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R Kano

Publications and source records attributed to R Kano.

At least 55 records · Page 3Linked to original sources
PubMed2001

[Advances in molecular biology of dermatophytes].

During the 44th meeting of The Japanese Society for Medical Mycology in Nagasaki, 2000, a forum was held entitled Advances in Molecular Biology of Dermatophytes. Based on the subject, target molecules and kind of approach, we selected seven presentations from over 100 of the poster abstracts. Six of them concerned identification and one concerned viability. Summaries of the 7 presentations are given in this article. Of presentations on the identification methods, 5 demonstrated their usefulness: 1) A sequence analysis of ITS 1 region in ribosomal DNA of several Microsporum species showed ITS 1 genospecies Arthroderma otae to be composed of A. otae, M. canis, M. equinum and M. audouinii. 2) RAPD may be useful for identifying isolates which are not clearly identifiable by conventional biological techniques. 3) Sequence analysis of CHS 1 was shown to be a rapid tool for species level identification of M. gypseum. 4) PCR-SSCP analysis was also useful for discrimination of dermatophytes with high reproducibility and sensitivity. 5) Strain identification of A. benhamiae isolates may be possible using RFLP analysis of NTS regions in ribosomal DNA. The other presentation concerning identification pointed out some important problems: RFLP of mitochondrial DNA and ITS sequencing of A. benhamiae showed that the results are sometimes in conflict with those obtained from biological techniques, or in some cases, between other molecular techniques. This implies that our concept of fungal species needs to be re-examined and perhaps amended. The presentation on viability introduced quantitative analysis of mRNA of ACT gene, a new application of a molecular technique. Since the mRNA expresses only in living cells, the method is highly useful as an indicator of fungal viability.

Actins↗
PubMed2001

Identification of Sporothrix schenckii based on sequences of the chitin synthase 1 gene.

Sporothrix schenckii is pathogenic to human and animals. To detect S. schenckii in the tissue, we designed specific oligonucleotide primers based on the chitin synthase gene. Amplification products were selectively obtained from S. schenckii DNA. Polymerase chain reaction analysis with the primer pair S2-R2 was able to detect 10 pg genomic DNA of S. schenckii with ethidium bromide staining. This detection system will be useful as a microbiological tool for the diagnosis of human and animal sporotrichosis.

Base Sequence↗
PubMed2001

Microsporum gypseum isolated from a feline case of dermatophytosis.

The 1- to 2-month-old female cross-breed cat presented with alopecia, erythema and many crusts were present on the tail. Microscopic examination of crusts from the tail disclosed epithelial debris, exudate, mycelium, and arthrospores. Microsporum gypseum which is rarely isolated from cats as a causative agent of dermatophytsis was cultured from the crusts on a 1/10 Sabouraud glucose agar at 27 degrees C for 1 week. The isolate of M. gypseum from the cat was examined by random amplification of polymorphic DNA (RAPD), chitin synthase 1 gene (CHS1) sequence and mating experiments. The RAPD band patterns of the clinical isolate of M. gypseum was identical to those of tester strains of Arthroderma gypseum. Nucleotide sequence analysis of the CHS1 gene fragments from the isolate and a tester strain of A. gypseum showed 100% similarity. The mating experiments on the clinical isolate of M. gypseum completely agreed with the results from RAPD and CHS1 gene sequence. The isolate from the cat was confirmed to be A. gypseum (-) mating type, which was consistent with the result of mycological examination by molecular analyses.

Animals↗
PubMed2000

First isolation of Exophiala dermatitidis from a dog: identification by molecular analysis.

The present study deals with the first isolation of Exophiala dermatitidis from a dog. The dog had a history of multicentric lymphoma for 4 months. On physical examination, 10-15 black or purple subcutaneous nodules were detected on the dorsum of the right neck. Microscopic examination of biopsy specimen from the nodules disclosed lymphocytes, neutrophils and moniliform hyphae. The colony of the clinical isolate was flat with black aerial hyphae and a wet margin after 2 weeks incubation on Sabouraud's dextrose agar at 27 degrees C. The microscopic examination of the clinical isolate revealed that hyphae were brown, septate, and smooth, producing branched or unbranched conidiospores laterally or on the apex. The conidia were one celled, subglobose, elliptical to cylindrical, smooth and hyaline. Nucleotide sequence analysis of the chitin synthase 2 (CHS2) gene fragments from the isolate and a reference strain of E. dermatitidis showed more than 99% similarity.

Animals↗
PubMed2000

In vitro study of neutrophil apoptosis in dogs.

In the present study, to investigate the apoptosis of the polymorphonuclear neutrophil (PMN) from healthy dogs, we carried out TUNEL assay and DNA analysis by electrophoresis on dog PMNs. The TUNEL assay indicated that apoptotic PMNs in dogs were 0.15+/-5% before incubation, 0.3+/-5% at 4h incubation, 1+/-6% at 8h, 9+/-4% at 12h and 28+/-5% at 24h, respectively. The ladder formation was much more clearly observed in DNA from PMNs after 24h incubation at 37 degrees C than that before incubation. The results in this study indicated that healthy dog PMNs undergo apoptosis spontaneously within hours to days, and that the apoptosis of PMNs might be related to the high turnover of these circulating cells in dogs.

