Behavior of mitochondria from normal and tumor cells in isopycnic sucrose gradients.

Rat and mouse liver mitochondria, when centrifuged in a sucrose density gradient (25--50%, w/w), showed the presence of heavy (H) and light (L) subfractions with buoyant densities 1.185 and 1.170--1.165 g/ml, respectively. Mild treatment with digitonin or EDTA (30 mM) shifted H-subfraction of mitochondria into the lighter zone of the gradient and as a result of this the mitochondria were distributed as a homogenous band with buoyant density 1.170--1.165 g/ml. Mitochondria isolated from both MD hepatoma and Zajdela rat hepatoma were characterized by a homogenous banding with buoyant density 1.160--1.165 g/ml. Regarding to this, the content and patterns of polyribosomes bound to outer membranes of mouse tumor mitochondria were studied. Analysis of polyribosomes as well as the results of RNA polyacrylamide gel electrophoresis indicate that the content of these polyribosomes in tumor mitochondria is less than that in normal liver ones. However, the decrease of cancer cell membrane-bound polyribosomes cannot account for the differences in buoyant densities of mitochondria from normal and tumor tissues.

Nucleic acids from subcellular fractions of N-nitrosodiethylamine-induced hepatoma in mice. II. Changes in gene expression during tumor progression.

The hybridization properties of in vivo labeled nuclear and mitochondrial ribonucleic acids of an transplantable hepatoma were studied. It was shown that during tumor progression the repression of nuclear genome revelead at its early stages was replaced by the de-repression at its later stages. The hybridizability of mitochondrial RNA with nuclear DNA was not changed.

Experimental hypoglycemizing tumor of B-cells of Langerhans islets produced by the combined action of streptozotocin annd nicotinamide in the rat.

Young male and female Wistar-Velaz rats were treated with streptozotocin-nicotinamide combination according to the method of Rakieten and examined periodically for 23 months for fasting and postprandial glycemia, by intravenous, intraperitoneal or intragastric glucose tolerance and tolbutamide tests with the aim to detect in vivo the experimentally produced nesidiomas. One male rat with severe hypoglycemia, apparent first on tolbutamide test after twelve months, later also on glucose tolerance test and fasting hypoglycemia associated with paraplegia with macroscopically and microscopically documented nesidioma is described. The exstirpation of this nesidioma was followed first by normalization and later by development of latent diabetes.

Effect of proteinase inhibitor in experimental tumors.

Aprotinin, a wide range proteinase inhibitor, was given alone to tumor-bearing mice and life span and several tumor growth parameters were recordered. Aprotinin showed anti-tumor effects in Hepatoma 22 and Lewis lung carcinoma, remaining ineffective in Sarcoma 37, Leukemia L1210 and Ehrlich ascitic carcinoma.

Effect of parathyroid hormone and insulin on extracellular cyclic adenosine-3',5'-monophosphate in patients with benign and malignant breast tumors.

Basal excretion of cyclic adenosine monophosphate (cAMP) and its basal level in blood plasma in breast cancer (BC) patients and those with fibroadenomatosis did not differ essentially. However, intravenous injection of parathyroid hormone (100 U) and insulin (0.08 U/kg body weight) was followed by a much less rise in urine-cAMP excretion and blood-cAMP levels in BC patients than in benign process in mammary gland. A substantial correlation between changes in plasma cAMP level and the degree of insulin-induced hypoglycemia was not observed. There was a negative correlation between reponse to parathyroid hormone and insulin and body overweight in BC patients. It was suggested that body fat content may influence the peculiarities of metabolism of extracellular cAMP in cancer patients considerably.

Effect of irradiation on enzyme activities of cyclic AMP system in the neuro- and adenohypophyses.

