The role of autosensibilization of lymphoid cells in genesis of antitumor resistance.

C3HA mice were inoculated with a mixture of syngeneic transplantable hepatoma 22a cells and syngeneic splenocytes of mice immunized with normal syngeneic tissues or subjected to partial hepatectomy. Control mice were inoculated with a mixture of tumor cells and splenocytes of intact or sham operated mice. A considerable inhibition of tumor growth was observed in the experimental series. Immunization of mice with normal syngeneic tissues as well as partial hepatectomy results in sensibilization of splenocytes not only to normal tissue antigens, but to the antigens of hepatoma 22a cells too. This was shown by the reaction of indirect inhibition of glass adherence of peritoneal cells. The data obtained are considered to be one more possible prove of a special form immune surveillance which controls the state of cytodifferentiation being active in the organism. The involvement of this form of immune surveillance in the genesis of antitumor resistance is discussed.

The passage of lymphatic leukemia L1210 cells injected into glass bead labyrinth: their flow and retention.

Leukemia L1210 cells and normal lymphocytes were injected into the glass bead packing perfused with a medium containing serum proteins in concentration sufficient to greatly reduce the surface adhesion. The passage of radioactively labeled cells introduced as a concentrated suspension, was studied by the analysis of transit-dilution curves. It was demonstrated that a fraction of cells was retained in the labyrinth; other parameters characterizing the cell flow were similar to those of the medium. The mechanism of the arrest of cells is discussed: it is related to geometrical features of the labyrinth and the hydrodynamic forces influencing the cell surface interactions. Intense washing of the glass bead column results in the release of only a minor fraction of cells which were previously arrested. When the L1210 cells which have passed through the labyrinth without being trapped are reinjected into another glass bead layer, their retention is similar to that observed after the first injection; it appears, therefore, that the L1210 cells are uniform as concerns the properties which are important for surface interactions under flow conditions.

Effect of chloro- and bromo-derivatives of isocrotonic acid of bioenergetic processes in Ehrlich ascites cells and isolated mitochondria.

The effect of nine chloro- and bromoderivatives of isocrotonic acid on some bioenergetic processes in both Ehrlich ascites cells and isolated rat liver mitochondria has been investigated. Substances studied in the concentration range 25--200 microM significantly inhibited incorporation of both 14C-adenine and 14C-valine into acid-insoluble material of Ehrlich ascites cells. The rate of 14C-precursors incorporation being directly related to the concentration of the inhibitor. Gamma,gamma-bis-4-ethylphenyl-alpha,beta-dichloroisocrotonic acid fully inhibits both aerobic glucose utilization and lactic acid formation at 200 microM concentration. At lower concentrations, however, glycolysis is stimulated. Maximal stimulation of rat liver mitochondrial respiration in state 4 with succinate as substrate was reached at concentrations as low as 10 microM. On the other hand, these substances were able to release by the oligomycin inhibited respiration of rat liver mitochondria. Our data suggest that cytotoxic and cancerostatic action of isocrotonic acid derivatives lies primarily in the exclusion of key processes in the energy metabolism of Ehrlich ascites cells and isolated mitochondria.

Excretion of melanogens and zinc during the growth of melanoma in hamsters.

Melanogens which are considered as probable precursors or their metabolites in the metabolic pathway toward melanin are excreted in an increasing amount during the melanotic melanoma growth in hamsters. Moreover, an increased excretion of zinc was observed. The evaluation of dependence between zinc and Thormählen positive melanogens excretions during the melanoma growth suggests an exponential type of dependence. Likewise as in the case of human melanoma the hyperzincuria proved to be a laboratory sign of tumor disease progression in Syrian hamsters as well. The cause of an increased zinc excretion during the melanoma growth seems to consist in an increased lysis of pigment cells. The presumed destruction of tumor cells was not accompanied by a higher excretion of magnesium, though. Concentration of creatinine proved to be a useful mean for expressing of standard excretion of various substances also in hamsters.

Stress and Ehrlich ascites tumor in mouse.

