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Biomedical subjects

Rui Kano

Publications and source records attributed to Rui Kano.

At least 55 records · Page 3Linked to original sources

Full-length cDNA cloning of Toll-like receptor 4 in dogs and cats.

In the present study, full length of canine and feline Toll-like receptor 4 (TLR4) cDNAs were sequenced, and the expression of canine and feline TLR4 mRNAs in dog and cat tissues were investigated. The full-length cDNA of TLR4 of dog and cat was 2709 bp encoding 637 amino acids and 3113 bp encoding 833 amino acids, respectively. The similarity of canine and feline TLR4 were 83.6% at the nucleotide sequence level and 77.6% at the amino acid sequence level. At the amino acid sequence level, canine and feline TLR4 showed sequence similarities of approximately 62-78% with those of Homo sapiens, Mus musculus, Bos taurus and Equus caballus, respectively. Southern hybridization analyses with TLR4 cDNA probes gave one distinct band in BamHI, EcoRI and HindIII digests of genomic DNA from dogs and cats, respectively, indicating the likely presence of a single TLR4 gene in each species. By RT-PCR analysis, mRNA of canine TLR4 was expressed highly in peripheral blood leukocytes (PBL), moderately in spleen, stomach and small intestine, at low levels in liver, with no expression in kidney, large intestine and skin. On the other hand, mRNA of feline TLR4 was expressed highly in lung, bladder and PBL, moderately in kidney, liver, spleen and large intestine and at low levels in pancreas and small intestine.

Amino Acid Sequence↗

Molecular taxonomy of dermatophytes and related fungi by chitin synthase 1 (CHS1) gene sequences.

In the present study, the nucleotide sequences of the CHS1 gene from dermatophytes and related fungi in the genera Chrysosporium, Epidermophyton, Microsporum and Trichophyton were investigated using molecular methods. About 440-bp genomic DNA fragments of the CHS1 gene from 21 species were amplified by polymerase chain reaction (PCR) and sequenced. The CHS1 nucleotide sequences of these fungi showed more than 83% similarity. The molecular taxonomy of the CHS1 gene sequences revealed that Microsporum was genetically distinct from Chrysosporium and Trichophyton, as classified by morphological characteristics.

Arthrodermataceae↗

Expression of RANTES mRNA in skin lesions of feline eosinophilic plaque.

One of the mechanisms of eosinophil infiltration is its induction by chemoattractants such as regulated upon activation, normal T-expressed and secreted (RANTES) which is a cysteine-cysteine chemokine that mediates chemotaxis and activation of eosinophils in humans and mice. Skin lesions of feline eosinophilic plaque are characterized by a predominant infiltration of eosinophils. The mechanism(s) of eosinophilic infiltration in the skin and/or mucosa of cats is unknown. It is possible that RANTES is involved. To investigate the presence of RANTES in the skin of cats with eosinophilic plaques and nonaffected skin, we cloned and sequenced the full-length feline RANTES cDNA gene, in order to determine whether it is present in the skin of cats with eosinophilic plaques and/or if it is present in normal adjacent skin. We were able to document the the expression of RANTES mRNAs in skin with feline eosinophilic plaque as well as in normal cat skin. The full-length cDNA sequence of the RANTES gene (742 bp) contained a single open reading frame of 276 bp encoding a protein of 92 amino acids. The amino acid sequence of feline RANTES shared 67 and 74% sequence identity with that of bovine and mouse RANTES genes, respectively. RT-PCR analysis on RANTES mRNA in the skin of cats with eosinophilic plaque revealed that its expression was higher in the eosinophilic plaque skin lesions than in the normal skin. The result suggested that RANTES might play a role to induce eosinophil infiltration in feline eosinophilic plaque lesions.

Amino Acid Sequence↗

Nocardia africana isolated from a feline mycetoma.

Nocardia africana was isolated from a subcutaneous nodule of a cat. The isolate developed orange, wrinkled colonies. The bacteria were rod shaped to coccoid (1 by 5 microm) and gram positive. Analysis of the 16S ribosomal DNAs of the isolate and a reference strain of N. africana showed 100% homology.

Animals↗

Aspergillus niger pulmonary infection in a dog.

A 6-month-old male golden retriever was presented with fever, bloody-watery diarrhea and mild cough. Parvovirus and Isospora canis infection was confirmed and successfully treated. Two weeks later, the dog had severe cough and mucopurulent nasal discharge. Aspergillus niger was cultured from endotracheal washings on blood agar at 37 degrees C. Treatment with itraconazole for about 10 weeks resolved the clinical signs.

Animals↗

Canine Bcl-xL gene and its expression in tumor cell lines.

The canine Bcl-xL gene was cloned and sequenced. Canine Bcl-xL cDNA clone was 1252 bp in length, and encoded 233 deduced amino acides. The predicted canine Bcl-xL amino acid sequence shared 99.6%, 97.0%, 97.9%, 98.7% and 98.3% homology with that of human, mouse, rat, sheep and pig Bcl-xL, respectively. RT-PCR analysis revealed that canine Bcl-xL mRNA was constitutively expressed in CL-1 (canine lymphoma) and GL-1 (canine B cell leukemia) cell lines.

Amino Acid Sequence↗

Detection of CAP59 gene in 2 feline cases of systemic cryptococcosis.