The enzyme activities of cyclic AMP system in the neuro- and adenohypophyses were studied, immediately after an irradiation by a single whole body exposure of 1600 R, in an attempt to find whether this intervention causes the changes in the responsiveness of the cyclic AMP regulatory system. In the irradiated rats the neurohypophyses revealed a reduced activity of adenylate cyclase, moderately increased activity of phosphodiesterase and slightly decreased activity of protein kinase, including the value stimulated by cyclic AMP. In the adenohypophyses the irradiation did not cause any significant changes in the enzyme activities of the cyclic AMP system, except of slightly decreased adenylate cyclase activity. The possible relationship of the plasma level of antidiuretic hormone immediately after irradiation and the enzyme activities of cyclic AMP system is discussed.

Ovarian tumors in childhood and adolescence. Histological types and malignancy with reference to the age and menstrual activity of patients.

Alterations in surface proteins of rat cells transformed by avian sarcoma virus B77.

Cell membrane proteins of avian sarcoma virus B77V--transformed cells LWF B55 and LWF B77 and uninfected rat embryo fibroblasts were analyzed by SDS acrylamide gel electrophoresis. Following alterations in cell surface proteins of LWF B55 and LWF B77 cells were found: one, a slight decrease of two high molecular weight proteins the larger of which corresponds presumably to "LETS" protein and a decrease of a protein with approximative molecular weight 50 000; two, increase in content of proteins with molecular weight of about 90--95 000 and 70--75 000 as well as a marked increase of a protein with molecular weight of 30--35 000.

Collagenolytic activity in leukocytes isolated from patients with leukemias and Hodgkin's disease.

Collagenolytic activity in leukocytes and plasma concentration of hydroxyproline were estimated in 80 patients with leukemias and Hodgkin's disease and in 20 healthy individuals. An increase of both studied parameters was found in chronic myelocytic leukemia and Hodgkin's disease, and a decrease was shown in acute myelocytic leukemia and acute lymphatic leukemia. The results obtained imply that metabolic changes occur in leukemia leukocytes, and indicate the presence of disturbances in the metabolism of connective tissue in the studied disorders.

Inhibition and promotion of growth of B77-virus-induced rat tumor with KCl-solubilized tumor cell components.

Rats were immunized with a single and repeated doses (1.0 mg of protein per dose per animal) of KCl-solubilized thymus and avian sarcoma virus B77-induced rat tumor cells LWF B77 prior to challenge with the same tumor cells. Results obtained showed that single administration of KCl-solubilized tumor cell material resulted in inhibition of growth of syngeneic tumor cells. Repeated inoculations of the same material, however, promoted tumor growth. Possible explanation of this dual effect is discussed.

Increased pathogenicity of avian sarcoma virus B77 in cyclophosphamide treated chickens.

The course of sarcoma development was studied in cyclophosphamide treated and control chickens injected with avian sarcoma virus B77 (B77V). It was found that the incidence of sarcomas was the same for both groups of birds. Progressive growth of sarcomas as well as high tumor mortality was observed in drug treated birds, whereas frequent regressions occurred in controls. The drug treatment after B77V infection did not change the response patterns and no cytostatic effect of the drug was observed. Cyclophosphamide treatment improved the rescuability of B77V genome from the transformed virogenic mouse and rat cells in vivo.

Protein synthesis in tumor host. II. Increased activity of peptide elongation factor 1 in experimental rat tumors and in host liver.

Both peptide elongation factors were purified from Zajdela and Walker tumors and from the host and normal liver. The activity of peptide elongation factor 1 from tumor tissues in promoting the binding of phenylalanyl-tRNA to ribosomes was significantly higher than that of normal liver. Also preparations from the host liver were markedly more active when compared with the corresponding factor from normal liver. Poly(U)-dependent phenylalanine polymerization was enhanced in subcellular systems containing elongation factor 1 from tumors or host liver. No differences were found between preparations of elongation factor 2 isolated from various sources. Tumor ribosomes showed an increased activity of both the acceptor and donor binding site of peptidyl transferase. In ribosomes from the tumor-host liver the activity of the acceptor site of this enzyme was decreased while that of the donor site remained unaltered. The enhanced activity of peptide elongation factor 1 from tumors and host liver is apparently the main reason of enhanced protein synthesis in these tissues. The enhanced activity of this factor is not specific for tumor growth as it occurs also in other pathological conditions.

