Effets de l'actinomycine D sur le développement normal et sur les modifications expérimentales de la morphogénèse de l'oeuf de l'oursinParacentrotus lividus

Summary In the presence of actinomycin D (20–40 µg/ml), the development of the eggs of the sea urchin,Paracentrotus lividus, is slowed from the late morula and stopped at the blastula stage. The development is immediately stopped in the blastula treated with actinomycin D (20–40 µg/ml). The inhibitory effects of actinomycin D are prevented by deoxyribonucleic acid. Actinomycin D does not exert animalizing or vegetalizing effects. However, the enhancing of vegetalizing action of lithium and the weakening of animalizing effects of zinc ions and Evans blue have been observed in the presence of actinomycin D. These observations may reflect some difference in the state of dependence of differentiation of entomesodermic and ectodermic structures towards the nucleus.     

Monoamine oxidase activity of the brain of Locusta migratoria in normal conditions and after intoxication by two insecticides: chlordimeform and dieldrin]

Our investigations have shown up an MAO activity in locust brain, by the use of a radio-isotopic method, as much at larval as at adult stage. This MAO activity is in a sample of 34,3 dpm/mg of brain tissue (wet weight). Investigations have also been undertaken on effects of dieldrin and chlordimeform poisoning on this MAO activity. Even at sublethal dosages, chlordimeform causes a significant inhibition of MAO activity in vivo. This finding is in accordance with Beeman and Matsumura's work in vitro. Moreover, acute poisoning by dieldrin produces more than 60% of inhibition of MAO, 3 hours after administration of 115 microng of this insecticide by injection in the hoemocelian cavity.     

Reversible depression of spontaneous brain electrical activity induced by actinomycin D and 7-aminoactinomycin D in rats

Summary Various types of actinomycin (C, D, S₂, I and V) and 7-amino-analogue of actinomycin D were injected into the right lateral ventricle of the brain through a chronically implanted cannula. In rats but not in mice actinomycin D, actinomycin S₂ and 7-aminoactinomycin D caused depression of EEG, while cardiac and respiratory activity were maintained. This effect of the EEG was reversible and a normal EEG pattern reappeared at least 3 h after administration.     

Effect of actinomycin D on Trypanosoma cruzi

Viable metacyclic forms of T. cruzi, Y strain, treated with an adequate dose of actinomycin D (50 micrograms Act-D/ml/10(7) parasites/ml for 72 h at 28 degrees C) showed the following properties: 1) they lost their ability to replicate in culture medium, in blood and in tissues of normal mice and were no longer able to incorporate tritiated thymidine; 2) they could not penetrate into Vero cells and could not replicate inside normal macrophages; 3) they retained their immunogenicity and the ability to protect mice against a virulent infection; 4) they did not induce histological lesions as described in chronic experimental Chagas' disease.     

Interferon-beta can induce the production of plasminogen activator by cultured human cancer cells

Three cultured human cell lines, renal cancer cells (ACHN), bladder cancer cells (EJ), and fibroblasts transformed in culture by Co-60 gamma rays (KMST-6), when treated with interferon-beta, produced 1.5 to 4 times as much plasminogen activator as the untreated control cultures. This enhanced production of PA was inhibited by cycloheximide or actinomycin D.     

Differential inhibitory effects of noradrenaline upon insulin secretion by the isolated perfused pancreas of the rat

Summary Responses of basal and glucose-induced secretion phases of the isolated perfused pancreas of the rat to graded doses of noradrenaline (NA) are described. Maximal inhibition of basal secretion was achieved at [NA]=40, ng/100 ml; at [NA]=80 ng/100 ml, the 1st phase of glucose induced secretion was abolished; at [NA]=100 ng/100 ml the 2nd phase was abolished. At [NA]40 ng/100 ml basal secretion was less sensitive to NA inhibition than glucose induced output.Acknowledgment. The authors are most grateful to Mr J. Stevenson for advice upon and assistance with statistical analysis of data.     

Binding of actinomycin D and divalent cations to lipopolysaccharides ofAgrobacterium tumefaciens as studied by fluorescence spectroscopy

Summary The binding of actinomycin D and divalent cations to lipopolysaccharides ofA. tumefaciens was studied. Fluorimetric titrations revealed 2 binding sites (low and high affinity sites) for divalent cations, and 1 high-affinity site for actinomycin D.Acknowledgment. This project was financed by the Department of Science and Technology, Govt. of India (grant No. D.O. No.11 (19)/77-SERC). To whom reprint requests should be addressed.     

