Potentiation of the thymosin immunoregulatory effect by hypocorticoidism in rats with DMBA-induced carcinogenesis in mammary glands.

Effect of adrenalectomy and or thymosin administration on the tumor growth and immunity system were investigated on the models of 7.12-dimethylbenza(a)anthracene(DMBA)-induced mammary neoplasms in rats. Adrenalectomy and thymosin administration resulted in antitumorigenic action, whereas maximal inhibiting effect was achieved after thymosin injection in adrenalectomized animals. The maximal antitumorigenic effect of thymosin in adrenalectomized rats was accompanied by synergistic immunopotentiating effect of thymosin and adrenalectomy. Immunorehabilitation with thymosin and simultaneous correction of hormone production in the adrenal cortex are believed to enhance the effectiveness of therapy in cancer patients.

Combined effect of cytostatic drugs and E-N-L-trimethyl lysine in healthy and transplantable tumor bearing mice.

E-N-L-trimethyl lysine (TML) decreases the toxicity of Vincristin, Cyclophosphamide and Doxorubicin (Adriamycin) when administered simultaneously to healthy mice. Simultaneous treatment of L1210 ascites leukemia bearing mice with 100 mg/kg TML and 2, 2.5, 3.2, 3.5, 4 mg/kg Vincristin or 10-15; 20 mg/kg Doxorubicin increases significantly the survival of the animals when compared with untreated and Vincristin or Doxorubicin treated mice. Repeated impulse treatment of S-180 subcutaneous sarcoma with 100 mg/kg TML and 50-100 mg/kg Cyclophosphamide results in a significantly higher surviving time and surviving rate than Cyclophosphamide treatment alone.

Some immunological and biochemical markers in chronic lymphatic leukemia patients.

The leukemic cells in chronic lymphatic leukemia (CLL) patients have been studied prior to theory with a panel of immunological markers. Cells were assayed for the presence of receptors for sheep erythrocytes (E active and total rosettes), C3d component of complement (EAC rosettes), mouse erythrocytes (M rosettes), some of them also for surface membrane immunoglobulins (SmIg). In vitro 24 h cultures without mitogen (detection of spontaneous DNA synthesis) or 72 h cultures with phytohemagglutinin (PHA) were also performed. These conventional immunological markers and functional lymphocyte characteristics have been correlated with enzyme activities of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP). Electrophoretic patterns of radiolabeled proteins under denaturing conditions (SDS-PAGE) have also been determined in some patients of this group. Phenotypic surface characterization of blood elements of all CLL patients studied revealed their B origin, with increased values of EAC rosette forming cells and especially increased values of M rosette forming cells. Significantly decreased values of both, ADA and PNP, were found in all the cases. Electrophoretic patterns of radiolabeled surface proteins from cells of CLL patients were essentially similar within the group with characteristically strongly radiolabeled glycoproteins gp44--HLA heavy chain and glycoproteins gp29, gp35--Ia-like or HLA-DR antigen.

Human melanoma cell lines: morphology, growth, and alpha-mannosidase characteristics.

Four human melanoma cell lines were characterized by evaluation of the morphology, ultrastructure, cytogenetics, plating efficiency, agar colony formation and alpha-mannosidase values. Biochemical studies demonstrate that all studied lines produced alpha-mannosidase even under condition of long-term in vitro growth. All respective lines were characterized by a great variations in biological markers studied and very few reliable comparisons could be obtained as regards the proliferation activity and capacity, cellular composition and clonogenic potential as well. Due to cellular selections during long-term in vitro cultivation, the range for using these experimental systems as models for the study of biology of human malignant melanoma seems to be rather limited.

Mutagenic effects of two nitrofuran food preservatives.

Two structurally related food additives, 3-(5-nitro-2-furyl)acrylic acid (5-NFAA) and 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2) showed a marked mutagenic effect on Salmonella typhimurium strains TA100 and TA98 and Escherichia coli WP2 uvrA+ and WP2 uvrA. On the molar basis 5-NFAA was about two orders of magnitude less effective than AF-2. In Salmonella typhimurium TA100 anaerobic conditions stimulated the mutagenic effect of 5-NFAA which was more pronounced in nitrofuran-reductase deficient strain of Salmonella typhimurium TA100 FR50. 5-NFAA increased the number of isoleucine revertants and induced mitotic recombination at tryptophan, threonine and adenine loci of the diploid strains Saccharomyces cerevisiae a/b and Saccharomyces cerevisiae SBTD. Activity of 5-NFAA was lower than that of AF-2. The test on sex-linked recessive lethal mutations in Drosophila melanogaster indicated that only 5-NFAA is mutagenic, increasing the mutation frequency about 10-fold above the control. Results with AF-2 fell within the control range.

