Method of separate bronchospirography with the use of bronchofibroscope and its clinical value in lung cancer patients.

In this paper the authors describe: a method of bronchospirography with the use of bronchofibroscope; a method of simultaneous performance of bronchospirography and bronchofibroscopy; criteria of indices and staging of each lung ventilation capacity; patterns of interrelation of the "healthy" and disabled lung according to the degrees of lung ventilation disturbance; indications and contraindications to operative treatment based on bronchospirographic data. For evaluation of the operability bronchospirographic investigations of the lung ventilation capacity were carried out in 225 lung cancer patients.

Risk evaluation of thromboembolic diseases according to functional test data in patients with advanced malignancies.

Evaluation of the risk of thromboembolic disease (TED) was based on the study of reactions of hemostatic and fibrinolytic systems in response to the influence of test-irritant such as twice applied local hypoxia of upper extremities. The study was made on 20 healthy subjects, 15 patients with chronic thrombophlebitis in acute condition and 45 advanced cancer patients. Cancer patients should be considered as having higher risk of TED due to the detected disturbances of hemostatic and fibrinolytic systems. The use of functional test offers useful information of the hemostatic situation.

Colorectal carcinoma and cholecystectomy.

In a group of 11,153 autopsied subjects a higher number of cholecystectomies was found in both sexes in individuals with large intestine tumors as compared with the rest of the autopsied persons without tumors (p less than 0.001). The results confirmed the hypothesis that cholecystectomy is a supporting factor in the process of carcinogenesis of the large intestine. We presume, in accordance to other studies, that the influence of cholecystectomy itself is not strong. For a development of a carcinoma, other carcinogenic factors must start or continue their effects. These factors are decisive for the genesis of a tumor.

Retrovirus like particles produced by human embryonal cells and cell lines derived from human malignancies. II. Protein structure.

Very small amounts of retrovirus-like particles were isolated from tissue culture media of various types of human cell lines derived from malignant as well as normal cells. The aim of the present study is the characterization of protein profiles of these isolates. The comparison of protein profiles of isolated human virus-like particles with the profiles of well characterized animal as well as human exogenous (LAV/HTLV-III) retroviruses revealed a 25k protein (p25) to be a major protein or at least one of the protein components of human retrovirus-like particles.

Modification by caffeine of acute cytotoxic response of cultured L5178Y cells to hydroxyurea treatment.

The effect of caffeine (CAF) on acute cytotoxic response of L5178Y lymphoblasts to hydroxyurea (HU) treatment was studied. The following events were examined: abnormal cell enlargement (giant cell formation), the rate of recovery of cell reproduction and DNA synthesis after releasing the cells from the HU blockage, parental DNA breakage and cell death. The presence of CAF at nontoxic concentration prevented giant cell formation, enhanced cell growth inhibition and cell killing. The effect of CAF was variable, dependent on the duration of exposure to HU and the time of exposure to CAF. To obtain maximal effect, the continuous presence of CAF during HU treatment and posttreatment time was necessary. Hydroxyapatite chromatography assay of single strand (ss) and double strand (ds) fractions in parental DNA and the measurement of the rate of post-treatment recovery of DNA synthesis indicated that CAF enhanced HU-induced DNA lesions. It is concluded that the results give further evidence that even short HU treatment can damage not only newly formed but also parental DNA. The lesions are normally, at least partly repaired and can be expressed under the conditions of DNA repair inhibition.

Further characterization of the leukemogenic activity of haloperidol in mice.

Haloperidol, a butyrophenon, is widely used for the treatment of psychotic disorders in man. Recently we reported that this drug causes, with high incidence, the development of monocytic-myeloid leukemias in male NMRI mice upon 5 X 5 mg/kg i.p. administration. Here we present evidence for the leukemogenic effect of haloperidol in two other strains of mice (XVII AKF1 hybrids, and the low leukemic BALB/c/BOM). The strain-dependent incidence of leukemias ranged both in males and females between 34% (AKR) and 69% (XVII AKF1) with average latencies between approximately 200 (AKR) and 600 (BALB/c) days. On the basis of cytological and cytochemical criteria the predominating type of leukemias was classified as monocytic-myeloid. These leukemic were serially transplantable. Cell-free extracts of leukemic tissues did not induce the disease indicating that no virus was activated by haloperidol. However, when the drug was administered to AKR mice after a suboptimal dose of nitrosomethylurea (NMU), a higher incidence of mixed-type leukemias was observed as with haloperidol alone. NMU alone induced lymphatic leukemias with proven viral involvement. The tumor promoter 12-0-tetradecanoylphorbol-13-acetate did not influence haloperidol-induced leukemogenesis.