L-asparaginase in treatment of acute leukemia in children.

L-asparaginase from Escherichia coli--Crasnitin was used in 14 children with acute leukemia unresponsive to conventional treatment: 11 acute lymphoblastic leukemias, 1 acute myeloblastic leukemia, 2 other forms of leukemia. The remission induction was obtained in 70% of applications. Median of remission duration was 90 days. Serious side effects were observed. The validity of L-asparaginase in therapy of advanced childhood ALL is stressed.

Chemoimmunotherapy of acute leukemia in children.

Comparative results of chemo- and chemoimmunotherapy with Soviet strain of BCG vaccine in children with acute leukemia are presented. The immunotherapeutical regimen includes 20 intracutaneous injections of 0.1 mg BCG vaccine given weekly during 5--6 months with subsequent 5--6 months intervals simultaneously with maintenance chemotherapy. In the control group children were only given conventional chemotherapy. The median duration of remission and survival in patients who had received BCG chemoimmunotherapy was 25.2 +/- 1.5 and 32.3 +/- 1.2 months respectively. In the group of patients with chemotherapy alone--13.2 +/- 0.9 and 21.8 +/- 1.1 months respectively (p less than 0.01). The best results were obtained in children who had received before BCG immunotherapy neuroleukemia prophylactic treatment.

Anti-A and anti-B isohemagglutinins in the course of children's leukemias.

The changes in titres of isohemagglutinins anti-A and anti-B were followed up in 48 leukemic children during relapses and remissions. The levels of titres were compared 1. with levels in healthy children in the same age group, 2. in the different phases of disease in the same child. Compared with healthy children, a higher frequency of low isohemagglutinin levels was found in the phases of the exacerbation of malignancy, no changes were observed in remissions. In some children a quite evident fluctuation of isohemagglutinin titres in relation to the phases of the malignant disease was found. The depression of blood group isohemagglutinins in the phases of activation of the malignant process is considered as a partial defect in IgM, analogical to the so called normoglobulinemic defect of the antibodies.

Prognostic significance of an enhanced number of T lymphocytes in chronic lymphatic leukemia.

Analysis of lymphocytes carried out by means of rosette tests in 24 patients treated for chronic lymphatic leukemia established the origin of malignant cells to be from Blymphocyte population. On the basis of a reactive T lymphocyte proliferation in patients with chronic lymphatic leukemia, a "coefficient of active T lymphocytes" has been deduced which proved to be a rapid indicator of a short-term prognosis. This coefficient may contribute in a substantial measure towards an objectivation of the indication of a repeated oncostatic therapy in relatively light leukocytosis or in otherwise clinically unresolved cases.

Mutual relationship between total and active T lymphocytes in patients with malignant tumors.

The values of B lymphocytes and of active and total T lymphocytes have been followed in 104 patients with malignant diseases and 35 controls of corresponding age, using the rosette tests with sheep erythrocytes. A statistical analysis of the changes of total and active T lymphocytes in three selected groups of the malignant disorder revealed the decline in total T lymphocytes to be parallel to that in active T lymphocytes. Therefore, a follow-up of the immunocompetence of oncological patients with the criterion determination of thymus-dependent lymphocyte population by the classical method of rosette formation was found to be equally informative in the three groups of patients studied here, as an evolution of active T lymphocytes in the active rosette test.

Hodgkin's disease in children: investigation of T and B lymphocytes.

A study has been made of twenty-five untreated patients with Hodgkin's disease (H. D.), ageing from 3 to 13. We have analyzed the correlations of T and B lymphocytes of peripheral blood and the involved lymph nodes and also the functional peculiarities of T-cells in PHA-response. It is shown that as well as adults children with H. D. have the depression of T-system of immunity, more marked in the presence of general symptoms and biological stage "b". No significant influence of the clinical stage on the level of immunological disorders has been found.

Treatment of solid Gardner lymphosarcoma with methotrexate sorbed on 2-hydroxyethylmethacrylate polymer in combination with leukovorin.