Carcinogenesis after sublethal ionizing irradiation and regular avoidance irritation.

160 mice of the BALB/C strain of both sexes, aged 3 months, were divided into four equal groups out of which two were regularly irritated by a combination of an optical signal and electrical stroke. After one month of irritation one nonirritated and one irritated group were whole body irradiated with a single dose of 6.65 Gy (1.83 Gy/min), the other two groups were sham irradiated. The mice lived until their spontaneous death or one year after irradiation, respectively, when the rest of the animals were sacrificed. The appearance of malignant tumors was noted. Irradiation shortened the survival time while the irritation had an appeasing, compensatory effect, more expressed in the males than in the females. After irradiation the number and assortment of the tumors increased and the latent period essentially shortened. In the irritated animals the number and assortment of the malignant tumors were reduced and a tendency for lengthening of the latent time period was seen; these differences, however, were not statistically significant. In spite of some differences the response in both sexes to irradiation and irritation or their combination was similar.

Lycurim (NSC 122402) in the treatment of chronic lymphocytic leukemia.

Effect of free and bovine fibrinogen-bound methotrexate and dibromoaminopterin on the dissemination of solid Gardner lymphosarcoma.

Solid Gardner lymphosarcoma transplanted subcutaneously to mice of strain C3H/Sumice does not grow solely at the site of inoculation, but disseminates also to remote tissues. An i. v. administration of fibrinogen-Methotrexate derivative or Methotrexate (8-15 mg antimetabolite per 1 kg) to mice with an early tumor, reduced the weight of the local tumor, prolonged their survival time and also the posttransplantation period during which the dissemination of malignant cells could not be detected. When the above substances were given to mice with advanced tumors whose dissemination could be reliably established, Methotrexate proved more effective than its fibrinogen derivative. 3.5'-dibromoaminopterin administered alone or bound to fibrinogen (14.6 mg of antifolate per kg) showed no effect both on survival rate and on tumor dissemination.

The ability of rat transplantable lymphosarcoma cells to synthesize immunoglobulins.

The hemolytic plaque assay of Jerne and Nordin has been used for the investigation of the ability of the transplantable rat lymphosarcoma cells to synthesize immunoglobulins. The data obtained indicate that in 3-4 days after antigen (sheep red blood cells) administration to the tumor-bearing rats, antibody (hemolysin) producers appear among lymphosarcoma cells. Most representative are the experiments on rats in which their own immune response was totally suppressed by X-ray irradiation. The induced hemolysis synthesis in rat lymphosarcoma cells persists at least 9 during successive transplantations of "primary immunized" tumors (45 cell generations). Total cytoplasmic RNA isolated from spleens of sheep red blood cells immunized rats induced hemolysin synthesis in lymphosarcoma cells in vitro. Such cells being inoculated into intact rats retained their ability to synthesize hemolysin up to 2-3 passages of tumor.

Some quantitative micromorphological characteristics of granulocytic leukemia in 239Pu-injected mice and untreated controls.

Cytologic and histologic evaluation of granulocytic leukemia was carried out in 22 plutonium-treated mice and in 17 controls. The contaminated animals were injected i. v. with 5.4 kBq 239Pu/mouse (180 kBq/kg) at the age of 70 days. Morphologic evidence of the disease was based on the identification of immature abnormal granulocytes in the bone marrow and spleen and on the histologically ascertained leukemic infiltration at least in one of the tissues examined (liver, lung, kidney, lymph nodes). Whatever further symptoms are thought to be of non-specific value. The leukemic process could be mostly evaluated as more florid in the spleen than in the bone marrow. The less active leukemic proliferation in 239Pu-treated mice than in diseased controls is the most important finding of this study. The results are discussed.

Clonal evolution of karyotype in blastic phase of CML.

A patient with chronic myelocytic leukemia (CML) had a Philadelphia chromosome--Ph1(t(9q +; 22q--)) in all evaluated bone marrow cells at the time of diagnosis. After 29 months of intermittent therapy (chemotherapy and immunotherapy) and 2 months before clinical signs of blastic phase developed, three additional cell lines in bone marrow and peripheral blood appeared: one line with extra chromosome Ph1, another one in which chromosome Y disappeared, and the third line with extra chromosome No. 13, evidently derived from the X-monosomie cell line. Five weeks before death a variable hypodiploidy was found in more than 50% mitoses. The patient died 47 months after the establishment of CML and seven months after the onset of the blastic phase.

