Abstract
DL-α-difluoromethylornithine (DFMO) inhibited ornithine decarboxylase (ODC) activity and arrested the growth of Plasmodium falciparum at the early trophozoite stage. The inhibition of ODC activity did not result in the formation of an alternative diamine such as cadaverine. When putrescine or spermidine were added to the parasites grown in culture, the arrest was reversed, and normal schizogony was completed even in the presence of DFMO. Some reversal of the inhibition was achieved with cadaverine at high concentrations, while 1,3-diaminopropane and spermine failed to restore the development. Resumption of growth could be detected when putrescine was added even after 67 h of DFMO treatment. Electron microscopy did not reveal any changes in the morphology of parasites treated for 47 h, while 73 h of treatment with DFMO induced massive accumulation of pigment. Death was observed a few hours later. These results suggest that DFMO acts as a cytostatic rather than as a cytocidal agent. The four carbon diamine restored cell growth while the shorter or the longer homologous compounds showed little activity.
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Alhonen-Hongisto L, JÄnne J (1980) Polyamine depletion induces enhanced synthesis and accumulation of cadaverine in cultured Ehrlich ascites carcinoma cells. Biochem Biophys Res Commun 93:1005–1013
Alhonen-Honigsto L, Seppanen P, HölttÄ E, JÄnne J (1982) Replacement of natural polyamines by cadaverine and its aminopropyl-derivatives in Ehrlich ascites carcinoma cells. Biochem Biophys Res Commun 106:291–297
Assaraf YG, Golenser J, Spira DT, Bachrach U (1984) Polyamine levels and the activity of their biosynthetic enzymes in human erythrocytes infected with the malarial parasite Plasmodium falciparum. Biochem J 222:815–819
Assaraf YG, Abu-Elheiga L, Spira DT, Desser H, Bachrach U (1986a) Effect of polyamine depletion on macromolecular synthesis of the malarial parasite, Plasmodium falciparum cultured in human eryhtrocytes. Biochem J (in press)
Assaraf YG, Golenser J, Spira DT, Bachrach U (1986b) Plasmodium falciparum: Synchronization of cultures with DL-α-difluoromethylornithine, an inhibitor of polyamine biosynthesis. Exp Parasitol 61:229–235
Bacchi CJ, Vergara C, Garofalo J, Lipschik GY, Hutner SH (1979) Synthesis and content of polyamines in bloodstream Trypanosoma brucei. J Protozool 26:484–488
Bachrach U (1973) Function of naturally occurring polyamines. Academic Press, New York pp 1–211
Bachrach U (1975) Cyclic AMP-mediated induction of ornithine decarboxylase of glioma and neuroblastoma cells. Proc Natl Acad Sci USA 72:3087–3091
Bachrach U (1984) Physiological aspects of ornithine decarboxylase. Cell Biochem Funct 2:6–9
Bachrach U, Shtorch A (1984) In: Caldarera CM, Bachrach U (eds) Advances in Polyamines in Biomedical Science. Editrice Press, Bologna, Italy pp 3–10
Bachrach U, Brem S, Werthman SB, Schnur LF, Greenblatt CL (1979) Leishmania spp: cellular levels and synthesis of polyamine during growth cycles. Exp Parasitol 48:457–463
Cohen SS (1971) In: Cohen S (ed) Introduction to the polyamines. Prentice- Hall, Englewood Cliffs, New Jersey pp 1–179
Dion AS, Cohen SS (1972) Polyamine stimulation of nucleic acid synthesis in an uninfected and phage-infected polyamine auxotroph of Escherichia coli K12. Proc Natl Acad Sci USA 69:213–217
Emanuelsson H, Heby O (1978) Inhibition of putrescine synthesis blocks development of the polychete Ophryotrocha labronica at gastrulation. Proc Natl Acad Sci USA 75:1039–1042
