Urinary system functions in children with intensive antileukemic therapy of acute lymphoblastic leukemia.

The urinary system functions were studied in 48 children with acute lymphoblastic leukemia who were given intensive therapy. At the beginning of treatment of signs of blastic renal infiltration were discovered for the majority of patients. The authors suppose that the patient having signs of serious renal injury at the beginning of acute lymphoblastic leukemia must be regarded as a high risk patient. There was no serious damage to urinary system by prolonged chemotherapy as showed by normal function tests after cessation of the treatment. Bacteriuria and alteration of radiorenogram were observed very often during chemotherapy.

Terminal hematological changes in mice bearing osteosarcomas induced by plutonium-239.

Terminal hematological changes were studied in female ICR mice injected intravenously with 181.1 kBq/kg of monomeric 239Pu. Cellularity and differential cell counts in femoral bone marrow and spleen, as well as peripheral blood counts were determined. The results were evaluated with respect to the quantitative hematological changes and to the diagnosis of proliferative disorders. As far as the nonstochastic response is concerned, evidently decreased cell counts, without abnormalities in myelogram, were observed in the femoral bone marrow of 239Pu-treated mice and only a mild leukopenia was present in peripheral blood. No significant difference was observed between the hematological response of tumor-bearing mice and the response of mice free of osteosarcomas.

Surface glycoproteins of human sarcoma- and fibroblastic cells.

A comparison was made of the cell surface glycoproteins of four human cell lines, namely a giant tumor of bone cell line, an osteosarcoma line, a fibrosarcoma line and a human fibroblast line. The cells were labeled by lactoperoxidase catalyzed iodination and the glycoproteins extracted by 0.5% Triton X-100 were bound to lentil-lectin and subsequently analyzed by SDS gel electrophoresis. While the cell lines examined shared a series of common glycoproteins, it was found that the giant cell tumor line and the fibrosarcoma lines exhibited a higher degree of homology than the other cell lines.

Some parameters of cell-mediated immunity in colon cancer patients.

16 colon cancer patients in the second clinical stage (according to the TNM classification), aged 34 to 71 years were studied before and 14 days after radical surgery. The in vitro cell-mediated immunity was evaluated using the rosette-forming test and blastogenic reactivity of blood lymphocytes to PHA. The percentage of total rosette-forming cells (T-cells) before and after surgery did not alter significantly (p less than 0.1). At the same time there was no significant difference between the number of rosette-forming cells in cancer patients compared to donors (p less than 0.1). Blastogenic reactivity of lymphocytes to PHA expressed as a stimulation index (S. I.) showed a significant decrease of that parameter in patients compared to donors (p less than 0.001). The S. I. was lower in patients 14 days after surgery than prior to treatment (p less than 0.01).

Spontaneous transplantable lymphatic leukemia in Lewis rat (KPH-Lw-I).

A spontaneous transplantable acute lymphatic leukemia in Lewis rats (KPH-Lw-I) is described. It is transplantable regardless of age and sex in inbred Lewis rats. The leukemic cells are of a normal karyotype. The progress of the disease is very rapid with many similarities to human acute lymphatic leukemia. Attempts to grow these leukemic cells in adult Wistar rats were unsuccessful, but they grow in newborn Wistar rats. During two years about eighty passages were done and the course of the disease was similar to that found in the original Lewis rat. No evidence for B-lymphocyte markers was found on the surface of leukemic cells.

On spontaneous skin tumours in amphibia.

In 7 from 320 Rana temporaria grass frogs and 16 from 978 Rana ridibunda pond frogs, the 31 and 47 spontaneous skin tumors respectively, were observed. The tumors were represented by cystadenopapillomas and adenocarcinomas, and developed from epithelium of skin mucous glands. The spontaneous pigment tumors (melanophoromas) were found in 2 axolotls Ambistoma mexicanum. These tumors developed from hyperplastic melanophores in pigment spots and focal proliferations of melanophores located deeply in derma.

Effect of tumor growth on migratory properties of T lymphocytes in rats.