Intensified proliferative activity of the CFU-S in vertebral bone marrow of 239Pu-treated mice as one of the factors involved in the induction of granulocytic leukemia.

Using the exocolonizing test, 59Fe-utilization technique and classical cytology, the authors observed 210 days after i.v. injection of 166.7 kBq of 239Pu/kg in about 30% of contaminated mice a proliferative activity in vertebral bone marrow, characterized with high relative numbers of pluripotent hemopoietic stem cells, significantly higher than in the seriously damaged vertebral bone marrow of the other 239Pu-treated mice and even higher than in the untreated corresponding controls. Simultaneously, the amount of cells in the granulocytic series was increased. After the transplantation to the heavily irradiated syngeneic hosts the stem cells differentiated into splenic colonies with higher iron utilization than in corresponding controls. Higher numbers of mature granulocytes were also found in the peripheral blood and spleen. The authors assume that this activity was an inadequate reparative response of the hemopoietic stem cell compartment to the damaging effect and they consider it to be the critical phase which not only preceded the induction of granulocytic leukemia but also afforded conditions favorable for leukemic transformation of the hemopoietic stem cells.

T-cell hybrids. II. BH2 line-derivation by fusion between two (BW5147 X EL-4R) thymoma lines.

Subpopulations of human T lymphocytes in patients with Hodgkin's disease before and after treatment.

Circulating T gamma and T mu cells enumerated in the peripheral blood of 51 untreated Hodgkin's disease (HD) patients and 66 treated HD patients tested after few months to more than 3 years of remission, brought about by radiation and/or chemotherapy. 53 age matched normal healthy individuals were studied as controls. Untreated HD patients showed significant increase in T gamma cells (p less than 0.001) and decrease in T mu cells (p less than 0.001) when compared to normal donors. The abnormal percentages of T cell subsets did not correlate with the severity of the disease. A progressive partial restoration in the proportion of these two subsets of T lymphocytes was seen in the disease free condition. However, even after 3 years of remission the recovery was not equivalent to controls. There was no correlation between the recovery of T gamma and T mu cells with the modality of treatment.

Polypeptide composition and gag gene-coded products of type-D oncovirus from HEp-2 cells.

The protein composition of type-D oncovirus HEp-2, isolated from cell-free medium of continuous human HEp-2 cell line, has been investigated using electrophoresis on gradient polyacrylamide gels with sodium dodecyl sulfate (SDS). Labeling with 14C-amino acids revealed five viral polypeptides with molecular weights of 70 000 (gp70), 27 000 (p27), 19 000 (p19), 15 000 (p15), 12 000-10 000 (p12-10). The 70 000 dalton protein was shown to be the only glycoprotein by incorporation of radioactive glucosamine. A polypeptide with molecular weight of 78 000 has been specifically precipitated from pulse-labeled type-D oncovirus producing HEp-2 cells with goat anti Mason-Pfizer p27 serum. This protein was shown to be gag gene-coded polyprotein precursor (Pr78gag) of the major virus polypeptide p27. Pulse-labeled HEp-2 and Mason-Pfizer infected Tu 197 cells were rinsed, lysed, clarified and precipitated with goat anti Mason-Pfizer p27 serum. In both cases Pr78gag was detected.

Distribution of normal T lymphocytes in tumor-bearing rats.

Changes in distribution properties of T lymphocytes from lymphoid organs of normal rats 3H-uridine labeled in vitro and i. v. injected to tumor-bearing recipients were studied at different stages of tumor growth and rejection. Comparisons were made with T cell migration from normal donors to syngeneic recipients. A temporary decrease in the ability of normal spleen T lymphocytes to migrate into the spleen of tumor-bearing recipients at early stages of tumor growth was in correlation with their enhancement in trapping by the liver of this recipient. At this stage the migrations into tumor-draining lymph nodes and peripheral blood were rather pronounced. The growing tumor evoked an increased migration rate of donor spleen cells into each of the recipient organs studied except for liver. Tumor rejection potentiated the distribution mainly in the draining lymph node and liver. Normal lymph node T lymphocytes migrated at the beginning of tumor growth into spleen, node and liver of tumor-bearing recipients. A sharp decrease of distribution in spleen at the time of maximum tumor growth was compensated by an enhanced migration into draining nodes, peripheral blood and liver. This enhancement was seen also during tumor rejection. Distribution of normal blood T lymphocytes into spleen and peripheral blood of tumor-bearing recipients was equal to normal values. There was only a transitional reduction in their migration rates into draining nodes caused by tumor growth. Migration rates of thymocytes in tumor-bearing recipients remained unaltered. The results presented indicate that the distribution pattern of i. v. injected labeled T lymphocytes from normal donors into lymphoid organs of tumor-bearing recipients was determined by changes in the immune status of the recipients brought about by tumor development.