Fozard JR, Prakash NJ, Grove J, Shechter PJ, Sjoerdsma A, Koch-Weser J (1980) L-ornithine decarboxylase: an essential role in early mammalian embryogenesis. Science 208:505–508
Fraser D, Mahler HR (1958) Effects of diamines on the protoplast-infecting agent derived from T2 bacteriophage. J Am Chem Soc 80:6456
Heby O (1981) Role of polyamines in the control of cell proliferation and differentiation. Differentiation 19:1–20
Hollingdale MR, McCann PP, Sjoerdsma A (1985) Plasmodium berghei: inhibitors of ornithine decarboxylase block exoerythrocytic schizogony. Exp Parasitol 60:111–117
Igarashi K, Sakamoto I, Goto N, Kashiwagi K, Honma R, Hirose S (1982) Interaction between polyamines and nucleic acids or phospholipids. Arch Biochem Biophys 219:438–443
JÄnne J, Poso H, Raina A (1978) Polyamines in rapid growth and cancer. Biochim Biophys Acta 473:241–293
Jensen JB, Trager W (1977) Plasmodium falciparum in culture: Use of outdated erythrocytes and description of candle jar method. J Parasitol 63:883–886
Jensen JB, Boland MT, Akood M (1982) Induction of crisis forms in cultured Plasmodium falciparum. Am J Trop Med Hyg 27:1274–1276
Langreth SG, Jensen JB, Reese RT, Trager W (1978) Fine structure of human malaria in vitro. J Protozool 25:443–452
McCann PP, Bacchi CJ, Hanson WL, Cain GD, Nathan HC, Hutner SH, Sjoerdsma A (1981) Effect on parasitic protozoa of α-difluoromethylornithine, an inhibitor of ornithine decarboxylase. Adv Polyamine Res 3:97–110
Metcalf BW, Bey P, Danzin C, Jung MJ, Casara P, Veret JP (1978) Catalytic irreversible inhibition of mammalian ornithine decarboxylase (E.C. 4.1.1.17) by substrate and product analogues. J Am Chem Soc 100:2551–2553
Peter HW, Gunther T, Seiler N (1979) Interrelations between polyamines and phospholipids in Escherichia coli. FEMS Lett 5:389–393
Prakash NJ, Schechter PJ, Grove J, Koch-Weser J (1978) Effect of DL-α-difluoromethylornithine an enzyme-activated irreversible inhibitor of ornithine decarboxylase on L1210 leukemia in mice. Cancer Res 38:3059–3062
Russell DH (1981) Ornithine decarboxylase: a key regulatory protein. Med Biol 59:286–295
Schnur LF, Bachrach U, Greenblatt CL, Ben Joseph M (1979) Polyamine synthesis and levels during the growth and replication of Leishmania tropica minor and Leishmania aethiopica. FEBS Lett 106:202–206
Seiler N (1970) Use of the dansyl reaction in biochemical analysis. Methods Biochem Anal 18:259–337
Tabor H (1981) Polyamine biosynthesis in Escherichia coli: construction of polyamine-deficient mutants. Med Biol 59:389–393
Trager W, Jensen JB (1976) Human malaria parasites in continuous culture. Science 193:673–675
Whaun JM, Brown ND (1985) Ornithine decarboxylase inhibition and the malaria-infected cell: a model for polyamine metabolism and growth. J Pharmacol Exp Ther 233:507–511
White E, Hart D, Sanderson BE (1983) Polyamines in Trichomonas vaginalis. Mol Biochem Parasitol 9:309–318
Williams-Ashman HG, Canellakis ZN (1979) Polyamines in mammalian biology and medicine. Perspect Biol Med 22:421–453
Yayon A, Vande-Waa JA, Yayon M, Geary TG, Jensen JB (1985) Stage-dependent effects of chloroquine on Plasmodium falciparum in vitro. J Protozool 30:642–647
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Assaraf, Y.G., Golenser, J., Spira, D.T. et al. Cytostatic effect of DL-α-difluoromethylornithine against Plasmodium falciparum and its reversal by diamines and spermidine. Parasitol Res 73, 313–318 (1987). https://doi.org/10.1007/BF00531084
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DOI: https://doi.org/10.1007/BF00531084