Further studies on liver glyoxalase I and glyoxalase II. Activity in mice bearing sarcoma 180 and L1210 leukemia.

Determinations were made of glyoxalase I and glyoxalase II acitivity in the liver of mice (BDF1 and DBA2 strains) bearing sarcoma 180 and L1210 leukemia in ascites form. A progressive decrease in the both glyoxalase I and glyoxalase II activities to about 40--60% of that of the control groups was observed within the developing period 8--9 days. Test results are interpreted in the light of the postulated role of this enzyme system in cell division and in the tumor development process.

Activity of N-acetyltransferase in patients with malignant lymphomas.

Activity levels of N-acetyltransferase and acetylation types in patients with malignant lymphomas, following sulfadimidine loading, were established. The ratio of "fast" and "slow" acetylators among patients with malignant lymphomas is shown to be similar to that in control group. The rate of acetylation in "slow"-acetylating patients, however, was higher than in healthy controls. A relationship of N-acetyltransferase activity, tumor progression and, to a considerable extent, effectiveness of therapy was shown.

Cell proliferation kinetics and nuclear morphology in endometrial cancer under progesteron treatment.

Cell proliferation kinetics by using the double labeling with 3HTdR--14CTdR, and nuclear morphology were studied in 20 patients with endometrial cancer who were given progesteron as a preliminary therapeutic measure. Results of these studies indicate that we have to face the fact that in all histological types of endometrial cancer considerable variations in the amount of tumor cells undergoing secretory conversion occur due to the primary heterogeneity of the tumor cell population.

Antilymphoid activity of leptospiral exoproducts.

Intraperitoneal application of supernatant fluid of L. patoc caused a pronounced and transient fall in peripheral lymphocyte count of Syrian hamsters. Pathogenic and possible cancerologic aspects of this findings are discussed.

Cytotoxic and cancerostatic effect of 1,4-dithiaanthraquinone-2,3-dicarbonitrile.

1,4-Dithiaanthraquinone-2,3-dicarbonitrile (DTA) has been found to exert a considerable cytostatic effect especially on some of the investigated types of eukaryotic cells, concretely on the HeLa cells, moulds, yeasts, protozoa and algae. In cells of the Ehrlich ascites carcinoma (EAC) DTA after a short exposition causes a parallel inhibition of incorporation of 14C-adenine and 14C-valine, in proportion to its rising concentration. The inhibition of biosynthetic processes thus made manifest, is probably a consequence of the primary DTA intervention into the energy metabolism of EAC cells, particularly in glycolysis. The effect of DTA in concentrations capable of bringing about full inhibition of glucose consumption or lactate formation in EAC cells also results in a loss of their transplantability. On the other hand, DTA also exerts a cancerostatic effect on the Ehrlich ascites carcinoma in mice.

Cellular mechanisms of the radiomodifying effect of hypothermia.

The irradiation of experimental tumors with a dose of 2000--2500 rad (20--25 J/kg) under hypothermia promoted an inhibition of the growth to a greater degree than the irradiation under normal conditions. In Guerin's tumor the inhibition of DNA and RNA synthesis was more expressed after the irradiation under hypothermic conditions than under the irradiation, and/or hypothermia alone. After the irradiation of the Guerin's tumor under hypothermia the cells were synchronized during the presynthetic phase of the cycle (block G1-S), and the effect of synchronization was more expressed in the tumor than in the normal tissue. The irradiation under hypothermia decreased the proliferative pool to a greater degree than the irradiation and/or hypothermia alone.

Some characteristics of karyotype changes in the NK/Ly ascites tumor during its growth.