Immunocytochemical studies of carcinoembryonic antigen (CEA) in bilharzial bladder.

Reactivity of dibenzopyrenes in UV- and gamma-radiation initiated oxidation reactions.

Potential anticancer agents. XXIII. 1. Qualitative structure--activity relationship in the "classical" antifolates area.

In order to obtain "classical" antifolates with improved therapeutic index, 32 new Methotrexate analogues were synthesized and studied. Their structure--activity relationship analysis led to the following conclusions: a) the replacement of glutamyl moiety with other amino acids led to compounds which are powerful inhibitors of DHFR but were devoid of activity against L1210 leukemia, b) the phenylic nucleus substitution with methoxy groups afforded potent inhibitors of DHFR and also effective derivatives against experimental tumors, c) the insertion of an extra amino acid between the phenylic ring and the terminal moiety proved to be an unfavorable event for the activity of such compounds, d) the MTX-analogues with the peptidic side chain grafted at C7 of the pteridine ring were ineffective against both DHFR and L1210 leukemia. From the investigated derivatives p[2,4-diamino-6-pteridinyl)-methyl-N10-methyl] aminobenzoyl-L-leucine 16 is twofold more potent as MTX by respect to DHFR and could be used in MTX-resistant (by impaired transport) cell lines being more hydrophobic.

Potential anticancer agents. XXIII. 2. Quantitative structure--activity relationships (QSAR) in the "classical" antifolates area.

Experimental approach for the search of effective drug combination in cancer chemotherapy.

A comparative assay of cytostatic combinations was carried out in a number of transplantable mouse tumors: L1210, Lewis lung carcinoma, Sarcoma 180 and others. The results suggest that the combination cyclophosphamide (CPH) plus procarbazine (PROC) enhances the antitumoral activity in comparison with monochemotherapy. The combination CPH + PROC can be used as basis for the elaboration of more effective polychemotherapy schemes. Combination of CPH + PROC + Prospidine and CPH + PROC + CCNU resulted in enhancement of the treatment effect in experimental tumors.

Leukemoid reaction induced by different transplantable tumors in three inbred mouse strains.

We have studied the induction of a granulocyte-associated leukocytosis (leukemoid reaction) in C3HA, C57B1/6, and DBA/2 mice by a number of transplantable tumors of different origin. Leukemia L1210, Hepatoma 22, a transplantable mammary carcinoma of spontaneous C3HA origin, and a L929 culture fibroblasts-derived rhabdomyosarcoma, all induced a leukemoid reaction in their specific mouse strain. Melanoma B16 and Lewis lung carcinoma gave no reaction; Adenocarcinoma 755 and Harding-Passey melanoma evoked a leukocytosis but not due to an increase in neutrophils. Some extratumoral factors can influence the hematological response; the intensity of final leukemoid reaction was higher in female mice than in males bearing the same tumor. On the other hand, Ehrlich ascites tumor transplanted in all three inbred mouse strains rendered different levels of leukemoid reaction; response was higher in DBA/2, intermediate in C3HA and lower in C57B1/6.

Recent trends in cancer incidence in Kiev 1969-1979.

The analysis of the trends of age-adjusted incidence rates of malignant tumors in the city of Kiev in the years 1969-1979 revealed a gradual increase of the majority of individual sites, mainly of the malignant tumors of rectum, lymphoid and hematopoietic tissues in both sexes and of breast in females. On the other hand a decrease of incidence rates of the malignancies of skin, lip and stomach in both sexes and of oesophagus, lung and uterine cervix in females was noted. These investigations formed the basis of the prognosis study of incidence rates of malignant tumors in Kiev until 1990. The importance of the study of incidence rates and trends of individual sites of malignant tumors for the investigation of their factors and causes in urban conditions and environment is emphasized.

Survival curve description for fractionated radiation therapy.

The methods of survival curve description are evaluated. New method of fitation of survival curve data is described. Representations of survival curve are compared and the shapes of different survival curve formula are shown. New simple formula is suggested for practical purposes.

Vascularization and radiocurability in cancer of the uterine cervix. A retrospective study.