Protective immunity by chemically modified tumor cell antigens extracted by 3 M KCl.

Tumor antigens from Sarcoma-180 ascites tumor cells were extracted by 3 M KCl before and after treatment with acriflavine. Following dialysis of the extracts against distilled water to remove KCl, the soluble material (antigen) and the precipitate formed during dialysis were collected from the dialysate. Protective immunity of the soluble antigen and the precipitate was evaluated by immunizing mice and subsequent challenge with native tumor cells. It was noted that the soluble antigen and the precipitate of the dialysate obtained from acriflavine-treated tumor cells induced significant protective immunity. The precipitate part appeared to be more effective. Interestingly, similar materials from untreated tumor cells did not show any such protective immunizing efficacy. Antigens extracted by 3 M KCl from chemically modified tumor cells assumed significance in respect to their antitumor immunity.

N-acetyltransferase phenotype of patients with cancer of the larynx.

In the present paper N-acetyltransferase phenotype has been evaluated in 128 patients with cancer of the larynx. Out of 128 patients, 86.7% were smokers. The corresponding control group consisted of 106 men who did not suffer from neoplastic diseases. 83.6% of the patients were slow acetylators. This is significantly different from the frequency of 60.3% of slow acetylators observed in the control group. The slow acetylators exhibited a greater susceptibility to the cancer of the larynx.

Combined chemotherapy and radiotherapy in the treatment of advanced head and neck cancer using cisplatin.

Between the years 1981-1984, 30 patients with advanced head and neck cancer (Stages III and IV) were treated by combined chemoradiotherapy. Chemotherapy with cisplatin was given before radiotherapy and three times after radiotherapy at the same dose (3 mg/kg body weight) monthly. Radiotherapy was administered with 60Co beam three times weekly in 17 fractions, at a total dose of 51 Gy. The results were summarized after completion of the combined therapy. Of the 30 patients 8 had complete remission (27%) and 13 patients had partial remission (43%). Adverse effects in this type of therapy were moderate and no serious complication occurred. The results were compared with a group of patients previously treated with methotrexate and radiotherapy. Optimal dosage of radiotherapy in combination with antineoplastic drugs is discussed.

Relationship of some biosocial factors to colon cancer in Belgrade (Yugoslavia).

Eighty-eight patients with histologically confirmed colon cancer and two control groups individually matched by age, sex and place of residence, were interviewed about demographic, biosocial and medical variables. The use of laxatives, past history of large bowel disease and coffee consumption for 20 or more years were significantly more frequent among colon cancer than in their controls. No significant differences were found between cases and controls with respect to education level, job activity, smoking habits, alcohol consumption and cholecystectomy.

A simple and sensitive method to measure radiolabeled antibody uptake by tumors in nude mice.

A method for detecting the uptake of 125I-labeled monoclonal antibody by human tumors xenografted into nude mice is described. After implantation of the tumor intramuscularly in one of the forelimbs, radioactivity was measured in the area of tumor inoculation and in the contralateral normal forelimb. For these measurements, a lead apparatus was constructed which shielded over 99% of the whole body radioactivity and permitted efficient counting of 125I in the exposed portion of each forelimb. For each tumor an uptake ratio (UR) was calculated for the radioactivity in the tumor compared to muscle. With this shield apparatus, reproducible, sensitive and relatively rapid determinations of UR values can be obtained. Antibody localization can be detected in some tumors which are nonpalpable or very small and for tumors of up to 2 cm3 in volume. Whole-body radioactivity ranging from less than 0.037 MBq to higher levels suitable for radioimmunoscintigraphy could be utilized in this procedure.

High yield of micronuclei and micronuclei premature chromosome condensation in a mouse tumor cell line cultured in vivo with prearrested mitotic metaphases.

It is generally argued that micronuclei and micronuclei premature chromosome condensation (MN PCC) in stable cell lines occur only after treatment with potent clastogens, mutagens or carcinogens. In the present paper we report on the occurrence of micronuclei and MN PCC (predominantly of S type) at a high frequency in a mixed in vivo culture of prearrested mitotic metaphases with an asynchronous population of mouse ascites tumor cells (S180) without the aid of any clastogen, mutagen or carcinogen.

Establishment of a tumor cell line and its use for an agar-free colony-forming assay.

