex LH-20. The amounts of nucleoside-bound adducts detected varied markedly with the different tissues and strains. These adducts were undetectable in liver DNA by 28 days. Although all lung preparations showed some reduction in adducts by 28 days, the amounts in A/J lung were always highest; this correlated with its high susceptibility to neoplastic transformation. In all preparations, radioactivity eluted from Sephadex LH-20 with the column void volume or with underivatized nucleosides. Tissue, but not strain differences were observed in these chromatographic profiles. The predominance of these early-eluting peaks in liver, rather than lung, suggests that they may represent noncarcinogenic lesions. This radiolabeled material remains uncharacterized, but some possibilities are discussed.


Direct inhibition of growth and antagonism of insulin action by glucocorticoids in human breast cancer cells in culture.

We have examined the interaction of dexamethasone with the ZR75-1 human breast cancer cell line to determine if glucocorticoids might directly inhibit growth of breast cancer cells. Growth of these cells in serum-free medium was stimulated significantly by physiological concentrations of insulin (0.1 to 1.0 nM). Pharmacological concentrations of dexamethasone (10 nM) reduced cell number below that found in controls and nearly abolished the effect of insulin after several days in culture. Thymidine and uridine, but not leucine, incorporation into macromolecules or acetate incorporation into fatty acids were similarly inhibited by dexamethasone in the presence of absence of insulin. Dexamethasone did not inhibit insulin effects by altering insulin receptor affinity or concentration, as determined by Scatchard analyses of insulin binding. Net thymidine uptake into the trichloroacetic acid-soluble fraction of the cell was stimulated by insulin and inhibited by dexamethasone also inhibited thymidine kinase activity multiple potential sites of glucocorticoid action that directly oppose the effects of insulin. They also suggest that glucocorticoids have a direct inhibitory effect on proliferation of human breast cancer cells, which may help explain breast tumor regression following pharmacological glucocorticoid therapy.


Role of adrenals and estrogen in regression of mammary tumors during postpartum lactation in the rat.

The effects of bilateral adrenalectomy or estradiol benzoate treatment were observed on growth of 7,12-dimethylbenz(a)-anthracene-induced mammary tumors during postpartum lactation. In the control and estradiol benzoate-treated postpartum lactating rats, the mammary tumors decreased approximately 40% in size by Day 5 postpartum and continued to regress to 50% of their average original diameter by Day 25 postpartum. Adrenalectomy on Day 3 postpartum prevented mammary tumor regression and resulted in renewed mammary tumor growth. By Day 10 postpartum, average mammary tumor size in the adrenalectomized rats reached prepartum diameter and continued to increase in size until Day 25. Although serum prolactin concentrations were significantly higher in the lactating rats with mammary tumors than in the nonlactating rats with mammary tumors, there were no significant differences in serum corticosterone values. Adrenalectomy resulted in a significant increase in serum prolactin levels and in a marked fall in serum corticosterone levels. It is concluded that in rats adrenocortical activity is primarily responsible for reduced mammary tumor growth during postpartum lactation.


Carcinoembryonic antigen-like substance derived from human prostate.

Carcinoembryonic antigen-like substance, previously detected in large amounts in the medium from cultures of human prostatic epithelial cells, also is present in extracts of benign and malignant human prostate. By column chromatography, the prostate-derived carcinoembryonic antigen-like substance derived from cultured prostate is the same as that in tissue extracts and is distinctly different from colon-derived carcinoembryonic antigen. The molecular weight of prostate-derived carcinoembryonic antigen-like substance is estimated to be greater than 5 x 10(5). Prostate-derived carcinoembryonic antigen-like substance may be a prostate-specific substance.


Zinc deficiency and growth of Ehrlich ascites tumor.

The growth rate of Ehrlich ascites tumors has been examined as a function of the zinc content of the diet of the host mice. Imposition of a diet containing a low amount of zinc (1 microgram/g) on the day of tumor transplant leads to a marked retardation in growth. Pretreatment of the mice with this diet slows the growth further so that the lifetime of the mice can be doubled. Cells in such animals are still viable and rapidly proliferate after the animals are placed on a diet containing zinc. Growth rate of the tumor is also recorded at levels of zinc (40, 80, 160, and 250 microgram/ml) in the drinking water. All of these results are examined in relationship to the zinc in the ascites fluid, which provides the zinc nutriture for the tumor. A direct correlation between growth rate and fluid zinc content is observed. The influence of diet and the tumor upon zinc content of the liver of the host is examined. The results indicate that the tumor essentially sequesters zinc from the animal under zinc-deficient conditions. Over a 10-fold range of fluid zinc values, there are no clear differences in the concentration of zinc within the ascites cells. This occurs despite the facile uptake and efflux of zinc ion by the Ehrlich cell.


Competitive binding radioassay for 5-fluorodeoxyuridine 5'-monophosphate in tissues.

A competitive binding radioassay has been developed for 5-fluorodeoxyuridine 5'-monophosphate, based on the tight binding of this potent inhibitor to thymidylate synthetase (EC 2.1.1.45). Unbound ligand may be separated from that bound to enzyme by precipitating the intact inhibitor-enzyme complex with trichloroacetic acid. Scatchard plot analysis using a two-site model for binding yielded apparent dissociation constants of 1.2 x 10(-11) and 1.7 x 10(-10) M from a least-squares computer fit of the data. 5-Fluorodeoxyuridine 5'-monophosphate could be detected in the range of 0.02 to 2.0 pmol with no apparent interference by other substances. Assay of 5-fluorodeoxyuridine 5'-monophosphate levels in L1210 ascites tumor following 5-fluorouracil in vivo revealed peak levels occurring within the first hr with a subsequent disappearance half-life of 3.9 hr. Close agreement was found between the previously described enzyme inhibition assay and the more rapid and sensitive competitive binding method.


Temporary disappearance ("eclipse") of LPC-1 plasmacytoma M component synthesis following tumor cell transfer.

The early phase of LPC-1 plasmacytoma development was studied by in vivo labeling with [6-14C]arginine using its M component (immunoglobulin G 2a,kappa) as marker. At a time when M component was not detected or faint by protein staining of electrophoretograms, significant labeling of M component was detectable by autoradiography. Labeling of the M component was fairly constant for the first 10 hr but was markedly decreased from Days 1 to 7. Nadir (0 to 3% of initial 30-min value) was observed on Day 3. Recovery of M component labeling to the 30-min level was complete by Day 13. This period of marked reduction or "eclipse" in newly synthesized M component was shortened by 2 days when mice were pretreated with pristane or cyclophosphamide prior to tumor cell transfer. The eclipse period was also 2 days shorter in athymic BALB/c-nu/nu mice than in normal BALB/c mice. The eclipse period corresponds to the classical "lag" following tumor cell transfer before tumor growth can be detected by conventional methods. The sensitivity of the [6-14C]arginine pulse permits the in vivo detection of small numbers of tumor cells (as few as 10(6) cells) over the early time periods after cell transfer. Modification of eclipse by manipulating host and/or tumor cells may elucidate the accompanying cellular and biochemical events.


Macromolecular binding and metabolism of the carcinogen 4-chloro-2-methylaniline.