The dynamics of changes was followed in the migratory properties of T lymphocytes from lymphoid organs of tumor-bearing rats, labeled in vitro with 3H-uridine and injected i.v. to normal recipients, a comparison being made with the migration of T cells from normal donors in syngeneic recipients. The progressive tumor growth was found to correlate with a decline of the ability of spleen T cells from a tumor-bearing rat to migrate to the spleen and lymph nodes of normal animals, following a transient striking migration in the early stages after inoculation with tumor cells. The site of an intensive migration was the liver. T lymphocytes of lymph node draining tumor migrated in correlation with its growth in an enhanced percentage into the spleen, and in a reduced ratio to the lymph node of a normal recipient. T lymphocyte migration of all the lymphoid organs followed became normalized in the stage of the tumor rejection. Peripheral blood T lymphocytes from a tumor-bearing donor showed a depressed ability to migrate into normal lymphoid organs, while the migration rate of thymocytes from such donors in normal recipients underwent no change with the exception of an increased migration into the liver during tumor regression. The results showed changes in the migratory properties of labeled T cells to depend on the immune status of the donor and the lymphoid organ source of the injected lymphocytes.

Comparison of LAI reactivity in B 77 tumor bearing rats between, before and after surgical removal of tumors.

Using a new modification of the tube LAI test, LAI reactivity was followed in rats of the inbred Lewis (LW) strain with transplanted syngeneic B 77 tumors before and one month after a surgical removal of the tumors. A significant decline in LAI reactivity was noted after a thorough extirpation of the tumors as compared to that prior to the surgical intervention. The results have been compared and are here discussed with a view to a possible application of the LAI test in studies of antitumor immunity in patients with a malignant disease.

LAI reactivity in rats immunized with tumor cells.

Leukocytes of peripheral blood of F1 hybrid inbred strain of rats LW X AVN and rats of inbred Lewis strain, immunized for three consecutive weeks with increasing doses of live or dead MC-1 or B 77 tumor cells, incubated for 20 hours with specific tumor extract, showed a lower adhering ability (LAI 32.8 +/- 16.6, 44.4 +/- 14.0, 43.1 +/- 7.4%) than that of the same cell population cultured without a specific antigen. The nonspecific tumor extract did not produce any LAI reactivity (4.4 +/- 5.9, 5.8 +/- 8.2, 6.7 +/- 5.9%). The values of LAI leukocytes of the controls tested by both the antigens were concordant with those found in samples of the same cell population tested without any antigens. The discussion bears on a possibility of applying the 20-hour modification of the LAI test in studies of cell immunity in immunized patients.

Activity of chromatin-bound protease in histone fractions from rat liver and Morris hepatoma.

Activity of chromatin-bound protease of rat liver and Morris hepatoma 7777 was studied. Proteolytic enzyme was copurified with histones during extraction of chromatin with 0.25 M HCl. Total histone was fractionated by Oliver's et al. method. Histone fractions were incubated in 0.01 M Tris-HCl buffer (pH 7.6) at 37 degrees C within different periods of time. The behavior of these fractions in polyacrylamide gel electrophoresis as well as the amounts of peptides soluble in 5% TCA released during incubation indicated that enzyme was coextracted with histone H2B only. It was shown that the activity of protease coextracted selectively with histone H2B was higher in tumor tissue than in normal liver.

Application of the indirect tube LAI assay in the study of cell-mediated immunity in rats immunized with B77 tumor cells.

The indirect tube LAI (leukocyte adherence inhibition) test has revealed that peripheral blood lymphocytes of rats of the inbred Lewis strain immunized with liver B77 tumor cells incubated in the presence of a specific tumor extract, release a factor into the culture medium that inhibits adherence of normal syngeneic peritoneal cells. Supernatants in which lymphocytes of healthy - control animals had been incubated in the presence of the same extract (B77), or lymphocytes of immunized donors incubated in the presence of another type of tumor extract (MC-1) were devoid of such a property. This soluble factor was found in the supernatants only after 20 hours of incubation of sensibilized lymphocytes with a specific tumor extract. Th percentage of adhering cells decreased in dependence on the concentration of the soluble factor. The results are discussed in connection with questions relevant to the mechanism of LAI reactivity and the application of this phenomenon in studies on cell immunity.

