Biomedical subjects

Rui Kano

Publications and source records attributed to Rui Kano.

At least 37 records · Page 2Linked to original sources

PubMed2005

Molecular cloning of canine mucosal addressin cell adhesion molecule-1 cDNA and its expression in normal tissues.

A cDNA encoding canine mucosal addressin cell adhesion molecule-1 (MAdCAM-1) was cloned. The entire open reading frame of canine MAdCAM-1 cDNA comprises 1137 bp, corresponding to 378 amino acid residues. The deduced amino acid sequence of canine MAdCAM-1 was 55.2%, 53.7%, and 52.4% identical to rat, mouse, and human MAdCAM-1, respectively. Canine MAdCAM-1 appeared to contain two immunoglobulin-like domains at the N-terminus, followed by a mucin-like domain and a third immunoglobulin-like domain. The structures of the dog, rat, and mouse proteins are likely similar because all of the cysteine residues in the immunoglobulin-like domains were conserved. Canine MAdCAM-1 mRNA was confirmed to express extremely in the mesenteric lymph node by RT-PCR.

Amino Acid Sequence↗

PubMed2005

VP2 gene of a canine parvovirus isolate from stool of a puppy.

VP2 gene of a canine parvovirus (CPV) isolate from the feces of a puppy which was diagnosed to be CPV infection was analysed. The result indicated that this clinical isolate was phylogenetically close to the isolate of wild-type CPV (strain CPV-T37) prevailing in Taiwan rather than isolates from Japan.

Amino Acid Sequence↗

PubMed2005

Enzyme-linked immunosorbent assay for the detection of canine Leptospira antibodies using recombinant OmpL1 protein.

OmpL1 is a 31-kDa outer membrane protein characterized in 1993 and known to be expressed only in pathogenic Leptospira spp. Recombinant OmpL1 (GST-rOmpL1) was expressed for use as an ELISA antigen for the detection of anti-Leptospira antibodies. In immunoblot analysis, the protein reacted with sera of dogs infected with three different serotypes of Leptospira interrogans, while did not react with sera of dogs both uninfected negative controls and infected with Borrelia burgdorferi, which is closely related to Leptospira spp. Moreover, in ELISA using GST-rOmpL1, the optical density (O.D.) values from the positive controls were very high (1.125 +/- 0.549). In contrast, the O.D. values from clinically healthy dogs and dogs with diseases other than leptospirosis were very low (0.109 +/- 0.046 and 0.089 +/- 0.046, respectively). These data suggest that the detection of anti-Leptospira antibodies by ELISA using the GST-rOmpL1 protein can be applied for diagnosis of canine leptospirosis.

Animals↗

PubMed2005

Antifungal activities of the combination of tacrolimus and itraconazole against Trichophyton mentagrophytes.

Tacrolimus was reported to have no antifungal activity against dermatophytes, but it might show a synergistic fungicidal activity with azoles against dermatophytes as in the case of pathogenic yeasts. Therefore, we examined the antifungal activity of tacrolimus combined with itraconazole (ITZ) against five strains of Trichophyton mentagrophytes by measuring cell growth and performing MIC (minimum inhibitory concentration) testing. The mean colony diameter of T. mentagrophytes on 1/10 diluted Sabouraud dextrose agar (dSDA) with tacrolimus combined with ITZ was significantly shorter than that on dSDA with tacrolimus or ITZ. MICs of tacrolimus, ITZ, and tacrolimus combined with ITZ against T. mentagrophytes were determined, respectively. The results revealed a synergistic activity of tacrolimus in combination with ITZ against T. mentagrophytes.

Antifungal Agents↗

PubMed2005

[Chitin Synthase 2 (CHS2) gene of Malassezia species].

