In vivo suppression of Ehrlich ascites carcinoma by human antibody-dependent splenocytes.

Inbred CFW/L1 mice were inoculated i. p. with Ehrlich ascites carcinoma (EAC) cells. Two and a half hours later they were given i. p. injection of human splenocytes or rabbit anti-EAC serum, or human splenocytes together with rabbit anti-EAC serum. When given alone splenocytes did not exert any antitumor effect. Anti-EAC serum given alone inhibited tumor growth in 50% of mice. Simultaneous application of splenocytes and antiserum inhibited tumor growth in 85% of mice. In vitro experiment suggests that antibody-dependent cellular cytotoxicity (ADCC) may be responsible for the observed antitumor effect in vivo.

Dysplastic lesions and abnormalities in amphibians associated with environmental conditions.

High incidence of developmental anomaly and tumor-like dysplasia of osteochondrous tissue of hind limbs has been detected in frogs and toads the larval development of which has taken place in water, contaminated with sewage. Correlation between frequency of dysplasia and the level of water oxidation has been noted. Study on histogenesis of the marked tumor-like dysplasia were performed. It is assumed that the major mechanism responsible for anomaly consists in cytotoxic effect of teratogenic agent and that the differentiation lesion of cartilaginous tissue is caused by blocking of maturation and death of chondrocytes.

Immunoglobulin profiles and blood variables in 50 patients with hairy cell leukemia. Effects of splenectomy.

The immunoglobulin profile in patients (50) with hairy cell leukemia (HCL) was assessed on the basis of quantitative estimation of IgG, IgA, IgM, IgD and IgE serum levels. In HCL patients IgG, IgA and IgE mean levels were found higher than in healthy persons. No significant difference was found in serum IgM and IgD levels as compared to the reference group. Considering three major immunoglobulins, in no class and no patient with HCL immunoglobulin deficiency has been recorded. The serum immunoglobulin profile might be useful in differentiating HCL from chronic lymphocytic leukemia or lymphocytic lymphomas. However, no difference in immunoglobulin profiles in all five classes under study could be found between splenectomized and nonsplenectomized patients. Four percent PEG precipitation technique was used for the detection and characterization of IgG immune complexes in sera. The percentage of IgG precipitated in 4% PEG from serum was significantly higher in patients with HCL than in healthy persons; similarly as in immunoglobulin study no significant differences between splenectomized and nonsplenectomized patients were recorded. The beneficial effect of splenectomy was documented by the improvement in hemoglobin concentration, platelet and neutrophil counts and erythrocyte sedimentation rate.

Late effects of fractionated irradiation of normal tissue.

The formulae derived in previous papers for cell tissue kinetics have been fitted to further animal data on late reactions. Equations for some human tissue late reactions after X-rays and neutrons have been derived and discussed. The presented analyses of clinical data have preliminary character and the suggested formulae should be further checked and precised before using in clinical radiotherapy.

Irradiation scheme using three daily fractions in the radiation therapy of pharyngeal and oral carcinomas.

A group of 34 patients were treated by irradiation using 3 daily fractions of 1 Gy each at 4 h intervals. The overall TD in all of the patients was 60 Gy administered within 26 days. The irradiation was carried out by telecurietherapy utilizing 60Co. Twenty-eight patients had the tumor in the oropharynx, 1 in the hypopharynx, 1 in the oro- and hypopharynx and 4 in the oral cavity. There was a prevalence of advanced clinical stages. At present 25 of the patients are living, 21 of them without clinical symptoms of the disease. The skin reacted to irradiation only with a slight erythema and hyperpigmentation. Irradiation reactions in the oropharyngeal and oral mucosa were grave and were accompanied by significant subjective complaints. No post-irradiation complications were observed.

Cholecystectomy and right-sided colon cancer.

The present study conducted in order to determine the association between cholecystectomy and colon cancer. Two groups of subjects were investigated: 478 patients who underwent cholecystectomy and 492 clinically healthy persons. The method chosen was the screening for colorectal cancer by means of detecting occult blood in the feces using Hemoccult test. In the cholecystectomy group two right-sided and in the control group two left-sided colon cancers were detected. These data support the hypothesis that cholecystectomy may be a predisposing factor in the development of right-sided colon cancer.