Animals↗
PubMed2000

Differences among chitin synthase I gene sequences in Trichophyton rubrum and T. violaceum.

Nucleotide sequences of chitin synthase 1 (CHS1) gene were analysed for the phylogenetic relation between Trichophyton violaceum and T. rubrum, including two isolates of T. raubitschekii and one isolate of T. rubrum var. nigricans. About 620-bp genomic DNA fragments of the CHS1 gene were amplified from these dermatophytes by polymerase chain reaction (PCR) and sequenced. The CHS1 nucleotide sequences of these dermatophytes showed more than 95% similarity between the species. The phylogenetic analysis of their sequences revealed that T. rubrum was genetically distinct from T. violaceum. The specific restriction endonuclease site for HinfI was present in the CHS1 gene sequence of T. rubrum but not in that of T. violaceum. A molecular analysis of CHS1 genes will provide useful information for the identification of these Trichophyton species and the understanding of their evolution.

Animals↗
PubMed2000

A case of black dot ringworm with a review of Japanese cases.

Black dot ringworm (BDR), caused by Trichophyton violaceum var. glabrum (T. glabrum), was observed in a 28-year-old Japanese female who had been treated with prednisolone (22.5 mg/day) for systemic lupus erythematosus. It was successfully treated with oral terbinafine (125 mg/day) for 12 weeks. The causative fungus was identified by molecular analysis as well as morphological and biochemical examination. The chitin synthase 1 (CHS1) gene cleavage pattern of the clinical isolate with restricted enzyme HinfI was identical to that of T. violaceum. We reviewed previous reports of BDR to determine the historical trend of this infection in Japan. Since 1974, 93 Japanese cases have been reported. The age distribution was bi-modal: the higher peak consisted of children (aged 0-15 years), and the lower peak was composed of the elderly (aged 60-75 years). In the elderly group, females were predominant (M:F=1:22, p<0.001). T. violaceum, including T. glabrum, was identified as the most common causative fungus of BDR (75.3%). Sixty percent of cases showed slight erythema. In 8 families, 16 cases were found to be intrafamilial infections. A history of previous steroid treatment was described in about 40%.

Adult↗
PubMed2000

Characterization of the cyclophilin of Trichophyton mentagrophytes.

A genetic approach to cyclophilins in a dermatophyte, Trichophyton mentagrophytes, was carried out. The nucleotide and deduced amino acid sequences of the cyclophilin of T. mentagrophytes shared about 70% sequence similarity with those of Schizosaccharomyces pombe, Saccharomyces cerevisiae and Candida albicans. However, the first 21 amino acid and the C-terminal amino acid regions of 188 to 226 of the T. mentagrophytes cyclophilin were distinct from those of the other fungal cyclophilins. The recombinant glutathione S-transferase (GST)-T. mentagrophytes cyclophilin fusion protein produced by Escherichia coli was purified. The protease digest of the fusion protein had a molecular weight of about 13 kDa and peptidyl-prolyl cis-trans isomerase (PPI) activity. This digest protein from T. mentagrophytes was confirmed to be cyclophilin by proving PPI activity.

Amino Acid Sequence↗
PubMed2000

Detection of Microsporum canis in the skin scrapings and hairs of dogs with dermatophytosis based on sequences of the chitin synthase 1 gene.

In the present study, to confirm Microsporum canis infection rapidly, we detected the chitin synthase gene 1 (CHS1) gene of M. canis in the hair and skin samples of four dogs with dermatophytosis. Amplification of the DNAs in the four samples with CHS1 primers yielded fragments of about 620-bp. Nucleotide sequence analysis of the CHS1 gene fragments from samples and a reference strain of M. canis gene showed more than 99% similarity. The method presented in this study can rapidly detect the DNA of M. canis in skin scrapings, and we anticipate that it will be a useful microbiological tool for the diagnosis of M. canis infections in animals and humans.

Animals↗
PubMed2000

First isolation of Stephanoascus ciferrii from a cat.

The present study deals with the first isolation of Stephanoascus ciferrii from a cat. A 2-year-old female Persian cat weighing 2.25 kg was referred to an animal hospital with a chief complaint of otitis externa of the left ear. Microscopic examination of specimen from the left ear disclosed yeast cells. The colony of the clinical isolate was cream-colored, rough, raised and wrinkled. The microscopic examination of the clinical isolate revealed abundant branched and septated mycelia with small ramified chains of oval blastoconidia, variable in size, and arranged alongside the hyphae. Amplification of the isolate DNA with LSU rDNA primers yielded a fragment of about 570 bp, whose nucleotide sequence of the isolate showed 100% similarity to that of Stephanoascus ciferrii in the GenBank database. Therefore, the isolate was identified as Stephanoascus ciferrii, confirming the result of mycological examination by molecular analysis.

