Dominant-lethal assay of selected cytostatics.

Dominant-lethal assays in male mice were made with the following cytostatics: Cyclophosphamide, TS-160, Edikron, Penberol, Cytembena, Mercaptopurine, Butocin, and Damvar. The cytostatics were administered mostly for 14-day periods, with the exception of Butocin (7 days) and high doses of Cyclophosphamide and TS-160 (single administrations). From the first day of administration on, the males were mated with intact females mostly at eight one-week intervals, and the quality of pregnancy was checked. Antifertility effects were found with TS 160, Penberol (at high dosage), and Mercaptopurine. Effects on permiogenesis with genetic risk were found with Cyclophosphamide, TS-160, Mercaptopurine, and less marked ones also with Cytembena. The effects were mostly manifested by increases in the numbers of early fetal resorptions, and less frequently by preimplantation loss of ova. No genetic risk was revealed by the assay in the cytostatics Edikron, Penberol, Butocin, and Damvar.

Subclasses IgG1--IgG4 in 84 sera with IgG paraprotein.

The distribution of subclasses IgG1-4 was determined in 84 sera with IgG paraprotein. The results were as follows: IgG1--58.3%, IgG2--25%, IgG3--11.9%, IgG4--4.8%. After division into the group of IgG myelomas (n = 47) and the group of non-myeloma IgG paraproteinemias (n = 37), the following distribution was found: IgG1--55.3%, IgG2--25.5%, IgG3--12.7%, IgG4--6.5% and IgG1--62.1%, IgG2--24.3%, IgG3--10.8%, IgG4--2.7%, respectively. The distribution of light chains in the individual subclasses of IgG paraproteins was also studied. In three double IgG and IgA myeloma paraproteinemias, subclass 1 IgG paraprotein was always demonstrated.

Infrared thermovision in tumors of the orofacial region.

The authors present briefly the physical principles of infrared thermovision and an overview of its utilization in stromatology. Among the 21 patients, suffering for the most part from a tumorous affection of the orofacial region, they describe in more detail the case of a carcinoma of the lower lip. A correlation of their clinical, histological and X-ray findings with those of infrared thermovision revealed agreement in every case insofar as the localization of the pathological process was concerned. They stress that infrared thermovision is not meant to replace histo-pathological investigation whose significance in the diagnosis of malignant tumors continues to be decisive.

Effectiveness of radiation therapy and combined radiation and surgical treatment in advanced breast cancer.

The effectiveness of two modes of combined radiation-surgery treatment and the correlation between the therapeutic results and the clinical type of primary breast cancer were followed in a group of 221 patients with Ca mammae in the III clinical stage. As criteria of evaluating, use was made of analysis of the temporal sequence of relapses and metastases formation following primary treatment. The higher effectiveness of the mode--surgery combined with pre- and postoperative irradiation as against that of surgery with only post-operative radiotherapy became manifest in the frequency of relapses but not in that of distant metastase formation. Simultaneously, a relationship has been established between the course of the disease after treatment and the various categories of T, or its dependence on the relation between the primary tumor and the skin and this within the IIIrd clinical stage of breast cancer.

Study on incorporation of 3H-DNA in the L-cells by means of electron microscopic autoradiography.

Uptake of isologous DNA (labeled with 3H-thymidine) by L-cells was investigated by means of electron microscopic autoradiography. It was found that exogenous 3H-DNA occurred not only in the cytoplasm, but, already from the 1st hr of incubation, permeated as far as in the host cell nuclei. During continuous cultivation, radioactivity established in the nuclei and cytoplasm increases with maximum value after 12 hr incubation. Beginning at the 3rd hr of incubation, in cytoplasma, grain clusters over loci of higher radioactivity were found. After 24 hr incubation, similar grain clusters in L-cell nuclei were observed. In the cytoplasma of the cells, the grains were often determined over or in the vicinity of lysosomes; in the nuclei they were localized predominantly over heterochromatine. The manner of 3H-DNA and 3H-TdR labeling was also compared.

Neutral lipids in nuclei and chromatin fraction of young and old Ehrlich ascites tumor cells.

