Malignant tumors in Cuban children. Fourth triennial 1973-1975 of the national cancer registry.

The incidence of malignant tumors in children is not as high as that in adults, though in Cuba and other countries they are the most frequent causes of death in this age group. Data of the fourth triennial of the Registro Nacional del Cáncer (Cuban Cancer Registry) in the period 1973-1975 are presented. In the age group 0-14 years, 1038 new cases were diagnosed and registered; this figure represented 2.5% of the total of tumors in the period studied; the mean annual rate was 11.3% per 100 000 population. Male patients were more frequently affected. Morbidity decreased in 0-4 year group and increased in 10-14 year group. As in the first, second and third triennials, malignant diseases as leukemia, lymphoma and tumors of the nervous system prevailed among children: these accounted for a little more than 2/3 of the total of tumors followed by tumors of the kidney and eye.

Autoradiographic study on the effect of hyperthermia (42.5 degrees C) on the proliferation kinetics of solid tumors in children.

Supranormal temperatures between 40 and 43 degrees C are not necessarily lethal to tumor cells, but lead to characteristic changes of the cell cycle. The parameters of the proliferation kinetics were studied in 35 solid tumors of children at a temperature of 42.5 degrees C with an autoradiographic in vitro method, in comparison to normothermia: 9 Wilms' tumors, 10 neuroblastomas, 8 osteogenic and soft tissue sarcomas, 6 non-Hodgkin-lymphomas, and 3 other tumors. 28 tumors showed significant prolongation of DNA synthesis times by an average factor of 1.27 (1.10--3.12). Mitosis times undergo an average prolongation by the factor 2.75 (1.07--8.87). Together with significant decrease of the 3H-thymidine labeling index the prolongation of the cell cycle time amounts to an average of 2.67 (1.05--8.30). The cause of the changes of the cell cycle are discussed. Probably, the heat sensitivity of tumors is correlated with the proliferation rate and with the degree of histological differentiation; but this cannot be confirmed statistically due to the small number of cases. 2 cases responded with a decrease of the duration of the cell cycle; in one case this was probably due to an exogenic thermotolerance. The changes of the cell cycle are of a particular importance for the therapeutic combination with radio- or chemotherapy. These relations are discussed.

A possible association between quality of drinking water and stomach cancer incidence among native and immigrant populations of a selected industrial city.

The city Zabrze (Upper Silesian Region, Poland) was divided into districts differing in respect to drinking water stations. Geographic correlation between the stomach cancer incidence in native and immigrant populations and the quality of water used for drinking were investigated. It has been shown that in the districts where water of greater hardness has been used for drinking, the stomach cancer incidence was much lower than in other areas, especially in native male populations.

Diagnosis of malignant tumors on the basis of the current generating capacity of malignant tumorous tissue. II. Study of the malignant tumors of female genital organs.

Description of an electrochemical measuring method for the in vivo quick-diagnosis of malignant tumors and dysplastic alterations of the cervical epithelium is given. The measuring results carried out in 660 cases on the surface of the cervical epithelium are reported. The current generating capacity of planocellular carcinoma of the cervical epithelium exceeds the upper limit of the normal value with 40% and is significantly higher than the values measured in dysplastic areas. With the aid of measuring, dysplastic alteration are demonstrable even in cases, in which the cytologic or coloposcopic examination is not indicative of dysplasia. Measurings in the benign and malignant tumorous tissue were carried out with needle electrode in 41 cases, during operation. The current generating capacity of malignant tumors exceeded with 150--170% the values measured in the healthy tissue, respectively in benign tumorous alterations.

Preliminary results with the new cytostatic Penberol in ovarian carcinoma.