Effect of actinomycin D onTrypanosoma cruzi

Summary Viable metacyclic forms ofT. cruzi, Y strain, treated with an adequate dose of actinomycin D (50 g Act-D/ml/10⁷ parasites/ml for 72 h at 28° C) showed the following properties: 1) they lost their ability to replicate in culture medium, in blood and in tissues of normal mice and were no longer able to incorporate tritiated thymidine; 2) they could no penetrate into Vero cells and could not replicate inside normal macrophages; 3) they retained their immunogenicity and the ability to protect mice against a virulent infection; 4) they did not induce histological lesions as described in chronic experimental Chagas' disease.     

Behavior and catecholamine levels: comparative study of gregarious insects, solitary insects, and those treated with carbon dioxide in Locusta migratoria (Insecta, Orthoptera)]

The dopamine (DA) and noradrenaline (NA) was measured in Locusta migratoria for 3 groups of individuals showing differences in their motility: gregarious are very active, solitary and animals in chronic treatment by CO2 (1 mn/day) show a very low motility. NA is present in small amounts (0,120-0,250 microng/g) in the 3 groups without significant differences. On the contrary, the quantity of DA is 5 times greater in gregarious than in the 2 other groups (1,78 and 0,31-0,39 microng/g). Thus it is suggested that DA is related to motility and must play its own role of a neurochormone, distinct from that of a metabolic intermediary between DOPA and NA. The question of a relationship between the metabolism of catecholamine and melanization is open.     

Change in the envelope of a murine retrovirus produced in the presence of toyocamycin]

Toyocamycin (TMC), an adenosine analog, impairs qualitatively and quantitatively virus production in a cellular system chronically infected by Friend Virus. Viral particles released by cell cultures treated with 0.2 microgram/ml of the drug have lost most of their glycoprotein (gp 70) content. This phenomenon is likely to modify the viral envelope and could explain the loss of infectivity of the virus.     

On the mechanism of pyrogenic action of ricin

Summary The incubation of rabbit white blood cells with ricin, the toxic protein of castor oil seeds, leads to the production of endogenous pyrogens. This induction can be inhibited by the antibiotics actinomycin D or cycloheximide. The results are discussed in terms of disturbed corticosteroid- and Mg²⁺-levels.Acknowledgments. The author is very grateful to the Uganda National Research Council for the Research Grant No. 60.660.00 (through Makerere University, Kampala) without which the completion of his inaugurated Ph. D. Thesis would not have been possible. This paper represents a part of this Thesis.     

Actinomycin-resistant antiviral activity associated with preparations containing chicken interferon.

Crude and purified preparations containing chicken interferon show a 2fold antiviral activity. One is inhibited by actinomycin and leads to total inhibition if interferon is added to cells before actinomycin, the other is insensitive to actinomycin and leads to partial inhibition if interferon is added simultaneously with or after actinomycin.     

Interferon-β can induce the production of plasminogen activator by cultured human cancer cells

Summary Three cultured human cell lines, renal cancer cells (ACHN), bladder cancer cells (EJ), and fibroblasts transformed in culture by Co-60 gamma rays (KMST-6), when treated with interferon-, produced 1.5 to 4 times as much plasminogen activator as the untreated control cultures. This enhanced production of PA was inhibited by cycloheximide or actinomycin D.     

Hydrogen peroxide and hydroxyl radical involvement in the activation of caspase-3 in chemically induced apoptosis of HL-60 cells

Apoptosis of HL-60 cells induced by actinomycin D, H7, or daunorubicin was shown to involve the activation of caspase-3-like protease, 2 h after the addition of these drugs, based on microassay of enzyme activity by high-performance liquid chromatography. Catalase and a spin trap, N-t-butyl--phenylnitrone, which effectively inhibited the apoptosis induced by these drugs, also inhibited the activation of caspase-3-like protease. These results suggest that hydrogen peroxide and the hydroxyl radical are common mediators of caspase-3 activation caused by these chemicals, with apparently different functional mechanisms. Based on mitochondrial activity determined by oxygen consumption, complexes I, II, and IV were inhibited by actinomycin D. H7 inhibited complexes I and IV, 1 and 1.5 h respectively, after the addition of the drug to HL-60 cells. Daunorubicin inhibited complex IV, 1.5 h after the addition of the drug to HL-60 cells. Inhibition of complex IV by actinomycin D, H7, and daunorubicin were almost fully restored by the addition of cytochrome c. The release to the cytosol of cytochrome c by these drugs was also demonstrated by Western blot analysis. Addition of catalase inhibited the depression of complex IV activity induced by actinomycin D and H7. These observations indicate a direct relationship between hydrogen peroxide and the release of cytochrome c during apoptosis caused by actinomycin D, H7, and daunorubicin. Received 24 November 2000; received after revision 2 January 2001; accepted 30 January 2001     

