An invertebrate infection model for evaluating anti-fungal agents against dermatophytosis.

Animal models of pathogenic infection are needed to evaluate candidate compounds for the development of anti-infectious drugs. Dermatophytes are pathogenic fungi that cause several infectious diseases. We established a silkworm dermatophyte infection model to evaluate anti-fungal drugs. Injection of conidia of the dermatophyte Arthroderma vanbreuseghemii into silkworms was lethal. A. vanbreuseghemii conidia germinated in liquid culture were more potent against silkworms than non-germinated conidia. Germinated conidia of other dermatophytes, Arthroderma benhamiae, Trichophyton rubrum, and Microsporum canis, also killed silkworms. Injection of heat-treated germinated A. vanbreuseghemii conidia did not kill silkworms, suggesting that only viable fungi are virulent. Injecting terbinafine or itraconazole, oral drugs used clinically to treat dermatophytosis, into the silkworm midgut had therapeutic effects against infection with germinated A. vanbreuseghemii conidia. When silkworms were injected with A. vanbreuseghemii expressing enhanced green fluorescent protein (eGFP), mycelial growth of the fungus was observed in the fat body and midgut. Injection of terbinafine into the silkworm midgut, which corresponds to oral administration in humans, inhibited the growth of A. vanbreuseghemii expressing eGFP in the fat body. These findings suggest that the silkworm infection model with eGFP-expressing dermatophytes is useful for evaluating the therapeutic activity of orally administered anti-fungal agents against dermatophytes.

[Evaluation of liaison-clinical pathway for patients with breast cancer undergoing adjuvant therapy after curative operation--data from questionnaire survey of 56 clinics and 105 patients].

To evaluate the liaison-clinical pathway for patients with breast cancer introduced since May 2008, the data from a questionnaires survey of 56 clinics and 105 patients were reviewed. Half of the clinics specialized in internal medicine. 93% of physicians recognized the utility of the pathway while 24% made the most of the pathway. About 40% of the clinics wished to enlarge both the patient number and treatment materials. Half of the patients were employed. 55% of patients valued the pathway as helpful. And 29% of patients used the patient booklet at all times. 8% of patients replied they had complaints went to clinics. There has been no serious problem in using the pathway. Countermeasures to electronic health records in clinics, and responses to requests from each patient will be needed.

[Introduction of a liaison-clinical pathway for patients with breast cancer undergoing adjuvant therapy after curative operation].

A liaison-clinical pathway for patients with stage I to III breast cancer undergoing adjuvant oral chemoendocrine therapy after curative operation has been introduced in our hospital since May of 2008. The form of this pathway was developed from the former cooperative treatment system between our hospital and local clinics. One hundred thirty four patients and 69 clinics have been using this pathway for 6 months. Our liaison-clinical pathway consists of a chart for the clinic-doctor, a leaflet with a checklist for the patient, and supplements. The aim of this pathway was care for patients with breast cancer in addition to total health care in a local clinic. Our pathway decreased patient anxiety and facilitated clinic-doctor acceptance for the cooperative cancer treatment system. A coordinator is a key person who constructs and manages this pathway. There has been no trouble in managing the pathway for 6 months. The liaison clinical pathway will be useful to reduce patient burden while maintaining treatment quality.

Isolation, characterization, and disruption of dnr1, the areA/nit-2-like nitrogen regulatory gene of the zoophilic dermatophyte, Microsporum canis.

A homolog of the major nitrogen regulatory genes areA from Aspergillus nidulans and nit-2 from Neurospora crassa was isolated from the zoophilic dermatophyte, Microsporum canis. This gene, dnr1, encodes a polypeptide of 761 amino acid residues containing a single zinc-finger DNA-binding domain, which is almost identical in amino acid sequence to the zinc-finger domains of AREA and NIT-2. The functional equivalence of dnr1 to areA was demonstrated by complementation of an areA loss-of-function mutant of A. nidulans with dnr1 cDNA. To further characterize this gene, dnr1 was disrupted by gene replacement based on homologous recombination. Of 100 transformants analyzed, two showed the results expected for replacement of dnr1. The growth properties of the two dnr1(-) mutant strains on various nitrogen sources were examined. Unlike the A. nidulansareA(-) mutant, these dnr1(-) mutants showed significantly reduced growth on ammonia, a preferred nitrogen source for fungi. These mutant strains were also able to utilize various amino acids for growth. In comparison with wild-type M. canis, the two dnr1(-) mutants showed reduced growth on medium containing keratin as the sole nitrogen source. This is the first report describing successful production of targeted gene-disrupted mutants by homologous recombination and their phenotypic analysis in dermatophytes.