The authors present results of a preliminary study on the efficiency of the preparation Penberol [VUFB Prague (cis-beta-4-pentoxy-benzoyl-beta-bromoacrylic acid) CHECO protocol number 4] in the ovarian carcinoma of the stage III and IV (FIGO) in which the drug was used after the failure of the first choice chemotherapy. Out of ten patients, complete regression of the lesions was achieved in 7 patients, only in two cases the regression was lower than 50% and in one case a progression was encountered during the therapy. The median of the duration of non-completed remission is 15+ months. As to the side effects, in half cases the dose of the preparation was reduced by 1/3 because of diarrhea induced by irritating action of the drug on the intestinal wall. No other side effects of the therapy were observed.

Survival of lung cancer patients treated with BCG and/or a soluble BCG fraction (F70) after surgery, radiotherapy and chemotherapy.

243 lung cancer patients treated with routine therapy (surgery 54, radiotherapy 46 and chemotherapy 143) in Plovdiv's District Oncologic Dispensary (PDOD) were submitted to an additional therapy with BCG or with its soluble fraction F70. The survival rate of these patients was compared with the survival rate of 305 lung cancer patients following routine therapy alone (surgery 78, radiotherapy 113 and chemotherapy 114). The two groups were not randomized. Between patients treated and non-treated with BCG(F70) a significant increase of the survival rate was found for patients submitted to BCG(F70) treatment after surgery or radiotherapy in the 2nd and 3rd year survival period and for patients submitted to BCG(F70) treatment after chemotherapy -- in the 1st year survival period of patients with limited disease only. The effect of the additional BCG(F70) treatment was less dependent on the preceding routine therapy than upon the clinical stage of the disease at the moment of BCG(F70) treatment beginning. More salient results were observed in patients with undifferentiated histological forms.

Drug sensitivity studies on clonal cell lines isolated from heteroploid tumor cell population. II. Sensitivity of clones growing in suspension cultures.

NK/S cell line and its clonal lines 6 and 10 were established in culture from NK/Ly mouse lymphoma. The clones differed from each other and from the parent line in chromosome pattern and growth rate. Clone 6 cells were polyploid and more sensitive to dianhydrogalactitol than the diploid clone 10 cells. However, clone 10 cells were more sensitive to adriamycin. A very small difference in sensitivity was found with vincristine. It was concluded that sensitivity of clones originating from the same parent population is drug-related.

Antitumor effect of N alpha-benzyloxycarbonyl-N,N-bis-(2-halogenethyl)-hydrazide derivatives of amino acids.

The antitumor effect of some N alpha-benzyloxycarbonyl-N,N-bis-(2-halogenethyl)hydrazide derivatives of lysine, glycine, cystine, phenylalanine and p-chlorophenylalanine, was studied. Six of eight newly synthesized compounds show considerable antitumor effect on solid Walker carcinosarcoma 256 (about 95% tumor growth inhibition). Three of these compounds under study increased the lifespan of mice with leukemia L1210. The investigation of the effect of N alpha-benzyloxycarbonyl,D,L-phenylalanine-N,N-bis(2-chloroethyl)hydrazine on various mouse tumors showed remarkable growth inhibition of the Ehrlich ascites carcinoma, sarcoma 37, colon adenocarcinoma akatol and lesser antitumor effect also on solid adenocarcinoma 755, Lewis lung carcinoma and melanoma B16. All investigated compounds exhibited depression of leukocyte count--their toxicity being, however, lower than that of sarcolysine in parallel experiments.

Inhibition of hematogenous metastatic dissemination by BCG-stimulated serum obtained from rabbits and mice.

A statistically significant metastasis-prophylactic effect was obtained by administration of sera originating from BCG-stimulated mice and rabbits. The benzo[a]pyrene induced sarcoma 276A served as experimental metastatic test model.

Influence of immunosuppressive pretreatment on the survival of mice challenged with Ehrlich ascites carcinoma (EAC).

Mice were pretreated with immunosuppressive doses of cyclophosphamide (CY), cortisone acetate (CA) or X-irradiation. 24 h later all mice were given 1 x 10(5) EAC cells intraperitoneally. In another experiment mice were splenectomized 1 h before injection of EAC cells. Neither CY nor CA pretreatment had significant influence on the survival of tumor-inoculated mice. On other hand, X-irradiation or splenectomy strengthened tumor resistance.