The establishment of a spontaneously transformed tumorigenic human fibrosarcoma line derived from the ovary of a breast cancer patient (NO-15) is described. This tumor cell line has been characterized by tumor growth in nude mice. The cells form colonies without the usually recommended agar technique and are sensitive to cis-dichlorodiammineplatinum(II) (cis-DDP), daunorubicin (DR) and bleomycin (BLM), three well-known chemotherapeutic agents. The cells were incubated in the presence of drugs using three different methods. These agents proved to be noncytotoxic to monolayers treated for three hours. Conversely dose-dependent cytotoxicity was seen in suspension cultures treated for three hours. Optimal correlation between the three methods for the evaluation of cytotoxicity was seen in monolayers exposed to these agents for 24 hours. The results of three different methods to evaluate drug efficacy (number of colonies, growth rate and 3H-thymidine incorporation) were in good correlation. The incorporation of the radiolabeled tracer into cellular DNA is advantageous for its rapidity and quantitative reproducibility.

Population changes of human somatic cells in culture during short-term treatment with drugs.

The human somatic cell line VUPT was treated with 8-azaguanine or 5-bromo-2'-deoxyuridine for a week, then the drug was removed and cultivation continued in a standard medium. During or after the exposure to the drug various population parameters, such as morphological variability, frequency of multinuclear cells, total mitotic index, index of atypical mitoses, dispersion of chromosomal counts were evaluated. Increase in both morphological and karyological variability during the treatment with drugs was found but this increase was temporary and during one or two weeks of cultivation in a standard medium the initial phenotypic equilibrium of the population was restored. The meaning of such population changes is discussed.

Possible prognostic significance of the assessment of dipeptidylpeptidase II in peripheral blood lymphocytes of patients with chronic lymphocytic leukemia.

In 115 patients with chronic lymphocytic leukemia (CLL), grouped according to the staging system as proposed by Rai et al., dipeptidylpeptidase II (DPP II) reaction was carried out in smears of peripheral blood using Lys-Pro-MNA and Lys-Ala-MNA as substrates and Fast Blue B (FBB) as the coupling agent in cacodylate buffer, pH 5.5. In patients with stable disease a significantly lower number of DPP II positive lymphocytes was found. On the other hand, a high number of DPP II positive lymphocytes indicated a progressive disease. Lys-Ala-MNA proved to be a better substrate for these purposes. When comparing these results with the stage of the disease proposed by Rai et al., a good correlation was found for Stages 0, I and II. However, in the Stages III, and particularly in the Stage IV, there was a wide range of DPP II values, although the average value calculated from the data of all patients of this group was high. Patients with low DPP II values had usually a stable disease in spite of thrombocytopenia which precluded them from allocation to Stage IV. It is concluded that the assessment of the number of DPP II positive lymphocytes represents a simple and reliable prognostic criteria in patients with B type CLL.

Factors associated with prognosis in human breast cancer. VII. A comparison between a Cuban and a Swedish study.

Prognosis in human breast cancer is important since therapeutic plans can be adjusted to individual probability of relapse. Comparative studies are not very frequent in literature. Prognostic factors are not assessed in the same way in different countries, thus contradictory results can be obtained. This paper summarizes the studies performed in two different populations of Swedish and Cuban patients. A group of prognostic factors was compared: age, tumor size, lymph node status, hormone receptors and DNA distribution. Some differences were obtained in these parameters and in short-term relapses. Lymph node status was more important in Swedish series, whereas ER content and DNA pattern were of more predictive value in the Cuban series. Biological interpretations of the results is suggested.

Effect of lomustin on some platelet functions in vitro.

The effect of lomustin on platelet functions was investigated in vitro using a wide battery of tests. Lomustin was found to act as a specific inhibitor of platelet aggregation, release reaction and clot retraction and to induce acquired thrombocytopathy. Thus, impairment of platelet functions might play a role in hemorrhagic complications accompanying lomustin therapy in some cases.

Role of levamisole immunotherapy as an adjuvant to radiotherapy in oral cancer. I. A three-year clinical follow up.

Eighty-two patients with squamous cells carcinoma of the oral cavity belonging to stages T1N0M0 and T2N0M0 were randomized to receive either levamisole or placebo therapy following conventional radiotherapy. Oral levamisole, at 150 mg daily doses for three consecutive days, was given once every two weeks. The patients were followed-up for three years and the results reported. Levamisole appears to have some beneficial effect in prolonging the disease-free interval of these patients (44% in the levamisole group compared to 32% in the placebo group after 30 months of therapy). This, however, did not have any effect of the metastatic potential of the tumors. The effects of levamisole on peripheral blood leukocytes and lymphocytes were more promising. The restoration of leukopenia and lymphopenia observed after radiotherapy was faster in the levamisole group when compared to the placebo group.

Risk factors of bladder cancer in Spain.