Two feline cases were diagnosed as systemic cryptococcosis due to Cryptococcus neoformans (teleomorph: Filobasidiella neoformans) by PCR assay with CAP59 gene primers using urine, serum and biopsy samples. The results of molecular analysis were consistent with the mycological findings.

Animals↗

Characterization of a monoclonal antibody against desmoplakin 1 for use in dogs.

Skin biopsy specimens from face, axilla, abdomen and thigh, mucocutaneous tissues from anus and vagina, and oral mucosa from six healthy Beagle dogs were examined for desmoplakin (Dsp) immunoreactivity using immunoblotting and immunohistochemical analysis. With immunoblotting using mouse antihuman Dsp 1 monoclonal antibody (DP2.17), a band was detected at 250 kDa in all the extracts as in normal human skin samples, although no band was detected at 210 kDa, suggesting that monoclonal antibody DP2.17 recognizes canine Dsp 1 but not Dsp 2. Moreover, the desmosome regions of all specimens were stained with DP2.17 using immunohistochemical analysis. From these results, DP2.17, developed for the examination of human skin, might be suitable for the investigation of Dsp-related skin disorders in dogs.

Animals↗

The first isolation of Nocardia veterana from a human mycetoma.

The clinical isolates from biopsy specimen human subcutaneous nodule developed orange-colored and wrinkled colonies on Sabouraud's dextrose agar at 24 C for 2 weeks. The isolates were aerobic and gram-positive. The bacteria were rod-shaped to coccoid and 1 x 5 microm in size. The assimilation tests revealed that the clinical isolate was identical to a reference strain of Nocardia veterana. A nucleotide sequence analysis of the 16S ribosomal DNA from the isolate and a reference strain of N. veterana showed 99.8% similarity. All data are consistent with the conclusion that the isolate in this human case of mycetoma is N. veterana.

Base Sequence↗

Interleukin-8 and tumor necrosis factor alpha production in human epidermal keratinocytes induced by Trichophyton mentagrophytes.

Production of interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) was confirmed by enzyme-linked immunosorbent assay in a medium where human epidermal keratinocytes were cocultured with Trichophyton mentagrophytes for 1 to 12 h. IL-8 and TNF-alpha mRNAs were also detected in the keratinocytes cocultured with T. mentagrophytes.

Enzyme-Linked Immunosorbent Assay↗

First report on Schizophyllum commune from a dog.

This report describes the first isolation of Schizophyllum commune from a granulomatous lesion on the neck of a dog. The biopsy specimen from the lesion disclosed granulomatous inflammation with branching fungal hyphae without clamp connections. The clinical isolate was identified as S. commune by mycological examination and analysis of ribosomal DNA sequences.

Animals↗

Detection and identification of the Candida species by 25S ribosomal DNA analysis in the urine of candidal cystitis.

Candida species in clinical urine samples were identified directly by the newly developed method of PCR analysis on 25S ribosomal DNA (rDNA). Two dogs were referred to the Animal Medical Center, Nihon University School of Veterinary Medicine, Fujisawa, Kanagawa, Japan for the examination of chronic cystitis. Microscopic examination of urine samples from these dogs revealed yeast cells. Urine culture on Sabouraud's dextrose agar at 27 degrees C for 5 days produced white to cream colored colonies. The isolates were identifical to Candida albicans and C. parapsilosis by mycological examination, respectively. The nucleotide sequences of 25S ribosomal DNA from these urine isolates showed 99% similarity to those of a reference strain of Candida albicans or C. parapsilosis. The nucleotide sequences of 25S rDNA obtained directly from urine samples were also identical to C. albicans and C. parapsilosis, respectively. Confirming the results on the isolates cultured from the same urine samples. This PCR analysis method could be available for the direct identification of Candida species in urine samples within 2 days.

Animals↗

Homogeneous cell suspension of Malassezia pachydermatis obtained with an ultrasonic homogenizer.

It is difficult to produce homogeneous cell suspensions of Malassezia pachydermatis, since yeast cells paste up and form many clumps. However, homogeneous fungal suspensions are required for susceptibility examinations and biochemical analyses. Although several types of trials have been carried out using glass homogenizers and many types of agents to obtain homogeneous fungal suspension. They have not yielded good results. We therefore attempted to use an ultrasonic homogenizer to separate clumps of yeast cells into separate individual cells. We succeeded in this fashion in producing homogeneous cell suspensions of M. pachydermatis. These results indicate that an ultrasonic homogenizer can be used to prepare homogeneous fungal suspensions of M. pachydermatis.

Animals↗

Parathyroid hormone-related protein (PTHrP) produced by dog lymphoma cells.

Parathyroid hormone-related protein (PTHrP) was investigated in a canine lymphoma case with hypercalcemia by means of immunoradiomentric assay (IRMA) and molecular analysis. The plasma calcium level of the patient dog was 13.7 mg/dl. The PTHrP concentration examined by IRMA was 6.1 pmol/L in the plasma sample from the dog, but it was undetectable (< 1.1 pmol/L) in plasma samples from 4 lymphoma cases without hypercalcemia or 5 normal dogs. The PTHrP concentration examined in the culture supernatant of the lymphoma cells from this case was 1.3 pmol/L, whereas those of the lymphoma cells from a lymphoma case without hypercalcemia was undetectable. PTHrP mRNA was clearly detected not only in the lymphoma cells from this dog with hypercalcemia but also in lymphoma cells from 4 lymphoma cases without hypercalcemia and 2 canine lymphoma cell lines.

Animals↗