Virus-like particles in long-term transplanted hamster tumors.

Among the studied long-term transplanted skin tumors in hamsters virus-like particles (VLP) have been disclosed by electron microscopy in two squamous cell carcinomas and one fibrosarcoma but not in three melanoma lines. The great majority of the particles have exhibited structure and intracisternal position typical for the R or H type VLP. Their amount have greatly varied in the different lines and within the period of study. In the fibrosarcoma cells A type particles have also been observed. The available literature on R type VLP have been reviewed and their possible role in oncogenesis was discussed.

Intragenomic distribution of 5-methylcytosine in various forms of human and murine leukemic cells.

The degree of 5-methylcytosine formation in DNA sequences differing in reassociation rate and susceptibility to DNase II digestion has been investigated in human chronic myelogenic leukemia and acute leukemia leukocytes, human PHA-stimulated lymphocytes and murine L5178Y lymphoblasts cultured in various phases of growth. The results indicate that in all forms of cells studied by us the general pattern of intragenomic 5-methylcytosine distribution is similar, with two preferentially methylated regions: the sequences fast reassociating and rendered Mg++-soluble after DNase II digestion of nuclei. The most variable fraction as regards the level of methylation seemed to be DNA of Mg++-soluble fraction of DNase II digest, which in acute leukemia leukocytes, PHA-stimulated lymphocytes and exponentially growing L5178Y cells revealed about twice greater relative proportion of methylated cytosines than in leukocytes of chronic myelogenic leukemia and L5178Y cells maintained at saturation density.

Application of gibberelic acid (GA) alone or together wih cytostatics in treatment of lung cancer.

Gibberelic acid (GA) was used alone or in combination with cytostatics (Vincristine, Cyclophosphamide, HN3), surgical treatment or radiotherapy, in treatment of 65 patients with lung cancer. The patinets were followed up from the end of 1972 until the end of 1977. It was found that the administration of GA, at the given doses, did not cause toxic effects in treated patients and it improved for shorter or longer period of time some metabolic functions and activity of hemopoiesis. Some analgetic effect(2) of GA was also registered.

Serum complement levels in patients with cancer of the uterine cervix before and after radiation therapy.

With single radial immunodiffusion method the level of the third component of the complement was investigated in 153 cervix cancer patients. In some cases the level of the fourth component of the complement and total hemolytic activity of the complement were parallely determined. The C'3 values elicited a polarized distribution pattern in contrast to the normal distribution of the controls. Mean C'3 level increased simultaneously with the progress of the disease up to Stage III, and decreased only in Stage IV. On the contrary, C'4 level was stable and practically equaled the normal value. Total hemolytic activity of the complement in cervix cancer patients usually exceed normal values. The C'3 changes during radiation therapy contradicted its role as an acute phase protein.

UICC postgraduate courses in clinical cancer chemotherapy: a teaching experience outside Europe.

Distribution of 65Zn in mice with melanomas and in the subcellular fractions of melanomas.

The tissue distribution of injected 65Zn into mice with Harding--Passey or B-16 melanomas was studied. Both types of melanoma took up remarkably large amounts of 65Zn (harding--Passey 17.4%; and B-16 almost 26.0% of the 65Zn dose administered). In the case of the B-16 melanoma, the tumor was the dominant site of incorporation even in terms of specific activity. On a subcellular level, melanosomes isolated by gradient ultracentrifugation were shown to be sites of preferred deposition of 65Zn. The uptake of 65Zn by melanomas and its specific binding by melanomes suggests that this isotope might be useful in detecting melanomas.

Growth characteristics and therapeutic response of transplantable colon tumors in animals.