Malassezia species have been recognized as members of the microbiological flora of human and animal skin; they are also considered to play an important role in the pathogenesis of folliculitis, atopic dermatitis and otitis externa. Therefore, the molecular characteristics were investigated to clarify the epidemiology and the pathogenesis of diseases associated with Malassezia species in human and animals. Molecular investigation was made of 105 clinical isolates of M. pachydermatis from dogs and cats by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD analysis and CHS2 gene analysis indicated that clinical isolates of M. pachydermatis were divided into four distinct genetic types (A, B, C and D). Type A was isolated from lesions of atopic dermatitis, flea allergic dermatitis, otitis externa, pyoderma and seborrheic (dermatitidis) in dogs and cats, and might be predominant on this. The phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of standard strains of 11 Malassezia species showed 11 distinct clusters of this species.

Animals↗

PubMed2005

Metalloprotease gene of Arthroderma gypseum.

The full-length cDNA sequence for metalloprotease (MEP) of Arthroderma gypseum (one of the teleomorphs of the Microsporum gypseum complex) was determined by the 5'-rapid amplification of cDNA ends (RACE) and 3'-RACE methods using cDNA as a template. The full-length cDNA sequence of the MEP (2,670 bp) gene was proved to encode 677 amino acids. The amino acid sequence of the A. gypseum MEP gene shared about 89 and 66% sequence similarity with the conserved region of the Microsporum canis MEP gene and Aspergillus fumigatus, respectively. Southern hybridization analysis of genomic DNA with an MEP probe gave many distinct bands in BamHI, EcoRI and HindIII digests of genomic DNA from A. gypseum. Reverse transcriptase-PCR analysis suggested that keratin might stimulate the expression of MEP mRNA in A. gypseum.

Amino Acid Sequence↗

PubMed2004

Expression of apoptosis-related gene mRNAs in feline T-cells infected with feline immunodeficiency virus (FIV).

In the present study, full length of feline bax, bcl-2, bcl-xL and caspase 3 genes were sequenced and the expression of these mRNAs were also investigated in FIV-infected lymphocytes. The full length cDNA sequence of bax (646 bp), bcl-2 (1423 bp), bcl-xL (1163 bp) and caspase 3 genes (1208 bp) contained a single open reading frame of 579 bp coding 193 amino acids, 708 bp coding 236 amino acids, 702 bp coding 234 amino acids and 834 bp coding 278 amino acids, respectively. Number of apoptotic Kumi-1 cells gradually increased after FIV infection and approximately 70% were apoptotic and 30% were viable in the cells infected with FIV after 8-day incubation, though approximately 80% were non-apoptotic and 20% were dead in non-infected cells. The expression of bcl-2 mRNA in lymphocytes of established cell line was increased by FIV. The amounts of mRNAs of bax, caspase 3 and bcl-xL in FIV-infected cells were not different from those in uninfected control cells.

Amino Acid Sequence↗

PubMed2004

First isolation of Cryptococcus magnus from a cat.

A 6-month-old male Japanese domestic cat with otitis externa due to Aspergillus fumigatus was treated with antifungal agents for 25 days and appeared to be cured. Many yeast colonies however developed from the ear canal samples on Sabouraud's dextrose agar at 27 degrees C for 5 days, instead of A. fumigatus. This yeast colony was cream-colored and slim in texture with smooth and highly glossy surface after 5-day incubation on Sabouraud's dextrose agar at 27 degrees C. The isolate was identified as Cryptococcus magnus by mycological analysis and 28S ribosomal analysis.

Animals↗

PubMed2004

Malassezia nana sp. nov., a novel lipid-dependent yeast species isolated from animals.

Five isolates of a novel species of the yeast genus Malassezia were isolated from animals in Japan and Brazil. Phylogenetic trees based on the D1/D2 domains of the large-subunit (26S) rDNA sequences and nucleotide sequences of the internal transcribed spacer 1 region showed that the isolates were conspecific and belonged to the genus Malassezia. They were related closely to Malassezia dermatis and Malassezia sympodialis, but were clearly distinct from these two species and the other six species of Malassezia that have been reported, indicating that they should be classified as a novel species, Malassezia nana sp. nov. Morphologically and physiologically, M. nana resembles M. dermatis and M. sympodialis, but can be distinguished from these species by its inability to use Cremophor EL (Sigma) as the sole lipid source and to hydrolyse aesculin. The type strain of M. nana is NUSV 1003(T) (=CBS 9557(T)=JCM 12085(T)).