Determination of lipid content of Ehrlich ascites tumor cell nuclei devoid of the nuclear envelope and chromatin isolated from them revealed considerable amounts of neutral lipids present in both preparations. The neutral lipid content always exceeded that of the phospholipid. An age dependent accumulation of neutral lipids could also be observed. The chromatin fraction always contained relatively higher amounts of neutral lipids than the respective nuclei with a predominance of cholesterol esters.

Cytofluorimetric measurement of DNA content in the cells of the transplantable melanotic and amelanotic Bomirski melanoma in golden hamster (Mesocricetus auratus, Waterhouse).

Cytofluorimetric measurements of the DNA content in transplantable melanotic and amelanotic tumor cells were carried out. The mean content DNA is higher in the amelanotic cells. The DNA distribution patterns are different in both tumors: there is a distinct stem line of the cells in the melanotic tumors whereas in the amelanotic cells there is a great variability in the DNA content and the stem line of the cells is absent. The correlation between these findings and some biological properties of the melanotic and amelanotic tumors are discussed.

Electrochemical properties of polycyclic compounds studied by the polarographic method in anhydrous systems. V. Oxidation potentials of carcinogenic hydrocarbons in acetonitrile.

The half-wave oxidation potentials of polycyclic carcinogenic hydrocarbons have been studied at the rotating platinum electrode in anhydrous acetonitrile with sodium perchlorate as the supporting electrolyte. The good agreement was obtained by the comparison of the carcinogenic activity of studied compounds with the values of the oxidation potentials. The values of the oxidation half-wave potentials were correlated also with the reduction potentials measured in acetonitrile and with the root xn of the Hückel secular equation for the highest bonding orbital.

In vitro cell proliferation studies in Hodgkin's disease.

A cytologic and proliferation analysis was performed in the lymph node samples of 30 nontreated Hodgkin patients using tritiated thymidine, the autoradiographic technique, and two different in vitro incubation schemes. A group of 9 inflammatory reactive patients was used as control. The results confirmed previous labeling data but showed that DNA synthesis can also be detected among the reticulum cells. The role attributed by some authors to the large rounded basophilic cells of lymphoid appearance found in Hodgkin's tissue is discussed in the light of the results in Hodgkin's and non Hodgkin's samples. Finally, some aspects dealing with the relationship and possible role of the Hodgkin and Sternberg-Reed cells are commented.

Effect of chemotherapy on the lymph node and bone marrow cell chromosomes in patients with Hodgkin's disease.

Chromosomal aberrations in bone marrow and lymph node cells have been investigated in 47 patients with lymphogranulomatosis 24--72 hr after administration of a single therapeutic dose of cyclophosphan and natulan. It is shown that the injurious action of cyclophosphan and natulan lasts more than 72 hr. An analysis of the dynamics of various types of chromosomal aberrations testifies that cyclophosphan and natulan affect cellular constituents found in the synthetic or post-synthetic phases of the cell cycle.

Importance of determinations of serum hexokinase, aldolase, and lactate dehydrogenase activities, and of the lactate/pyruvate-quotient in the diagnosis of malignant tumors.

Relation between the chemical structure and acute toxicity of 1-phenyl-3-methyltriazenes as activated metabolites during the carcinogenic action of 1-phenyl-3,3-dimethyltriazenes.

The mol LD50 values were correlated with Hammett sigma constants in terms of the Hansen empiric equation 6 for para substituted derivatives of 1-phenyl-3-methyltriazene. Good agreement was achieved and the line slope of the correlation, 0.3, is the same as that found during an identical experiment with derivatives of 1-phenyl-3,3-dimethyltriazene [1]. This is an indirect evidence that the considered mechanism of the interaction with biological substrate goes through the "activated carcinogen" - derivatives of 1-phenyl-3-methyltriazene, formed through the oxidative N-demethylation of the initial 1-phenyl-3,3-dimethyltriazenes.

The effects of continous irradiation on histological picture and deoxyribonucleoprotein content in spleen of rats.

