Cytogenetic observations in chronic myeloid leukemia (CML) correlations of the cytogenetic findings with morphological cytochemical features of blast cells in various types of blastic phase.

Forty-nine cases of chronic myeloid leukemia have been investigated using cytogenic methods. Twenty-six patients were studied in chronic phase, twenty-three during the transformation of the disease. Out of the second group of the patients, nine were myeloid, five myelomonocytic, two promyelocytic, two megakaryoblastic and five lymphoid in their morphological and cytochemical appearance. Some correlation could be observed between the clinical, morphological, cytological and cytogenetic findings of different subgroups of the blastic phase in CML. The results demonstrate the divergent clonal evolution of transformed blast cells, and emphasize the importance of chromosome investigations -- in addition to the early diagnosis of metamorphosis -- in distinguishing the different subgroups of the terminal stage of CML.

Inhibition of colony-forming cells from bone marrow of leukemic patients by 3-oxauracil.

An in vitro system of colony forming cell inhibition was used for evaluation of 3-oxauracil (2,3-dihydro-1,3-6H-oxazine-2,6-dione) effectivity against bone marrow cells from 18 leukemic patients. The highest inhibitive activity of the 3-oxauracil was found in cases of acute lymphoblastic leukemia, comparing with bone marrow cells from patients with acute myeloblastic and chronic myeloid leukemia. According to the morphological characteristics of the formed colonies, lymphoblastic leukemia cells could be discerned from the myeloblastic ones in this system.

Differentiation of Friend erythroleukemic cells in media lacking fetal calf serum.

The results presented in this paper provide evidence that the erythroid differentiation induced with n-butyric acid (BA) in F4N Friend cells depends mainly on the ability of different media to support an active cellular proliferation. No significant differences in the percentage of hemoglobin-positive cells were observed when a comparative induction was performed in media, supplemented with either calf or fetal calf serum; the most satisfactory results were obtained in Dulbecco's minimum essential medium (DMEM), lacking fetal calf serum but containing calf serum instead. The great sensitivity of cells to BA in the presence of calf serum allows us to suggest that there is not any component, specific for the fetal calf serum, critically required for the induction of erythroid differentiation in this clone.

Some immunological peculiarities in generalized plasmacytoma and their possible clinical interpretation.

According to the immunological classification, plasmacytoma is assigned to the group of secretory immunocytomas. The designation "immunoproliferative hemoblastosis" implies certain peculiarities of immunological reactivity in its malignant transformation. The study deals with some of the more recent concepts on the immunochemical properties and morphological differentiation of plasmatic or plasmacytoma cells. Mention is also made of the results of some tests for evaluation of the function of T and B lymphocytes in patients suffering from a generalized plasmacytoma, during the course of chemotherapy.

Immune status in untreated cervical cancer patients.

Eighty untreated cervical carcinoma patients were tested for their immune status by evaluating levels of circulating T and B lymphocytes and lymphocyte reactivity to PHA. Significant reduction in T lymphocytes and depressed lymphocyte reactivity to PHA was observed following the progression of the disease. However, B lymphocyte levels remained unaffected. The general impairment observed in cell-mediated immunity was found to be more pronounced in the disseminated disease than in the localized group.

Effect of tumor-related factors on the in vivo and in vitro colony-forming ability of normal mouse marrow cells.

The effect of normal mouse marrow preincubation with Ehrlich tumor fluid (EAF) and Ehrlich tumor cell-conditioned medium (ET-CM) was investigated both on the in vivo assay for pluripotential stem cells (CFU-S) and on the in vitro assay for granulocytic-macrophagic progenitors (CFU-C). A significant reduction of the number of CFU-S was found when bone marrow cells were incubated with EAF. The morphological analysis showed a relative increase in the granulocytic type colonies besides an absolute increase in undifferentiated ones and an absolute decrease in erythroid colonies. The in vitro data show a decrease in CFU-C production, by stimulation with standard Colony Stimulating Factor (CSF), when the marrow cells are preincubated with EAF and ET-CM. When standard CAF (obtained from L-cells) is used at the higher concentrations, the CFU-C in the ET-CM treated marrow increases to the control values. EAF and ET-CM incubation also affects the CFU-C differentiation by increasing the number of pure granulocytic clones. The experimental data were discussed in order to explain an hypothesis about the activity of tumor-related factors.