The NK/Ly ascites tumor has been studied with respect to the karyotype and its changes during tumor growth. The numeric stem line of the tumor cells is 43. The chromosome markers are as follows: a large metacentric chromosome, a large submetacentric chromosome with the long arm divided or, in a few cases, not divided by achromatic space, a large telocentric chromosome with a well-visible achromatic space which divides its long arm into two sections, and 1--2 extremely small chromosomes. The combination of the large metacentric chromosome, the large submetacentric chromosome with achromatic space, and of 1--2 extremely small chromosomes is modal. With the NK/Ly tumor growth, the number of modal cells decreases, while the variability in the number of chromosomes in the cells increases, i.e. their number is diminishing, and so is the percentage of cells with the modal combination of chromosome markers. On the 12th day of tumor growth, no clearly visible stem line can be noticed. The observed karyotype changes may be considered, alongside with other causes, to be due to longer period of the cell cycle during the NK/Ly ascites tumor growth.

Growth characteristics of the RL-67 lung tumor -- a new model for experimental therapy of metastatic processes.

Implantation of the RL-67 tumor in to the limb muscle of C57Bl strain mice resulted in the tumor growth at the site of implantation and the metastatic process became manifest predominantly in the lungs of all animals. A direct correlation was established between the primary tumor and its metastases. The number of metastases on the 20th day after the transplantation amounted to about 60. The mean survival time of the tumor-bearing mice was approximately 24 days. Seven different substances were administered intraperitoneally to tumor-bearing animals and their effect on the tumor and its metastases was established. It was found out that heparin and carboxymethyldextran led approximately to a 50% decrease in the number of lung metastases in comparison with the controls. A certain "prophylactic" effect on metastases was achieved with Bleomycin. The data of the biological and histological studies suggested that the RL-67 lung tumor may be used as a suitable model for investigation of the factors influencing the tumor metastases.

Protein synthesis in tumor host. I. Enhanced peptide elongation in transplantable tumors and host liver.

Protein synthesis was significantly enhanced in subcellular systems containing ribosomes and cytosol from the liver of Walker tumor-bearing rats from the second week following the tumor transplantation and this enhancement persisted for the whole period of tumor growth. Homologous systems from Zajdela hepatoma and host liver showed a markedly increased poly(U)-dependent peptide elongation when compared with normal liver tissue. A stimulation of polyphenylalanine synthesis resulted from the addition of cytosols from tumors or host liver to ribosomes from normal rat liver. Similar results were found for the binding of phenylalanyl-tRNA to ribosomes. Ribosomes from tumors and host liver are more active in peptide elongation than particles from normal liver tissue. A more than 10-fold stimulation of phenylalanine polymerization resulted from the addition of poly(U) to ribosomes from Zajdela hepatoma whereas only less than 2-fold enhancement was found when using ribosomes from normal or host liver. Hepatoma ribosomes apparently contain only a low proportion of polyribosomes carrying natural message. Enhanced protein synthesis in tumors and host liver is apparently due, in particular, to an increased activity of soluble factors required for protein synthesis and less due to an increased activity of ribosomes.

Differences in uptake of aminoacids by patients with various forms of cancer.

The serum concentrations of alphaaminonitrogen (AAN), lysine, valine and leucines were determined before and within the period of 24 hr after the administration of Trophysan (10% solution of glucose containing a mixture of aminoacids) in 100 patients with cancer (17 with gastrointestinal carcinoma, 34 with uterus carcinoma, stages I to III; 8 with breast carcinoma, stages II and III; 15 with bronchogenic carcinoma, 10 with various localizations and 15 with metastatic cancer) and in 22 patients with benign tumors. A significant decrease in the serum content of AAN, valine and lysine was noted in patients with cancer (stages I to III) at 24 hours after the administration of Trophysan. This effect was absent for the patients with benign tumors. The enhanced uptake of aminoacids found in patients with cancer is probable the result of the negative nitrogen balance associated with the malignant state.

Effect of cytostatics on some platelet functions in vitro. VI. Vinblastine.

