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Biomedical subjects

R Kano

Publications and source records attributed to R Kano.

At least 37 records · Page 2Linked to original sources

Differentiation of Malassezia furfur and Malassezia sympodialis by glycine utilization.

The genus Malassezia has been revised to include six lipophilic species and one nonlipophilic species. These Malassezia species have been investigated to differentiate their morphological and physiological characteristics. However, assimilation of amino acids as a nitrogen source by these species was not well elucidated. In the present study, isolates of Malassezia species were examined with a glycine medium (containing 7-266 mmol glycine, 7.4 mmol KH(2)PO(4), 4.1 mmol MgSO(4)7H(2)O, 29.6 mmol thiamine, 0.5% Tween-80 and 2% agar) and a modified Dixon glycine medium (0.6% peptone, 3.6% malt extract, 2% ox-bile, 1% Tween-40, 0.2% glycerol, 0.2% oleic acid, 7 mmol glycine and 2% agar). All M. furfur isolates developed on the glycine medium, assimilating glycine at concentrations of at least 7 mmol l(-1). However, the other six Malassezia species were unable to grow on the glycine medium. Also, many colonies of M. furfur grew rapidly, within 2-3 days on the modified Dixon glycine medium, although the other six species showed slow and poor development. From these results, it was suggested that M. furfur might be able to utilize glycine as a single nitrogen source, which the other Malassezia species could not. Therefore, glycine medium was recommended for the differentiation of M. furfur from other species of Malassezia.

Animals↗

Chitin synthase 1 gene of Arthroderma benhamiae isolates in Japan.

In the present study, the chitin synthase 1 (CHS1) gene of eight clinical isolates of Arthroderma benhamiae in Japan was investigated. Nucleotide sequence analysis of the CHS1 gene fragments from clinical isolates of A. benhamiae and from standard strains of Americano-European race and African race A. benhamiae indicated more than 90% similarities among these dermatophytes. An especially high degree of similarity was noted in nucleotide sequence CHS1 gene fragments, with more than 99% among eight clinical isolates and standard strains of the Americano-European race of A. benhamiae. The phylogenetic analysis of their sequences revealed that the eight clinical isolates and the standard strains of the Americano-European race of A. benhamiae were included in the same cluster, and that the African race of A. benhamiae formed a cluster that was distinct from the Americano-European race of A. benhamiae, A. simii and A. vanbreuseghemii.

Animals↗

Molecular identification of Trichophyton rubrum isolate from a dog by chitin synthase 1 (CHS1) gene analysis.

A nonsporulating isolate from a dog with dermatophytosis was identified as Trichophyton rubrum by molecular analysis. The nucleotide sequence analysis of the chitin synthase 1 (CHS1) gene from the isolate indicated more than 99% sequence similarity with other human and canine isolates of T rubrum. The molecular typing suggested that isolates of T. rubrum from human and canine sources were genetically identical.

Animals↗

Chitin synthase 1 (Chs1) gene sequences of Microsporum equinum and Trichophyton equinum.

Chitin synthase 1 (Chs1) genes from Microsporum equinum and Trichophyton equinum were compared with those of the other dermatophytes. The Chs1 nucleotide sequences of these dermatophytes from horses showed more than 80% similarity to those of Arthroderma benhamiae, A. fulvum, A. grubyi, A. gypseum, A. incruvatum, A. otae, A. simii, A. vanbreuseghemii, Epidermophyton floccosum, T. mentagrophytes var. interdigitale (T. interdigitale), T. rubrum and T. violaceum. Especially high degree of nucleotide sequence similarity of more than 99% was noted between the Chs1 gene fragments of M. equinum and A. otae, and those of T. equinum, T. interdigitale and A. vanbreuseghemii, respectively. The phylogenetic analysis of their sequences revealed that M. equinum was genetically very close to A. otae and T. equinum to A. vanbreuseghemii. A molecular analysis of Chs1 genes will provide useful information for the genetic relatedness of M. equinum and T. equinum and confirm the value of DNA sequencing in identification of these two dermatophytes.