The aim of the study was to investigate the interaction of the psychosomatic stress and the Ehrlich ascites tumor (EAT) growth in mice. The stressor consisted of combination of a light signal followed by a mild electric shock. The first experiment was performed on CBA mice irritated for 0, 2 and 4 weeks respectively, prior to intraperitoneal transplantation of the EAT. In the second study, mice of BALBc strain were used. Stress was applied 4 weeks before the tumor transplantation and continued throught the experiment. Both the irritated and the nonirritated animals were subjected to either intraperitoneal or subcutaneous inoculation of the EAT. In both experiments, mice were left to live until their spontaneous death. In the first experiment, after a 2-week irritation the experimental animals showed a significantly longer survival time as compared to the controls. Longer or shorter duration of the irritation had no significant effect on the results obtained. Results yielded by the second experiment showed no significant difference in the time of survival of the irritated and nonirritated animals after the i. p. transplantation, whereas after the s. c. inoculation of the EAT, the irritation significantly increased the survival period. The EAT in irritated mice was observed to have invaded the vitals later and less frequently than in the nonirritated animals. Quantitative histological analysis of some endocrine and lymphatic organs revealed signs of stress in the experimental animals. The EAT transplant, per se, had a stressogenic effect too.

Invasive thymoma (a case report).

A 10-year old Nigerian boy was admitted to hospital with a history of swelling in the neck, change of voice, dysphagia and dyspnoea of 2 weeks duration. He died on the 2nd day of admission. Autopsy revealed a thymoma infiltrating the thyroid, trachea and neck muscles and transforming the tracheal lumen into a slit like space. The condition is considered worthy of record on account of its rarity and of the short clinical course terminating in death. The authors feel that the most important factor in determining the prognosis of thymoma is the presence or absence of gross invasion.

Cell surface adhesiveness of mouse sarcoma lines evaluated by latex particle adherence assay: correlation with growth behavior and electrophoretic mobility.

Using the latex particle adherence assay and five mouse sarcoma cell lines of the identical origin, etiology and genotype but differing in malignancy we attempted to correlate the degree of cell surface adhesiveness with growth behavior and electrophoretic mobility of cells. Higher tumorigenicity of four of the cell lines (Mc11--Mc14) was associated with lower cell surface adhesiveness and, conversely, lower malignancy of the fifth line (Mc15) with higher cell surface adhesiveness. No simple correlation or causal relationship was found among the electrophoretic mobility of the lines and other cellular characteristics.

Human lymphoblastoid cells in long-term cultures: electron microscopic study.

Fine structure of lymphoblastoid cell lines (LCL) derived from leukocytes of three patients with acute myelosis and one patient with undifferentiated acute leukemia were studied repeatedly by electron microscope. The majority of cells in all lines was of the same primitive lymphoblastic type as refered to in literature regardless to their origin. Morphology of the cells did not change with time of cultivation for several months, by freezing and thawing and by new recultivation after prolonged cryopreservation. No morphologic signs of contamination by viruses were found. No evident morphologic signs of differentiation to mature granulocytes or "mature" lymphocytes or plasmatic cells were observed.

Some cytochemical reactions in the cells of long-term cultures derived from the blood of leukemic patients.

Combined therapy of L1210 leukemia with Damvar and cytostatics.

Damvar, the delta-(2-amino-6-hydroxy-3,4-dihydro-4-oxo-5-pyrimidinyl) valeric acid, administered alone s. c. or i. p., has no effect on L1210 leukemia. In DBA2 mice with L1210, Damvar administered in combination with 5-fluorouracil, or with 6-mercaptopurine, or with cyclophosphamide, in optimal doses, enhances or intensifies the antileukemic effects of the above cytostatics.

Modification of adriamycin cardiotoxicity by Damvar.

In experiments performed in mice, Adriamycin (15 mg/kg i. p.) produced serious damage of the heart muscle. Damvar (2 x 200 mg/kg orally) a new cytostatic drug combined with Adriamycin decreased conspicuously its cardiotoxicity.

Effects of cyclophosphamide on the prenatal development of the Swiss strain mice.