The influence of vinblastine upon principal functions of blood platelets in vitro (adhesion, aggregation, platelet release reaction, membrane stabilization) has been followed using an extended system of laboratory examinations. The authors have found that the concentration of vinblastine which is higher than the average therapeutic dose (10(-5)M) inhibits the functional ability of platelets so that it may cause a thrombopatic disorder.

Influence of heparin and defibrase on transplantability of syngeneic tumors in mice.

It had been previously demonstrated that fibrin clot might have an important role in the growth of malignant tumors and in metastasis formation. For this reason an attempt had been made to find out whether inhibition of fibrin formation by heparin or fibrinolysis induced by defibrase could limit tumor growth. Male mice of inbred strain were challenged with suspension of tumor cells. For anticoagulation and/or fibrinolytic therapy two different schedules were used. The treatment was started either simultaneously with the tumor transplantation, or one week prior to the injection of tumor cells. The size of tumors was bigger in the group of treated animals, when compared with the controls. However, the difference was not statistically significant. On the other hand, frequency of implanted tumors was significantly higher in animals with heparin and defibrase treatment or pretreatment.

Estrogenic effect in vaginal smears in cancer of the uterine cervix.

Values of karyopyknotic index (KPI) in a series of 37 uterine cervix cancer patients were significantly increased when compared with the same groups of 61 healthy women. Radiosensitivity of the tumors was accompanied by subsequent decreasing in the KPI values while in all radioresistent cases as well as in all early recurrences persistence of initial high KPI values could be proved. Significant differences in the kinetics of KPI changes during and after radiation in the group of praemenopausal and postmenopausal patients support suggestion for non ovarian origin of this estrogen like activity.

Carcinogenicity and polarographic behavior of dibenz[a,h]anthracene, dibenz[a,h]acridine and dibenz[a,h]phenazine.

In a simultaneous test of carcinogenicity the authors have studied the activities of dibenz-[a,h]anthracene, dibenz[a,h]acridine and dibenz-[a,h]phenazine. These compounds were dispersed in paraffin disks and subcutaneously implanted in rats. Each experimental group consisted of 30 animals. The number of sarcomas induced by the above-mentioned compounds was 20, 5 and 3 respectively. Tumorigenicity of the aza analogues of dibenz[a,h]anthracene was proportional to their electron donation and inversely proportional to their electron acceptance in the performed polarographic test. The authors discuss the possibilities of using polarography as a screening test for the exclusion of carcinogenic polycyclic compounds.

Combined chemoimmunotherapy of L5178Y lymphoma.

The antitumoral effect of chemotherapy and immunotherapy with BCG and/or irradiated L5178Y cells has been compared to antitumoral effect of chemotherapy alone. Significantly better results were noted in a transplantable L5178Y lymphoma grafted on DBA/2-J mice treated with methotrexate and BCG than in any other group which was not treated with BCG. The effect of irradiated lymphoma cells was similar to effectivity of methotrexate alone.

Electrochemical properties of polycyclic compounds studied by the polarographic method in anhydrous systems. VI. The influence of proton-donor on reduction of carcinogenic aromatic hydrocarbons in anhydrous environment.

The authors have studied the electrochemical behaviour of a series of carcinogenic and inactive aromatic hydrocarbons in anhydrous environment in the presence of a proton-donor. As anhydrous environment the authors used water-free dimethylformamide and dimethylsulfoxide; as proton-donor phenol. Phenol gave rise to quantitative changes in the number of polarographic waves when the carcinogenic aromatic hydrocarbons were undergoing reduction. Between the two original waves there originated in the presence of phenol a new polarographic wave in these hydrocarbons. In inactive analogues of the investigated hydrocarbons no such effect of phenol has been ascertained. In their present study the authors discuss the potential mechanism of electroreduction of carcinogenic and inactive aromatic hydrocarbons in anhydrous environment in the presence of a proton-donor and compare the values of half-wave potentials of the investigated compounds measured in water-free dimethylformamide, dimethylformamide + phenol and in ethylenglycolmonomethylether.