A case-control study was performed on 406 (353 males and 53 females) bladder cancer patients and 406 controls matched in age, sex and date of admission to the hospital. The questionnaire included questions about socio-economic status, residence, personal and familial histories of diseases, professional occupations, and habits such as cigarette smoking and consumption of coffee, artificial sweeteners, alcoholic and nonalcoholic beverages, and water. Up to three different exposures to occupational risks were taken into account. Data were studied by a discriminant analysis technique. The results suggest a multifactorial etiology for this disease. The factors increasing the risk of bladder cancer, listed in order of importance, were: total number of cigarettes smoked, history of urologic disease, second exposure to an occupational risk, use of artificially sweetened beverages, low consumption of water, family history of cancer, third occupational exposure to a risk, use of artificial sweeteners, and years of coffee drinking.

The role of proteinases, interferons and hormones in proliferative activities of nonmalignant and malignant cells.

Proliferative responses, in form of colony forming ability, of nonmalignant Chinese hamster fibroblasts (V79) and malignant human laryngeal carcinoma cells (HEp) were examined after treatment with proteinases, proteinase inhibitors, interferons or hormones, in G0-phase, early G1-phase and G1/S point of the cell cycle. These phases of the cell cycle are of particular importance from the point of control and regulation of the proliferative activity of cells, and the effects of proteinases and other agents examined in this study occurred predominantly at these points. Among the agents tested, cysteine proteinase inhibitor (CPI) and calf liver neutral proteinase acted differently on malignant cells as compared to nonmalignant cells: they inhibited the proliferative commitment of malignant HEp cells in early G1-phase, while stimulated colony formation of V79 cells at the same point of the cell cycle.

Alpha-fetoprotein, carcinoembryonic antigen and various biochemical tests in patients with tumorous and inflammatory liver diseases.

Over the period of the past 9 years (1980-1988), 320 patients (mean age 60.9 +/- 13.2 years) suffering from various liver diseases have been examined. There were three main groups of patients: (1)--24 patients with primary liver cancer (PLC), 19 of them with hepato- and 5 with cholangiocellular carcinoma, (2)--153 patients with metastatic liver tumors (MLT), and (3)--143 patients with inflammatory liver diseases (ILD). The results of examination of alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), alkaline phosphatase (ALP) and gamma-glutamyl transpeptidase (GMT) in these patients have been analyzed with the aim to evaluate their contribution to the differential diagnostics of tumorous and inflammatory liver processes. For the diagnostics of malignant hepatoma AFP appeared to the most specific test. The significance of other tests for diagnostics of malignant hepatic diseases is obviously limited. These tests are recommended to be considered (in the case of their increase) in close connection with the clinical image and additional examinations. The importance of correlation between cirrhosis and malignant hepatoma is also to be noticed. In spite of all this, we believe that in the case of positivity of the above tests the patients have to be carefully examined and followed up, and that the clinical course and the dynamic of the mentioned tests has to be thoroughly observed. Because of the specificity of values of the AFP-test with malignant hepatoma, we find it useful to perform this test in all patients with chronic liver diseases.

Comparative study of cyclocytidine and arabinosylcytosine disposition in rats.

The stability of cyclocytidine (II) in aqueous solutions decreased with increasing pH and temperature. After intravenous injection to rats the biphasic decline of cyclocytidine in the serum was accompanied by simultaneous formation of arabinosylcytosine. Since under physiological conditions (37 degrees C, pH 7.2) the half-lifetime of cyclocytidine dissolved in water was 5 hours, the very early appearance of arabinosylcytosine in the serum of rats suggests that various hydrolytic enzymes may also be responsible for the conversion of cyclocytidine in to arabinosylcytosine in vivo. An equimolar dose of arabinosylcytosine (I) injected to rats produced serum levels which were several times higher than those of cyclocytidine under the same conditions. Since the rate constant of overall elimination was similar for both compounds, this difference in serum levels may most probably be explained by a much higher distribution volume of cyclocytidine resulting also in a higher plasmatic clearance of this compound.

Pharmacokinetics of a spin-labeled rubomycin analog.

Pharmacokinetics of a spin-labeled analog of rubomycin (ruboxyl) was studied. Differences were found in ruboxyl pharmacokinetics in normal and tumor-bearing animals. Most of the drug was excreted within 6 h. The differences in pharmacokinetics of ruboxyl and nitroxyl radical were established.

Suppression of tumorigenicity in clones of hamster cells transformed with avian sarcoma virus.