Balkan nephropathy and malignant tumors.

The question of whether the cancer death risk is higher in Balkan nephropathy (BN) foci has been approached by a comparison of 25 BN endemic municipalities with the same number of the group matched ones. It came out that the total cancer mortality was considerably higher, and mortality of nonurinary cancer slightly higher in BN endemic municipalities. The absence of a significant difference for the non-urinary cancer sites the authors explain by the fact that in municipalities designated as endemic only a part of the population lived in actual BN foci. They re-evaluated data published for Bulgarian BN endemic foci and concluded that, apart of a higher total cancer mortality, there was a significantly higher (although unrecognized) risk of nonurinary deaths in BN endemic settlements.

Early paraneoplastic syndrome in medical oncology: clinicopathological analysis of 1694 patients treated over 20 years.

During the period of 1963-1982 we treated at our hospital 1694 patients with diagnosed internal malignancies. Of them, paraneoplastic syndrome as the first manifestation of the disease was found in 127 (7.4%). In 79 patients (4.6%) this first symptoms has also been an early manifestation of the malignoma (good general condition, without typical clinical symptoms of cancer). The paraneoplastic syndrome as the first symptom of the disease was manifested mainly as rheumatologic (58 patients; 3.4%) and thrombophlebitis (33 patients; 1.9%) symptoms. While thrombophlebitis as the first symptom of the disease was at the same time only in 48% also an early symptom, rheumatologic syndromes were also early symptoms in as many as 74% of cases. Hemoblastoses, lymphomas, and plasmacytomas are diseases which were accompanied by a paraneoplastic syndrome not only as the first but also as an early manifestation of the disease.

Enhancement of leukemia and mammary tumor development in ICRC mice with subcapsular graft of neonatal thymus.

Weanling ICRC mice were given a syngeneic graft of neonatal thymus. The control (C) and experimental mice (IT) were observed for their natural death. Neoplasms of the reticular tissue developed in 80% (19/23) mice of IT group and 21% (5/23) in control. Mammary tumors developed in 52% (12/23) in IT group and 35% (8/23) in controls. Cell of origin of the leukemia was reticular in 9/23, lymphocytic in 7/23 and mixed lesion in 3/23 in IT group; while in controls it was 2/23 and 3/23, respectively and mixed lesion was absent. All mammary tumors were of adenocarcinoma series.

Differences in the blast cells sensitivity to cytostatic drugs in various acute leukemia types and their clinical implications.

In 80 patients with AML, ALL, BC-CML and other proliferative blood diseases, the sensitivity of blast cells to cytostatics (vincristine, cytosine arabinoside, daunorubicin, l-asparaginase, hydrocortisone) was determined. The studies were made with the use of liquid scintillation measuring the inhibition of 3HTdR incorporation induced by the cytostatics tested. It has been shown that the method makes it possible to demonstrate individual reactivity of acute leukemia blasts to cytostatics. Variations in the individual sensitivity to cytostatics were mostly seen in acute myeloblastic leukemia, blastic crisis of chronic myelocytic leukemia, less often in acute lymphocytic leukemia.

Lactate dehydrogenase isoenzymes in human malignant tumors.

For estimation of the value of Chemetron method for LD isoenzyme separation on cellulose-acetate membranes in cancer tissues taken from corpses, malignant tumors revealed by autopsy were investigated. Overall stability of LD, as well as the maintenance of various forms of isoenzymes in the extracts with regard to post-mortem autolysis in corpse and long periods of freezing, have been demonstrated. No correlation was found between the degree of isoenzyme changes and the histologic grading of tumors. LD isoenzyme pattern in cancer metastasis has been considered with respect to vascular and metabolic role of the environmental normal tissue. In order to point out the possible role of LD isoenzyme pattern in cancer diagnosis a lot of preliminary studies has been reviewed. The clarification of the specific distribution of LD activities in different tissues has been considered.