Establishment of cell line derived from human malignant melanoma.

A new human cell line of malignant melanoma (MJM) was established with the use of the in vitro fragment technique. It has been maintained over 34 months of continuous cultivation. Three types of cells can be recognized by light microscope. The epitheloid elements predominate, less frequent are fibroblastoid and giant multinuclear cells. The pigment production is not macroscopically visible. Over 60 per cent of analyzed metabphases showed hyperdiploid number of chromosomes, the rest was mostly tetra and hexaploid. No marker chromosomes were detected. The growth studies indicate the MJM cells have 63-hr doubling time. Cytochemistry revealed positive pigment or propigment granules in 36 per cent of cells. Ultrastructural studies did not detect melanin granules but some particles resembling atypical premelanosomes and melanosomes were recognized in some sections.

Cytologic criteria in diagnostics of esophageal dysplasia.

On the basis of exfoliative cytology material of 62 patients with precancerous esophageal diseases 3 stages of esophageal dysplasia were determined: mild, moderate and severe. In order to establish the stage of dysplastic alteration the author used morphological, functional, morphometric and cytochemical criteria. Cytologic criteria suggest by the author are objective indicies to find out early malignancy of esophageal epithelium. The usage of the above mentioned criteria enable to distinguish earlier forms of cancer of the esophagus and permits to lower the number of advanced cases.

Immunoglobin levels in patients with Hodgkin's disease.

The authors examined a course of immunoglobulin values (IgA, IgG, IgM) in 40 patients with the Hodgkin's disease and repeatedly in 11 healthy persons. In the patients they followed the levels of the immunoglobulins during the disease, up to 5 years. In untreated patients there were statistically significant increased IgA levels and insignificant increase of IgG. In the course of remission the immunoglobulin values were normalized. The radiotherapy did not affect significantly the value of immunoglobulins, the combined chemotherapy (COPP) led to a drop of this level. The levels of immunoglobulins are recovered after chemotherapy in the case of successful treatment. In terms of the prognosis the authors consider as unfavorable high increase in the immunoglobulin levels in untreated patients and their rapid drop during the chemotherapy. They observed an average preterminal drop of the IgA by 27%, IgG by 23% and IgM by 44% with respect to controls.

Delayed hypersensitivity reactions in patients with breast cancer.

Fifty six patients with metastatic cancer of the breast (stage IV) were treated with Cyclophosphamide, 5-Fluorouracil and Cyclophosphamide + 5-Fluorouracil. Tests for delayed hypersensitivity to homologous tumor antigen before treatment were positive in 83.9% and negative in 16.1%. Response to DNCB was positive before treatment in 51.8% and negative in 48.2%. Following chemotherapy the skin reaction, to homologous tumor antigen remained positive, only in 12.6% and negative in 87.4%. The reaction of DNCB remained positive after treatment only in 17.8%. In the remaining 82.2% suppression of the reaction occurred. These data show that chemotherapy may suppress, to a certain excent, immune responses. It is established that among patients who have shown a positive reaction to homologous tumor antigen 55.3% of all cases have displayed objective response to the treatment, and among these with negative skin reactions objective responses were observed in 22.22% of all cases. In patients with positive DNCB reactions objective responses were observed in 79.3% and among the negative ones--in 37.4%.

The response rate at chemotherapy in advanced Hodgkin's disease reflected by peripheral lymphocyte count.

Forty patients with advanced Hodgkin's disease stage III and IV were treated with combination chemotherapy. Absolute lymphocyte count in the peripheral blood was determined before start to chemotherapy. Higher levels of lymphocytes in the peripheral blood were associated with higher chemotherapy response rates. This difference was statistically significant (P less than 0.05).

Metabolic turnover of pyridine nucleotides in ascites cells of sarcoma Sa 180 and in the liver tissue of rats before and after ionizing radiation.