Biochemical investigations relating to the mechanism of action and mechanism of activation have been made for the carcinogen, 4-chloro-2-methylaniline. Radioactivity from 4-chloro-2-[methyl-14C]methylaniline became extensively bound to protein, DNA, and RNA of rat liver, but macromolecules of some of the other tissues examined contained little radioactivity. Enzymatic activity dependent upon reduced nicotinamide adenine dinucleotide and leading to irreversible binding to radioactivity from labeled 4-chloro-2-methylaniline to macromolecules in the reaction system was present in microsomes from rat liver. The activity was inducible by phenobarbital. Two soluble products of microsomal enzymes were identified by mass spectral analysis and chemical synthesis as 5-chloro-2-hydroxylaminotoluene and 4,4'-dichloro-2,2'-dimethylazobenzene. The hydroxylamino compound appears to be a more activated form of 4-chloro-2-methylaniline.


Bacillus Calmette-Guérin enhancement of colony-stimulating activity and myeloid colony formation following administration of cyclophosphamide.

Pretreatment of mice with Bacillus Calmette-Guérin (BCG) enhances recovery of the peripheral granulocyte count from cyclophosphamide (CTX)-induced leukopenia. In the present study, in vitro bone marrow culture was used to assess the effect of BCG pretreatment on serum levels of myeloid colony-stimulating activity and on regeneration of bone marrow myeloid colony-forming units (CFU-c) following CTX. C57BL/6 mice received either BCG or 0.9% NaCl solution i.p. 8 days prior to the administration of CTX (300 mg/kg i.p.). Following CTX, peak serum levels of myeloid colony-stimulating activity were strikingly higher in the BCG-pretreated mice [1320 +/- 30 (S.E.) units] than in the 0.9% NaCl solution-pretreated mice (764 +/- 37 units). BCG pretreatment also resulted in higher numbers of marrow CFU-c during recovery from CTX (e.g., 43.9 +/- 1.8 versus 20.0 +/- 0.9 myeloid colonies/10(5) marrow cells, Day 3 post-CTX). However, neither the rate of decline nor the absolute nadir of CFU-c, nor CFU-c cell cycle characteristics were affected by the pretreatment. The initial effect of i.p. BCG pretreatment on recovery from CTX-induced granulocytopenia is an augmentation of serum myeloid colony-stimulating activity which precedes the enhanced regeneration of CFU-c and the accelerated recovery of the peripheral granulocyte count.


Increased in vitro toxicity to mouse bone marrow with 1,3-bis(2-chloroethyl)-1-nitrosourea and hyperthermia.

Hyperthermia has been shown to increase the cytotoxic effect of certain cancer chemotherapeutic agents in vitro and in vivo. It is not known whether the combination of hyperthermia plus these chemotherapeutic drugs will also increase the toxicity of the drugs. This study was undertaken to evaluate the toxic effects of 1,3-bis(2-chloroethyl)-1-nitrosourea combined with hyperthermia on normal mouse bone marrow in vitro. An increased effect of the combination was seen when the marrow was heated at 42 degrees in the presence of the drug for 15 min (p less than 0.025) and for 1 hr (p less than 0.005). Incubations for 2 hr produced a significant decrease of control growth, such that no increased effect of the combination of drug plus hyperthermia could be seen. In order for the increased effects to be seen, heating had to be done simultaneously in the presence of the drug. Clinicians using the combination of whole-body hyperthermia plus 1,3-bis(2-chloroethyl)-1-nitrosourea should be aware of possible increased marrow toxicity.


Dose response for benzo(a)pyrene adducts in mouse epidermal DNA.

The dose dependency of the binding of benzo(a)pyrene (BaP) with DNA of mouse epidermis was investigated. BaP-conjugated epidermal DNA was isolated and enzymatically degraded to deoxyribonucleosides. The BaP-DNA adducts were separated by Sephadex LH-20 column or high-performance liquid chromatography. Two major BaP-DNA adducts were found. One was in the region of the elution profile that contained polycyclic aromatic hydrocarbons adducted to deoxyribonucleosides. The other adduct was eluted from Sephadex LH-20 and high-performance liquid chromatography columns before the deoxyribonucleosides and after deoxyribonucleotides. Both adducts of BaP in epidermal DNA reached a maximum 7 hr after a single skin application, and subsequently little, if any, loss of adducts was observed for 49 hr. Both adducts varied as a linear function for topical doses in the range from 0.01 to 300 microgram/mouse. The formation of DNA adducts by BaP occurred in proportion to dose at doses several orders of magnitude below those that are feasible to test for carcinogenicity.


Cervicovaginal and mammary gland abnormalities in BALB/cCrgl mice treated neonatally with progesterone and estrogen, alone or in combination.

Neonatal female BALB/cCrgl mice (mammary tumor virus unexpressed) were given a daily injection of estradiol and/or progesterone for 5 days, beginning within 36 hr after birth. About one-half of each group was ovariectomized when 40 days old, and all mice were killed when between 18.5 and 26 months of age. Neonatal progesterone leads to ovary-dependent persistent vaginal cornification and hyperplasia. In addition, 16 of the 24 progesterone-treated mice had genital tract lesions, and 4 of these showed predominantly glandular features. No such lesions were observed in either oil-treated or untreated mice. Lesions were also observed in both intact and ovariectomized mice treated with estrogen-progesterone combinations, but most of the lesions were not as severe as those seen in mice treated neonatally with progesterone alone, and they were predominantly squamous in appearance. Although mammary tumors were not observed in either the control or the neonatally steroid-treated intact mice, many in the latter groups possessed hyperplastic alveolar-like mammary nodules and other abnormalities.


Mechanism of action of tetra-mu-carboxylatodirhodium(II) in L1210 tumor suspension culture.

The effect of tetrakis-mu-methoxyacetato, tetra-mu-acetato, tetra-mu-propionato, and tetra-mu-butyratodirhodium(II) on the proliferation and macromolecular synthesis of leukemia L1210 cells in suspension culture was evaluated. The cytotoxicity of these dimeric rhodium(II) complexes to tumor cells in suspension culture follows the same trend as observed in vivo, i.e., butyrato greater than propionato greater than acetato greater than methoxyacetato. The cellular synthesis of DNA and protein was found to be strongly inhibited by tetra-mu-propionatodirhodium(II), whereas minimal inhibition of RNA synthesis was observed. Flow microfluorometric analysis of the drug-treated cells revealed an arrest of cellular development during the G2 phase of the cell cycle. The inhibition of DNA synthesis was attributed at least in part to the arrest in G2 which is consistent with the observed inhibition of protein synthesis.


Transplantability and sex steroid hormone responsiveness of cervicovaginal tumors derived from female BALB/cCrgl mice neonatally treated with ovarian steroids.