Local eosinophilia in stroma of tumors related to prognosis.

The stromal reactions of oral, gastric, breast, and cervical carcinomas were uniformly examined for the presence of eosinophilic granulocytes. Their numbers were expressed in terms of a three-grade scale. While in the carcinomas of the uterine cervix a positive correlation between the quantity of the reaction and survival rate was found, in the other groups of patients eosinophilic granulocytes occurred only as a small admixture in a limited number of cases.

Comparability of cancer mortality data in different regions of Yugoslavia.

The standardized cancer mortality ratio based on well-defined causes of death (MSMR) was analysed. It was shown how the use of MSMR gave wrong results just because differences related to the age structure of compared populations were not eliminated. In this paper, the presented calculation pointed out that, actually, MSMR only standardizes age distribution of well-defined causes of death. The age standardization can be obtained with MSMR only if compared populations have the same age specific rates from all causes of death. At the same time standard population must also have the same age specific rates from all causes of death or have the same or similar ratios of age specific rates from all causes of death as populations under comparison.

Tumor promoting activity of Euphorbia striatella (Boiss) and skin irritant activity of some Euphorbia species.

Out of seven plants selected due to their use in folk medicine, five possessing ingenol as diterpene parent, exhibited mild skin irritancy where as E. striatella latex was highly skin irritant and produced papilloma, in initiation-promotion experiment on mouse back skin. This tumor promoting activity has been demonstrated due to the presence of ingenol-3-O-decanoate.

Effects of N4-methyl and N4,N4-dimethyl derivatives of 5-azacytidine on liver RNA synthesis and gastric secretion in rats.

Of different N-alkyl substituted derivatives of 5-azacytidine tested for their action on gastric secretion in rats with ligated pylorus the most effective beside 5-azacytidine were N4-methyl and N4,N4-dimethyl derivatives. The drugs blocked gastric secretion, gastric acidity, the extent of hemorrhage and the number and size of gastric defects. Simultaneously a higher synthesis of RNA in the liver of drug-treated animals was observed.

Inhibition of phytohemagglutinin-induced DNA synthesis and of adenosine deamination by 5-azacytidine and 5-aza-2'-deoxycytidine in intact human cells.

5-Azacytosine nucleosides block phytohemagglutinin-induced thymidine incorporation into DNA in intact human blood lymphocytes in a dose-dependent manner at 1-100 micrograms/ml. Maximal inhibition was observed when mitogen and 5-azacytidine or 5-aza-2'-deoxycytidine were added to the culture in admixture. Both analogues also inhibit deamination of exogenous adenosine in stimulated cells without affecting the synthesis of nucleotides from adenosine, adenine or hypoxanthine. Similar inhibitory effect of 5-aza-2'-deoxycytidine on the metabolism of adenosine was observed in normal (WI-38) and SV40 virus-transformed (VA-13) human fibroblasts.

Adjuvant and curative chemotherapy of the gastric cancer.

The authors report on the results of combined chemotherapy using cyclical alternations with several non-crossed resistant combinations in patients with gastric carcinoma in the adjuvant and palliative indication. The report includes also a small group of 6 patients with locally inoperable carcinoma, treated by radio-chemotherapy, followed by subsequent second-look resection and postoperative chemotherapy. Prolongation of survival in these patients indicates that possible benefit of the treatment regimens administered may be expected. Prospective randomized study is recommended to check the obtained results.

Expression of retrovirus-related functions in the methylcholanthrene-induced rat myelogenous leukemia.

The rat MR-leukemia (MR-Le) induced in Wistar rats by methylcholanthrene and whole-body irradiation, has been shown to be transmitted by means of cell-free filtrates of spleen and liver extracts. These earlier results lead us to determine the expression of retroviral functions by MR-Le myeloblasts in vivo and in vitro. DNA polymerase activity associated with particulate material purified from plasma of rats carrying MR-Le and from MR-Le tissue culture fluid, increased with the number of MR-Le myeloblasts. Interspecies-specific antigenic determinants, common to some mammalian retroviruses, are expressed on the surface of MR-Le cells. Electrophoretic pattern of the MR-Le viral material purified from leukemic plasma and tissue culture fluid revealed the presence of polypeptides with molecular weights related to proteins of mammalian retroviruses which were not identical with main structural proteins of the endogenous rat C-type helper virus (RaLV). The results strongly support the proposition that RaLV-unrelated rat retrovirus is involved in the onset of MR-Le.