Metabolic turnover of NADP+ labeled with 14C in the ribose moiety of their molecules was determined in the ascites cells of sarcoma Sa 180 and in the rat liver tissue. The half-lives of NAD+ and NADP+ in the Sa 180 sarcoma cells were 60 min (NAD+) and 90 min (NADP+) and 120 min (NADP+). Experiments were done on animals to study the time-dependent effect of ionizing radiation on the values of pooled NAD+ and NADP+ after 0.129 C/kg (500 R) and 0.387 C/kg (1500 R) whole-body irradiation, and the metabolic turnover of these nucleotides 5 h after whole-body irradiation with 0.387 C/kg (1500 R). Exposure to 0.129 C/kg (500 R) whole-body irradiation induced no apparent changes compared to the controls. Within 5 h of irradiation the whole-body dose of 0.387 C/kg (1500 R) produced changes in rat liver cells characterized by a reduction of the nucleotide biological half-lives (NAD+ from 80 to 60 min and NADP+ from 120 to 70 min). No such changes in the pyridine nucleotide turnover were detected in Sa 180 ascites sarcoma cells.

Mitochondrial ATPase of Zajdela hepatoma. VI. Effect of extramitochondrial ATP and pH on uncoupler-sensitivity of mitochondrial ATPase activity.

Coupled Zajdela hepatoma mitochondria were reported to exhibit uncoupler-insensitive ATPase activity. The results of this study show that under specific conditions the ATPase activity of Zajdela hepatoma mitochondria can be stimulated by uncouplers. These conditions include (a) the addition of ATP to the mitochondria before the uncoupler in the ATPase activity assay or (b) elevation of pH (above 8.5) of the ATPase activity assay medium.

Serum enzymes in ovarian carcinoma.

The following enzymatic activities were measured in serum of patients with benign and malignant ovarian tumors before treatment: alkaline and acid phosphatases, aspartyl (AspAT) and alanyl (AlAT) aminotransferases, leucyl (LAP) and alanyl (AAP) aminopeptidases, lactate dehydrogenase (LDH), gamma-glutamyl transpeptidase, cathepsin, alkaline ribonuclease (RNase) and beta-glucuronidase. It was shown that at least three determinations (phosphatases and LAP) are practically useless in a discrimination between the examined groups. RNase in combination with AspAT (AlAT) or RNase with AAP and LDH were found to give the best results as marker enzymes.

CEA-like antigens in different histologic types of human lung carcinoma.

Comparative studies on the presence and immunological reactivity of CEA and CEA-like antigens in lung and digestive tract cancers were performed. Specific goat and rabbit anti-CEA immune sera were used. Immunofluorescence studies revealed in histologically different types of the lung carcinomas the presence of the cross reacting antigen with classical CEA. Absorption procedures, and double diffusion tests have shown the existence of two at least antigenic determinants in CEA molecule: 1. the determinant specific for the lower parts of the digestive tract cancers not found in the lung carcinomas, and 2. the determinant common for the lung, and the digestive tract cancers.

Carcinomas of the liver in rats ingesting kepone.

Young male and female albino rats ingested 0, 1, 5, 10 or 25 ppm Kepone, an organochlorine pesticide, in the diet for two years. Carcinomas of the liver, as well as hyperplastic nodules and moderate and severe diffuse hyperplasia were observed in Kepone-treated rats. Such hepatic lesions were not seen in control rats. Female rats ingesting Kepone were more susceptible than male rats to hepatic carcinogenesis. Rats ingesting 50 or 80 ppm Kepone developed severe diffuse hepatic hyperplasia and did not survive beyond 26 weeks.

Relationship between alpha-fetoprotein positive cells and precancerous lesions in rat liver during 3'-methyl-4-dimethylaminoazobenzene carcinogenesis.