Groups of pregnant mice were given cyclophosphamide at daily doses from 6.4 to 40.8 mg/kg from the 11th to the 15th day of gravidity. Significantly lower weight and length of fetuses were noted after each of the above dosis as against the controls, while the volume of the amniotic fluid was significantly increased. The number of resorptions rose significantly with the growing dose, the embryotoxicity proving 100-percent effective in both the highest doses. Malformations were followed in live fetuses. Evident malformations resulting from cyclophosphamide application were those of the head (hydrocephalus, exencephalus, exophthalmus, median cleft palate, micrognathia), of the extremities (syndactylia, polydactylia), caudal malformation (stunted and crooked tail) and an overall developmental retardation of the fetuses. Skeletal anomalies comprised synostosis, retarded ossification, deficiency or redundance of some bones of the head, trunk and limbs.

White and red spleen pulp recovery in mice after continuous irradiation.

The process of spleen recovery after the continuous irradiation with higher daily doses up to the total dose 9570 mGy was studied in histological slices. Histological picture seen immediately after the irradiation gives evidence that significant changes occur in spleen white and red pulp. The lymphopoiesis recovery is slow and lasts till day 60, resp. 90 after irradiation. Erythropoiesis and myelopoiesis recovery of the red pulp is more intensive and is completed within 14-28 days in all three examined groups.

Tumor differentiation and immunocompetence in cervical cancer patients.

The authors investigated cell-mediated and humoral immunity parameters of 161 and 131 cervical cancer patients in connection with the histological differentiation of the tumor. As regards the carcinoembryonic antigen and acute phase proteins no difference was found between differentiated and undifferentiated types of tumors. Significantly higher levels of IgA, IgM, 3rd component of the complement and of skin reactivity to phytohemagglutinin were observed in patients with differentiated tumors.

Mental changes in women with gynecologic cancer.

In 47 women with gynecologic cancer a questionnaire method was used for investigation mental changes in these patients. Alteration of value orientation, self-image, sexuality, attitude to cancer, etc., were analyzed in the group of the followed patients.

The influence of 5-nitro-2-furyl acrylic acid on human fibroblasts cultivated in vitro.

The authors studied the in vitro effects of 5-nitro-2-furyl acrylic acid on growth activity of EUE human fibroblasts. They investigated the course and intensity of macromolecular syntheses, mainly DNA synthesis in these cells. Going by the results obtained it can be concluded that the cytotoxic effects of 5-nitro-2-furyl acrylic acid are also connected with its genotoxic effects, since that substance will induce damage to DNA in human cells.

Cell proliferation in the thymus of Ehrlich ascites tumor bearing mice.

Thymic involution of Ehrlich ascites tumor bearing Swiss and NMRI mice was associated with an increased incorporation of 3H-thymidine into thymus DNA and an enhanced activity of adenosine deaminase. The apparent depletion time constants, which were determined by the following time course of the retention of 3H-DNA, indicated that the rate cell depletion was increased in the thymus of tumor bearing mice. The results suggest that the thymic involution was correlated with an increased proliferative activity and an accelerated depletion of thymic cells.

The prognostic value of serum innumoglobulin levels in Hodgkin's disease.

Serum immunoglobulin concentrations have been measured in 31 patients with Hodgkin's lymphoma and related to disease activity and clinical stage. The patients with active untreated disease, the mean IgA concentration was significantly above the normal mean level and the mean IgM concentration was normal. No significant correlation was found between high IgA concentrations and liver involvement. There were decreased mean IgA and IgM concentrations during remission. The changes in immunoglobulin concentrations were found to be dependent on the clinical stage. The data suggest that in certain patients with active Hodgkin's lymphoma, the humoral-mediated immunity may be affected in addition to the cellular-mediated immunity.

Does lymphography promote dissemination of tumorous disease?

In two groups of patients with malignant melanoma on the lower limb and with metastases in the inguinal lymph nodes the authors followed the time of appearance of lung metastases and the survival time. In one group lymphography of the extremity was performed, the other one served as a control. Tabulated time data were mutually evaluated between the two groups by the parametric T-test and with the aid of the nonparametric Me-test. No significant difference was found between the two groups, not even with 20% probability. Thus, lymphography is not likely to have an influence on deissemination of malignant melanoma.

Combination of lymph node and lung metastases in thyroid cancer.

In thyroid cancer patients with pulmonary metastasis lymph nodes involvement was found in 62.6%. The incidence of lymph nodes metastases is discussed in relation to the age of patients and to the stage of the disease.

Electrochemical properties of polycyclic compounds studied by the polarographic method in anhydrous systems. VII. Polarographic oxidation of carcinogenic hydrocarbons in dimethylformamide.

