Self-labeling of human polymorphonuclear leucocyte myeloperoxidase with125iodine

In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with¹²⁵Iodine: chloramine T, lactoperoxidase, and an original technique of self labeling based on the ability of the enzyme to oxidize and bind¹²⁵I in the presence of H₂O₂. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 Ci/gmg MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits.     

The phytotoxins ofMycosphaerella fijiensis,the causative agent of Black Sigatoka disease of bananas and plantains

Black Sigatoka is the most costly to control disease of bananas and plantains in the world. Currently, a worldwide search is underway either to find or to produce cultivars that are disease-resistant or-tolerant. Phytotoxins isolated from the pathogen might facilitate the discovery of such cultivars. Several aromatic compounds from liquid cultures ofMycosphaerella fijiensis, the causal agent of Black Sigatoka disease of bananas and plantains, have been isolated. The most abundant and phytotoxic of these compounds is 2,4,8-trihydroxytetralone, which induces necrotic lesions at 5 g/5 l in less than 12 h on sensitive cultivars of bananas. This compound exhibits host-selectivity that mimics that of the pathogen. Other phytotoxins isolated from this fungus, in lesser amounts, were juglone, the novel compound 2-carboxy-3-hydroxycinnamic acid, isoochracinic acid and 4-hydroxyscytalone. Some of the phytotoxins isolated are melanin shunt pathway metabolites, which makes this fungus unique among plant pathogens.