The histological picture and deoxyribonucleoprotein (DNP) content in the spleen of the white male Wistar rats continuously irradiated with the daily dose rates 9.57 mGy (1 R), 95.7 mGy (10 R), 478.5 mGy (50 R) and 657.0 mGy (100 R) were studied. The animals were examined within the day until 60 of irradiation. The number of lymphocytes permanently decreased at the daily dose rates 95.7 mGy and higher ones. The increased activity of reticulum with the simultaneous multiplication of the erythroid cells in the red pulp of spleen was observed at day 10 and 60 of irradiation with 9.57 mGy and 95,7 mGy. Simultaneously DNP content in spleen increased in accordance with the increased activity of reticulum and extramedullar erythropoiesis. The significant decrease in DNP content alter the onset of irradiation with the daily dose rates 478.5 mGy and 957.0 mGy was caused mainly by the fall of lymphocytes. The incidence of eosinophilic leukocytes increased from day 40 irradiation with the daily dose rates 9.57 mGy, 95.7 mGy and 478.5 mGy and decreased with the daily dose rate 957.0 mGy. The plasma cells tended to increase in accordance with the increasing accumulated dose of irradiation with all daily dose rates.

Inhibition of leukocyte migration by tumor-associated antigen and its modification by serum; IgA as a blocking factor.

Based on the investigations of patients the migration inhibition assay proved to be a useful method for the detection of antitumorous cellular immunity. Out of the twenty patients tested 15 gave migratory inhibitory effect against specific tumor antigen. In 8 cases out of the above 15 the blocking activity of sera has been proved. This blocking activity is bound to IgA subclass. The complement played an "unblocking" role. The modifying role of serum factors in cellular immunity against tumors was discussed.

Individual serum proteins and acute phase reactants in monoclonal immunoglobulinopathies (a study in patients with IgG myeloma).

In 92 patients with multiple myeloma and IgG monoclonal proteinemia concentrations of seventeen different serum proteins were specifically determined. Prealbumin, albumin, alpha, HS-glycoprotein, alpha-macroglobulin, transferrin and immunoglobulins IgA, IgM and IgD were significantly decreased in patients with IgG myeloma. On the contrary the means found for the typical acute phase proteins i.e. haptoglobin, orosomucoid and CRP were significantly elevated. No significant differences were demonstrated for less typical acute phase protients, i.e. alpha1-antitrypsin, ceruloplasmin and C3-component as well as for hemopexin and beta2-glycoprotein I. CRP values were strongly elevated in some sera, however in majority of patients they were within the normal limits. Negative correlation was found between monoclonal IgG and the most of the studied proteins inclusive immunoglobulins IgA, IgM and IgD. No correlation was demonstrated between the monoclonal IgG and the triad of typical acute phase proteins. Positive correlation was found between monoclonal IgG and the total serum protein and further among the proteins negatively correlated with monoclonal IgG as well as among the individual acute phase proteins. Explanation of the correlations reported has been suggested.

Pharmacokinetics of butocine.

The pharmacokinetics of Butocine (glycine, N-[1-oxo-5-(1H-purin-6-ylthio)pentyl], ethyl ester) was studied in 8 individuals with malignant tumor after a single oral dose of 500 mg. Butocine is quickly absorbed from the alimentary tract. Highest serum levels were found 45 minutes after administration. Its urinary excretion is very low (2.5 per cent in 15 hours), 1.7 per cent being eliminated 3 hours after administration. Its renal clearance is likwise very low, i.e. 1.5 ml/min compared with endogenous creatinine clearance of 81.2 ml/min.

Changes in the activities of glycolytic serum enzymes, and of lactate and pyruvate levels in the blood during antineoplastic treatment.

Drill biopsy in the diagnosis of metastic and primary neoplasms of lymph nodes.

The author analyses the results of 300 drill biopsies, performed on peripheral lymph nodes of 266 patients. The usefulness of this method in the histological diagnosis of metastatic and primary neoplasms of lymph nodes is assessed. Drill biopsy proves to be especially valuable in the diagnosis of metastatic cancer. By means of the first biopsy, 90.4% of cases of metastatic cancer were diagnosed. In the great majority of cases, the histological type of metastatic neoplasms was precisely defined. Accuracy in the diagnosis of malignant lymphomas was lower. Drill biopsy of peripheral lymph nodes can be performed in outpatients.

Anaplastic transformation of medullary thyroid cancer.

The paper presents a detailed microscopic study of a case of medullary thyroid carcinoma with the loss of amyloid production and of argyrophilic cellular granules combined with the prevalence of giant multinucleated cells in a part of the primary tumor and namely in metastatic deposits. These changes are believed to give evidence of anaplastic dedifferentiation. Similar cases to that reported are reviewed with the conclusion that not only differentiated thyroid cancers but medullary thyroid cancers as well are capable of anaplastic transformation. However rare the medullary thyroid cancers are, they should be diagnosed and radically treated so as to prevent their fatal anaplastic transformation.