The electrooxidation of polycyclic carcinogenic hydrocarbons in dimethylformamide solutions was studied by polarography. The results generally have confirmed that a correlation exists between oxidation potentials and carcinogenic activity of the studied polycyclic aromatic hydrocarbons. The half-wave potentials of the polarographic curves have been found to be directly related to the root xn of the Hückel secular equation for the highest bonding orbital and to the reduction potentials measured in dimethylformamide.

Serum immunoglobulin levels and survival rates in bronchogenic carcinoma patients.

Serum immunoglobulin IgG, IgA and IgM levels were estimated in 112 patients with bronchogenic carcinoma, and the obtained values were as follows: IgG = 1762 +/- 595 mg/100 ml; IgA = 332 +/- 104 mg/100 ml; IgM = 157 +/- 76 mg/100 ml. Subsequently the patients were divided in a group of 74 patients with epidermoid carcinoma, and a group of 38 patients with small cell anaplastic carcinoma of the bronchus. In patients with epidermoid carcinoma, in cases with disseminated disease the mean survival was shorter (7.9 months) when compared to the mean survival of patients with localized disease (20.8 months). Similarly, a depression of lymphocyte counts was observed in cases with disseminated disease. In cases with IgA concentrations ranging from 300 to 410 mg/100 ml longer survival rates were observed. In patients with small cell anaplastic carcinoma variety, the survival rates in patients with localized disease were almost identical with those in patients with disseminated disease (11.4 months versus 9.9 months). The longest survival rates were observed in cases with IgG concentrations ranging from 1600 to 2100 mg/100 ml, and IgM concentrations ranging from 190 to 320 mg/100 ml of serum.

Natural changes in the cell division rate in normal tissues and the possible role of adrenalin concentration.

Mitotic incidence (MI) and DNA labeling index (LI) were measured in the liver, lung and kidney of rats at different times throughout the day. There were significant variations in MI and LI in all organs and there were also significant differences between organs in respect of the pattern of variation. It was found that adrenalin had no consistent inhibitory action on the MI and LI of these tissues maintained in organ culture. These results are not consistent with the view that diurnal (or circadian) rhythms in mitotic activity are directly mediated by changes in adrenalin secretion. In organ culture the rate of entry into DNA synthesis was higher than the rate of mitosis, leading to significant increases in the proportions of cells with reduplicated DNA content.

Production of plasminogen activator and inhibition of embryonic cell aggregation by cultured human normal and neoplastic cells.

The plasminogen activator (PA) production and the capacity to inhibit embryonic neural retina (NR) cell aggregation by human normal and neoplastic cell lines have been studied. The PA production was detected by both iodinated fibrin and casein lysis assays, and by changes in cell morphology at the presence of activated PA, using dog serum. Since the casein lysis assay and morphological changes proved to be less sensitive than 125I-fibrin lysis assay, a good correlation between these three assays could be observed provided that PA production measured by fibrinolysis exceeded 10--20%. The neoplastic cell lines exhibited the PA production to quite a large extent. The highest fibrinolytic activity (78%) was found in the case of bladder carcinoma cells T24, while the B-5GT cells from giant cell tumor of bone failed to produce any detectable amount of the PA. The cells from synovial sarcoma and both glioma lines exhibited fibrinolytic activity of about 10% and four sarcoma cell lines over the range 20--50%. Out of 13 normal cell lines tested, 7 were negative or exhibited very low fibrinolysis not exceeding 3% of total radioactivity. Four cell lines derived from kidneys, lungs, intestines, and from mixed embryonic tissues showed a marked fibrinolytic activity of about 10--37%, a slightly elevated fibrinolysis being found in embryonic lung cells LEP and cells from fetal skin tissue only at the presence of dog serum. The fibrinolysis detected in the neoplastic cloned cell populations showed considerable differences in the PA production between individual cell clones isolated from the same parental cell line. Unlike the normal fibroblastic cells B-41FB derived from bone, all neoplastic cell lines tested possess the capability to inhibit embryonic NR cell aggregation significantly. The results suggest the effect not to be dependent upon the PA production.

Changes in blood flow of growing experimental tumor determined by the clearance of 133Xe.