The growth characteristics and the effect of clinically available chemotherapeutic agents on two transplantable colon tumor lines were studied. These are subcutaneously transplanted undifferentiated carcinoma AKATOL, originating from tumors "spontaneously" appearing after foetal colon implantation, and moderately differntiated carcinoma No. 173 obtained likewise with additional treatment by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Some basic kinetic parameters of tumor growth are determined. The tumors show a relatively slow growth, the median survival time of animals being approximately 50 and 38 days, respectively. The investigation of the sensitivity of tumor lines shows that they are sensitive to many standard antitumor drugs. In the case of AKATOL a high responsiveness to antibiotics and to a smaller degree to other groups agents was observed excluding sarcolysine, CCNU, alexan (cytosine arabinoside) and vinblastine. In the case of colon tumor No. 173 strong antitumor effect for CCNU, 5-fluorouracil, cyclophosphamide, methotrexate and vinblastine was observed. The possibilities to use these tumor systems for screening and evaluation of antitumor agents are discussed.

Potential anticancer agents. XX. 1. Steric fit analysis in aromatic nitrogen mustards area.

A series of 31 aromatic nitrogen mustards was treated by means of the Minimal Tropological Difference (MTD)-receptor site mapping procedure by respect to toxicity (LD50) and efficacy (TGI%) against Jensen sarcoma. The correlation equations obtained for the two cases are: (Formula: see text), The low correlation coefficients indicate a not too high dependence upon steric factors of TGI% and LD50. However these results suggested that: a) the ortho-substitution represents a favorable event for both toxicity and antitumor effectiveness (j = 1 and 18 being good vertices in both corresponding hypermolecules), b) despite this effect, the steric features involved in these two biologic parameters seems to be different.

Potential anticancer agents. XX. 2. Quantitative structure--activity relationships (QSAR) in aromatic nitrogen mustards area.

Quantitative structure--activity relationships have been fomulated for 27 aromatic nitrogen mustards derived from benzoic acids (9 monosubstituted and 18 disubstituted derivatives). Their toxicity (LD50) and antitumor activity against Jensen sarcoma were correlated with hydrolysis rate (log k66) lipophilicity constants (pi) and steric parameters (MDT). The chemical reactivity of the nitrogen mustard moiety (expressed as log k66) seems to be of main importance in determining the biological properties of these derivatives. The favorable effect of ortho-substitution was pointed out. Generally these results are in good agreement with those obtained by HANSCH et al. [7] on a different series of nitrogen mustards.

Chemotherapeutic activity and histopathologic feature of BCNU + 5-FU effect on Lewis lung carcinoma.

In this study we investigated the effects of BCNU + 5-FU on the intramuscularly implanted Lewis lung carcinoma and on its spontaneous lung metastases. Chemotherapeutic agents were given i. v. either alone and in combination seven days after transplantation of 2.5 x 10(5) viable tumor cells, when the average weight of the primary tumor was 500 mg. The analysis of the following parameters: T/C, T--C, of %-metastases reduction, indicates that the simultaneous administration of BCNU + + 5-FU induces a more relevant response on the Lewis lung carcinoma than single drug therapy, especially evident at level of lung metastases.

Relationship between different cell surface markers in chronic lymphocytic leukemia.

Peripheral blood lymphocytes from 15 patients with chronic lymphocytic leukemia (CLL) and 13 normal controls were examined for the presence of surface markers: CLL antigen, surface immunoglobulins, receptors for sheep erythrocytes, receptors for the IgG Fc, C3b, C3d and receptor(s) for mouse erythrocytes. Leukemic lymphocytes were characterized by the presence of CLL antigen and a significant increase in the capacity to bind mouse erythrocytes. CLL antigen was correlated with surface immunoglobulins and receptor of IgG Fc, however, the receptor for mouse red cells was not correlated with other surface markers. Neuraminidase treatment of CLL lymphocytes significantly increased proportion of mouse erythrocyte rosettes. The routine use of mouse erythrocyte rosettes in the immunodiagnostics of B lymphoproliferative disorders is postulated.