Animals↗

PubMed2004

Decreased apoptotic polymorphonuclear leukocyte rate in dogs with pyometra.

Polymorphonuclear neutrophil (PMN) apoptosis was examined in three dogs with pyometra by TUNEL assay in a 24-hr incubation period and compared with that in healthy control dogs (n=5). The incidence of apoptotic PMNs in dogs with pyometra was 26.4 +/- 5% and that in healthy dogs was 54.3 +/- 7%. The results indicated that apoptotic PMN rates in dogs with pyometra were significantly lower than those in control dogs (p<0.05), suggesting the prolongation of PMN survival.

Animals↗

PubMed2004

Severe calcification of mucocutaneous and gastrointestinal tissues induced by high dose administration of vitamin D in a puppy.

A three-month-old male Bull Terrier was referred to the Animal Medical Centre, Nihon University with chief complaints of subacute emesis and lethargy. Severe leukocytosis, high CRP, hypercalcemia and hypochloremia were detected. Moreover, severe calcification of gingival mucosa and abdominal skin, and abnormalities of the skeletal system were discerned. Abdominal X-ray and endoscopic examination revealed ulcer and hemorrhage on the mucosal membrane of the stomach. This might have been due to injections of high dose vitamin D at 3 and 2 weeks ago by another practioner, according to the detailed history of medication. After two months, a gastrointestinal and skin disorder disappeared, although calcification of the stomach membranes remained and abnormality of the skeletal system had worsened. Therefore, vitamin D should be carefully administrated to a puppy.

Animals↗

PubMed2004

Thiazole orange positive platelets in a dog with Evans' syndrome.

We examined transition for the percentage of reticulated platelets (RP%) and platelet count in a canine case of Evans' syndrome. The result demonstrated that measurement of the RP% can be useful in evaluating platelet production in the bone marrow and response to treatment.

Anemia, Hemolytic↗

PubMed2004

Molecular cloning of canine thrombomodulin cDNA and expression in normal tissues.

Thrombomodulin (TM) is a glycoprotein localized mainly on endothelial cell surfaces, and is a major regulator of vascular thromboresistance. The entire open reading frame of canine TM cDNA comprises 1737 bp, encoding 578 amino acid residues. Comparison of the deduced amino acid sequence from canine TM with those of human, mouse, rat, rabbit and bovine (partial) TM sequences revealed 73.1%, 69.1%, 65.8%, 74.3% and 69.5% identity, respectively. Canine TM mRNA expression was confirmed by RT-PCR analysis in lung, liver, spleen, kidney, pancreas and lymph node, and was relatively low in heart, cerebrum, urinary bladder and uterus. The present results provide valuable data for research into canine coagulation disorders.

Amino Acid Sequence↗

PubMed2004

Characterization of canine caspase-3.

The canine caspase-3 gene was cloned and sequenced. The canine caspase-3 cDNA clone was 1473 base pairs in length and encoded 277 amino acids. The predicted amino acid sequence showed 88.4%, 88.0%, 85.9%, 65.9% and 60.6% homology with that of human, pig, mouse, chicken and zebra fish caspase-3, respectively. The caspase-3 mRNA was confirmed to express in skin, lymph node, bone marrow, small intestine and lung from a healthy dog by RT-PCR analysis.

Amino Acid Sequence↗

PubMed2004

Molecular cloning of canine Mcl-1 gene and its expression in tumor cell lines.

The canine Mcl-1 gene was cloned and sequenced. Canine Mcl-1 clone was 2694 base pairs in length and encoded 350 amino acids. The predicted amino acid sequence was 87.7%, 77.1% and 75.7% homologous to predicted human, mouse and rat Mcl-1, respectively. RT-PCR analysis revealed that canine Mcl-1 mRNA was expressed in PBMCs (peripheral blood mononuclear cells), bone marrow cells, MDCK (Madin-Darby canine kidney) and GL-1 (canine B cell leukemia) whereas undetectable in CL-1 (canine T cell lymphoma) cell line.