A comparison of the macrophage electrophoretic mobility test and the leukocyte migration inhibition test in the detection of histocompatibility antigens on tumor cells.

Lymph node cells from mice sensitized by rejection of skin allografts were incubated in vitro with tumor cells carrying the sensitizing alloantigens. Products of the lymphocyte--antigen interaction released into the culture medium were simultaneously assessed by the macrophage electrophoretic mobility test and the leukocyte migration inhibition assay. Both macrophage slowing factor (MSF) and a factor which inhibited leukocyte migration (LIF) were produced. The release of both factors was detectable after comparable periods of incubation; their production was transient and ceased within 24 hours after it started.

Comparison of the hemocytometer and tube modifications of the leukocyte adherence inhibition assay -- II. Application of both modifications for detection of anti-tumor immunity in man.

The hemocytometer and the tube modifications of the leukocyte adherence inhibition (LAI) assay were compared under identical experimental conditions in patients with carcinoma of the larynx. In the direct test, both modifications showed adherence inhibition if patients peripheral blood leukocytes (PBL) were incubated with a larynx cancer antigen preparation. Cell-free supernatants obtained by cocultivation of patients PBL with the antigenic extract yielded positive results only in the hemocytometer modification. Those supernatants inhibited the adherence of both normal human PBL and guinea pig peritoneal cells. Human encephalitogenic protein used as antigenic preparation in both direct and indirect hemocytometer modifications caused changes in adherence too. The presented results confirm the opinion that the hemocytometer and tube LAI assays base on different mechanisms.

The inhibitory effect of vinylfurans on the glycolysis in tumor and yeast cells.

Most of the eighteen vinylfurane derivatives studied fully inhibit the glycolysis of both Ehrlich ascites carcinoma (EAC) cells and respiratory deficient yeast Saccharomyces cerevisiae at concentrations lower than 0.5 mmol/l. The inhibition of glycolysis is a consequence of some thiol enzymes inactivation. This concerns namely hexokinase (EC 2.7.1.1), glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12) and especially 6-phosphofructokinase (EC 2.7.1.11). Interference of vinylfurans with energy metabolism resulted in the depression of biosynthetic processes followed (14C-precursors incorporation into proteins and nucleic acids) and finally in the loss of EAC cell transplantability.

Immunoperoxidase studies on a tumor-cell-associated antigen(s) in rat renal neoplasms induced by dimethylnitrosamine.

Renal neoplasms were induced in rats of Wistar line by a single, intraperitoneal injection of dimethylnitrosamine (60 mg/kg). The histological investigation indicated that these tumors were entirely mesenchymal in nature. In some of them (16.6%) small adenocarcinomas were seen. Immunological studies were carried out by light and electron microscope, using the immunoperoxidase "bridge" technique. Tumor cell-associated antigen(s) was detected in 62% of the neoplasms examined. This antigen was individual as it did not react with sera from the other tumor bearing rats. It was established that this antigen was located mainly on the plasma membrane and also, even though in a lesser degree, on the endoplasmic reticulum and nuclear envelope of the tumor cells of mesenchymal and epithelial origin. Except the existence of tumor cell-associated antigen(s) in these neoplasms, the presence of antibodies produced against them in the sera of the diseased rats were also detected by these investigations.

Treatment of adult acute lymphoblastic leukemia: results of two trials.