The proportion of vascular elements in relation to the parenchymal and stromal components was determined by morphometric analysis of the histologic preparations from biopsies of uterine cervix cancers. In all cases, the material was obtained from tumors in Stages Ib and IIa before any therapy was initiated. In the 23 cases in which radiation treatment of the lesions resulted in a survival longer than 5 years, the proportion of blood vessels in the neoplastic tissue was found to be larger, and especially the stromal components were richer in vascular elements than in the 22 cases in which survival was shorter than 5 years. It was concluded that vascular density at least in cervical cancers of the stages studied can be a diagnostic parameter of a prognostic value as well as of a therapeutic usefulness.

In vivo malignant transformation of cells from human oral lichen planus lesions.

When incubated in presence of patient's serum, but not normal serum, and complement, oral mucosa cells from patients with oral lichen planus (OLP) clumped together, the cell membrane was destroyed and the cells lysed. Whereas, cells from healthy volunteers maintained their morphology. Sixty to seventy days after inoculation into a thymic Nu/Nu BALB/c nude mice, cells from OLP lesions produced lesions in the recipient animals. Histological examination of these lesions showed hyperplasia with increased number of mitotic figures and dyskeratosis. These lesions resembled severe dysplasia and early squamous cell carcinoma.

The effect of ring-substituted and quaternary chlorpromazine derivatives on the survival of the NK/Ly ascites tumor bearing mice.

The effect of quaternary chlorpromazines like-methiodide, allylammonium chloride, -octylammonium iodide, benzylammonium chloride, -phenacylammonium bromide, 9-phenanthryl-methylammonium bromide, trimethylene-bis-chlorpromazine-ammonium bromide and among the ring-substituted derivatives the 7-hydroxy-, 7,8-dihydroxy-, 7,8-dioxo-, 6,9-dihydroxy-, 6,9-dioxo-chlorpromazines and the chlorpromazine-5-oxide on the prolongation of survival time of NK/Ly ascites tumor bearing mice was studied. The intraperitoneally administered quaternary chlorpromazine derivatives in 10-25 mg/kg daily dose did not increase the life span of the animals, and did not prevent the development of the ascites tumor. However some ring system substituted chlorpromazines such as the 7,8-dioxo- and 6,9-dioxo-derivatives markedly increased the survival time of the tumor bearing mice and reduced the ascite volume in 10 mg/kg body weight dose.

Pharmacokinetics of Damvar.

The pharmacokinetics of granulated Damvar (delta-(2-amino-6-hydroxy-3,4-dihydro-4-oxo-5-pyrimidinyl) valeric acid) after a single oral dose of 1000 mg was studied in 10 subjects with neoplastic disease. The rate of Damvar absorption from the digestive tract is not very fast. Maximal serum levels (13.5 micrograms/ml) were recorded 3 h after administration with minor interpersonal variations. The time course of Damvar serum concentrations coincides with its distribution in a two-compartmental pharmacokinetic model with biological half-life of 9.72 +/- 0.84 h. Only a small amount of Damvar is eliminated in urine during a period of 24 h (3.1% of the administered dose). Its renal clearance is also low (0.05 ml/s). The analysis of Damvar excretion in urine shows that kidneys play a minor role in its elimination from the body. Therefore the attention should be concentrated on the effect of Damvar administration in patients with disturbed metabolic functions.

Natural killer lymphocyte function in cervical cancer patients.

Authors investigated the natural killer and antibody dependent cellular cytotoxicity functions of lymphocytes in cervical cancer patients. According to the results the natural killer function showed remarkable changes related to the extent and the course of the disease. On the contrary, radiation therapy did not influence this function. The antibody dependent cellular cytotoxicity function of the same lymphocytes did not show any alteration under circumstances mentioned above.

Neoplastic progression evidenced in the L929 cell system. I. Selection of tumorigenic and metastasizing cell variants.

Starting with a L929 cell strain of low tumorigenicity, and through in vivo and combined in vivo--in vitro manipulation, we have isolated a number of related culture cell lines with different tumorigenicity and metastasis ability in C3HA/H mice. Within our system, tumorigenicity can be greatly enhanced without the simultaneous development of metastasis ability, the latter arising only after long-term in vivo serial transplantation. Metastatic variants express better the ability to undergo to cascade of metastasis than the primary tumor. The selection of metastatic variants from a tumor, using its subcutaneous inoculation, renders cell lines with different behavior. Finally, viable cells are recovered from lungs of animals inoculated with metastatic variants that would not be expected to show true metastatic foci; the results are commented in relation to the phenomenon of metastasis dormancy. Our experiments are discussed under the scope of the hypothesis of neoplastic progression as a multistep process.