Relationship between peroxidase activity and steroid receptors in human mammary tumors.

The interrelationships between levels of estradiol receptors (ER) and estrogen-dependent proteins, peroxidase and progesterone receptors (PgR) were examined in 54 histologically proven primary breast tumors in Indian women. ER and PgR were determined by the agar gel electrophoresis and dextran coated charcoal methods, respectively. Peroxidase activity (PA) was estimated by the guaiacol method. Our results showed a lack of correlation between ER and PA as well as between concentrations of PgR and PA in the tumors. It was concluded from this preliminary investigation that, in addition to ER and PgR, estimation of PA does not contribute significantly to more accurate assessment of hormone dependence of human breast tumors.

Schistosomiasis, metaplasia and squamous cell carcinoma of the prostate: histogenesis of the squamous cancer cells determined by localization of specific markers.

Histogenesis of squamous cell carcinoma in two prostates heavily affected by schistosomiasis was determined immunohistochemically by localization of two prostatic specific markers and keratin. The demonstration of prostatic specific antigen and keratin served to differentiate between metaplasia and squamous cell carcinoma associated with prostatic schistosomiasis from other prostatic and urinary bladder neoplasms.

Ammoniacal silver staining of lymph node cells. II. Reactive follicular hyperplasia and its relationship with Hodgkin's disease.

Lymph node biopsy specimens from 150 cases of reactive follicular hyperplasia, 52 cases of Hodgkin's disease and 30 cases of chronic lymphadenitis were studied. Clinical follow-up of cases for a period of two years at least was also considered. Formalin fixed and paraffin embedded tissues were stained with hematoxylin and eosin, and ammoniacal silver. Cytoplasmic accumulations of basic proteins were evidence in 30 out of 150 cases, but only 27 of these cases presented new lymph nodes with histological setting of Hodgkin's disease. The data suggest possible relationship between reactive follicular hyperplasia and Hodgkin's disease. However this relationship is not absolute as documented by two cases that being negative for the ammoniacal silver staining, were later diagnosed as Hodgkin's disease. Different types of atypical cells occurring in lymph nodes of reactive follicular hyperplasia were also studied. The potential use of ammoniacal silver staining in the early diagnosis of Hodgkin's disease is discussed.

Partial purification and characterization of tumor associated antigen in cervical carcinoma.

An identical component of tumor associated antigen (TAA) was detected in all clinical stages of the carcinoma of cervix (CaCx), in both premenopausal and postmenopausal patients, using heterologous antisera against the cancer tissues by immunodiffusion and immunoelectrophoretic tests. The TAA appeared to be cross-reacting specifically with carcinoma of other gynecological organs. A second TAA component was observed to be present only in CaCx, Stage II of premenopausal patients. The TAA component of CaCx common for all clinical stages, irrespective of climacteric states, was partially purified by subjecting postmeno CaCx, Stage II to gel filtration on Sephadex G-200. The antigen was found to be glycoprotein in nature, enzyme sensitive, highly thermostable and antigenically active at a pH range of 2.0-10.7. The approximate molecular weight of the component was found to be 67 000.

Determination of type specific antigenic reactivities and quantitation of murine mammary tumor virus by ELISA.

Type specific differences associated with the antigenic determinants of exogenous and endogenous murine mammary tumor viruses (MuMTVs) isolated from strain C3H(Jax) and ICRC were demonstrated by competition enzyme linked immunosorbent assay (ELISA). In this assay, highly purified intact MuMTV from the milk of strain C3H(Jax) was conjugated to the enzyme penicillinase using glutaraldehyde. IgG fraction of antiserum prepared against disrupted C3H (Jax) MuMTV was used in the competition ELISA. The assay was highly reproducible, specific to MuMTV antigens and detected 6-9 ng of the viral proteins. For competition, MuMTV preparations from corresponding endogenous MuMTV carrying subline C3H(Mect) and ICRCf were also used. The assay demonstrated the type specific antigenic reactivities inherent in the MuMTVs isolated from C3H(Jax) and ICRC strains of mice. The competition ELISA was also used to quantitate MuMTV specific proteins in tissue extracts and milk plasma.

