Interaction between radiation and drug damage in mammalian cells. II. The effect of actinomycin-D on the repair of the sublethal radiation damage in plateau phase cells

The effect of actinomycin-D (AMD) on radiation damage repair was studied in plateau phase V79 Chinese hamster cells. Sublethal radiation damage repair, as demonstrated by survival fluctuations following two x-ray exposures separted by time, was observed in our plateau phase cells. Plateau phase cells exposed to 0.01-0.04 mug/ml AMD (a nontoxic regimen to 8 hours) between x-ray exposures were less able to repair sublethal damage. If plateau phase cells were plated at low dilutions into fresh medium (conditions for resuming exponential growth) immediately after the first x-ray dose, and exposed to 0.01--0.04 mug/ml AMD until the second dose, inhibition of sublethal damage repair and additional cell killing were observed particularly at 0.04 mug/ml AMD. It is suggested that radiation-drug damage interactions should be studied in plateau phase cells and in cells resuming exponential growth after plateau phase (possibly analogous to "recruitment"), as well as in exponential phase cultures.     

Specific cytotoxicity in vitro of lymphocytes sensitized in culture against tumor cells

The production of tumor-specific cell-mediated cytotoxicity following in vitro sensitization of C57BL spleen cells against a syngeneic 3LL Lewis lung carcinoma was studied. Lymphocytes were sensitized on monolayers of the tumor cells for 4-5 days. The cytotoxicity was assayed by measuring the reduction in 3H-leucine and 3H-thymidine incorporation by target cells after interaction with the sensitized lymphocytes. Spleen lymphocytes sensitized on monolayers of 3LL tumor cells caused a high extent of lysis; such cells tested on C57BL or C3H fibroblast targets evoked only a low level of cytotoxicity. C57BL spleen cells sensitized on C57BL fibroblasts caused a low level of cytotoxicity when tested on a 3LL target. Thus cytotoxicity appeared to be tumor specific. The reduced incorporation into protein and DNA of target tumor cells caused by the sensitized lymphocytes was a measure of cell injury, which was more sensitive than direct cell count or uptake of 51CR. Lymphocytes from syngeneic tumor-bearing mice, tested 13-25 days after tumor inoculation, did not manifest in vitro cytotoxicity. On the contrary, such lymphocytes sometimes appeared to have a promoting effect on the tumor cells.     

Bird breeder's hypersensitivity pneumonitis: progress studies of lung function after cessation of exposure to the provoking antigen

Lung function and immunologic studies have been performed in 9 patients with bird breeder's hypersensitivity pneumonitis at the time of diagnosis and at intervals from 8 to 30 months after they ceased exposure to the provoking antigen. All 9 patients had evidence of small airway obstruction at the time of diagnosis. Complete recovery of lung function occurred in 4 patients. The other 5 all had evidence of interstitial damage; 3 of them had progressive increase in the degree of airway obstruction, and one had progressive loss of elastic recoil. The patients were nonsmokers, and these progressive changes could not be explained by occult exposure to antigen because the tests of immunologic sensitivity to relevant avian antigens became negative. Neither the nature or degree of lung function abnormality nor the form of clinical presentation was related to the development of residual damage. The period of exposure to antigen after symptoms developed and the age of the patient appeared to be the most important factors determining recovery of lung function.     

Healing of deep venous thrombosis: venographic findings in a randomized study comparing streptokinase and heparin

Sequential ascending venographic studies were used to assess the healing of deep venous thrombosis in 50 patients randomly assigned to streptokinase or heparin therapy. Various degrees of thrombolysis and/or recanalization were demonstrated by venograms performed on the fourth and tenth days of treatment. Late follow-up studies (mean, 7 months after treatment) showed three basic patterns of resolution: (1) return to normal, (2) complete recanalization, and (3) incomplete recanalization and/or collateralization. Loss of valves or their function was associated with recanalization. The character, speed, and outcome of healing reflected the nature and extent of thrombosis, prior thrombotic disease in the extremity, and the type and timing of treatment. Streptokinase was highly effective and preferable to heparin in patients with deep vein thrombosis when therapy was begun within 4 days of onset of symptoms. In later stages of acute or recurrent deep vein thrombosis, the effectiveness of both drugs was significantly reduced.     

Gonadoblastoma: histologic, ultrastructural, and histochemical observations in five cases

In this series of gonadoblastomas it appeared that the germ cells were the motivating force underlying either tumor proliferation or regression. The ultrastructural morphology confirmed the presence of undifferentiated gonadal cells with active steroid synthesis by the interstitial cells. The Call-Exner-like bodies which showed extensive calcification were composed of basement membrane material containing a sulfated mucosubstance. A histochemical comparison with the noncalcifying Call-Exner bodies of a granulosa-cell tumor differed only in their glycoprotein content. The calcium deposits were identified as oriented hydroxyapatite crystals by electron diffraction, and it is proposed that the basement membrane material serves as a nucleation site for calcification. High serum testosterone levels were correlated with the presence solely of interstitial cells in one case. The finding of a gonadoblastoma without the presence of a Y chromosome contradicts earlier proposals concerning the requirement of a Y chromosome for germ cell proliferation.     

Direct effect of endotoxin on the gastric mucosal microcirculation and electrical gradient

The effects of intra-arterial infusion of E. coli endotoxin at 1.0 mg. per minute on the gastric total and mucosal blood flows, electrical potential difference, and ionic fluxes across the gastric mucosa were studied in an exteriorized, chambered preparation of canine fundic stomach. Gamma-labelled microsphere technique was used in addition to venous drainage and plasma aminopyrine clearance for the measurement of total and mucosal blood flow, respectively. In spite of normal systemic blood pressure throughout the experiment, E. coli endotoxin infusion caused a significant decrease in total gastric blood flow and in the fractional distribution of flow to the mucosae. There was no significant arteriovenous shunting of microspheres. Significant reduction in potential difference and hydrogen-ion back diffusion also was noted after endotoxin infusion, possibly as a consequence of reduced mucosal blood flow. The results indicate that significant gastric mucosal ischemia can occur and may represent a mechanism in the development of gastric erosions in endotoxemia, even in the absence of systemic hypotension.     

Changes in the cell surface of the dimorphic forms of Candida albicans by treatment with hydrolytic enzymes

The release of acid phosphatase and polysaccharide-peptide complexes by hydrolytic enzymes from the surface of the blastospore and mycelial forms of Candida albicans has been examined in cells from 4 h and 18 h cultures and the results correlated with the appearance of the treated cells in the electron microscope. Treatment with dithiothreitol was necessary for the degradative action of the enzymes to occur. Material released by all the treatments used had a similar qualitative composition, but the proportions of mannan, glucan, peptide and acid phosphatase varied with different treatments and with the type of cell examined. I,3-beta-Glucanase was required for major changes in the cell wall to be effected, but a significant amount of material was released with a chitinase preparation containing some protease activity. Protoplasts were obtained from all types of cell using Cytophaga lytic enzyme L1 which had I,3-beta-glucanase and protease activity, but the purified I,3-beta-glucanase and protease prepared from Streptomyces violaceus cultures required the presence of a chitinase before protoplasts were released. The bonding association between the major components which comprise the cell wall, and the spatial distribution of these macromolecules, varies appreciably between the two dimorphic forms and with the age of the culture.