Thirty eight patients with acute lymphoblastic leukemia were treated protocol 0171 (VCR, PRED, MTX, cyclophosphamide +/- +/- 6-MP) and protocol 0276/A (VRC, PRED, L-ASP, MTX, 6-MP, cyclophosphamide). Overall complete remission rate in both studies was 84--85%, and additional treatment in protocol 0171 resulted in complete remission rate of 92%. Median duration of complete remission in protocol 0171 was 23 months and median survival of all patients was 33 months. Six patients randomized to regimen "A" (without 6-MP in intensification) had median duration of complete remission 8 months and media survival was 13 months. Seventeen patients treated with regimen "B" (with 6-MP in intensification) had median duration of complete remission 25 months and median survival was 39 months. Median survival of patients allocated on protocol 0276/A in 21+ months and median duration of complete remission is 23 months at present. Twelve percent of patients treated with the best regimen have survived more than 66 months in continuous complete remission. The incidence of drug related death in complete responders was 6%. The relapses were most frequent during the first two years of remission. Extramedullary leukemia as the initial site of relapse was observed in 9% of patients.

Circulating immune complexes in patients with melanoma.

Sera from 51 melanoma patients and from 55 healthy donors were examined for the presence of immune complexes (ICs) by the EA-rosette forming cell inhibition assay. Using a cut-off point of 5% rosette inhibition, ICs were found in 144 out of 213 sera from patients with melanoma (67.6%) and in 11 out of 55 control subjects (20%, p less than 0.001). The percentage of positive results was significantly lower in patients with no evidence of disease (60%) than in patients with clinical symptoms of their tumor (localized disease -- 80%, p less than 0.02, metastatic disease -- 78%, p less than 0.05). The mean inhibitory rate in patients with metastatic disease (21%) was higher than in patients with localized tumor (15.1%, the difference not significant) and than in patients with no evidence of disease (10%, p less than 0.01). The high levels of ICs or a tendency for their increasing in most cases accompanied the dissemination of the neoplasm. The stabilization of ICs on the low levels was found to be a prognostically favorable sign.

Application of tube LAI assay in larynx cancer patients.

The tube LAI assay was used for long-term follow-up of specific antitumor immunity in larynx cancer patients. A high percentage of patients examined before surgery exhibited positive LAI reactivity, and the success of treatment was reflected in the disappearance of positive LAI reactions. The tube LAI assay may be useful in diagnosing the precancerous states. Finally, positive LAI reactions were observed in hospital staff members exposed to contact with cancer patients over several years.

The Eker renal tumor rat. General immune status in relation to a varying tumor burden.

Evidence was obtained that the response of blood lymphocytes to phytohemagglutinin was reduced in some but not all rats with massive hereditary renal tumors (less than 50 mm mean diameter). Erythrocyte-antibody sensitized, complement dependent (EAC) rosette formation by sensitized spleen cells was also depressed in these animals, but plaque forming cell activity was essentially normal. Lesser tumor growth was associated with general cell-mediated and humoral immune responses which were within the range found for non-tumor-bearing syngeneic rats.

Stress and radiation carcinogenesis in mice.

In the present experiment irritation consisting of a combination of an optic signal followed by a mild electroshock administered at regular intervals was started in 2 groups of animals at the age of 3 months. At 4 months of age, one of the irritated and one of the nonirritated groups were exposed to whole-body gamma irradiation at 20 daily doses of 0.5 Gy (50 rad(, 1.4 Gy/min (140 rad/min), while the other 2 groups were sham-irradiated. The animals were autopsied and the specimens were microscopically studied for the presence of malignant tumors. Malignant tumors involving particularly the testes and lungs and leukosis were found in 29% of males, whereas in females the tumor incidence with mammary, pulmonary and ovarian involvement and leukosis was 39%. The irradiation decreased the minimum latency time in the irritated males and both female groups. In males, the irritation lowered the cumulative prevalence of malignant tumors, a significant decrease being noted at the age of 15 months. In females, it was increased, with a significant rise observed to occur at the end of the experiment. The opposite effects of irritation on the radiation carcinogenesis in males and females can be attributed to the irradiation-induced specific alterations of the gonads in females and, in part, to a longer survival time observed in the irradiated females.