Twenty-eight cervicovaginal tracts from approximately 2-year-old female BALB/cCrgl mice neonatally exposed to ovarian steroids were cut into small segments and transplanted into syngeneic hosts. Within six months, six of 28 host animals developed tumors. Three tumors were from progesterone-exposed mice, two were from estrogen-exposed mice, and one was from estrogen-progesterone-exposed mice. These tumors have been maintained by serial transplantation for approximately two years. The progesterone-induced tumors are mixed tumors with both squamous cell and glandular components. The estrogen-induced tumors are squamous cell carcinomas. The estrogen-progesterone-induced tumor was originally a squamous cell carcinoma, which now resembles a basal cell carcinoma. The other tumors have maintained their original morphological characteristics. All tumors have proven to be hormone independent. No control cervicovaginal tracts developed tumors after transplantation, even after 24 months in the host animals.


Two-dimensional electrophoresis of membrane proteins from normal and transformed cells.

We have compared cell surface proteins and glycoproteins from normal and transformed cells by two-dimensional electrophoresis. The tumor lines studied include viral and spontaneous transformants which cause tumors that either regress, or grow progressively and kill the host. After the cells were labeled, crude membranes were prepared, and their protein components were resolved by two-dimensional electrophoresis. Cells were labeled with either [35S]methionine, [14C]glucosamine, or 125I-lactoperoxidase. This allows comparison of cell-cell differences in surface structure as well as membrane proteins and glycoproteins. We have demonstrated that not all transformed cell lines give a characteristic "transformed" protein pattern. Many of the differences between the normal and transformed cell protein patterns may be related to alterations in the carbohydrate of membrane glycoproteins. A correlation can be made between the patterns obtained for cell lines and the immunogenicity and tumorigenicity of these cells in vivo.


Increased progesterone receptor concentrations in bladder lesions of estrogen-treated Syrian hamsters.

During studies of renal tumorigenesis induced by estrogen in Syrian hamsters, we have observed that about 15 to 20% of animals develop bladder lesions with an increased wet weight of tissue from 0.2 g to 0.4 to 1.7 g. Histological examination of the lesions showed a spectrum of changes from inflammatory reactions to squamous metaplasia and intense hyperplasia of the transitional epithelium. The concentration of progesterone-binding sites was increased in the bladders with lesions. No specific progesterone-binding sites could be detected in the cytosol of bladders from hamsters not treated with estrogen. The affinity constant for the progesterone-binding sites in cytosol from bladders with lesions was 10(9) M-1, the same as that reported for progesterone receptors in other target tissues for estrogen. The binding sites are specific for progesterone and are not competed for by 17 beta-estradiol, 5 alpha-dihydrotestosterone, or aldosterone.


Stimulation of differentiated functions in human melanoma cells by tumor-promoting agents and dimethyl sulfoxide.

Treatment of cultured human HO melanoma cells with the mouse skin tumor promoter phorbol-12-myristate-13-acetate (PMA) at 5 x 10(-10) to 5 x 10(-7) M resulted in a dose-related inhibition of growth and a stimulation of differentiated functions. These included melanin synthesis and formation of dendrite-like structures. Higher doses of phorbol dibutyrate, a less potent tumor promoter, were required to produce an effect comparable to that of PMA for dendrite induction. Phorbol and two other phorbol esters, which lack tumor-promoting activity, were either inactive or elicited a poor response. In addition to morphological changes, treatment with PMA altered glucosamine incorporation into membrane gangliosides. After PMA treatment, glucosamine incorporation increased 8- to 10-fold in the GM3 ganglioside and decreased 2-fold in the GM1 ganglioside, as compared to phorbol or untreated control. Inhibition of cell growth and stimulation of melanin synthesis were also observed after treatment of the HO cells with dimethyl sulfoxide. Unlike the tumor-promoting agents, dimethyl sulfoxide did not induce the formation of dendrite-like structures in the cells. These findings indicate that HO melanoma cells can be stimulated into terminally differentiated cells after treatment with tumor-promoting agents such as phorbol diesters.


Response of exponentially growing, stationary-phase, and synchronized cultured human colon carcinoma cells to treatment with nitrosourea derivatives.

The lethal effects of two nitrosourea derivatives, 1,3-bis(2-chloroethyl)-1-nitrosourea and 4-[3-(2-chloroethyl)-3-nitrosoureido]-cis-cyclohexanecarboxylic acid, on a continuous line of human colon carcinoma cells (LoVo cells) were investigated. The survival response of exponentially growing and stationary-phase LoVo cells to both drugs were of the threshold exponential type. Survival was identical whether drugs were dissolved in medium or in Hanks' balanced salt solution. In contrast to previous results obtained for human lymphoma cells, 1,3-bis(2-chloroethyl)-1-nitrosourea exerted a progressively greater killing effect on LoVo cells as the incubation time was prolonged, while 4-[3-(2-chloroethyl)-3-nitrosoureido]-cis-cyclohexane-carboxylic acid, under similar circumstances presented decreased lethality in comparison to both 1,3-bis(2-chloroethyl)-1-nitrosourea and the effect previously observed for the lymphoma cells. Although no recovery from potentially lethal damage was noted for both exponentially growing and stationary-phase cells treated with each agent, in split-dose experiments, LoVo cells were able to recover from sublethal damage. No significant cell cycle stage dependent differences in killing ability were observed for the two agents.


Neutral fucolipids and fucogangliosides of rat hepatoma HTC and H35 cells, rat liver, and hepatocytes.

Neutral fucolipids and fucogangliosides of normal rat liver, normal rat hepatocytes, and rat hepatoma H35 and HTC cells have been compared. H35 cells were characterized by accumulation of a relatively large quantity of a few neutral fucolipids and a major fucoganglioside. HTC cells were characterized by accumulation of several neutral fucolipids and by the absence of fucogangliosides. Those fucolipids which accumulated in hepatoma cells were absent in normal rat liver and normal hepatocytes. Normal hepatocytes and normal rat liver contained a low quantity of fucolipids which migrated differently on thin-layer chromatography from those which accumulated in hepatoma cells. In addition to these qualitative differences, the quantity of total fucose in purified glycolipid fraction was much higher in H35 and significantly higher in HTC cells than that in rat liver. A fucoganglioside which accumulated in H35 cells was tentatively characterized as a fucosylated GM1 ganglioside with the following structure: (formula: see text). The results indicate that anomalous fucosylation on glycolipid may take place in hepatoma cells.


Nuclear retention of all steroid hormone receptor classes in the hamster renal carcinoma.

The estrogen-induced hamster renal carcinoma contains appreciable amounts of all cytosolic steroid receptor classes sedimenting as 7 to 8S moieties following sucrose gradient centrifugation. Their relative concentrations, expressed in fmol/mg protein +/- S.E. are progesterone (1496 +/- 23) greater than estradiol (218 +/- 3) greater than 5 alpha-dihydrotestosterone (154 +/- 7) greater than dexamethasone (138 +/- 3) greater than aldosterone (40 +/- 2). No cross-competition is apparent for either estrogen or progesterone receptors, but progesterone competes for both androgen and adrenocorticoid binding. These hormone-receptor complexes undergo nuclear translocation using purified tumor nuclei and in tissue minces at elevated temperatures. Salt-extractable nuclear receptors for estrogen (5S), androgen (3.2S), progesterone (2.7S), and corticosteroid (3.5S) have been identified. The existence of five specific steroid receptors within a single tissue is unique and provides a novel model for studying the interrelated actions of all steroid hormones and their therapeutic responses in a hormone-dependent neoplasm.