Growth of mammalian cell lines in a spinner culture.

Several mammalian cell lines, derived from embryonic, normal adult and malignant tissues were adapted for long-term growth in agitated fluid medium. A simple method of spinner culture is described, employing a portable magnetic stirrer. Adaptation of cells for growth in spinner culture took a relatively short time, usually a few weeks, depending on the type of cell line used. Characteristics of cell lines maintained in spinner culture are described in details.

The tumorigenic effect of low dose levels of N-methyl-N-formylhydrazine in mice.

Solutions of 0.002 and 0.001% N-methyl-N-formylhydrazine (MFH) were given separately daily in drinking water to 6-week-old randomly bred Swiss mice for life. The treatments resulted in induction of tumors of the lungs, blood vessels, liver and gall bladder. The 0.002% solution gave rise to tumors of the above tissues in incidences of 76, 27, 28 and 11%, respectively, while the 0.001% solution induced tumors in incidences of 75, 23, 7 and 6%. Histopathologically, the neoplasms were adenomas and adenocarcinomas of lungs, angiomas and angiosarcomas of blood vessells, benign hepatomas, liver cell carcinomas and adenomas and adenocarcinomas of the gall bladder. MFH is an ingredient of the edible mushroom, the false morel Gyromitra esculenta. The present study proves the carcinogenicity of MFH administered at low dose levels. The possible environmental significance of the findings to the human population is discussed.

The use of Ficoll gradient centrifugation to obtain synchronously growing L5178Y cells.

Linear Ficoll gradient centrifugation has been used to separate exponentially growing cells according to their position in the cell cycle and to obtain synchronously growing cells. Analysis of the distribution of 3H-thymidine prelabeled cells revealed the optimal separation of the cells in the G1 phase in the gradient prepared from 3--9% solutions. Such G1-rich cell fraction collected from the gradient has proved useful to obtain fairly good synchronization of cell growth as evaluated by the measurement of cell number, 3H-thymidine incorporation and mitotic index.

Immunogenicity of methylcholanthrene-induced tumors tested by draining lymph node assay in syngeneic rats.

The immunogenic properties of MC-induced and not metastasizing tumors were tested in inbred Wistar and August strains by the aid of lymphocyte stimulation in syngeneic draining lymph nodes in the comparison with the opposite ones. The lymphocyte stimulation was estimated in vitro by the spontaneous incorporation of 3HTdR. It was found that the peak of the first response of draining lymph nodes was noted on the 7th day after injection of syngeneic tumor cells. This antitumor response was specific because: 1. the second immunological response of draining lymph nodes to the same tumor cells developed earlier and was higher than the first response and 2. the rats previously immunized with MC-induced tumor, and then challenged with syngeneic sarcoma or syngeneic fibroblasts exhibited the lack of the second response and only some cross-reactivity was observed by the use of the other MC-induced sarcoma. MC-induced tumors appeared earlier and more frequently in August than in Wistar strain, the level of syngeneic draining lymph node response to immunogenic tumors was higher in August than in Wistar rats despite of the lack of difference between the percentages of immunogenic tumors arising in these two rat strains.

The influence of hepatocarcinogens on the central effects of phenobarbital an D-amphetamine.

The influence of two hepatotropic chemical carcinogens - 2-acetylaminofluorene (AAF) and N-nitrosodiethylamine (NDEA) on the processes of adrenergic activation of central synaptic structures and narcotic effect of phenobarbital was investigated. AAF and NDEA modified the stimulating effect of D-amphetamine which was demonstrated by the decrease of stereotype behavior interval and increase of its toxicity. Both carcinogens potentiated the narcotic effect of phenobarbital. The investigation of the terminal component of hepatic mixed-function oxidases system (cytochrome P-450) during carcinogen injections allows to exclude the great importance of D-amphetamine and phenobarbital metabolic variation in modification of their effects. The results of experiments assume the concept of chemical carcinogen effects on the functions of central and peripheral nervous system synaptic structures related to control of the proliferation and differentiation as initial step in hepatic carcinogenesis.