Gyromitrin as a tumor inducer.

Acetaldehyde methylformylhydrazone (gyromitrin) was administered in propylene glycol as 12 weekly subcutaneous injections of 50 micrograms/g weight to randomly bred Swiss mice. The treatment induced lung and preputial gland tumors in incidences of 51 and 0% in females and 46 and 28% in males, respectively. In the propylene glycol injected control groups, the corresponding tumor incidences were 28 and 0% in females and 32 and 0% in males. Histopathologically, the tumors were classified as adenomas and adenocarcinomas of the lungs and squamous cell papillomas and carcinomas and adenocarcinomas of preputial glands. Gyromitrin is an ingredient of the wild edible false morel mushroom Gyromitrin esculenta. The environmental significance of the findings is discussed.

Dominant-lethal test of 6-mercaptopurine: dependence on dosage, duration and route of administration.

A dominant-lethal test of 6-mercaptopurine (Spofa, CSSR) was carried out in male mice with four intraperitoneal, two oral, and one subcutaneous dosage levels. Furthermore, the drug was administered either in single doses or repeatedly for 7 and 14 consecutive days. With all of the dosage levels as well as administration durations and routes a genetic risk was proved at the periods of meiotic division and late premeiotic stages of spermatogenesis, manifested by increased percentages of fetal resorptions. In relationship to the total dose administered, irrespective of the route of administration, the most marked effect increasing the frequency of resorptions within the specified period of spermatogenesis was found in the groups that had received, by repeated administrations, more than 5.5 mg of 6-mercaptopurine per one male. Fertility reduction was apparent mainly after intraperitoneal injection; the differences were significant in weeks 3--5 of the test. The prenidation loss figures brought no univocal information about the interrelationship between the genetic effect and the stages of spermatogenesis.

Chemical carcinogens: screening, testing, risk assessment for man.

Testing of chemicals for carcinogenic activities is an important part of a comprehensive toxicological test program. Screening tests (short-term tests) can be utilized as methods to select suspicious chemicals which should or must be further tested in long-term animal experiments. Up to now long-term animal bioassays, even when taking into account all limitations of evaluation and extrapolation of results to man, rank prominent in predicting carcinogenicity of a compound in man. Any substance which is shown to cause tumors in animals should be considered carcinogenic and therefore a potential hazard for man.

Therapeutic evaluation of liposome-encapsulated Daunoblastin in murine tumor models.

Daunoblastin in free and liposome-encapsulated form was tested in the L1210 murine leukemia and the intramuscularly transplanted 276A sarcoma. Both therapeutic (% ILS, tumor volume inhibition) and toxicologic (leukocytes, body weight difference) parameters were evaluated. The liposome preparations showed similar therapeutic effects as the free substance but caused a lower toxicity with a lower mortality rate, higher leukocyte values and smaller body weight reduction. Longer sonication time with the output of more smaller unilamellar vesicles had no influence on the parameters in the solid model, but resulted in shorter ILS values in the L1210 model. Administration of empty liposomes immediately before liposomal Daunoblastin did not result in better antineoplastic activity but yielded higher leukocyte values.

X-irradiation and the incorporation of precursors for salvage and de novo synthesis of DNA in hepatomas and liver.

The effect of X-irradiation 15 Gy (1500 rad) on precursor incorporation into DNA was studied in transplanted Morris hepatomas and liver of the tumor-bearing rats. Precursors were studied which are utilized by salvage mechanisms ([3H] thymidine and [3H]deoxyuridine) or in de novo synthesis of DNA ([14C]aspartate and [14C]formate). The data suggested that salvage mechanisms are more sensitive to inhibition by X-irradiation and that DNA repair activity is greater in liver than hepatomas.