Models for development of nonreceptor methods for distinguishing androgen-sensitive and -insensitive prostatic tumors.

From the original Dunning R-3327 rat prostatic adenocarcinoma, several distinct sublines have been obtained. These sublines include a well-differentiated, slow-growing, androgen-sensitive tumor (R-3327-H); a well-differentiated, slow-growing, androgen-insensitive tumor (R-3327-HI); and a fast-growing, androgen-insensitive, anaplastic tumor (R-3327-AT). These three sublines were compared in order to develop new model methods for the prediction of the androgen sensitivity and the degree of differentiation of prostatic adenocarcinomas. The R-3327-AT was very distinct in all parameters examined except the tissue protein electrophoretic patterns which contained a uniform pattern in all tumors. The significant differences between R-3327-H and -HI sublines were (a) the inability of testosterone to stimulate DNA synthesis in the R-3327-HI tumor and (b) the difference in the enzymatic profiles of these sublines. The specific activity of three enzymes (3 alpha-hydroxysteroid dehydrogenase, leucine aminopeptidase, lactic dehydrogenase) increased while the activity of another three enzymes (6 alpha,7 alpha-hydroxylase, 5 alpha-reductase, alkaline phosphatase) decreased in the sublines which are androgen insensitive and less differentiated. An arbitrary index was constructed, based upon these enzyme differences, which clearly discriminates the degree of androgen sensitivity and differentiation of these R-3327 rat prostatic adenocarcinomas.


Differences in the removal of N-methyl-N-nitrosourea-methylated products in DNase I-sensitive and -resistant regions of rat brain DNA.

The present study with N-methyl-N-nitrosourea and rat brain DNA was performed in order to study the distribution of alkylated products and the difference in the removal of these products from DNase I-sensitive and -resistant regions of DNA. Nuclei were isolated from N-[3H]methyl-N-nitrosourea-treated rats and incubated in the presence of DNase I (5 microgram/ml). Digested DNA was further hydrolyzed in 0.1 N HCl, and the alkylated products were identified by chromatography on a cation-exchange column. Resistant DNA was isolated, hydrolyzed, and again the alkylated products were determined. At 4 hr, the specific activity of all the alkylated products in the sensitive regions of DNA was several times higher than the resistant fraction. The rate of loss for the products was greater in the sensitive than the resistant fractions. O6-Methylguanine was removed from the sensitive regions but was more stable in the resistant regions. These results suggest that DNase I-sensitive regions of the DNA are preferentially alkylated by N-methyl-N-nitrosourea and that the alkylated products, including O6-methylguanine, are selectively removed from the DNase I-sensitive regions of the DNA.


Metabolism of the liver carcinogen N-nitrosopyrrolidine by rat liver microsomes.

This report represents a study of the total metabolism of the hepatocellular carcinogen, N-nitrosopyrrolidine (NO-PYR), by rat liver microsomes and postmicrosomal supernatant. [2,5-14C]NO-PYR, which is totally extractable from aqueous solution with methylene chloride, is converted to radioactive nonmethylene chloride-extractable products by these fractions. The initial rate of conversion to nonmethylene chloride-extractable products follows simple Michaelis-Menten kinetics with an apparent Km of 3.6 x 10(-4) M NO-PYR. The major products of NO-PYR metabolism by rat liver microsomes and postmicrosomal supernatant have been isolated and identified. One product of metabolism of NO-PYR is 2-hydroxytetrahydrofuran formed by alpha-hydroxylation by the microsomes. In the presence of postmicrosomal supernatant enzymes, this compound exists only as a transient intermediate which is rapidly converted to 1,4-butanediol or gamma-hydroxybutyrate. These compounds may be cycled into general cellular metabolism resulting in the production of CO2. Two minor pathways of metabolism have also been found.


Migration of intraperitoneally injected thyroid cells in the Amazon molly, Poecilia formosa.

We have previously reported the development of an extensive invasive growth of the thyroid gland of the gynogenetic teleost, Poecilia formosa (the Amazon molly), following i.p. injection of UV- or gamma-irradiated thyroid cells. This result was surprising by comparison with mammalian work, in which the thyroid is rarely the site for tumor metastases, but the anatomy of the circulation of fish is different from mammals, and in fish the gills and thyroid gland would be among the first tissues in which injected cells might be arrested. Techniques using a fluorescent dye, 125I membrane label, or [3H]thymidine label were used to follow the distribution of i.p. injected cells in the Amazon molly. Fish sampled as soon as 30 min after injection had some labeled cells dispersed in the connective tissue around the ventral aorta and in the bases of the gills, and by 1 to 4 hr large numbers of cells had moved into the thyroid region. A few cells still persisted there 200 hr later. Experiments on the distribution of heat-killed cells indicated that the initial distribution of the cells was largely governed by mechanical factors. Injected cells would appear to be disseminated in fish by mechanisms similar to those in mammals.


Gastrointestinal carcinogenesis in germ-free rats given N-methyl-N'-nitro-N-nitrosoguanidine in drinking water.

This study was designed to clarify the role of gut microflora in tumorigenesis by a comparison of tumor production between male germ-free and conventional Wistar rats given N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 100 microgram/ml in drinking water. Ninety-one % of conventional MNNG-treated rats that died or were killed by Day 314 of the experiment developed tumors in the gastrointestinal tract, whereas only 17% of germ-free treated rats developed such tumors. In addition, large tumors, some 5 cm or more in diameter, were frequently observed in the conventional rats, whereas only small tumors 0.4 to 1.2 cm in diameter were present in the germ-free rats. Furthermore, multiple tumors including double tumors were often found in the conventional rats, while such tumors never appeared in the germ-free rats. The results suggest that gut microflora might exert a promoting influence on tumorigenesis by MNNG in the gastrointestinal tract. The promoting influence of the microflora in conventional rats might not be of a simple nature, since the influence of a variety of factors modified by the micorflora on tumorigenesis by MNNG p.o. is unavoidable.


Calcium transport and translocation of adenine nucleotides in mitochondria from Morris hepatoma 3924A.

The interaction of Ca2+ with Morris hepatoma 3924A mitochondria and its effect on the adenine nucleotide translocation have been studied. The characteristics of the Ca2+ transport process in mitochondria from Morris hepatoma are not significantly different from those of normal liver mitochondria. The Km for Ca2+ is 2 to 3 microM, and the rate versus concentration curve exhibits hyperbolic kinetics. A lower activity of the adenine nucleotide translocation was found, probably due to the high endogenous Ca2+ content of Morris hepatoma mitochondria (123 +/- 15 nmol Ca2+ per mg protein). No further inhibition of the translocase activity was observed after isolated mitochondria had accumulated extra amounts of Ca2+. The total amount of adenine nucleotides in tumor mitochondria is one-half those present in control liver, and a significantly lower percentage of the pool is present as adenosine 5'-monophosphate.


Potentiating effect of beta-aminopropionitrile on the DNA synthesis induced by isoproterenol.