Animals↗
PubMed2000

Molecular analysis of CAP59 gene sequences from five serotypes of Cryptococcus neoformans.

The nucleotide sequences of CAP59 genes from five serotypes of Cryptococcus neoformans were analyzed for their phylogenetic relationships. Approximately 600-bp genomic DNA fragments of the CAP59 gene were amplified from each isolate by PCR and sequenced. The CAP59 nucleotide sequences of C. neoformans showed more than 90% similarity among the five serotypes. By phylogenetic analysis, their sequences were divided into three clusters: serotypes A and AD, serotypes B and C, and serotype D. In addition, the results of reduced amino acid sequences were similar to the nucleotide sequence data. These data revealed that serotype AD was genetically close to serotype A rather than serotype D, although it had been considered to be a mixed type of serotype A and D by serological analysis. Furthermore, the nucleotide sequences of the serotype B and C isolates of C. neoformans were very similar to each other. These results indicated that serotype B and C isolates belonging to C. neoformans var. gattii were genetically homogeneous and closely related. The molecular analysis of the CAP59 gene will provide useful information for the differentiation of serotypes of C. neoformans and for an understanding of their phylogenetic relationships.

Animals↗
PubMed2000

Molecular characteristics of cutaneous papillomavirus from the canine pigmented epidermal nevus.

To investigate the relation between the canine pigmented epidermal nevus (PEN) and cutaneous papillomavirus, we cloned and sequenced the L1 gene of papillomavirus from the canine pigmented epidermal nevus (PEN). Amplification of DNA sample with the L1 consensus primers yielded an expected fragment of approximately 450-bp. The nucleotide sequences of the fragment showed about 64% of sequence similarity to the L1 region of human papillomavirus isolate CP6108 and less than 57% sequence similarity to those of canine oral papillomavirus (COPV). In situ hybridization determined the presence of papillomavirus DNA mainly in the upper stratum granulosum of skin in this case. The results indicated that the canine cutaneous papillomavirus from the PEN lesion was genetically close to human papillomavirus rather than COPV.

Animals↗
PubMed2000

A study on therapeutic teams in psychoanalytical hospital treatment. Diversity and agreement within the team.

The author evaluates the group function of therapeutic teams in psychoanalytical inpatient treatment, by examining the association between perception of factors which affect treatment in 4 groups comprising the therapeutic team: the patients, attending physicians, supervisors, and nurses. The survey was conducted prospectively following the course of hospitalization on 98 patients and staff groups using 8 rating scales. The following findings were obtained: 1) The only significant correlation between all 4 groups was in evaluations on treatment progress upon discharge and one item pertaining to collaboration. 2) Among the many factors examined, the patient having clear therapeutic goals and striving for their attainment, difficulty of treatment, and certain negative emotions were the only items for which significant correlation was seen between all 3 staff groups. 3) No evidence was found of splitting in the patient's view of different staff. These findings indicated that disagreement in perception within the therapeutic team may be a phenomenon peculiar to team therapy. As such, the need for open discussion on disparities in perception, difficulty in treatment, and negative emotions among staff, the sharing of clear treatment goals between patients and staff, and recognition of staff and team meetings as therapeutic means were emphasized.

Affect↗
PubMed1999

Repeated ubiquitin genes in Trichophyton mentagrophytes.

Using Trichophyton mentagrophytes cDNA as a template, we performed PCR amplification with the UB1S and UB1R primers for isolation of a partial fragment of T. mentagrophytes ubiquitin gene. We screened the T. mentagrophytes cDNA library prepared with Uni-ZAP phage vector by hybridization with the T. mentagrophytes ubiquitin fragment, UB1S-UB1R. The nucleotide and deduced amino acid sequences of T. mentagrophytes ubiquitin gene encoded two ubiquitin repeats in 162 bp to 624 bp. The deduced amino acid sequences of ubiquitin from T. mentagrophytes shared about 98% similarity with those of Saccharomyces cerevisiae, Neurospora crassa, and Cryptococcus neoformans.

Amino Acid Sequence↗
PubMed1999

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.

Animals↗
PubMed1999

Phylogenetic analysis of Leptospira strains of pathogenic serovars using 23S rDNA gene sequences.

The 23S ribosomal DNAs were amplified from 11 strains of Leptospira interrogans sensu lato by polymerase chain reaction (PCR) and sequenced. The PCR products of about 290-bp DNA fragments indicated more than 97% sequence similarity to each other. The phylogenetic tree based on the 23S ribosomal DNAs obtained in this study revealed that 11 strains of L. interrogans examined composed a cluster distinct to that of L. weilii and L. borgpetersenii, confirming that these strains were similar to strain Moulton of L. interrogans serovar canicola in 23S rDNA sequence.

Base Sequence↗