Implantation of 2-hydroxyethylmethacrylate microporous carrier with sorbed methotrexate (Hema-Hex-MTX) proved more toxic on C3H strain mice with a solid Gardner lymphosarcoma than on tumor-free mice of the same strain. If, during the first week of the experimental disease, the Hema-Hex-MTX implantation was followed by administration of leukovorin in course of next 24 hours the toxicity of MTX was abolished and the treated mice survived significantly longer than control animals.

Stages of the development of immunologic response in regional lymph nodes draining breast cancer.

During the studies of regional lymph nodes originating from 50 patients with breast cancer, there were distinguished four stages in development of immune response. The first, stage of induction of the response concerned patients with non-infiltrating carcinoma. The second stage of the active immune response was observed in 1/3 of the patients with infiltrating cancer without metastases. In these cases, in the pattern of regional lymph nodes predominated medium sized lymphocytes within the thymus-dependent inner cortex, while in outer areas there was an increased number of small active follicles. Within the sinusoidal structures there predominated proliferating prohistiocytes growing in interaction with small lymphocytes. The third stage of immune response was characterized by weak activity and progressive cortical strophy of the lymph nodes and it was observed in 2/3 of the patients with invasive cancer. The fourth stage of immunologic response concerned the patients with minute-metastases within the regional lymph nodes. In these cases one group of the lymph nodes showed unstimulated pattern while the other a highly stimulated pattern with the presence of giant follicles.

Evidence of cell-mediated antitumor immunity in rats bearing syngeneic tumors by the leukocyte adherence inhibition test.

A modified form of the leukocyte adherence inhibition (LAI) test has been used to follow cell-mediated immunity (CMI) in rats of the Lewis inbred strain (LW), F1 hybrids inbred strains LW x AVN with transplanted syngeneic tumors B77, MC-1, and the random-bred Sprague Dawley strain with spontaneous REF tumor. Peripheral blood leukocytes of animals incubated witha specific tumor extract showed an adherence capacity by 30--40% lower than the same cell population cultured without this specific extract, or with a foreign nonspecific extract. The results are commented in connection with a possible application of this modified form of LAI test in studies of the immunological state of patients with a malignant disorder.

Studies on T and B lymphocytes in rats bearing methylcholanthrene-induced tumor.

The authors have studied the influence of primary, methylcholanthrene-induced tumor on T and B lymphocytes of spleen, thymus, draining lymph nodes and peripheral blood of rats. Differences in weight of tumors were found to correlate with changes in proportionality of T and B lymphocytes of followed organs. Small tumors induced but insignificant changes. There was increased trapping of T lymphocytes in spleen and lymph nodes with simultaneous decrease in peripheral blood. The authors noted a high percentage of blasts. B lymphocytes showed a tendency to compensate for the loss of T cells. Large, progressively growing tumors caused evident exhaustion in the number of both cell types in lymph nodes and peripheral blood. In the spleen there was slower exhaustion. Reduction in the number of B lymphocytes correlated with the size of tumor. Blasts disappeared. Proportionality of T and B lymphocytes in thymus did not appear to be influenced by the size of tumor.

Immediate and remote results of dermatoplastic operations in patients with malignant skin neoplasms.

The results of dermatoplasty made in 698 patients suffering from skin neoplasms are reported. The plastic operations do not negatively affect the course of the disease; at the same time they enable the surgeon to perform more radical resection of a tumor.

The influence of extracts from the skeletal muscle of guinea-pigs infected with Myco bovis BCG-Praha and/or with Myco fortuitum on the viability of Ehrlich ascites tumor cells.

Description of results obtained by following the influence of muscle from guinea-pigs infected with Myco bovis BCG-Praha and/or Myco fortuitum on the viability of Ehrlich ascites tumor cells is presented. The muscle extracts were prepared from the striated muscles of the upper and lower extremities as well as the sternal muscle of guinea-pigs killed on the 1st, 15th, 35th and 70th days after infection with the above-mentioned mycobacterial strains. It follows from the results obtained that only muscle extracts from guinea-pigs infected with BCG-Praha will exhibit a considerable potential inhibiting the viability of Ehrlich ascites tumor cells. This inhibitory factory could not be eliminated from muscle extracts either through saturation by a homologous mycobacterial strain or by homogenates of striated muscles gained from healthy, non-infected guinea-pigs.