Animals↗

Molecular epidemiology of Arthroderma benhamiae, an emerging pathogen of dermatophytoses in Japan, by polymorphisms of the non-transcribed spacer region of the ribosomal DNA.

In Japan, several isolates of Arthroderma benhamiae, a teleomorphic member of Trichophyton mentagrophytes complex, which were not found by earlier mating studies, have recently been recovered from human and animal dermatophytoses. In the present study, intraspecies polymorphism of A. benhamiae isolated in Japan was investigated using restriction fragment length polymorphisms (RFLP) of the non-transcribed spacer (NTS) region of the ribosomal DNA (rDNA), a method introduced to detect intraspecies polymorphisms of other dermatophyte species, such as T. rubrum. Based on their restriction profiles, there were five DNA types out of eight strains of A. benhamiae isolated in Japan. None of the five DNA types were found among the registered tester strains of A. benhamiae. Therefore, several different strains of A. benhamiae may have been brought into Japan separately.

Arthrodermataceae↗

Feline ubiquitin fusion protein genes.

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs. The recombinant glutathione S-transferase (GST)-feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum. The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay. These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.

Amino Acid Sequence↗

Case report. First isolation of Trichosporon domesticum from a cat.

The present study deals with the first isolation of Trichosporon domesticum from the urine of a cat with chronic cystitis. A 5-year-old domestic short hair cat weighing 5.6 kg was referred to an animal hospital with the chief complaint of haematuria having persisted for 3 weeks. The isolate was identified as T. domesticum, confirming the result of mycological examination by molecular analysis.

Animals↗

The effects of Malassezia yeasts on cytokine production by human keratinocytes.

Yeasts of Malassezia, members of the microbiologic flora of the skin, cause pityriasis versicolor and have also been implicated in the pathogenesis of other superficial dermatoses; the most important ones are seborrheic dermatitis, folliculitis, and atopic dermatitis. The mechanisms by which the yeasts cause these dermatoseş however, are not yet clear, and there have been no studies on the interaction between fungi and keratinocytes, especially the effects of fungi on the production of cytokines by human keratinocytes. Recently, the genus Malassezia has been expanded to seven species based on molecular data. In this study, we estimated the effects of Malassezia yeasts on cytokine (interleukins 1beta, 6, and 8, monocyte chemotactic protein-1, and tumor necrosis factor-alpha) production by human keratinocytes in order to examine whether the pathogenicity of the respective Malassezia yeasts is different from each other and to elucidate the mechanism by which Malassezia yeasts cause the dermatoses with different clinical and pathologic manifestations. Variable levels of interleukin 6 and 8, and tumor necrosis factor-alpha in the supernatants in response to Malassezia yeasts (except M. furfur) increased from 1 to 24 h co-culture, but the monocyte chemotactic protein-1 was undetectable. Furthermore, cytokine levels in the supernatants were undetectable 1-24 h after the keratinocytes were harvested with only supernatants of Malassezia. These results indicate that Malassezia stimulates cytokine production by keratinocytes, the cytokine production needs the presence of Malassezia, and there are differences in ability to induce cytokine production by human keratinocytes among Malassezia yeasts. These differences may reflect the different inflammatory responses in Malassezia-associated dermatoses, resulting in different clinical and pathologic manifestations.

Cell Survival↗

The genetic diversity of clinical isolates of Malassezia pachydermatis from dogs and cats.

Molecular investigation of 110 clinical isolates of non-lipid-dependent Malassezia pachydermatis from dogs and cats was carried out by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD analysis indicated that the clinical isolates of M. pachydermatis constituted four distinct genetic types (A, B, C and D). Moreover, the results from CHS2 gene analysis completely agreed with those from the RAPD analyses. The clinical isolates of M. pachydermatis were obtained from normal external ears, lesions of atopic dermatitis, flea allergic dermatitis, otitis externa, pyoderma and seborrheic dermatitidis in dogs and cats. Type A consisted of 93 clinical isolates as well as the ex-neotype strain of M. pachydermatis. The isolates of type A M. pachydermatis originated from lesions of all kinds of diseases. They were predominant on dog and cat skin. The other types, B, C, and D were isolated mainly from otitis externa.