Chromatin structure, number, volume and position of nucleoli in rat hepatocyte after treatment with benzo(a)pyrene and N-acetylaminofluorene.

In Wistar rat liver, after the acute treatment with two carcinogenic substances - benzo(a)pyrene and N-acetylaminofluorene - changes involving the nucleus are observed 2, 4 and 6 hours later. The electron-microscopic aspect of the chromatin, as well as the number and volume of nucleoli are altered. The parameters considered (number, mean volume and total nucleolar volume related to nuclear number) showed a movement of the nucleoli inside the nucleus from and towards the nuclear membrane, as a reaction to the chemical oncogenic stress. Thus, there is a relative variation of the number of nucleoli which come into contact with the nuclear envelope or occur within the nucleus. The number and volume are increased 2 hours after the treatment in the case of the nucleoli placed close to the nuclear membrane; the same process is observed after 4-6 hours in nucleoli located centrally. In the experiments described, the reversibility of most of these changes 24 hours after the treatment appears as being the rule.

A prospective study of lymphocyte responses to phytohemagglutinin in melanoma patients. (lack o prognostic value of correlation with minimal tumor burden).

Clinical findings in 63 patients with malignant melanoma were compared with the results of PHA-induced lymphocyte transformation tests. The patients had been followed up for 1 to 60 months (average 16 months) and repeatedly tested at regular intervals. Reactivity of lymphocytes in stage I patients was not altered if compared with that of controls. The frequency of lowered values increased in more advanced stages. Only in stage II patients a significantly impaired PHA-lymphocyte transformation was demonstrated in comparison with controls. In stage I and II patients, there was no significant difference in lymphocyte transformation between those with the presence of tumor and those without clinically evident tumor. Prognostic value of PHA transformation tests as regards the appearance of distant metastases within two years after the first examination, could not be proven in patients at initial stages of melanoma.

The blood volume and splenic red cell pool in hairy cell leukemia.

Investigating the splenic red cell pool in 7 patients with hairy cell leukemia we have found unusually high values of retained erythrocytes (means 527 ml, SD 264) with a range from 300 to 998 ml. This amount represents on average 26% of the total red cell mass. The blood volume in these patients ws 26% greater than predicted by NADLER et al. Following splenectomy, the hemoglobin improved with 49.3% (SD 29.1).

Skin reactivity to dinitrochlorobenzene in cancer patients.

Delayed skin hypersensitivity to DNCB was evaluated in 179 patients with cancer and in 71 control subjects. Cancer patients had a lower incidence of positive reactions than controls (50.2 vs. 84.5%, p less than 0.001). Incidence of impaired skin reactions in cancer patients varied according to the tumor type. Reactivity was most depressed in patients with lymphatic tumors (23.3% positive results) and least depressed in those with male genitourinary cancer (64.3% positive results). The number of positive results was significantly higher in cancer patients with localized tumor than in those with disseminated disease (60.2% vs. 44.1%, p less than 0.05). DNCB reactivity correlated well with the subsequent course of disease. Incidence of positive results in patients who had remained free of disease after 12 months of follow-up (64.3%) was significantly higher than in patients who had progressive disease (33.3%) or who died of disease (44.8%).

Thyroidectomy as a pathogenic factor in the evolution of thyroid cancer.

In a group fo 860 patients with thyroid cancers (TC), 66 were found having had thyroid surgery for benign disorder more than 5 years before the diagnosis of TC. More than 70% of these patients were operated upon 10 or more years ago. The incidence of cancers developing in thyroid postsurgical remnants could thus be estimated to 5.5-7.7% of all thyroid cancers. These patients had predominantly follicular cancers and usually were older than 40 years at the age of diagnosis of TC. Unusually high incidence of distant metastases (pulmonary or osseous) was proved compared to other patients with TC, while the increase of patients having both pulmonary and osseous metastases was not found. It seems that the risk of developing TC late after thyroid surgery is somehow connected with endemic goitre areas, as all such reports arise from the central European area. Whichever the pathogenic mechanism may be, all patients following surgery for benign eumetabolic goitres should be checked up in regular intervals with regard to the possible benign or malignant recurrence. Patients operated upon because of hyperthyroidism do not seem to carry any similar risk.