The effect of repeated BCG vaccine administration associated with surgery upon the growth of lung metastases in rats with transplantable Morris 5123 hepatoma.

Eighty four male Buffalo rats bearing transplantable Morris 5123 hepatoma were used for the experiment. Half of the animals were given into the tumor BCG vaccine (final dose 2.2 X 10(6) living mycobacteria), the other 42 rats after BCG underwent surgical removal of the hind leg together with the tumor. Though the immunotherapy reduced the size of lung metastases of the tumor it did not, however diminish the survival rate of the animals. Multiple BCG doses associated with the surgical procedure prevented dissemination of the tumor cells and prolonged the survival of rats up to the 100th day of the follow-up.

Prognostic significance of reactive changes in regional lymph nodes in cancer of the oral cavity.

Investigated were lymph nodes in a group of patients with spinocellular carcinoma of the oral cavity. As regards patient survival, lymphocyte predominance and sinus histiocytosis manifest prognostically the most favorable changes. A predominance of germinal follicle centers in our group of patients cannot be interpreted as indicator of an improved prognosis. Sinus histiocytosis has never been found in nodes with metastases.

Evaluation of immunological reactivity on the basis of spleen white pulp morphology in patients with far advanced urological malignancy.

The histological alterations in the white pulp of the spleens collected from sixty-one patients who had died of widespread urological carcinomas were assessed by using a standardized reporting system previously introduced by the author. As a control material the same number of age, and sex-matched patients who had died on myocardial infarction without signs of any malignancy was used. Special attention was focused on the evaluation of the lymphocyte populations (T- and B-cell areas) responsible for the immunological reactivity of the host. Morphological characteristics suggesting to reflect the activity of both the cell-mediated and humoral immune responses were found to be within normal limits in the control spleens, whereas in the carcinoma series both the T- and B-cell areas showed signs at cellular depletion. The results were interpreted, although with some preservation, to suggest that some degree of impairment of both the humoral and cell-mediated immunity exists in patients dying of far advanced urological malignancies.

Changes in skeleton, bone marrow reactivity and in immunity of patients with tumors of testis during long-term complete clinical remission.

A series of various tests were carried out in 19 patients treated for germinal tumors of the testis, with a long-term survival rate (7 to 28 years) in a complete clinical remission and the results compared with those of a previous investigation made 6 years earlier [3]. The tests involved: radionuclear investigation of the skeleton (85Sr--test), determination of bone isoenzyme of serum alkaline phosphatase, urinary hydroxyproline excretion, cytological examination of bone marrow, assay of serum levels of IgM, IgG and IgA as also of C3 complement, determination of the percentage and absolute number of peripheral T-lymphocytes, the test of blastic lymphocyte transformation after PHA and the skin test with DNCB. A chronic restructuralization of bone tissue (positive and suspected 85Sr-test in 74% of patients) was noted even after the relatively long time span. Likewise, an enhanced activity of bone isoenzyme of serum alkaline phosphatase was found in the majority of the patients, as also nonspecific reactive changes of bone marrow in the form of total hyperplasia and hyperplasia of various cellular elements, particularly of plasmacytes, eosinophils and lymphoid reticulum. Immunological tests revealed a depressed cell-mediated immunity -- especially a significant drop in the absolute number of peripheral T-lymphocytes.

Trends in cancer mortality in Czechoslovakia 1949-1978.

The analysis of the trends of the principal causes of deaths in Czechoslovakia indicated leading second position of malignant neoplasms in the whole mortality over the period 1949 through 1978. The rapidly growing all sites cancer mortality trends in males together with their unimportant changes in females resulted in the steadily increasing overmortality of males. Among digestive organs important decrease of stomach and in lesser extent also of oesophageal cancers with simultaneously increasing mortality trends of malignant neoplasms of colon, pancreas and rectum in both sexes were observed. The trends in the cancer mortality of respiratory organs as well as the increasing rates of the whole male cancer mortality were strongly influenced with dramatical increase of lung cancers while these of larynx showed slow increase in males and decrease in females. The rapidly growing mortality trends showed also breast cancer, reaching at the end of the investigated period the first position in the whole female cancer mortality, while the increasing mortality from the malignant neoplasms of the different genital and urinary organs of females were less or more expressed. The relatively high average annual increase was characteristic for the mortality trends of the malignant neoplasms of bladder and other urinary organs and of prostate gland and testis in males. Among other sites the increasing mortality trends from malignant neoplasms of brain and leukemias were of interest.