Localization of characteristic precancerous lesions was compared with that of alpha-fetoprotein (alpha-FP) positive cells during hepatocarcinogenesis in Wistar rats fed 0.06% 3'-methyl-4-dimethylaminoazobenzene in basal diet. Proliferative activity of liver cells was simultaneously followed by deoxycytidylate deaminase. The first alpha-FP positive hepatocytes were found after 3 weeks of carcinogenesis in certain parts of the capsule 0.1--0.2 mm thick. In the seventh week of carcinogenesis their number increased and they were present in groups also subcapsularly. At this stage, hyperplastic islets began to be formed in the liver parenchyma. After 14 weeks of carcinogenesis, alpha-FP positive cell clusters and islets were also found in the deeper layers of liver parenchyma in regions with proliferation of transitory cells. Hyperplastic basophilic foci and islets occurred regularly, however, these were not identical with alpha-FP positive sites and their incidence was more frequent.

Preferential replication of murine xenotropic type-C virus in human lymphosarcoma-derived cell lines.

Murine xenotropic virus, designated 698/X, was recovered by implantation of human lymphosarcoma-derived cells U-698M into nude mice of Giovanella's colony. The budded and extracellular particles revealed typical type-C morphology, the latter possessing reverse transcriptase (RT) activity and exhibiting a buoyant density 1.17 g/ml in sucrose gradient. In competitive radioimmunoassay using iodinated p30 of Rauscher MuLV, the 698X viral concentrate and cell extracts of both implanted lymphosarcoma cells (U-698M-N-1 and U-715M-N-1) were as effective as Gross MuLV, thus indicating the murine origin of the virus. The propagation of the 698/X virus in five human, four mouse (permissive for N- and B-tropic MuLV), two rat and one bovine cell lines was followed by RT, XC syncytia assays and EM investigations. The replication of the 698/X virus seems to be restricted mainly to both human lymphosarcoma-derived cell lines U-698M and U-715M. The new recovery of the virus from the nude mouse by implantation of U-175M cells has asserted its high tropism to human lymphosarcoma cells and its murine origin. The comparative response of mouse, rabbit and rat cells exposed to both NZB and 698/X xenotropic murine viruses exhibited host range differences between these viruses. The rabbit SIRC and rat embryonic cells REC were fully permissive for the murine xenotropic NZB virus, while low viral production was detected by RT assay only in 698/X virus infected SIRC cells.

Demonstration of alpha 2-pregnancy associated-glycoprotein (alpha 2-PGA) in serum of men with different localization of cancer.

By counterimmunoelectrophoresis in agarose gel the presence of alpha 2-PGA in the sera of 28 untreated patients with different localization of cancer was demonstrated. In 17 (60.7%) a precipitation line between the well with patient serum and that with monospecific anti-alpha 2-PGA serum, diluted 1 : 7 was obtained. At 1 : 7 dilution, with the sera from 28 healthy persons precipitation lines were not obtained. The determination of alpha 2-PAG can be made quickly and contributes to cancer diagnosis in men.

Combined regional chemotherapy and radiation therapy in the treatment of epidermoid carcinoma in the oro-facial region.

Followed treatment of oro-facial epidermoid carcinoma by combined chemo- and radiation therapy with eventual surgical intervention. Cytostatic drugs were applied intraarterially. After a monocytostatic treatment trial with Methotrexat (MTX), a combined cytostatic program was worked out utilizing two cytostatic drugs, viz. MTX and Bleomycin (BLM) in a cyclic alternation. Usefulness of chemotherapy and its inclusion into the treatment of epidermoid carcinoma in the oro-facial region was found justified principally in the case of combined therapy. However, the selected intraarterial mode of cytostatic administration is attended by certain drawbacks, mainly of a technical nature and is not suitable for routine application. For this reason, the authors have abandoned the i. a. mode of cytostatic administration in epidermoid oro-facial carcinoma and have adopted systemic application and this in combination with radiation, eventually surgical therapy with the utilization of chemotherapy.