Amino Acid Sequence↗

PubMed2004

[Cytokine production and dermatophytosis].

The characteristic pathological feature of dermatomycosis is numerous neutrophilic infiltrates within the epidermis. However, the precise mechanism of this infiltration remains unknown. In this study, interleukins 1 beta, 6, and 8, monocyte chemotactic protein-1 (MCP-1), and tumor necrosis factor (TNF)-alpha levels in the medium where keratinocytes were co-cultured with Candida albicans, Malassezia and Trichophyton mentagrophytes, were determined by enzyme-linked immunosorbent assays (ELISAs) in order to estimate the effect of these fungi on the cytokine production from human keratinocytes. The IL-8 level in the supernatants increased with 1 to 14 hours of co-culture in response to live C. albicans, but the other cytokines were undetectable. Furthermore, the mRNA of IL- 8 in keratinocytes was also confirmed to increased. This data suggested that C. albicans directly induce interleukin 8 production from human keratinocytes without activated macrophages. The IL-6, IL-8, and TNF-alpha levels in the culture supernatants increased with 1 to 24 hours of co-culture with keratinocytes and Malassezia species but the MCP-1 level was undetectable. The IL-8 and TNF-alpha levels in the culture supernatants increased with 1 to 24 hours of co-culture with keratinocytes and Trichophyton mentagrophytes but the other cytokine levels were undetectable. The ELISA analysis of cytokine production by human keratinocytes will provide useful information in understanding the pathogenesis of dermatomycosis.

Candidiasis↗

PubMed2004

[Chitin synthase (CHS) gene analysis of dermatophytes].

About 620-bp genomic DNA fragments of CHS1 genes were amplified from 13 species of dermatophytes by polymerase chain reaction (PCR) and sequenced. The phylogenetic analysis of CHS1 gene fragments of these dermatophyte species revealed that 3 genera of Epidermophyton, Microsporum and Trichophyton were genetically different from each other. The molecular analysis of CHS1 genes will provide useful information for the identification of dermatophytes. The species-specific primers were designed from the nucleotide sequences of CHS1 gene in 3 teleomorphs of T. mentagrophytes. Using these primers the PCR analysis identified the clinical isolates of T. mentagrophytes from rabbit as A. benhamiae. By PCR analysis with the dermatophyte specific primer pair, dermatophyte DNA could be diagnosed directly and rapidly in clinical skin samples. The full length of CHS1 and CHS2 genes of Arthroderma benhamiae (one of the teleomorphs of T. mentagrophytes) was sequenced by 5'-RACE and 3'-RACE methods using cDNA as a template. The full length cDNA sequences of CHS1 gene (3158bp) and CHS2 gene (3392bp) were proved to encode 890 and 419 amino acids, respectively. The amino acid sequences of A. benhamiae CHS1 and CHS2 in the conserved regions shared, respectively, about 70% and 80% sequence similarity with those of the other filamentous ascomycetes registered in the data base of the GeneBank. RT-PCR analysis suggested that chitin synthase inhibitors (nikkomycin Z and polyoxin D) might stimulate the expression of CHS1 mRNA in A. benhamiae, and not the expression of CHS2 mRNA.

Arthrodermataceae↗

PubMed2004

Isolation of a promoter region of a secreted metalloprotease gene from Microsporum canis.

MEP3 of Microsporum canis encodes a 43.5 kDa extracellular keratinolytic metalloprotease, which is thought to be one of the virulence-related factors in dermatophytosis. In order to analyze the system underlying the regulation of MEP3 gene expression, the 5'-upstream region was isolated by inverse PCR. The nucleotide sequence of a DNA fragment of about 2.1 kb containing the coding region contains putative transcription factor binding sites and transcriptional initiation points. Further analyses of the regulatory sequence may be useful for understanding the molecular basis of the coordinated expression of the various genes involved in dermatophytosis.

Amino Acid Sequence↗