Cellular reaction to autologous tumor tissue followed by "skin window" technique in orofacial tumors.

In a sample of patients with pseudotumors and benign and malignant tumors of the orofacial region there has repeatedly been investigated, after radical surgery, the cell reaction to autologous tumor tissue applied at indicated time intervals on a small skin lesion. The prognostically favorable picture of a cellular-type hypersensitivity reaction with massive participation of lymphoid cells and basophil granulocytes was only found in some basaliomas and leukoplakias. Conversely, spinocellular carcinomas are, irrespective of localization, characterized by areactivity.

Effect of M.bovis BCG and M.microti on sarcoma 180 growth in mice.

The antitumorous efficacy of the Czechoslovak BCG 725, Czechoslovak BCG 901, the Danish SAS maintained, the Japanese SAS and SH maintained BCG strain and the Czechoslovak OV 247 (M.microti) strain was studied. (SAS--original Sauton's medium with asparagine, SH--Sauton's medium with the asparagine replaced by enzymatic casein hydrolysate.) Ascitic Sa 180 cells were used as the experimental tumor and 800 mice as experimental animals. All the strains employed displayed an anticarcinogenic effect in a protective assay (immunization 8 weeks before inoculation of Sa 180 cells). In a therapeutic test (vaccine administered 24 hr after inoculation of Sa 180 cells and then at weekly intervals), Czechoslovak BCG 725 and Czechoslovak OV 247 were used and proved effective.

Radioprotective effect of Aspergillus flavus.

A study designed to determine the extent and intensity of radioprotective effects of the mold conidium A. flavus in white mice revealed a higher survival rate with irradiation doses in the range of LD70-LD90; with irradiation with supralethal doses there is no difference in survival by stimulated and control mice. The radioprotective effect is independent of whether the strain A. flavus had been isolated from environment with enhanced radioactivity or not. It depends on a structure of the wall of conidia and not on the toxin produced by mold cells.

Time dependence of radioprotective effects of A. flavus conidia.

No essential difference in radioprotection results when conidia of A. flavus are applied to mice within a time lapse of a few minutes up to 48 hours prior to irradiation. When applied 72 hr before irradiation, they fail to affect mouse resistance. Application of conidia provide no protection to mice irradiated for 18-20 hr at the dose rate of 1 R/min; similarly ineffective are the chemoprotectors cistafos and mexamine. A combined administration of conidia and the two protective substances increases survival in mice and prolongs the mean survival period.

Distribution and pharmacokinetics of methotrexate in localized chemotherapy of solid Gardner's lymphosarcoma.

Pharmacokinetics and organe distribution of Methotrexate (MTX) in Gardner lymphosarcoma bearing C3H mice was investigated following two ways of drug administration: 1. intraperitoneal injection, 2. intratumoral implantation of 2-hydroxyethylmethacrylate gel with sorbed Methotrexate (localized chemotherapy). The highest level of MTX in blood appeared 2 hours after intrperitoneal injection but 7 hr after localized intratumorous application. Following intraperitoneal application the drug level in tumor reached its maximum two hours after injection; after 7 hr the drug could not be detected any longer. The localized chemotherapy led to six times higher concentration of the drug in the tumors as compared with the intraperitoneal application. This high level persisted for 17 hr and decreased moderately for 48 hr. MTX was accumulated in liver after both modes of administration with a half life 6.1 hr after intraperitoneal injection and 12.4 hr after the localized chemotherpay, respectively.

An autoradiographic study of the NK/Ly tumor transplanted into the brain of the mouse.

The NK/Ly ascites tumor transplanted into the brain of mice is well accepted and the growth is similar to that of malignant lymphomas in the human brain. The authors studied the incorporation of the tritiated thymidine in the malignant lymphoma cells transplanted into brain of the mouse by autoradiographic method.