The replication of Nodamura virus, type species of the family Nodaviridae, takes place principally into the interfibrillar spaces of the sarcoplasm in close association with mitochondria in the infected muscles of both G. mellonella and suckling mice. The most striking event in infection of G. mellonella muscle cells was the aggregation and shape modification of numerous mitochondria (elongation, interdigitation, and vesiculation) at an early stage of infection. These clusters of mitochondria were cemented by a thick electron-dense material at the periphery of which viral particles undergo maturation process. In a later stage, degenerated and dilated mitochondria showed a clear assembling of virus particles on their outer membrane and occasionally on some inner membranes. These facts and the localization of RNA by RNase-gold technique in the assembling sites of viruses around and inside the mitochondria allowed us to postulate a role for the mitochondria as supports and/or energy suppliers for viral RNA synthesis and translation.

Two murine monoclonal antibodies (MAbs) against AspergUlusfumigatus were produced and characterized. Splenocytes from cell wail-immunized BALB/c mice were fused with SP2/0 myeloma cells. The hybridomas were screened with a cold alkali (CA) extract of mycelium containing protein, mannose, and galactose, and two MAbs of the immunoglobulin M class were purified from ascites fluid. MAbs 1 and 40 were characterized by double immunodiffusion against CA antigen, indirect enzyme immunoassay with mannans of Candida albicans serotypes A or B or Candida tropicalis, indirect immunofluorescence with C. albicansor A. fumigatus-infected tissues, indirect immunofluorescence with smears of other pathogenic fungi, Western blotting (immunoblotting) with the lectin concanavalin A or BS-1 from the seeds of Bandeiraea simplicifolia, and immunoelectron microscopy. MAb 1 did not cross-react with Candida mannan and recognized a periodate-sensitive, pronaseand heat-resistant epitope in CA antigen and three mannose-and galactose-containing components (80, 62, and 49 kilodaltons) of a mycelial homogenate. Immunoelectron microscopy demonstrated binding of MAb 1 to the inner cell wail and intracellular membranes of hyphae and conidia of A. fumigatus. Circulating antigen was detected in experimental invasive aspergillosis by inhibition enzyme immunoassay with MAb 1 and CA antigen. MAb 40 was a nonprecipitating antibody cross-reactive with Candida species, and competition for an epitope located diffusely in the cell wall of A. fumigatus hyphae was demonstrated by incubating MAb 40 with mannan of C. albicans serotype A. These results suggest that MAb 1 recognizes immunodominant oligogalactoside side chains of A. fumigatus galactomannan, while MAb 40 binds to mannopyranosyl side chains common to A. fumigatus galactomannan and C. albicans mannan.

The fungicidal effect of lysozyme on Candida albicans involves ultrastructural modifications previously described (G.

To further define the ultrastructural events associated with the killing of Candida albicans by human neutrophils, four methods were used: (1) the periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) staining of vicinal-glycol-containing complex carbohydrates; (2) the localization of thermostable immunodeterminants of the yeast cell wall, mannans or mannoproteins, using monospecific antibodies and a protein A-gold complex (monAb-gold); (3) the localization of mannose residues with concanavalin A labeled with gold particles (Con A-gold); (4) the localization of chitin oligomers using wheat germ agglutinin and ovomucoid labeled with gold particles (WGA-gold). The mannan-rich cell wall layers were progressively lost as shown by altered PA-TCH-SP reactivity and a diffuse pattern of staining with Con A-gold and monAb-gold. The de novo appearance of conspicuous amounts of glycogen-like particles near the plasmalemma and in the cell wall was interpreted as evidence of a reparative process of the yeast cell wall. Chitin was seemingly unaltered and readily demonstrated by the WGA-gold in the wall remnants of ghost cells.

Fischer 344 rats on a prolonged low-iodine diet developed thyroid nodules which were transplanted subcutaneously into 33 thyroidectomized rats. One graft took and gave a papillary carcinoma. Serially transplanted syngeneically at 3to 4-month intervals, it became autonomous in third generation hosts. With electron microscopy examination it was microfollicular in year 8 and anaplastic, with totally unpolarized epithelial cells, in year 15. Basement membrane present in year I was no longer visible in specimens of years 8 and 15. Tight and gap junctions still present in year 8 were absent in year 15, although a few desmosomes persisted. From years 8 to 15, free ribosomes and microfilaments became more frequent, whereas the number of mitochondria and the development of the Golgi complex declined. male rats were fed a low-iodine diet (modified Remington) for several months according to a method previously described by Axelrad and Leblond. ' The animals developed enlarged thyroids showing hyperpla~ia'-~ of follicular epithelium and eventually nodules. I-' Fragments of these thyroids were transplanted by Mandavia and Nadler6.' under the skin of 33 thyroidectomized rats of the same inbred strain. Only one graft took, but it was successfully transplanted in all second generation animals. These were either normal rats placed on a lowiodine diet for 4 weeks prior to transplantation, or rats thyroidectomized 48 hours prior to transplantation, both groups being kept on a low-iodine diet following inoculation.6 Tumors from nonthyroidectomized rats on the low-iodine diet were successfully transplanted into all third generation hosts whether the recipients From the