Production of butyrylcholinesterase by Caco-2 cells: lack of relationship with triglyceride production

Elevated levels of butyrylcholinesterase activity occur under a number of hypertriglyceridemic conditions, including diabetes and obesity. This study examines whether butyrylcholinesterase activity has a direct effect on triglyceride production, using Caco-2 cells, a human intestinal adenocarcinoma cell line. Caco-2 cells were incubated with 500 μM oleate to stimulate triglyceride production, and butyrylcholinesterase activity was measured in the cellular homogenate. Butyrylcholinesterase activity was approximately 3 × 10^-3 mmol/min per milligram protein. Although triglyceride production increased by almost five-fold after 18 h of stimulation with oleate, butyrylcholinesterase activity was not increased. Furthermore, inhibition of butyrylcholinesterase activity using 1 mM tetraisopropylpyrophosphoramide did not significantly affect triglyceride production or secretion. Human insulin (100 μU/ml) increased the production of butyrylcholinesterase without increasing triglyceride production. This demonstrates that stimulation of fatty acid production and butyrylcholinesterase activity occur by independent mechanisms and suggests that their correlation in hyperlipidemic conditions is not due to a direct relationship in production in situ. Received 23 April 2001; received after revision 25 May 2001; accepted 20 June 2001     

Effects of mistletoe (Viscum album L.) extracts on cultured tumor cells

Bacterially fermented mistletoe preparations (BFMP) were tested on rat hepatoma tissue culture (HTC) cells and human leukemia Molt 4 cells. A dose-dependent inhibition of the growth rate of the cells was observed. For both cell lines, cytostatic concentrations, expressed in weight of fresh plant, were 0.5 mg/ml culture medium for oak BFMP and 1 mg/ml for apple tree BFMP. However, the action of the two preparations was markedly different on each cell line. Non-viable HTC cells were not stained by trypan blue while non-viable Molt 4 cells were fully colored by this reagent. A lysis of cellular membranes of HTC cells was observed by electron microscopy. Furthermore, oak BFMP inhibited the growth of virus transformed 3T3-SV40 cells more than that of non-transformed 3T3 cells. In contrast to BFMP, non-fermented extracts and a purified mistletoe lectin showed a greater inhibition of the growth of Molt 4 cells than of HTC cells. Samples withdrawn at different times during fermentation gradually lost their inhibitory effect on the growth of Molt 4 cells while their action on HTC cells increased up to the 4th day of fermentation. These results are discussed in relation to the cytotoxic substances of mistletoe already characterized.     

Murine primitive neuroectodermal tumor: nucleolar isolation and RNA polymerase inhibition.

A nucleolar isolation procedure was developed for the murine primitive neuroectodermal tumor originally induced with methylcholanthrene. This procedure utilizes sonication for breakage of the nuclei at a magnesium concentration which disperses the extranucleolar chromatin. The nucleoli retain RNA synthetic activity which is inhibited to about 90% by actinomycin D and the chemotherapeutic drug adriamycin.     

Aplysianin-A, an antibacterial and antineoplastic glycoprotein in the albumen gland of a sea hare, Aplysia kurodai

Aplysianin-A, an antibacterial and antineoplastic factor in the albumen gland of the sea hare Aplysia kurodai, was isolated. It had a molecular weight of approximately 320 kD and consisted of subunits with a molecular weight of 85 kD. It contained 9.8% neutral sugar. Aplysianin A showed 50% inhibition of Bacillus subtilis growth at a concentration of 4 microgram protein/ml and 50% lysis of murine MM46 tumor cells at 14 ng protein/ml. A partial identity of antigenic specificity of the purified specimen with an antineoplastic factor from Aplysia eggs was observed in immunodiffusion tests.     

Actinomycin-resistant antiviral activity associated with preparations containing chicken interferon

Summary Crude and purified preparations containing chicken interferon show a 2fold antiviral activity. One is inhibited by actinomycin and leads to total inhibition if interferon is added to cells before actinomycin, the other is insensitive to actinomycin and leads to partial inhibition if interferon is added simultaneously with or after actinomycin.Work supported by the Swiss National Foundation, grant 3.399.70.The authors wish to thank Miss E. Browse, Miss U. Kohler and Mr U. Rohrer for excellent technical assistance.