Young mice chronically treated with beta-aminopropionitrile (BAPN) were subsequently given injections of graded doses of isoproterenol, and the response of various tissues to IPR in synthesizing DNA was analyzed. The results indicate that chronic administration of BAPN potentiates the effect of IPR for inducing DNA synthesis in the parotid, submandibular gland, kidney, and urinary bladder. The effect of BAPN was most pronounced in the submandibular gland. Namely, acinar cells in the submandibular gland normally respond less to IPR than do those in the parotid but respond more when mice have been treated with BAPN. It was suggested that the structural modification of collagen may counteract the blockage of events leading to DNA synthesis in epithelial cells in vivo.


Comparative metabolism of 2-[bis(2-chloroethyl)amino]tetrahydro-2-H-1,3,2-oxazaphosphorine-2-oxide (cyclophosphamide) and its enantiomers in humans.

The comparative metabolism of the enantiomers of cyclo phosphamide and of the racemate has been studied in humans. Four patients were each given, sequentially, the racemate, the (+)-enantiomer, and its (-)-antipode. The plasma levels of parent drug and the urinary output (24 hr) of unchanged drug and of two enzymatically produced metabolites, 4-ketocyclophosphamide and carboxyphosphamide, were determined using mass spectrometry-stable isotope dilution. There was no significant difference between the three forms of cyclophosphamide with respect to plasma half-life (beta phase) or in the urinary outputs of the drug or of carboxyphosphamide. The output of 4-ketocyclophosphamide after administration of (+)-cyclophosphamide was significantly greater than that produced from the racemate. Cyclophosphamide recovered from the urine of patients given the racemate was either racemic or only slightly enriched in the (-)-enantiomer. The two enantiomers were almost equally bound to plasma protein. Based on these metabolic studies alone, there is little reason to predict that the enantiomers will differ from each other or from the racemate in their therapeutic effects in humans, but there are other factors, e.g., stereoselective uptake of the intermediary 4-hydroxylated metabolites by neoplastic cells, which could elicit such differences.


Characterization of rat hepatoma glucosamine 6-phosphate synthase and its relation to liver and fetal forms.

Glucosamine 6-phosphate synthase (EC 5.3.1.19) purified from various rat tissues by a procedure involving chromatography on diethylaminoethyl Sephadex and hydroxylapatite were characterized by means of isoelectric focusing. The non-hepatic isozyme, previously reported to be present in Yoshida sarcoma, has a pI of 4.1 and is distinguished from the hepatic isozyme, with a pI of 5.0. The pI 4.1 form is the major one in all of the fast-growing, transplantable hepatomas studied. Although not detectable in 19-day fetal liver or normal adult liver, the pI 4.1 form has been observed in the whole 12-day fetus and adult brain as almost the sole form of glucosamine 6-phosphate synthase.


Glucosamine 6-phosphate synthase of regenerating rat liver.

When rats were subjected to partial hepatectomy, glucosamine 6-phosphate synthase (EC 5.3.1.19) of the remaining liver underwent alterations both in activity and in molecular form. To study the molecular alterations, glucosamine 6-phosphate synthase was purified from regenerating as well as control liver and was analyzed by isoelectric focusing. Although control liver exhibited only one form of glucosamine 6-phosphate synthase with a pI of 5.0, sequential and transient appearance of three other forms, with pI values of 4.3, 4.8, and 4.5, respectively, was observed for regeneration liver within 72 hr following partial hepatectomy. Laparotomy, on the other hand, induced in the liver only the pI 4.8 form, and injection of a mixture containing triiodothyronine, amino acids, glucagon, and heparin induced only the pI 4.3 and 4.5 forms. It therefore appears that the pI 4.3 and 4.5 forms, but not the pI 4.8 form, are associated with hepatic DNA synthesis. The pI 4.8 form is induced in the liver in response to surgical stress.


DNA strand scission in vivo and in vitro by auromomycin.

The mechanism of action of auromomycin, a new tumor-inhibitory antibiotic, was studied in a growing culture of mouse lymphoblastoma L5178Y cells and with isolated viral DNA. Auromomycin prevented growth of L5178Y cells completely and irreversibly at antibiotic concentrations higher than 0.03 microgram/ml. DNA synthesis was preferentially inhibited by the antibiotic, whereas RNA and protein syntheses were not significantly affected. In synchronous cultures of L5178Y cells, results indicated that limited auromomycin-induced inhibition of DNA synthesis may occur independently of a much stronger inhibition of mitosis. In a short incubation period, a marked strand scission in cellular DNA of auromomycin-treated L5178Y cells was observed by an analysis of alkaline sucrose gradient centrifugation. In vitro, the antibiotic also induced strand breaks in linear duplex T-7 phage DNA and in the supercoiled circular duplex of SV40 DNA. 2-Mercaptoethanol neither enhanced nor was required for strand scission of isolated DNA by auromomycin. These data indicate that the mechanism of the antitumor activity of auromomycin is different from that of bleomycin, neocarzinostatin, or macromomycin.


Age-related decrease of ultraviolet light-induced DNA repair synthesis in human peripheral leukocytes.

The capacity for ultraviolet light-induced DNA repair synthesis, studied in peripheral leukocytes from 58 healthy subjects 13 to 94 years old, was found to vary greatly between individuals. A negative, statistically significant correlation was obtained between age and this synthesis, indicating a decrease in repair capacity with age. An age-related decrease in DNA repair may increase the susceptibility of cells to agents causing DNA damage, i.e. carcinogens and certain cytostatic drugs.


Glycosaminoglycans and proteoglycans of normal and tumoral cartilages of humans and rats.

Differences in the glycosaminoglycans and proteoglycans synthesized by "young," "adult," and tumoral chondrocytes are reported. Young cartilage and human chondrosarcoma contain chondroitin 4- and 6-sulfates, whereas adult human cartilage contains almost exclusively chondroitin 6-sulfate. High keratan sulfate content is reported in adult cartilage, whereas it is almost absent in young and tumoral cartilages. The electrophoretic pattern and keratan sulfate content in these proteoglycans from adult cartilage are clearly distinct from those of the young and tumoral cartilages. The high molecular weight is the distinguishing property of the glycosaminoglycan synthesized by tumoral chondrocytes.


Immunotherapeutic response of concanavalin A-bound L1210 vaccine enhanced by a streptococcal immunopotentiator, OK-432.

Immunotherapeutic response to concanavalin A (Con A)-bound L1210 murine leukemic vaccine and immunopotentiators was examined in histocompatible animals bearing a small burden L1210 leukemic cells. When combined with Con A-bound vaccine, a streptococcal immunopotentiator, OK-432 (NSC B116209), prepared from Streptococcus pyogenes, was potent in antitumor therapy and resulted in a number of cured animals. Administration of either Con-A-bound vaccine or OK-432 alone did not produce any beneficial effect on leukemic animals. The enhanced therapeutic response was dependent on the effectiveness of the dose and timing of the administration of OK-432 when given after vaccination. Combined modality of Con A-bound L1210 vaccine and OK-432 was not effective in animals bearing P388 murine leukemic cells, which indicates specificity of therapeutic response. In enhancing the therapeutic potency of Con A-bound leukemia vaccine, pyran copolymer (NSC 46015) was as effective as OK-432, whereas Bacillus Calmette-Guérin and Corynebacterium parvum were far less effective. When combined with OK-432, therapeutic response to Con A-bound L1210 vaccine was much greater than response to glutaraldehyde-, mitomycin C-, or Vibrio cholerae neuraminidase-treated L1210 vaccine.