Humoral and cellular immunity in long-term surviving patients with malignant lymphoma.

Humoral and cellular immunity was followed in a group of long-term surviving patients with malignant lymphoma, viz. serum immunoglobulins quantitatively, T lymphocytes by the E rosette and B lymphocytes by the EAC rosette methods. A single examination of this group was compared with data from a control group of normal humans and a smaller group of patients with malignant lymphoma at an advanced stage. A significant increase of IgA (3.73 g/l onthe average) was noted in the surviving group as against both the controls (mean 2.31 g/l in the healthy and 1.23 g/l in the patients with advanced disease). IgM values were found to be significantly decreased in the latter patients (mean 0.85 g/l) in comparison with both the other groups. The rosette tests yielded lower absolute values of E rosettes in the same patients with advanced lymphoma (mean 725/ml) as against the healthy subjects (mean 1525/ml) and the long surviving patients (mean 1277/ml). The absolute value of active E rosettes in the two groups of patients was lower (741 and 580/ml) than that in the healthy group (mean 1067/ml). A percentage determination of rosette values proved to be of low statistical significance.

Spleen white pulp morphology related to immunological reactivity of women died of widespread breast carcinoma.

The histologic changes in the white pulp of the spleens collected from thirty women died of widespread adenocarcinoma of the breast were assessed by using the standardized reporting system previously introduced. As a control material, the same number of age-matched women died of myocardial infarction or cerebrovascular accident without signs of any malignancy was used. Special attention was focused on the evaluation of the lymphocyte populations (T- and B-cells) responsible for immunological reactivity. Histological characteristics suggested to reflect the activity of both the cell-mediated and humoral immune reactions were found to be within normal range in the control patients, whereas in the cancer series both these elements seemed to be profoundly deranged. The significance of these observations was discussed with regard to the background of the previously demonstrated immunological reactivity against the cells of human breast carcinoma, and the conclusion was drawn that an impairment of both the humoral and cell-mediated immune responses must exist in women dying of widespread adenocarcinoma of the breast. The applicability of the standardized reporting system used in the assessment of spleen white pulp morphology was emphasized.

Cobalt-activated acylase and aminoacylase I in human malignant tumors.

The activities and isoenzyme pattern of cobalt-activated acylase and of aminoacylase I were estimated in carcinomas of bronchi, lung, thorax, stomach, colon and uterus. In all cancer tissues the activity of cobalt-activated acylase was markedly increased as compared with the normal tissue. Alterations of the isoenzyme pattern of cobalt-activated acylase were found in the carcinoma of stomach and of uterus, with the increased expression of the form-1 of the enzyme. No regularity in the aminoacylase I changes in tumor tissues has been observed.

Epidemiological evaluation of incidence of lung cancer in inhabitants of Warsaw in the period 1966--1975.

Lung cancer is most frequent in men and occupies the first place among the causes of cancer deaths. In the period 1956--1975 in Poland the incidence rate of lung cancer increased about ten times in men and seven times in women. Analysis of age-specific incidence shows a significant increase in the group age of 35--39 years in men and in 45--49 in women. The absolute increase of incidence of lung cancer, the poor prognosis and the high mortality should call the attention of the whole population to canecerogenic factors, especially the noxious influence of smoking.

Perinatal subglottic and hepatic hemangiomas as potential emergencies: effect of radiotherapy.

The authors analyze 5 cases of perinatal hemangiomas in internal localizations respresenting relative or immediate emergencies. Three cases were subglottic proliferating hemangiomas with threatening suffocation, the other two were large hemangiomas of the liver causing conspicuous hepatomegaly. In 3 children there were also hemangiomas of the skin and/or in the oral cavity seen as important signs in the diagnostic reflections on the nature of the urgent clinical pictures. Moreover one child with subglottic hemangioma suffered from hematological disorders characterizing the syndrome Kasabach--Merritt. In all the children transcutaneous radiotherapy was performed (ranging from 12 Gy in two weeks up to an exceptional dose of 25 Gy over 3 months). This was followed by recession of subjective complaints and, eventually, by complete regression of the irradiated angiomatous lesions, both subglottic and hepatic. The disorders of hemocoagulation disappeared also quickly and completely. At present, i. e. after 3 to 21 years, there are no undersirable post-irradiation changes in any of the patients. Nevertheless, in view of possible post-irradiation effects, particularly on the thyroid gland, the patients continue to be regularly followed up at the respective clinical departments.