Effects of tumor cell culture supernatants on macrophages.

It is proved that supernatants of various tumor cell cultures, as well as cell-free ascitic fluid and sera from tumor bearing animals induce the detachment of macrophages from glass, the loss of their ability to form pseudopodia, the inhibition of their migration and the permeability alteration of their membranes. The cytotoxic effect is not produced by normal tissues, excepting placenta. The effect of tumor cell culture supernatants depends on the characteristic evolution time of each tumor; irradiation diminishes this effect. The involvement of macrophages in immune surveillance and response makes the results interesting for the tumor-host immune relation.

Twenty-four-four rhythm in DNA synthesis, mitotic activity and mitosis duration in ascites tumor NK/LY.

The suitability of different quantitative methods for determination of the cytotoxic activity of agents in cell cultures.

Studies related to the measurement of growth inhibition of HeLa cells by 6-Thioguanine (TG) and Vermiculine (V) have been described. Control and treated cell populations were enumerated as uniform suspensions of whole cells, prepared by standard treatment with trypsin. Determination of cell number, measurements of protein, DNA and RNA as well as of glucose consumption from the culture medium were made throughout the treatment with the studied drugs. It was apparent that the total cell number in control cultures was directly proportional to the total cell protein or nucleic acids. However, in both treated cultures the relationship between these parameters was disturbed as consequences to unbalanced growth which was an integral part of the cytotoxic reaction of the studied agents. From the results presented it followed that only the direct cell enumeration was suitable for the detection of agents' cytotoxicity.

5-Hydroxy-6-methoxy-2-indolylcarboxylic and 6-hydroxy-5-methoxy-2-indolylcarboxylic acids in the urine of hamsters with melanotic melanoma.

The isomeric metabolites, 5-hydroxy-6-methoxy-2-indolylcarboxylic and 6-hydroxy-5-methoxy-2-indolylcarboxylic acids have been identified in the urine of hamsters bearing transplantable melanotic melanoma. Their identification was carried out by spectral and chromatographic comparison of the two isomers isolated from urine with the same compounds achieved synthetically.

Quantitative parameters of melanomas differentiation.

Melanin concentration which represents one of biochemical parameters expressing the degree of pigment cell differentiation was studied by new gravimetric procedure in human, Bomirski hamster, Harding-Passey mouse and horse melanomas and also in several non-tumor pigment tissues. In amelanotic varieties of human and hamster melanomas relatively high melanin level was discovered. This finding (together with known electron microscopic evidence of pre-melanosomes presence in some amelanotic melanomas) suggests that the term "amelanotic" is not correct. Comparison of melanin content between melanomas and normal pigment tissues clearly demonstrates in quantitative way that as for melanomas their proliferation does not decrease the degree of their biochemical differentiation.

The cell-mediated immunity in patients with malignant melanoma.

Cellular immunity in 22 patients with malignant melanoma and the blocking effect of their sera was followed by means of the leukocyte migration inhibition test. The migration of peripheral leukocytes from malignant melanoma patients was significantly inhibited in the presence of malignant melanoma antigen in 50% of examined patients and only in 2 cases in controls. The blocking activity of serum on migration inhibition was observed in 3 melanoma patients. From the results of the tests performed related to the two-years-course of the disease the conclusion may be drawn that the prognosis in patients with proved cellular hypersensitivity to malignant melanoma antigen is more favorable than in anergic individuals.

Prognostic factors in gastric carcinoma.

One hundred and thirty eight gastric carcinomas were studied histologically with special reference to the morphology of the tumor, its surrounding tissues and the regional lymph nodes. A special search was focused on the morphologic manifestations of possible host factors in association with gastric carcinoma. The most prominent findings were as follows: 1. The nuclear grade of the tumor was positively correlated with the 5-year survival rate of the patients. 2. The content of tumor-derived mucus was not a prognostic determinant. 3. The intensity of the stromal lymphocyte and plasma cell reactions did not affect the prognosis but was inversely related to the frequency of nodal metastases. 4. Sinus histiocytosis and nodal mast cell reactions were an important determinant of whether nodal metastases appear or not. 5. An active paracortical area of the lymph node was almost incompatible with the appearance of nodal metastases.

