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Influence of lodgement site on the proliferation of metastases of Walker 256 carcinoma in the rat.

The growth of s.c. Walker 256 carcinoma was found to be independent of secondary growths induced by i.v. injection. Tumour cells injected i.v. lodged mainly in the lungs, with small clusters of cells in the lymph nodes. The rate of cellular proliferation of these secondary growths of Walker carcinoma was significantly higher than that observed in the s.c. tumour. In addition, host lung tissue was found to inhibit the development of metastases, and it is postulated that the host tissue may produce a diffusible inhibitor and that differences in the effectiveness of these humoral factors may account, in part, for locational differences in tumour growth patterns.

Animals

Paramagnetic changes in cancer: growth of Walker 256 carcinoma studied in frozen and lyophilized tissues.

Samples of Walker 256 carcinoma grown in muscles of Sprague-Dawley rats were studied at low temperatures before and after lyophilization. The effects of lyophilization on the ESR spectra were different for tumours and normal muscle. Prior to lyophilization of a tumour sample, there was a decrease in free radicals, while after the lyophilization, there was an "increase". The "increase" was due to the lyophilized tumour having a narrower line, producing a greater peak-peak height measurement than in muscle, without an increase in the total number of spins. Exposure of lyophilized samples to air produced an increase in the intensity of the spectra and a change in line shape; also these effects differed for tumour and muscle. Mn++ levels were lower in tumour than in muscle, a difference eliminated by lyophilization. Poor growth conditions in tumours increased the occurrence of ESR spectra due to NO complexes with both heme and non-heme iron proteins. These results may help to resolve the principal controversies about experimental findings in ESR of tumours. At least part of the signals seen after lyophilization do not reflect free radicals in vivo. The signals after lyophilization reflect biochemical differences between tumour and muscle; spectroscopic data indicate that it is feasible to determine the molecular basis of these differences.

Animals

[Biochemical investigations of cancer cachexia. II. Depletion of glycogenolysis and stimulation of gluconeogenesis in Walker carcinoma 256 bearing rats (author's transl)].

150-200 g heavy, Walker-carcinoma bearing, male Sprague-Dawley-rats showed rapid, tumour weight dependent, loss of liver glycogen until complete depletion in tumour groups heavier than 40 g/animal. Simultaneously the glycogen mobilization after massive glucagon stimulation, was successivly diminished and finally abolished in different groups with increasing tumor weight. Concomitantly the spontaneous and stimulated activity of liver phosphorylase a was found markedly reduced in advanced tumour cachexia, the extent of stimulation of liver phosphorylase a activity by intracardial injections of epinephrine not being altered. Tumour induced inhibition of glycogen mobilization thus appears to have been excluded. To account for the relative late pronounced hypoglycemia in peripherial rat blood in face of the early loss of liver glycogen, accelerated gluconeogenesis has been postulated. In accord with this spontaneous rise in liver tyrosine amino transferase was found in tumour bearing rats along with a doubled maximal stimulation value after medrol injection as compared to control groups. This behavior could not be shown for liver alanine aminotransferase and liver fructose 1,6-di-phosphatase. The former showed no differences between control and tumour groups neither of spontaneous nor of stimulated activity. The latter showed only a very reluctant rise after massive stimulation by triamcinolone for 3 days in the control groups, the tumour bearing groups showing no deviation from spontaneous control values.

Alanine Transaminase

Blood flow and heat transfer in Walker 256 mammary carcinoma.

True thermal conductivity of 13 Walker 256 mammary carcinomas in noninbred Sprague-Dawley rats averaged 3.2+/-0.9 mW/cm/degrees C under physiologic conditions. A comparison of the effective thermal conductivity in 4 tumors with and without blood flow revealed large differences ranging from 14 to 132%. When the blood supply to the tumor was doubled or reduced to one-half, the effective thermal conductivity varied proportionally to the square root of the perfusion rate. The values of thermal conductivity were obtained from a tumor preparation in which blood flow was monitored continuously during temperature changes. These changes were measured by thermistors and produced by thermal probes incorporated by the growing tumor, not surgically inserted within the tissue at the time of measurement. Inasmuch as tissue necrosis was not a dominant factor, the data are interpreted to reflect the degree of difference in local perfusion of the neoplastic tissue.

Animals

Antineoplastic agents. 2. Structure-activity studies on N-protected vinyl, 1,2-dibromoethyl, and cyanomethyl esters of several amino acids.

Previously reported work on N-protected activated esters of phenylalanine has been extended to include N-protected vinyl, dibromoethyl, and cyanomethyl esters of several other amino acids. These compounds have been synthesized and evaluated in Ehrlich ascites carcinoma, Walker 256 carcinosarcoma, and and P388 lymphocytic leukemia tests. Among compounds tested were derivatives of tyrosine, tryptophan, glycine, leucine, proline, aspartic acid, glutamic acid, 4-aminobutyric acid, and 6-aminocaproic acid. Compounds of greatest potential interest from this study are N-carbobenzoxyglycine 1,2-dibromoethyl ester and N-carbobenzoxy-L-leucine 1,2-dibromoethyl ester. Both compounds were highly active in Ehrlich ascites test systems (33 mg/kg/day). The glycine derivative was also active in the Walker 256 test (2.5 mg/kg/day. Values for LD50's in mice were 148 mg/kg (0.37 mmol/kg) and 225 mg/kg (0.50 mmol/kg) for glycine and leucine derivatives, respectively; therefore, these compounds do not appear to be toxic at effective dose levels.