Immunological studies in breast cancer. Modifying activity of the serum (MSA).

Cancer patients' sera can modify results obtained with cell mediated immune reactions in vitro. We have investigated the modifying activity of breast cancer patients' sera on the inhibition elicited in a leukocyte migration inhibition test with two cellular extracts of breast cancer hepatic metastases (CaMa Hu-2 and CaMa Hu-3). These extracts elicited positive delayed hypersensitivity reactions in vivo in 80% of the patients studied with at least one of the two preparations, and 40% of the patients showed positive results with both preparations. In vitro similar quantitative results were obtained with the leukocyte migration inhibition test. With these antigenic extracts significant statistical differences were obtained between the cancer group compared to the controls with benign breast lesions (p < 0.01) and to the normal controls (p < 0.001). The modifying activity of the sera (MSA) showed three effects: blocking , potentiation and induction of inhibition of leukocyte migration in vitro in 80% of samples of sera tested. Blocking activity was associated mainly with not too advanced clinical stages, while potentiation and induction effects were associated with more advanced clinical stages. These MSA may be explained by assumed presence of tumor antigenic material, antibodies and immune complexes in breast cancer patients' sera.

Isolation and partial characterization of plasma membranes from chicken liver and from Mc-29 virus induced transplantable hepatoma.

Plasma membranes have been isolated from chicken liver and from Mc-29 virus induced transplantable hepatoma. The purity of membrane preparations has been checked by electron microscopy and by determination of the activity of some enzymes: 5'-nucleotidase, Na+, K+-ATP-ase, Mg2+-ATP-ase, alkaline beta-glycerophosphatase and glucose-6-phosphatase. In hepatoma membranes the activity of 5'-nucleotidase, Na+, K+-ATP-ase and Mg2+-ATP-ase was lower, that of alkaline phosphatase higher, than in liver membrane preparation. The incorporation rate of glucosamine-14C into UDP-N-acetylglucosamine and into plasma membrane glucosamine have been studied as well. The rate of synthesis of UDP-N-acetylglucosamine was faster in liver than in tumor cells. The labeling of hepatoma plasma membranes with glucosamine-14C occurred more slowly than that of liver ones. The rate of transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to membrane-bound glucosamine is lower in hepatoma, than in liver cells.

Strandquist's graphs and the new mathematical formula for cumulative biological effect.

The new formula for cumulative biological effect (CBE) derived on the basis of main characteristics of Strandquist's graphs has been used for an analysis of experimental fractionation data. Some new aspects concerning fractionated irradiation have been mentioned.

Some problems in using density gradient centrifugation for synchronization of L5178Y-S cells.

The application of sucrose gradient sedimentation for synchronization of murine lymphoma L5178Y-S cells was investigated. Centrifugation of the cells in a linear 2--10% sucrose gradient in Fischer's medium allows to obtain populations enriched in (I) young cells, (II) DNA-synthesizing cells, and (III) old cells. Population I showed highest degree of synchrony and contained at least 70% G1 cells. However, only a proportion of cells from population I progressed normally (approximately 30% of cells from this population remained in G1 during post-separation incubation). It was shown that the sucrose concentrations used for separation did not affect growth of L5178Y-S cells. On the other hand, relatively gentle mechanical treatment severely inhibited proliferation of these cells. It is suggested that excellent adaptation of cells to the culture medium and complete stability of the medium's composition are the prerequisites for a successful synchronization of L5178Y cells by gradient centrifugation.