Hamster cells transformed with avian sarcoma virus, which had been selected for resistance to 8-azaguanine, were cloned. Several single cell clones were isolated which differed significantly in their tumorigenicity in comparison to the parental highly tumorigenic cells. The nontumorigenic cell clones contained rescuable avian virus. Both parental and resistant cells possessed transformed phenotype. Comparative studies with parental highly tumorigenic cells and nontumorigenic cell clones failed to detect any difference in the expression of p60src, in this phosphorylation and in phosphokinase activity. The isolated nontumorigenic cells represent cell mutants in which the viral src gene is controlled by a suppressor gene. The cells might be useful for further characterization of a putative antioncogene.

Association between the occurrence of antibodies to simian vacuolating virus 40 and bladder cancer in male smokers.

An association between the occurrence of antibodies to simian vacuolating virus 40 and bladder cancer in male smokers was found in a case-control study conducted in 200 male patients with bladder cancer and 400 control men. The risk of bladder cancer increases with the years of cigarette smoking.

Restoration of immune response by levamisole in B-77 virus induced tumor-bearing rats.

The immunorestorative effect of levamisole was observed in Lewis rats inoculated with a syngeneic B-77 virus induced tumor and immunized with sheep erythrocytes. In tumor-bearing rats the cell-mediated cytotoxicity, the delayed footpad reaction and the antibody production were suppressed. While the cytotoxicity suppressed in tumor-bearing rats was not restored by levamisole, delayed footpad reaction and antibody production were enhanced as compared with the normal reaction, or were restored to normal levels by levamisole, respectively. Tumor growth was significantly suppressed in rats treated with levamisole. The presence of tumor changed the uptake of 3H-uridine labeled lymphocytes in the spleen and peripheral blood. Restorative effect of levamisole on T lymphocytes was different in the studied organs.

Structural changes in DNA of human breast carcinoma.

Rearrangement and loss of genetic material in DNA of human ductal breast carcinoma was analyzed by restriction enzyme digestion, agarose gel electrophoresis and Southern blot hybridization using 32P labeled c-Ha-ras-1 and multilocus, minisatellite 15.1.11.4 as a probe. In several cases an allelic loss of c-Ha-ras-1 locus in heterozygous patients were noticed. Rearrangement of the genetic material in neoplastic tissues was also observed after hybridization with a minisatellite probe. These results provide evidence that loss and redistribution of the genetic material does occur in human ductal breast carcinoma.

BrdU-induced changes in the provirus of PR-RSV in mammalian cells.

Previously we demonstrated differences in the organization and methylation pattern of integrated Rous sarcoma proviral sequences in helper-dependent virogenic rat TWERC cells. In the present study we attempted to induce changes in the integrated viral genome of TWERC cells using 5-bromodeoxyuridine (BrdU). Four clones (A, B, C, and F) derived from the parental cell line were treated for 10 months with different concentrations of BrdU. Restriction enzyme analysis of the parental cell line and its clones showed that the cells contained in their genomic DNA two copies of deleted provirus. In TWERC cells, the PR-RSV provirus lost the whole env gene and part of the 3' end of the pol gene. The proviral sequences in DNA from the TWERC-derived clones were found to be hypermethylated. A slightly different situation was seen in clone C where demethylation of the provirus in the 3' region and in the 5' end of the genome was found. The level of mRNA expression both in the parental cells and in the clones correlated with the methylation pattern of the PR-RSV provirus. Clone C was less methylated and expressed more virus-specific RNA. The possible role of BrdU in these events is discussed.

Concentration of retroviruses by low-speed centrifugation.

Concentration of retroviruses from volumes of up to 6 liters of medium by low-speed centrifugation is described. In contrast to pelleting, no damage or aggregation of particles was observed. Surface glycoproteins were also fully preserved. This method enables simple handling of relatively large volumes of medium. Highly purified mouse mammary tumor virus (MMTV) was obtained and its transframe protein p30 in SDS-PAGE was recognized as a double band.

Mucopolysaccharides in gastric cancer.

The histochemical properties of mucus in 48 diffuse type gastric carcinomas and in 46 intestinal type gastric cancers were investigated. In the diffuse type there was a prevalence of intracellular secretion of mucopolysaccharides with a significantly larger portion of sulfated mucopolysaccharides. Acid mucopolysaccharides occurred in the tumor cells only along with neutral ones. In the intestinal type of this cancer, in turn, extracellular secretion prevailed, along with the occurrence of purely acid mucopolysaccharides and a smaller portion of sulfated ones. The secretion of pure neutral mucopolysaccharides in both types of gastric cancer was very low. In the marginal areas of intestinal type carcinomas, incomplete intestinal type IIb metaplasia was frequently found.