Cell surface labeling of proteins and glycoproteins of some human cultured lymphoid cells.

Cell surface proteins and glycoproteins of some human lymphoblastoid- and neoplastic hematopoietic cell lines were labeled by three different techniques: a) lactoperoxidase catalyzed iodination, b) galactose oxidase-tritiated sodium borohydride and c) reductive methylation of free amino groups by tritiated sodium borohydride--a modification of the technique known for the radiolabeling of soluble proteins. Electrophoretic analysis (SDS-PAGE) of labeled surface proteins and glycoproteins resulted in comparable and complementary electrophoretic patterns obtained by these three techniques. Electrophoretic patterns of studied lymphoblastoid cell lines were essentially similar, with an intensively labeled group of glycoproteins (gp44, gp31, gp24). These glycoproteins were markedly reduced on studied neoplastic cell lines. Further differences in several large proteins were observed between lymphoblastoid- and neoplastic cell lines.

Immunological reactivity of glycoprotein antigens from human lung cancers.

Glycoprotein antigens were extracted with perchloric acid (PCA) from the 14 individual lung anaplastic cancers and 4 lung adenocarcinomas and their antigenic activity was compared with carcinoembryonic antigen (CEA) isolated from liver metastases of colon cancer. The studies were performed by double immunodiffusion, immunoelectrophoresis and radioimmunoassay, using the specific immune sera against PCA-extracted glycoproteins of lung anaplastic cancer (anti-80 PCA-CaLu), and also anti-CEA, anti-AFP, and anti-alpha 1-acid glycoprotein sera. The existence of at least three groups of antigens in lung cancers was demonstrated, i. e. CEA-like antigen, group antigen characteristic for most of the lung anaplastic cancers and lung adenocarcinomas non-related to CEA and antigen(s) detectable only in some of anaplastic cancers. Moreover antigen of alpha 1-acid glycoprotein reactivity was detected in all lung cancers studied.

Specificity of non-histone proteins from hamster Kirkman-Robbins hepatoma.

Nuclear phosphoproteins from Syrian hamster liver and Kirkman-Robbins hepatoma were obtained by phenol method of TENG et al. and their chemical composition was investigated. Amino acid composition of phosphoproteins from both tissues resembled each other to a great extent. There was almost twice more phosphorus in the preparations from hepatoma than in the ones from liver. Using two electrophoretic techniques: SDS-polyacrylamide gel electrophoresis and isoelectric focusing some differences between the examined proteins of neoplastic and normal tissue were found. Three additional fractions (molecular weights 29 000, 89 000 and 93 000, respectively) could be observed in case of hepatoma proteins in comparison with the ones from liver. The phosphoproteins of Kirkman-Robbins hepatoma revealed three additional bands with isoelectric points 6.6, and 7.1.

Single versus combination chemotherapy of L1210 leukemia.

Thirteen anticancer agents, ten dual-agent combinations and five three-drug combinations were tested for treatment of L1210 leukemia in dBA/2-J mice. Data obtained form each three-drug regimen were compared with those obtained after administration of each drug alone and each two-drug combination. Cure (greater than 60 days survival) was observed in most of animals treated with VP-16 213 and VM-26. Certain regimens produced 90-100% cure rates (cyclophosphamide plus VP-16 213 or cytosine arabinoside). Inclusion of second or third agent in the treatment schedules produced improvement, deterioration or no effect on median survival time and cure rates, depending on the choice and sequences of evaluated agents. Eighty per cent of mice inoculated even with 10(7) L1210 can be cured by administration of modified schedule of VP-16 213 plus cyclophosphamide.

Cytotoxicity and the effect of combined treatment with X-rays of an anticancer 1-nitro-9-aminoacridine derivative in L5178Y-R and L5178Y-S cells.