Cancer overview.

Cancer is a very common group of diseases. There are 700,000 cases of the disease reported each year, and about 350,000 persons die of these diseases. There is extensive literature and documentation relative to alcohol as a probable factor in the development of head, neck, and esophageal cancers among others. However, there is little in the literature relative to the interrelationship of alcohol and cancer in the area of cancer treatment. Dignity of the dying patient and his family is maintained, and home care is a vital component. While alcohol use is not generally written about, it is used as desired by the patient. A "Brompton's-type cocktail," a mixture of morphine, cocaine, and alcohol, forms a basis of pain control in English and many United States' hospices. One purpose in today's conference is to look at another lifestyle factor, alcohol, to determine its role alone or in conjunction with other factors in the causation of cancer. Are there high risk groups which need attention in alcohol cancer prevention programs?


The family physician and confrontation in alcoholism.

Identification of alcoholism may be crucial for ensuring the most appropriate treatment for patients suffering with cancer. In this setting, the family physician can best provide the leadership in coordinating the critical resources necessary for successful treatment by utilizing the family, the minister, the employer, and most important, alcoholism counselors and Alcoholics Anonymous. To avoid a realistic assessment of alcoholism in a cancer patient will only perpetuate individual sorrow and suffering. The physician may win the battle against cancer, but the war can be lost by the self-destruction resulting from alcoholism.


The disease of alcoholism.

A pharmacological view of narcotic addiction and alcoholism, or sedative addiction, serves as a basis for the further examination of these complex illnesses. The disease called alcoholism (sedativism) consists, however, of much more than just a chronic or periodic addiction to a sedative drug. The urgent need to understand the psychological and societal parameters of this illness is underscored by the likelihood that alcoholism represents the major factor in the cause of death of young adults ( less than 45 years) in the United States today. Its further role as a carcinogen is only recently achieving adequate examination.


Epidemiology of alcohol and cancer.

There is still insufficient knowledge of the distribution of drinking habits in human populations, and more descriptive surveys are needed. Both prospective and retrospective epidemiological studies indicate that alcohol consumption is a cancer hazard. Prospective studies on excessive drinkers have shown an increased risk for cancer of the mouth, pharynx, larynx, esophagus, liver, and lung. Retrospective studies have confirmed this excess risk. For cancers of the buccal cavity, pharynx, larynx, and esophagus, the effect of drinking has been shown to be associated with the effect of smoking. In the case of esophageal cancer, these two effects are independent, and the observations made are consistent with a multiplicative model. Primary liver cancer is also associated with alcohol consumption, probably by a less direct action; the importance of the impact of alcohol on primary liver cancer is probably underestimated. Animal experiments have not shown that ethanol alone has a carcinogenic effect, and the mechanisms by which alcoholic beverages act on humans remain unknown. The proportion of cancer cases at sites known to be associated with alcohol consumption is approximately 8% in most population groups in the United States. This indicates that a sizeable proportion of cancers is potentially preventable if appropriate action is taken.


Epidemiological opportunities in alcohol-related cancer.

Alcohol combined with tobacco smoking is an established risk factor for cancers of the oropharynx, esophagus, and larynx. It should be possible to clarify further the role of alcohol itself, the modifying effects of tobacco, dose-response relationships, and nutritional cofactors. Studies are also needed to delineate the steps by which alcohol consumption leads to liver cancer and to resolve the suggestion that certain beverages may predispose to other cancers including those of the pancreas and rectum. Epidemiological investigations should be combined with experimental work to identify hazardous fractions in alcoholic beverages and to delineate the mechanisms by which alcohol promotes carcinogenesis. Epidemiologists and biometricians may also contribute toward the development of programs aimed at primary prevention and early detection of cancers related to alcohol and tobacco. Incorporation of research questions into data collection systems deserves serious questions into data collection systems deserves serious consideration as a means of obtaining additional valuable information for etiological studies.


Rehabilitation and continuing care of cancer patients.

Health professionals are responsible for helping cancer patients and their families readapt to society. Frequently, coping with alcoholism is an integral facet of the rehabilitative process, and in this respect a patient's family often presents more of a problem than does the patient himself. Physicians have shown a lack of knowledge in detecting and dealing with the alcohol problems of their patients. Health professionals involved in the management of patients with cancer must learn to recognize alcoholism to ensure that the problem is referred to an organization or individual who will cope with it adequately. Volunteer groups such as Alcoholics Anonymous, as well as the clergy, some psychiatrists, psychologists, and sociologists, have demonstrated expertise in dealing with alcoholism.


Continuing care of the cancer patient as a social engineering problem.

Care of the cancer patient has become fractionated toward maintaining the physiological integrity of the patient's life support systems, e.g., the hematopoietic or neurological systems. The concept of total continuing care for the cancer patient is illustrated in this description of a pilot hospice project in a community medical center. The project also illustrates the concept of team support by the health care staff and volunteers. The total continuing care unit described provides support and care which makes living not just possible, but worthwhile, for cancer patients.


Possible relationships of alcohol in membranes to cancer.

Ethanol can be used as a chemical tool to alter membrane fluidity or composition, or both, and to study the effects on induction, growth, spread, or treatment of cancers. Ethanol rapidly equilibrates with total body water and enters all cell membranes. Ethanol molecules are intercalated between the lipids of the bilayer membranes. This expands membranes and increases their fluidity, which in turn affects cell agglutination, phagocytosis, membrane transport, membrane enzyme activities, and many other membrane functions. After 3 to 5 days of continuous ethanol administration, the original membrane fluidity is restored by the incorporation of "stiffening" lipids, such as cholesterol, into the bilayer and by the increase of the chain length and saturation of fatty acids. The desired membrane effects (increased fluidity or altered membrane composition) can be obtained by adjusting time-dose relationships of ethanol administration. There may be an important role of moderate alcohol consumption in cancer biology that is not presently recognized by epidemiological studies because both cancers and moderate alcohol consumption are very prevalent in the general adult population. Moderate, social alcohol use could potentially either suppress or enhance the induction, growth, spread, or therapy of cancers. Such potential roles of alcohol in cancer biology could easily be tested in animals by incorporating the feeding of alcohol-containing diets into experiments that follow standard cancer protocols.


Phase II study of adriamycin, 5-fluorouracil, levamisole, and irradiation in carcinoma of the bladder.

Since July 1976, 19 patients with carcinoma of the bladder have been treated with adriamycin, 5-fluorouracil, and levamisole combined with radiotherapy (60 Gy [6000 radsA1/24 fractions/6 weeks). Chemotherapy and radiotherapy were initiated simultaneously, with the entire treatment lasting 7--8 months. Three months after the completion of radiotherapy, 14 of the 18 patients in whom cystoscopy was performed were found to be in complete remission. Overall, 17 of the 19 patients have responded to the treatment and 15 patients have at some time shown complete remission. The toxic effects seen were myelosuppression, nausea, vomiting, diarrhea, loss of weight, and alopecia. Thirteen patients received the entire treatment as outpatients while six patients had to be hospitalized for a period of 8--14 days because of severe side effects, especially in Weeks 3--8. Serious complications such as bowel perforation were not seen, but one patient died with septicemia as a result of agranulocytosis, which was attributed to the treatment with levamisole.