133Xe clearance method has been employed for measurement of the blood flow in 6C3HED lymphosarcoma implanted in flank of C3H mice. In small tumors (9-day old) the mean rate of blood flow was 9.4 +/- 5.0 ml/100g/min. but with progressing growth of tumors the rate of blood flow significantly decreased. In large tumors (14-day old) the mean rate of the blood flow was only 5.4 +/- 2.8. Our results demonstrate that this type of tumor, similarly as most of transplanted tumors are not well perfused and, that there is an inverse relationhip between the blood flow and the tumor size.

Changes in hemopoiesis of mice of the C3H strain following transplantation of Gardner lymphosarcoma and infection with LDH-virus. I. Circulating blood.

The values of red and white blood count, of spleen and liver weight were determined in mice of the C3H strain after transplantation of Gardner solid lymphosarcoma contaminated with LDH-virus and after infection of LDH-virus, and compared with those found in normal intact mice. Special attention was devoted to early post-transplantation period and the final stage of tumor growth. The second day after infection of mice with LDH-virus, leukopenia with marked lymphopenia was observed, together with a reduced number of reticulocytes and spleen enlargement. The same changes became more pronounced in tumorous mice on the second posttransplantation day. The changes--with the exception of spleen enlargement--following LDH-virus infection became normalized within the period of the final stage of tumor growth. Contrarily, in mice with tumors in the final stage of the disease besides spleen enlargement also the reduced erythrocyte counts, leukopenia with pronounced lymphopenia and thrombocytopenia were found.

Gamma-glutamyltransferase in Morris hepatomas 5123D passaged in Buffalo and in F1 (Buffalo x Wistar) rats.

Comparative biochemical and immunological investigations were performed on purified gamma-glutamyltransferase from the Morris hepatoma 5123D and from the 5123D/AS variant passaged in F1 (Buffalo x Wistar) rats. The enzyme was also localized in hepatoma sections by immunofluorescent technique. The obtained results confirm the hypothesis that low gamma-glutamyltransferase activity in Morris hepatoma 5123D/AS is caused by the enzymatic protein defficiency and not by the inhibition of the enzyme.

Phase I study using combination of hydroxyurea and 5-azacytidine (NSC-102816).

Thirteen patients were treated with a hydroxyurea -- 5-azacytidine combination in an attempt to show an increase in therapeutic efficacy of 5-azacytidine without increased bone marrow toxicity. Leukemic and nonleukemic patients were selected in order to compare the effect of a combination therapy on bone marrow. There was no clear-cut tumor response observed in evaluable patients. Overall activity of the combination, as judged by the onset of myelosuppression, appears to be higher than that of 5-azacytidine given alone.

Inhibition of leukocyte migration by soluble tumor extracts in human malignant melanoma.

Leukocytes from 63 melanoma patients and from 38 control donors were examined by a leukocyte migration inhibition (LMI) assay. The antigens were three soluble extracts of fresh melanomas and one of lung cancer. A total of 62 out of the 99 (62.6%) blood samples from melanoma patients reacted with allogeneic extracts of melanoma in comparison with 11 out of the 48 (22.9%) in the controls (the values significantly different; P less than 0.001). The migration inhibition of melanoma patients' leukocytes by the lung cancer extract was markedly less than inhibition by any of the melanoma extracts. LMI in patients with distant metastases before treatment (9 out of the 11--81.8% positive results) was higher than in patients with regional lymph node metastases (5 out of the 9--55.5%) and than in patients with localized disease (3 out of the 7--47.8%). The reactivity of patients after surgery was higher than in patients before surgical resection of the tumor. Positive reactions were less frequent in patients with distant metastases receiving chemotherapy or/and immunotherapy than in patients before treatment.

5-Aza-2'-deoxycytidine: preclinical studies in mice.

5-Aza-2'-deoxycytidine administered to normal mice repeatedly on daily schedule affects mainly the number of blood leukocytes and bone marrow myeloid cells. Using the cell-free extract from mouse spleen phosphorylated intermediates of 5-aza-2'-deoxycytidine-3H have been isolated and kinetics of their formation both in vitro and in vivo in mouse spleen cells have been studied. The administration of the drug affects the synthesis of DNA in mouse spleen resulting in its initial depression followed later by the enhancement of thymidine uptake.