The macrophage electrophoretic mobility assay in patients with renal cell carcinoma.

An antigen-induced release of a macrophage slowing factor (MSF) by peripheral blood lymphocytes was used to evaluate lymphocyte sensitization to various antigens in 21 patients with renal cell carcinoma (RCC) and in 14 control subjects. Sixteen of 21 patients with RCC, but no controls, were found to be sensitized to a soluble antigen prepared from an allogeneic kidney tumor by 3 M potassium chloride extraction. Peripheral blood lymphocytes from some patients with RCC displayed sensitization to protein isolates from fetal kidney (4 of 19), control "normal" kidney (4 of 21) and urinary bladder carcinoma (3 of 19) tissues. The results of macrophage electrophoretic mobility (MEM) measurements indicate that the MEM assay may provide a means of monitoring immune responses in patients with RCC. Further studies are needed to correlate the immune response detectable by the MEM test with the patient's prognosis.

Detection of anti-tumor immunity in man by the indirect macrophage adherence inhibition assay using guinea pig peritoneal cells as indicator cells.

Supernatants obtained after incubation of tumor patient lymphocytes with the homologous tumor-associated antigen (TAA) elicit adherence inhibition in normal guinea pig peritoneal cells. From 37 tested patients with different carcinomas, the supernatants of 30 caused more than 10 per cent adherence inhibition (breast cancer 11/11, lung cancer 7/8, colon cancer 6/7, stomach cancer 6/11). Besides 40 healthy blood donors were tested to a panel of TAA. There was a reactivity more than 10 per cent only in 2 cases. The supernatants of 14 patients with nonmalignant disorders produced adherence inhibition more than 10 per cent in 2 cases. By incubation of tumor patient lymphocytes with the human encephalitogenic protein the supernatants of 19/27 patients elicited a positive reaction. The results suggest that anti-tumor immunity can be detected in an indirect heterologous adherence inhibition system.

Antitumor activity of flavonoids on NK/Ly ascites tumor cells.

Among the various flavonoids, rutin and quercetin increased the survival time of mice inoculated with NK/Ly ascites tumor cells. The best results were obtained when the mice were given 2.0 mg rutin twice daily for 8 days. The O-silyl-substituted rutin and quercetin were less effective than rutin or quercetin themselves. Besides rutin, quercetin and morin two other flavonoids, luteolin and pelargonidin also exerted growth inhibitory effects on NK/Ly ascites tumor cells cultures in vitro.

The preoperative diagnosis of malignancy of ovarian cysts.

The diagnostic difficulties of ovarian carcinoma are well known. In the present investigation efforts were made to improve the diagnosis of malignant ovarian cystic tumors by utilizing their proliferative growth with high vascularity. Under the guidance of ultrasound, fine-needle aspiration biopsy specimens were obtained, and the presence of fibrin/fibrinogen degradation products and the DNA synthesis were determined in the aspirated cystic or ascitic fluid, besides cytology. With the combination of these diagnostic tools, the possibility of a true preoperative diagnosis is nearly one hundred per cent, which improved the chances of an effective therapy.

A Fortran program for the calculation of estrogen receptor contents in human breast cancer.

A computer program in Fortran-IV for the processing of data from estradiol receptor assays in human breast cancer is described. The program prints the results in two tables together with the Saturation graph and the Scatchard's Plot. It includes a systematic, operator-independent, method for the elimination of "deviated points".