Acetonitriles

Decreased hepatic nuclear L-triiodothyronine receptors in rats and mice bearing transplantable neoplasms.

The concentration of hepatic nuclear T3 receptors was measured in isolated nuclei from athyreotic mice bearing thyrotrophic tumors and intact rats with Walker 256 carcinoma. Receptor concentration was reduced in all tumor-bearing animals. The mean receptor capacity of the Walker tumor-bearing rats [0.31 +/- 0.05 (SEM) ng/mg DNA] was significantly decreased from simultaneously assayed controls (0.47 +/- 0.04 ng/mg DNA; P less than 0.01). No change in the apparent equilibrium association constant was observed. In individual rats, the magnitude of the decrease in nuclear T3 receptor concentration was highly correlated with the decrease in tumor-free body weight. Additional studies showed that the decrease in nuclear receptors was not due to delayed equilibration of added T3 with nuclear sites in vitro or to an increase in endogenous hepatic T3 concentration. The plasma concentration of total and free T4 and T3 was decreased in tumor-bearing rats. Plasma TSH concentration, however, remained unchanged. Thus, these transplantable neoplasms seem to be associated with decreased hepatic nuclear receptors and low concentrations of plasma thyroid hormones. The unchanged plasma TSH suggests that the animals remained euthyroid.

Animals

Tumor reoxygenation and postirradiation vascular changes.

Postirradiation reoxygenation kinetics of the Walker 256 carcinoma were examined and related to radiation-induced changes in tumor vascular functions. Changes in the hypoxic cell component of irradiated Walker tumors may occur by two mechanisms. Reoxygenation of previously hypoxic cells may be due to an increased rate of blood flow in some parts of the tumor. Hypoxic tumor cells may also be eliminated by cell death due to prolonged or severe hypoxia in other parts of the tumor as a result of substantial damage to the tumor vasculature as seen in radiation-induced changes in vascular volume and blood vessel permeability. Reoxygenation of experimental tumors following single, massive doses of radiation may be different from that occurring in human tumors during the course of multifraction radiotherapy.

Animals

Ornithine as a possible marker of cancer.

The nonprotein amino acid ornithine is the major source of polyamines in mammalian physiological systems. Increased urinary polyamine levels have been demonstrated in humans with varied types of cancers. The metabolism of DL-[1-14C]ornithine monohydrochloride in rats with either Walker 256 carcinoma or chemically induced methylcholanthrene tumors was studied. Following the i.p. injection of 3 muCi[14C]ornithine per 100 g body weight, the decarboxylation of ornithine-yielding 14CO2 was monitored by utilizing the vibrating reed electrometer-ionization chamber model of Davidson and Schwabe. Tumor-bearing animals showed significant increases in ornithine metabolism as compared to controls; for Walker 256 the tumor-bearing animal to control ratio rose from 1.16 to 1.78, for methylcholanthrene implants it rose from 1.19 to 1.82, and for methylcholanthrene paintings it rose from 1.00 to 2.20. With tumor regression ornithine levels of metabolism in the tumor-bearing animals returned to base line or nearly base-line levels. These results encourage us in our attempt to develop ornithine as a biological marker of cancer.

Animals

Biosynthesis of cholesteryl 14-methylhexadecanoate in the liver of rats bearing transplantable tumors and during chemical carcinogenesis.

Biosynthesis of cholesteryl 14-methylhexadecanoate, cholesteryl palmitate and cholesteryl stearate was studied in the liver of rats bearing the Walker 256 carcinoma. Zajdela hepatoma and during chemical carcinogenesis following the administration of benzo[a]-pyrene. An up to 9-fold enhanced production of all these esters was found in liver homogenate during the 10--16th day after Walker tumor transplantation. Only the enzyme system esterifying cholesterol in the cytosol at pH 6.5 was stimulated while the activity of similar enzymes in mitochondria, microsomes and cytosol at an acid pH were not affected. Activity of the cytosol enzyme esterifying cholesterol at pH 6.5 was also enhanced during the active growth of Zajdela hepatoma and during the period of chemical carcinogenesis characterized by the appearance of first palpable subcutaneous tumors. Enhanced activity of cholesterol esterifying enzymes in the liver exactly coincided with periods of elevated levels of cholesteryl 14-methylhexadecanoate in the liver and blood plasma as described earlier. An increased demand of the tumor-bearing host for this cholesteryl ester utilized as a co-factor for enhanced protein synthesis is obviously met by its stimulated production in the liver tissue.

Animals