Biochemistry of drugs. XXIV. Substances with antineoplastic effect. LXII. Pharmacokinetics of gamma,gamma-bis-4-ethylphenyl-alpha,beta-dibromoisocrotonic acid-3H (Edikron-3H) in experimental animals.

After oral administration of Edikron-3H to mice with mammary gland adenocarcinoma HK, within 24 h after ingestion almost 50% of administered radioactivity was excreted (16% in urine, 33% in faeces), and within 4 day it was almost 80% (27% in urine, 50% in faeces). Like in mice, also in dogs the faecal excretion preponderated. In mice, the levels in most tissues and blood culminated at hour 6 after administration of the substance. Relatively high levels were found in the liver, the levels in other tissues and blood being substantially lower in comparison. The levels in the tumor were initially low, later on they rose, and culminated at hour 24. Besides, it was found that the transfer of radioactivity by milk into the sucklings' bodies was relatively low, and radioactivity penetration into fetuses was negligible. In urine, unchanged parent substance was present in very slight amounts only.

Biochemistry of drugs. XXV. Substances with antineoplastic effect. LXIII. Pharmacokinetics of delta-/2-amino-6-hydroxy-3,4-dihydro-4-oxo-5--pyrimidinyl/valeric acid-14C (Damvar-14C) in experimental animals.

In experiments on H mice and Wistar rats it was found that radioactivity was excreted after subcutaneous injection of Damvar-14C preponderantly in urine, and after oral administration, in faeces. After subcutaneous injection the urinary excretion set on very fast. After subcutaneous injection of Damvar-14C to H mice with Crocker's ascitic sarcoma S 180, peak radioactivity values were found in the majority of organs, blood, and ascitic fluid at hour 1 after injection; in the kidneys the peak value appeared as late as at hour 6. The renal levels were conspicuously higher than the other ones. Relatively high radioactivity levels were found in the ascitic fluid, skin, and blood. After oral administration the radioactivity levels in all organs, blood, and ascitic fluid culminated at hour 3 after ingestion. The highest levels after oral administration were likewise found in the kidneys and ascitic fluid. With the exception of the kidneys, blood, and skin, the peak values of specific radioactivity in organs were equally high or even higher after oral than after subcutaneous administration of the labeled substance. Furthermore, it was found that after administration of Damvar-14C to rat mothers the transfer of radioactivity into the milk and then into the sucklings was very slight, and so was the penetration of radioactivity into fetuses.

Some cytostatic and pharmacological properties of 2,3-dihydro-1,3-6H-oxazine-2,6-dione ("3-oxauracil").

Cytostatic effects and some pharmacological properties of a new metabolic inhibitor "3-oxauracil" were studied. The cancerostatic effect was examined on 7 experimental tumors in mice and on two types of tumors in rats. After the i. p. application of 20 mg/kg, there was both a statistically significant decrease of tumor weight and increase of animals' survival time in NK lymphoma of mice. Significant changes in one of both parameters followed occured in all experimental tumors after the i. p. application but only in the Krebs ascitic carcinoma after the oral application of "3-oxauracil". The acute toxicity of the substance in water was 322 mg/kg i. p. and 850 mg/kg p. o. The ethanol solutions were more toxic. The distribution of the 3H- and 14C-labeled substance was followed up in blood, urine, liver, brain and kidney. After the p. o. application, the radioactivity peak was reached after 2 hr in blood and high radioactivity levels were found in kidney followed by brain and liver. 96 hr after the drug was applicated perorally, only 60% of radioactivity was found in urine.

The significance of some modifying factors during transplacental blastomogenesis by ethylnitrosourea in rabbits.

The development of the transplacental blastomogenesis induced by ethylnitrosourea (ENU) was studied in rabbits under the influence of modifying factors (organospecific immunity, chronic irradiation of the peripheral nerve, injection of methylnitrosourea -- MNU). Statistical analysis of the results showed that the postnatal immunization by the antigen prepared from homologous tissue of the peripheral nerve or its chronic irritation always stimulates the frequency of the tumors development. The injection of MNU inhibits the tumorigenesis in the tested animals. According to the data different modifying factors distinctly act upon the transplacental blastomogenesis in rabbits.