Animals↗

Cluster analysis of human and animal pathogenic Microsporum species and their teleomorphic states, Arthroderma species, based on the DNA sequences of nuclear ribosomal internal transcribed spacer 1.

We performed a cluster analysis of human and animal pathogenic Microsporum species and their teleomorphic states, Arthroderma species, including A. otae-related species (M. canis, M. audouinii, M. distortum, M. equinum, M. langeronii, and M. ferrugineum) and M. gypseum complex (A. fulvum, A. gypseum, and A. incurvatum) using DNA sequences of nuclear ribosomal internal transcribed spacer 1 (ITS1). The dendrogram showed the members of A. otae-related species to be monophyletic and to construct an extremely closely related cluster with a long horizontal branch. This ITS1-homologous group of A. otae was organized in 6 unique genotypes, while sequences of the members of the ITS1-homologous group of M. gypseum complex are more diverse. This ITS1-based database of Microsporum species and their teleomorphic states will provide a useful and reliable species identification system: it is time-saving (takes two to three days), accurate and applicable even to strains with atypical morphological features or in a non-culturable state.

Animals↗

Expression of ubiquitin gene in Microsporum canis and Trichophyton mentagrophytes cultured with fluconazole.

The expression of the ubiquitin (Ub) gene in dermatophytes was examined for its relation to resistance against the antifungal drug fluconazole. The nucleotide sequences and the deduced amino acid sequences of the Ub gene in Microsporum canis were proven to be 99% similar to those of the Ub gene in Trichophyton mentagrophytes. Expression of mRNA of Ub in M. canis and T. mentagrophytes was enhanced when the fungi were cultured with fluconazole. The antifungal activity of fluconazole against these dermatophytes was increased in the presence of Ub proteasome inhibitor.

Amino Acid Sequence↗

Molecular identification of Candida parapsilosis from crop mucosa in a cockatiel.

A 2-month-old cockatiel was evaluated for diarrhea, dyspnea, and death. Histologic examination of lesions in the crop mucosa revealed hyperkeratosis and the presence of blastoconidia and hyphae. Positive immunohistochemical staining of the organisms was achieved with an antibody directed against Candida spp. Polymerase chain reaction amplification of DNA from crop lesion material with internal transcribed spacer 2 (ITS2) primers yielded fragments of approximately 300 bp, which demonstrated 95% DNA homology with the corresponding sequence from a strain of Candida parapsilosis deposited in the GenBank data base. The Candida species in the lesion of the crop mucosa was therefore identified by DNA sequence analysis as C. parapsilosis.

Animals↗

Immunohistochemical analysis of cytokeratin expression in dog skin.

The expression of cytokeratins and involucrin was analyzed to identify the skin cells which compose the epidermis of dogs. The distribution of cytokeratins and involucrin in normal dog skin was immunohistochemically examined with 27 commercial monoclonal antibodies for human use. Antibodies, No.4. OV-TL12/13, 35betaH11, 4.1.18, CAM5.2, NCL5D3, Ks.13.1, Ks.18.04, Ks.19.1, 170.2.]4 and Ks.20.8 stained hair follicles and/or the sweat gland duct, but not the epidermis. Antibodies, 34betaB4, AE3, 34betaE12. LP34, RCK102, MNF116, AE1, KLI, DE-K10 and DE-K13 reacted with every layer of the epidermis, hair follicles and the sweat gland duct. These results were similar to those reported in the human skin. No positive staining, however, could be detected in the epidermis, hair follicles and the sweat gland duct with commercial antibodies, 6B10, Ks.7.18, Mu146-uc, E3, RCK108 and involucrin. Therefore, immunohistochemical investigation with these commercial antibodies developed for human skin examination might be available for investigating the origin of skin tumors in dogs.

Animals↗

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.

Animals↗