Clinical controlled trial in advanced breast cancer: CMF (cyclophosphamide, methotrexate, 5-fluorouracil) versus CMF + T (cyclophosphamide, methotrexate, 5-fluorouracil, tamoxifen). CMEA Cooperative Treatment Group.

A controlled study with two arms was performed in 153 evaluable patients with advanced breast cancer. All patients were postmenopausal and untreated. The first combination included cyclophosphamide, methotrexate, 5-fluorouracil (CMF, 82 patients) while the second one contained the same cytostatic combination and tamoxifen (zitazonium) (CMF + T, 71 patients). Patients were prospectively stratified according to the dominant sites of metastases and randomized to the respective treatment schedule. In CMF combination the response rate (CR + PR) was 33/82 (40%) and complete response (CR) 8/82 (10%), respectively. In the CMF + T arm the corresponding values were 29/71 (41%) and 14/71 (20%), respectively. From the analysis of the data the superiority of the CMF + T arm could be established concerning CR in soft tissue and CR + PR in bone localization.

In vitro transport of F1-ATPase beta-subunit into mitochondria of Zajdela hepatoma and rat liver.

Transport of precursor of F1-ATPase beta-subunit into isolated mitochondria of Zajdela hepatoma and rat liver was examined. The hepatoma mitochondria were more active in the process than the liver organelles indicating that the relative F1-ATPase deficiency in the tumor mitochondria does not result from an impaired transport of F1-ATPase subunits into the tumor organelles. Similar results were obtained using digitonin-treated rat hepatocytes and Zajdela hepatoma cells instead of isolated mitochondria. The suitability of the digitonin-treated cells in the study of protein transport into mitochondria in vitro is demonstrated and the advantages of this system over isolated mitochondria are discussed.

Regulatory role of macrophages in tumor allograft development.

In the study of the role of macrophages in antitumor immune response the model of Sa I (H-2a) allograft development in B10 mice (H-2b) treated with xenogenous antithymocyte serum (ATS) was used. In the treated recipients tumor growth was enhanced as compared to untreated mice, subsequently in some of the mice the tumor permanently regressed while in the rest the temporary regression was followed by a secondary progressive growth. To determine the role of macrophages at different periods of tumor development, both silica (which is known to damage the function of macrophages) and double-stranded RNA (dsRNA; which, in turn, stimulated their function), were used. The administration of silica on the day of tumor cell transplantation slightly promoted tumor regression, whereas silica administered on days 7 or 14 after the cell transfer enhanced the secondary growth of the tumor. On the other hand, administration of dsRNA on the day of tumor transplantation promoted tumor growth, whereas, when administered on days 7 or 14, it enhanced the regression of the tumor. An administration of both substances at later periods had no effect on the growth of the tumor. From these results it can be assumed that macrophages at the time of induction of antitumor immune responses participate in the events enhancing the subsequent growth of the tumor allograft, whereas at later times they have an antitumor activity.

The study of the cytotoxic effect of anticancer drugs on human lung tumors transplanted to mice by use of diffusion chambers.

The effect of chemotherapeutic drugs (adriamycin, rubomycin, nitrosomethylurea, cyclophosphamide, vincristine, methotrexate) on the proliferative activity of cells of 19 human lung cancer (adenocarcinoma--8, squamous cell carcinoma--10 and small cell cancer--1) transplanted to mice in diffusion chambers was studied. Rubomycin and adriamycin showed highest activity while methotrexate was least effective. The cyclophosphamide, vincristine and nitrosomethylurea had the moderate and similar activity. The tumors of different histological types hade close sensitivity to the tested drugs. The 3H-thymidine incorporation inhibitory effect of drugs differed for squamous cell carcinoma and adenocarcinoma probably reflecting certain differences in cellular mechanisms of drug action in lung cancers of different histological types. The data on the cytotoxic activity of anticancer drugs against lung tumors were closed to known efficacy of these drugs in treatment of patients with lung cancer. The high activity of rubomycin in human lung tumors allow to hope for its high activity in patients. The results suggested that human lung tumors in diffusion chambers may be used in preclinical tests of antitumor drug activity against lung tumors.

