Trail antigen in Eimeria stiedai sporozoites associated with a thrombospondin-related motif and the entry of cultured cells

In order to examine the antigenic similarity and specificity of the trail antigen of Eimeria stiedai and Etp 100, a microneme protein of Eimeria tenella, monoclonal antibodies to the trail antigen of E. stiedai sporozoites were selected by an indirect immunofluorescent antibody method. The monoclonal antibody of one clone, 3D10, reacted with the anterior portion of non-fixed sporozoites. By immunoblotting, the monoclonal antibody was found to react with a 100 kDa antigen of E. stiedai sporozoites, and a 117 kDa antigen of E. tenella sporozoites and merozoites. It was also found to react with a recombinant protein with thrombospondin-/properdin-like motifs homologous to E. tenella microneme protein Etp 100. The monoclonal antibody significantly inhibited the penetration of E. stiedai sporozoites into cultured rabbit hepatobiliary epithelial cells. These results suggest that E. stiedai sporozoites have a trail antigen, located in the anterior region on the outer surface of the sporozoites, which has an epitope with thrombospondin-/properdin-like motifs similar to E. tenella microneme protein Etp 100. This protein may play an important functional role in the process of penetration of host cells.     

Arthroderma benhamiae infection in a rabbit

The isolate from the rabbit with dermatophytosis which was transmitted to the owners was proved to be Arthroderma benhamiae (-) by mating experiments as well as by chitin synthase 1 (CHSI) gene analysis.     

Identification of Trichoderma SKT-1, a biological control agent against seedborne pathogens of rice

Trichoderma SKT-1 was previously reported as a powerful biological control agent against seedborne pathogens of rice, but the taxonomic disposition of the fungal isolate was not clear. Trichoderma SKT-1 produced irregular pyramidal warts on conidia and had an optimum growth temperature of 30°C. Morphological characteristics and colony growth were identical to those of known species of Trichoderma, including the newly recognized species T. asperellum. The 5.8S rDNA with the internal transcribed spacer (ITS) region (ca. 514 bp) of the fungus was compared with those of known species to determine the phylogenetic placement of the fungus. The length and sequence of the regions from Trichoderma SKT-1 were completely identical to those of an isolate of T. asperellum NRRL 5242 (AJ230669). On the basis of these results, we concluded that Trichoderma SKT-1 was T. asperellum.     

Molecular heterogeneity in clinical isolates of Malassezia pachydermatis from dogs

Molecular investigation of 16 strains, conventionally identified to be Malassezia pachydermatis, isolated from dogs in Japan was carried out by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD band patterns of 13 clinical isolates were identical to that of standard strain of M. pachydermatis (CBS-1879). The other three clinical isolates were different from the standard strain of M. pachydermatis in RAPD patterns, and two of the three isolates were identical. About 620 bp genomic DNA fragments of the CHS2 gene were amplified from the same 16 clinical isolates of M. pachydermatis by polymerase chain reaction (PCR) and sequenced. The phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of the 16 clinical isolates revealed that the 13 strains were genetically very close to the standard strain of M. pachydermatis and the other two isolates were genetically close to the standard strain of M. furfur rather than M. pachydermatis. The remaining one isolate was phylogenetically distinct from all the seven Malassezia species reported so far.     

Ultrastructural Changes in Calvarial Osteoblast Cytoskeleton after Prostaglandin E2 Administration in Rats

Recent studies on the cytoskeleton of osteoblasts have been made mainly using cultured cells. However, the morphology of cultured cells may be altered during subculture. Therefore, cytoskeletal changes of calvarial osteoblasts were investigated in situ by electron microscopy using the detergent perfusion method to preserve cell morphology as well as selectively observing the cytoskeleton in the presence of a high concentration of prostaglandin E2 (PGE2) in the calvarial periphery in rats. Rats were perfused with a mixture of Triton X-100 and glutaraldehyde, then the cytoskeleton was observed by transmission electron microscopy. In osteoblasts of the control group, thick bundles of microfilaments ran parallel to the long axis of the cells immediately below the cell membrane adjacent to the osteoid layer. In contrast, in the osteoblasts of the PGE2-administered group, the external morphology was changed to an asteroid or cubic shape, and thick bundles of microfilaments immediately below the cell membrane adjacent to the osteoid were not observed, although microfilament fibres, with a diameter of 5-6 nm, were observed in the cytoplasm.     

Synthesis and insecticidal activity of benzoheterocyclic analogues of N'-benzoyl-N-(tert-butyl)benzohydrazide: Part 3. Modification of N-tert-butylhydrazine moiety

Nineteen analogues were synthesized by modifying the tert-butylhydrazine moieties of N'-tert-butyl-N'-(3,5-dimethylbenzoyl)-5-methyl-2,3-dihydro-1,4-benzodioxine-6-carbohydrazide and N'-tert-butyl-N'-(3,5-dimethylbenzoyl)-5-methylchromane-6-carbohydrazide (chromafenozide), and the synthesized analogues were evaluated for their insecticidal activity against Spodoptera litura F. While all of the synthesized analogues had insecticidal activity inferior to those of the lead compounds, several of the analogues nonetheless showed high insecticidal activity. Chromafenozide has shown very high selectivity toward lepidopteran species.     

Medical dissolution of struvite nephrolithiasis using amino acid preparation in dogs

A new chemolytic therapy with amino acid preparation was tried for use on struvite renal stones in two dogs, which were admitted to Azabu University Veterinary Teaching Hospital. No other special diet, for the purpose of dissolving the stones, was fed during the treatment. Dissolution of the stones began at week 1, and no stone was detected on radiography at week 4 in both cases. The amino acid preparation thereafter has been administered every 2-3 days to prevent formation of the stones. After 6 months from treatment, complete dissolution was confirmed, and no relapse has been observed. Therefore, it is concluded that chemolytic therapy with amino acid preparation might be useful for the treatment of nephrolithiasis with struvite renal stones in dogs.