Phase II study of 5-fluorouracil, methyl-CCNU, and daunorubicin in colorectal cancer: a Cancer and Leukemia Group B study.

The three-drug combination of 5-fluorouracil, methyl-CCNU, and daunorubicin was evaluated in 38 patients with unresectable or metastatic carcinoma of the colon. There were five partial responses and one complete response for an overall response rate of 16%. Although toxicity was tolerable, daunorubicin failed to add to the 5-fluorouracil and methyl-CCNU combination.


Role of cytoreductive surgical treatment in the management of advanced ovarian cancer.

Twenty-eight abdominal operations were performed on 26 consecutive patients with stage III and IV ovarian carcinoma over a 3-year period. The goal of each operation was to excise all tumors greater than 1 cm in diameter. This goal was achieved in 12 of 15 primary operations, in seven of nine operations after induction chemotherapy, and in three of four operations performed for tumor recurrence. There were two major complications but no postoperative deaths. Analysis of survival and disease status indicated that patients having operations followed by chemotherapy fared the best. Analysis of prognostic variables suggested that the administration of combination chemotherapy was the most important determinant of survival once the surgical goal had been accomplished. In this latter group, all nine patients who were evaluable by laparoscopy had responded to adriamycin-cyclophosphamide and eight of the nine had complete responses.


Different chemotherapeutic sensitivities and host factors affecting prognosis in advanced ovarian carcinoma versus minimal residual disease.

Treatment of patients with advanced ovarian carcinoma (stages IIIB and IV) using either cyclophosphamide alone (1 g/m2) or cyclophosphamide (500 mg/m2) plus adriamycin (40 mg/m2) by iv injection every 3 weeks each produced partial regression in approximately one third of the patients. Survival curves and time-to-progression curves for the two regimens were nearly identical in these patients with advanced disease. These same regimens produced different results when used monthly in patients who had minimal residual disease (stages II and IIIA). In patients with minimal residual disease the therapeutic index of the combination regimen was superior to that of cyclophosphamide alone. Prognosis was better overall among patients with minimal residual disease than among patients with advanced disease. Within the minimal-disease group grossly complete excision of tumor prior to chemotherapy was associated with still better prognosis. Among patients with advanced disease, prognosis was significantly better for older patients despite their generally less favorable performance scores. Much of this prognostic superiority appeared to be related to menopausal status and presumably to the depletion of endogenous estrogens in the older patients.


Chemotherapy for murine ovarian cancer: a rationale for ip therapy with adriamycin.

The metastatic spread of a trnasplantable murine ovarian cancer is similar to the spread of ovarian cancer in patients with advanced disease, making it a useful model to investigate novel experimental therapies. The ip inoculation of 10(6) tumor cells into C3HeB/FeJ mice leads to the formation of ascites, sub-diaphragmatic tumor deposits, intra-abdominal tumors, and death within 25 days. Adriamycin (ADR) was found to be an active agent against this murine ovarian cancer. The effects of ADR were dependent upon the route of administration. A single ip LD10 dose of ADR (5 mg/kg) administered 2 days after inoculation with 10(6) tumor cells produced long-term survival (greater than 60 days) in 70% of the mice. An iv LD10 dose had no effect on survival. The survival advantage of ip ADR (compared to the iv route) was found to be related to: (a) a greater suppression of DNA synthesis in the tumor; (b) a rapid penetration of ADR into the nuclei of ascites tumor cells and into sub-diaphragmatic tumor deposits; and (c) significantly higher levels of ADR in tumor cells following ip administration. The ip route may also be less cardiotoxic since the peak levels after an iv dose were three times greater than after an equal ip dose. If local toxicity does not prove to be a major problem, then ip ADR may be a useful mode of therapy in patients with intra-abdominal tumors.


Chemoimmunotherapy in the management of primary stage III ovarian cancer: a Gynecologic Oncology Group study.

The effectiveness of chemoimmunotherapy was evaluated in 45 previously untreated stage III ovarian cancer patients. Response rate, progression-free interval, and survival were considerably better than in a similar group of 63 patients treated with a single alkylating agent alone.


Phase II trial of dianhydrogalactitol in metastatic malignant melanoma: a Southwest Oncology Group study.

Dianhydrogalactitol given iv in a schedule of 30 mg/m2/day for 5 consecutive days every 4 weeks was administered to 27 patients with metastatic malignant melanoma. All patients had received extensive prior therapy including chemotherapy and had progressive disease at the start of the study. Of 24 patients evaluable for response, 21 demonstrated progressive disease and three had stable disease for periods of from 4 to 11 months. No objective responses were observed. Two of the remaining three patients died 6 and 10 days after entry in the study, while the third refused to return after one drug course. Adverse effects included myelosuppression in eight patients, nausea and vomiting in five patients, and alopecia in one patient. Dianhydrogalactitol is considered to be insignificantly active in the secondary treatment of metastatic malignant melanoma at the dose and schedule studied.


Experimental skin necrosis produced by adriamycin.

Skin ulceration caused by extravasation of Adriamycin follows a severely protracted course accompanied by considerable morbidity. To develop an animal model of Adriamycin ulceration, we compared intradermal injection of Adriamycin to injection beneath the panniculus carnosus with varying drug volumes and concentrations. Injections beneath the rat panniculus carnosus caused only irregular ulcerative lesions. Intradermal injection produced predictable, uniform skin necrosis and ulceration. Both increasing volume and increasing concentration of Adriamycin caused proportionate increases in skin ulcer size and in time required for healing. A critical concentration range for Adriamycin necrosis is 0.010--0.020 mg/ml, suggesting that the drug would have to be greatly diluted to reduce clinical skin ulceration. Adriamycin-induced skin necrosis heals at a slower rate than surgically created skin defects of similar size, indicating a reduced rate of wound contraction. Removal of the necrotic skin allows faster healing, although still slower than normal, due to removal of splinting effect. Histology shows early skin necrosis, with acute inflammation developing after 1 week. Epidermal hypertrophy is present at the edges of the necrosis. The small vessels remain patent. Multiple small vesicles of unknown etiology are seen in the necrotic dermis.


Exposure to chemicals, physical agents, and biologic agents in mycosis fungoides and the Sézary syndrome.