The response to nitracrine, 1-nitro-9-[3'(N,N-dimethyl)aminopropyl] aminoacridine, an anticancer agent, was examined in two strains of murine lymphoma L5178Y inversely cross-sensitive to X-rays and UV light. A marked difference was found in sensitivity to the drug between L5178Y-R and L5178Y-S strain (D0 values 2.1 and 13.9 ng/ml, 3 h, 37 degrees C); survival was related to the yield of chromatid aberrations. Enhanced sensitivity of L5178Y-R cells to nitracrine as well as to UV light and an antitumor platinum complex has been ascribed to the previously found defect in post-replication repair in these cells. Combined treatment with nitracrine and X-rays under oxic conditions gave additive effects in both L5178Y strains.

New potential cytotoxic and antitumor substances. III. In vitro effect of 3-benzazepine derivatives on P388 cells.

3-Benzazepine derivatives manifested cytotoxic effects in in vitro tests on lympholeukemia P388 cells. The most efficient derivative (QF 1), i. e. 7,8-dihydro-3,4,12-trimetoxy-7-dimethylamino-10,11-methylenedioxy-5H-indolo [1,2-b][3]benzazepine-5-on in a concentration as low as 5 microliter/ml, considerably inhibited only the incorporation of labeled uridine into P388 cell fractions. In in vitro experiments, this substance blocked cell proliferation. After its prolonged action, the number of dead cells increased and this also when its interaction with the cells lasted but a short time--the substance being removed from the medium. In all probability, the substance retains the cells in the G1/S phase. A marked synergistic effect of tubercidine was attained in the presence of the substance.

Nucleoli of lymphocytes in the peripheral blood of patients with gastric cancer and gastric ulcer.

Nucleoli in lymphocytes were investigated in the peripheral blood of patients with gastric cancer and gastric ulcer. The number of "active lymphocytes" (lymphocytes with compact nucleoli or nucleoli with nucleolonemata as functionally dominant nucleoli) was increased in the peripheral blood of patients with gastric cancer as well as gastric ulcer. In contrast, the number of "resting lymphocytes" (lymphocytes with ring shaped nucleoli as functionally dominant nucleoli which under proper conditions can be stimulated with respect to the blastic transformation) was decreased in the peripheral blood of patients with gastric cancer. The decrease of these lymphocytes in number in the peripheral blood of patients suffering of gastric cancer might represent one of factors responsible for the spread of the malignant process.

Results of radiotherapy in Hodgkin's disease.

An analysis was made of the results of irradiation therapy applied during the period 1965-1976 to 131 patients with Hodgkin's disease. The results of radiotherapy only of clinically positive lymphogranuloma localizations were compared with those of irradiation of both positive and negative nodes. The evaluation criteria used were the numbers of 3-year long primary remissions, 3-year long survival rates, frequency of onset and localization of relapses. Better therapeutic effects were achieved by radiotherapy as regards duration of primary remission, survival rate and frequency of relapses in originally clinically negative nodes proximate to positive nodes. However, the number of trans-diaphragmatic and extralymphatic relapses did not decline. A 5-year survival rate regardless of the irradiation method and clinical stage in 131 patients (1965-1974) was attained by 70.2%. The discussion bears on problems of an optimum radiotherapy and combined radiochemotherapy, and the significance of diagnostic laparotomy in lymphogranuloma.

On the problem of gastric carcinoma cluster determination.

In the present study the author describes a method for the determination of a limited area (group of population) for carrying out epidemiological investigations into the occurrence of gastric cancer. The area thus determined is referred to as a focus or a cluster of gastric carcinoma occurrence. These clusters are derived from the comparison of the minimum population number (N-) in which there may develop, with 99% probability, kappa cases of gastric cancer at the given incidence rate, with the actual population number. All the necessary calculations were made using specially designed tables. If the actual population (with a rural Soviet) taken as a territorial unit) was less than N- at kappa cases of the disease, the author concludes that the occurrence of the given number of the disease cases at the corresponding Soviet exceeds the limit of chance and that in such a case it would be justified to undertake detailed analytical epidemiological investigations to review the hypotheses as to the genesis of gastric carcinoma.