The in situ immunological reactivity and its significance in the clinical behavior of the cervical human papillomavirus lesions.

The in situ immunocompetent cell (ICC) infiltrates in 286 cervical punch biopsies derived from the women prospectively followed-up since 1981 (16 +/- 14 months, M +/- SD) for an established human papillomavirus (HPV) infection with or without coexistent cervical intraepithelial neoplasia (CIN) were characterized using the histochemical ANAE (acid alpha-naphthyl acetate esterase) staining. Only minor fluctuations in the relative proportions of ANAE T+, ANAE M+, and ANAE- cells were found between the three types of HPV lesions (flat, inverted and papillomatous condylomas), as well as between the lesions with varying degree of HPV-CIN. The percentage of ANAE- cells (mediators of humoral immune response) increases in parallel with the increasing density of the ICC infiltrate. The intensity of the local infiltrate was inversely correlated with the progression, but not with regression or persistence of the HPV lesions. The percentage of ANAE T+ cells was highest in the 32 (18.7%) HPV lesions progressed during the follow up, almost identical in the 88 (51.5%) persistent lesions and lowest in those 51 (29.8%) lesions showing clinical regression. The results are discussed in terms of the proposed immune surveillance functions attributed to the local ICCs according to the MALT (mucosal-associated lymphatic tissue) concept. Conclusion is drawn that no major imbalance in the ANAE- definable main populations of ICCs exist in the cervical HPV lesions, which readily could explain their divergent clinical behavior. Other factors (like HPV type, synergistic action of cocarcinogens or a deranged balance between the ICC subsets) should be searched for to elucidate this issue.

No radiosensitivity-related change in plasma membrane properties in X- or gamma-irradiated L5178Y-R and L5178Y-S cells.

Two strains of murine lymphoma cells, L5178Y-R (LY-R) and L5178Y-S (LY-S) differ in radiosensitivity (D0 ca 1 and 0.5, respectively), in Na+/K+ and Mg2+-ATPase activities and susceptibility to heat; their fatty acid composition is also slightly different. Nevertheless, neither Na+-dependent amino acid uptake, nor membrane fluidity change after X- or gamma-irradiation (10 Gy under aerobic conditions, 27 Gy under extreme hypoxia). Although in LY-S cells there is a decrease in partition coefficient in a two-phase system, which indicates a late (24 h after irradiation) change in surface charge, this change is not related to viability, membrane fluidity, amino acid transport and survival.

The etiology of underlying liver lesions in 70 autopsied cases of hepatocellular carcinoma.

In 70 patients with hepatocellular carcinoma without history of antineoplastic chemotherapeutic drugs, anabolic and contraceptive steroids, representative sections of nonneoplastic liver tissue were examined for the presence of etiological markers. Hepatitis B surface antigen-positive hepatocytes were found in 16 (22.8%), alpha-1-antitrypsin globules in 3 (4.2%), Mallory bodies in 9 (12.8%), acicular inclusions in 1 (1.4%), diffuse giant mitochondria in 2 (2.8%), copper-binding protein in 25 (35.7%), greater amount of hemosiderin in 9 (12.8%) cases. Thorotrast was not detected. One or more markers were seen in 38 (54.3%) cases, most frequently in association with liver cell dysplasia and alcoholism. The presence of hepatitis B surface antigen in livers with dysplastic foci was highly significant as compared to organs showing no dysplasia. Only the mentioned antigen and the alpha-1-antitrypsin globules were found to indicate the etiology of the underlying liver lesion. The value of the other markers was found inconsistent in etiological diagnosis.