Amplification of bleomycin-induced DNA cleavage by pyrrole triamide.

We investigated the amplification of bleomycin-induced DNA cleavage by synthetic pyrrole triamide (PyPyPy) using 32P-labeled DNA fragments obtained from human genes. Peplomycin, a kind of bleomycins, plus Fe(II) caused DNA cleavage at the 5'-GC-3' and 5'-GT-3' sequences (damaged bases are underlined). The addition of PyPyPy enhanced the cleavage at cytosine and thymine residues 3' to consecutive guanines, particularly at the 5'-GGGGC-3' and 5'-GGGGT-3' sequences. These results suggest that PyPyPy binds to DNA to induce its conformational change, resulting in alteration of the site specificity and amplification of DNA cleavage. The present study on amplifiers of antitumor drugs would show a novel approach to the establishment of more effective chemotherapy.     

Phorbol esters stimulate spermidine/spermine N1-acetyltransferase activity in mitogen-stimulated bovine lymphocytes

Phorbol 12-myristate-13-acetate (PMA) is shown to induce spermidine/spermine N1-acetyltransferase, a rate-limiting enzyme of polyamine biodegradation, in bovine lymphocytes. When PMA and phytohemagglutinin (PHA) were added simultaneously, the enzyme activity was stimulated synergistically. The ability of phorbol esters to stimulate the enzyme activity was consistent with their tumor-promoting ability. Phorbol, which is not a tumor promotor, was incapable of stimulating the enzyme activity. Phorbol diacetate weakly stimulated the activity of the acetylase. Phorbol dibutyrate had a similar stimulatory effect to PMA. These results suggest that the spermidine/spermine N1-acetyltransferase may play an important role in changes in polyamine levels in phorbol ester-treated cells and that the increase in the enzyme activity may have some relationship to the control of cell growth and differentiation by phorbol esters.     

Thymus-independent anti-DNP antibody responses to DNP-casein and DNP-gelatin.

Although studies on the molecular nature of thymus-independent antigens suggested that the polymeric structure with repeated antigenic determinants and slow metabolism are responsible for thymus-independence, we found that anti-DNP antibody responses to DNP-casein and DNP-gelatin were thymus-independent as well as macrophage-independent. These antibody responses were not affected by in vivo treatment with carrageenan or anti-thymocyte serum. In addition, responses of athymic nude mice to both antigens did not show any significant differences when compared with heterologous nu/+ mice. The findings were confirmed by in vitro experiments; non-adherent spleen cells or T cell-depleted spleen cells responded well to both antigens to the same extent as normal spleen cells. Since both casein and gelatin are polyclonal B cell activators and are not presumed to be high polymer or slow-metabolizing substances, we suggest that thymus-independence in many kinds of antibody response should be reconsidered.