The history of occupational, environmental, and/or iatrogenic exposure to potential carcinogenic agents was obtained at the time of onset of skin disease in 43 of 44 patients with cutaneous T-cell lymphoma (mycosis fungoides and the Sézary syndrome) entering a National Cancer Institute therapeutic trial. A history of multiple exposures to these agents was common, the two most frequent being chemicals (91% of patients) and drugs (86%). Mean duration of expsosure was 13 years for chemicals and 18 years for drugs. The most common chemicals were air pollutants (39%), pesticides (36%), solvents and vapors (30%), and detergents and disinfectants (14%). Increased severity of disease was seen with increaed duration of chemical exposure in stage V cutaneous T-cell lymphomas only. The most frequent drugs besides tobacco (86%) were analgesics (20%), tranquilizers (18%), and thiazides (14%). Second cancers occurred in four patients, including two renal cell carcinomas, and a family history of cancer was present in 11. Fourteen of 22 patients questioned had recurrent herpes simplex. Patients with chronic skin disease who have long-term exposure to combinations of chemicals, physical agents, and biologic agents, are heavy smokers, or have recurrent herpes simplex appear to be prime candidates for developing mycosis fungoides or the Sézary syndrome.


Mycosis fungoides: epidemiologic observations.

An analysis of cases from a multi-hospital, pathologically verified clinical series and of deaths from US mortality statistics available at the county level for 1950--1975 (excluding 1972) was made in order to obtain information on the etiology of mycosis fungoides (MF). Despite the absence of an appropriate comparison group, the cases seemed to have a high frequency of antecedent allergies, fungal and viral skin infections, sun sensitivity, familial aggregation of lymphoma and leukemia, and employment in a manufacturing occupation (especially petrochemical, textile, metal, and machinery industries). The mortality survey revealed a predominance among males, nonwhites, and residents of the northeastern US, the latter due partly to an association between MF mortality and population size. The influence of occupational exposures was suggested by the excessive MF mortality rates in counties where petroleum, rubber, primary and fabricated metal, machinery, and printing industries were located.


Methoxsalen photochemotherapy for mycosis fungoides.

Methoxsalen photochemotherapy (PUVA; psoralen plus ultraviolet light) is effective in the treatment of mycosis fungoides (MF). The mechanism of this beneficial effect is unknown but probably involves covalent photo-binding of methoxsalen molecules to pyrimidine bases in DNA at the cellular level and impaired T-cell function or survival at the tissue level. Eleven patients (seven with plaques and four with erythroderma) were referred for PUVA therapy because of poor response to conventional therapy. Seven patients had complete clearing of skin lesions and three improved markedly. All ten of these patients experienced good to excellent control of the disease while receiving maintenance therapy, resulting in a virtual remission lasting for greater than 2 years in four patients. Of the six patients who discontinued PUVA during the followup period, three died within 1 year and three experienced progression of MF despite conventional therapy. The very promising results in this small patient group should encourage further studies of PUVA for cutaneous T-cell lymphomas.


Photochemotherapy for mycosis fungoides: long-term followup study.

A 3-year followup on ten of 12 patients with mycosis fungoides (MF) treated with photochemotherapy (PUVA; psoralens + ultraviolet light) reveals eight patients whose diseases have remained clear while receiving maintenance PUVA. An additional 35 patients with MF have been successfully treated with PUVA. Eight cases of parapsoriasis en plaque have cleared with PUVA. PUVA is indicated in early eczematous and plaque stages of MF. It may be used as adjunctive therapy in tumor-stage MF treated with X-ray therapy or chemotherapy.


Direct effects of dopamine, orciprenaline and norepinephrine on the right and left ventricle of isolated canine hearts.

In 31 canine heart-lung preparations the direct cardiac effects of dopamine were compared to those of norepinephrine and orciprenaline. All three agents increased heart rate by 50-60% to a maximum of 240-250 beats/min. The inotropic effects of the three drugs were not significantly different. Even with the highest doses cardiac arrhythmias were rarely observed. To achieve maximal chronotropic and inotropic effects, dopamine required four times higher doses than norepinephrine and orciprenaline. All three drugs increased right ventricular dP/dt max by about twice as much as left ventricular dP/dt max. Peak systolic pressure (P max) in the right ventricle was increased 4-5 times more than P max in the left ventricle while mean aortic pressure was elevated by 12-16 mm Hg and mean pulmonary pressure by 6-14 mm Hg. The positive inotropic effect of all three sympathomimetics and of CaCl2 was substantially more pronounced on the right than on the left ventricle.


Effect of inosine on ventricular regional perfusion and infarct size after coronary occlusion.

In 16 dogs inosine was infused at 0.5 mmol/min i.v. for 5 min beginning 15 min after coronary occlusion. Tracer microspheres were used to estimate flow in subepicardium and subendocardium of nonischemic, central ischemic, and borderline ischemic muscle. Estimates of flow before occlusion, 5 min after occlusion, during inosine infusion, 30 min after infusion and 60 min after infusion were obtained. Coronary occlusion reduced flow in the central ischemic regions by 75-95%. The reduction in flow was greatest in subendocardium. In the borderline regions subendocardial flow was reduced by 30% while subepicardial flow was unaffected. The major effects of inosine were seen in nonischemic and borderline ischemic regions. Flow in borderline subendocardium returned to its pre-occlusion value, and flow in nonischemic myocardium increased by approximately 60-80%. However, only in the ischemic regions was the increase in flow sustained for the entire 60 min. In 20 dogs infarct size was determined using nitro blue tetrazolium stain. In 10 controls infarct size was 20.1%, while in 10 inosine-treated dogs infarct size was 15.2% of left ventricular weight (p less than 0.01). Thus, following coronary occlusion inosine infusion was associated with an increase in perfusion of ischemic myocardium and a reduction in infarct size.


Influence of some hypotensive drugs on the effects of hypothalamic stimulation.

Clonidine, L-alpha-methyldopa, propranolol as well as noradrenaline, when injected directly into the hypothalamic paraventricular (PV) nucleus area, enhance the activity of this center. Lidoflazine appears to be inactive. Since the electrical, chemical and pharmacological stimulation of the PV area elicits the same depressor cardiovascular reactions, a specific differentiation of this nucleus as a depressor center is proposed. The above-mentioned drugs, when introduced into the  ventromedial nucleus area, enhance the pressor effects produced by stimulation of this nucleus. Thus, the hypothalamic depressor activity is not linked with an inhibition of the ventromedial pressor activity.


Return to work after myocardial infarction.

The period of inability to work after acute myocardial infarction was examined in follow-up studies. Without organized rehabilitation, 7.5% of patients past myocardial infarction returned to work after 6 months. After early mobilization in the hospital and intensive follow-up treatment in a rehabilitation center, 18% of the patients returned to work within 6 months. After the establishment of coronary groups in the vicinity of the patients' homes for the continuation of therapy, the percentage of patients able to return to work within 6 months rose to 42.5%, as shown in a later study. A further study checked the reliability of the rehabilitation clinic's prediction as to the expected date of return to work; only 24% of these predictions were correct. But this percentage rose to 82% after prolonged talks between physician and patient as to the motivation for return to work, modification of the conditions and a detailed report to the local physician.


Comparative dilator effect of verapamil and sodium nitroprusside in forearm arterial bed and dorsal hand veins in man: functional differences between vascular smooth muscle in arterioles and veins.

Verapamil and sodium nitroprusside induce a dose dependent dilatation in the forearm arterial bed and preconstricted dorsal hand vein in man. Verapamil is about twenty times more effective in dilating arterioles than veins; sodium nitroprusside is about ten times more effective in veins than arterioles. These differences can be related to functional differences between the two types of vessel.