Biomedical subjects

A Kuwano

Publications and source records attributed to A Kuwano.

At least 19 recordsLinked to original sources

PubMed2005

Quantities and types of ceramides and their relationships to physical properties of the horn covering the claws of clinically normal cows and cows with subclinical laminitis.

Quantities and types of ceramides and their relationships to physical properties of the horn covering the claws of clinically normal cows and cows with subclinical laminitis were investigated. Total ceramide content of the horn covering the sole and wall from cows with subclinical laminitis was 872.2 +/- 146.6 microg/g and 528.6 +/- 61.3 microg/g, respectively, and was significantly (P < 0.01, 0.05) lower than that from clinically normal cows. The mean moisture content in the claws from cows with subclinical laminitis (43.5% +/- 4.3%) was significantly (P < 0.05) higher than that in the claws from clinically normal cows. The hardness of claws from cows with subclinical laminitis (35.2 +/- 3.5) was significantly (P < 0.05) less than that of claws from clinically normal cows. Significant correlations between ceramides and moisture content (P < 0.001) and between ceramide and hardness (P < 0.001) were found in clinically normal cows and cows with subclinical laminitis. Our results indicate that decreases in ceramide contents may be related to changes in physical properties of the horn covering the claw in cows with subclinical laminitis.

Animals↗

PubMed2004

Prevalence of superficial digital flexor tendonitis and suspensory desmitis in Japanese Thoroughbred flat racehorses in 1999.

REASONS FOR PERFORMING STUDY: Overstrain injuries to the superficial digital flexor tendon (SDFT) and suspensory ligament (SI) are among the most common musculoskeletal injuries which contribute to the considerable wastage of racing Thoroughbreds. Many epidemiological studies have demonstrated the prevalence of and risk factors for tendon injury when racing but have not included those injuries sustained during training. However, since tendon injury during training is seen commonly in clinical practice, it is appropriate to determine the overall prevalence of tendon injury sustained during both training and racing. OBJECTIVE: To determine the prevalence of overstrain injury to the SDFT and SL during training and racing among Thoroughbred flat racehorses in Japan in 1999. METHODS: A retrospective study was performed using a sample population of 10,262 Thoroughbred racehorses. The medical information database of Thoroughbred racehorses registered by the Japan Racing Association (JRA) in 1999 was analysed for SDFT and SL overstrain injury diagnosed by a veterinarian employed by JRA during training and racing. Jump racehorses were excluded from this study. RESULTS: The prevalence of forelimb SDFT tendonitis and SL desmitis was 11.1% (1130 cases) and 3.61% (370 cases) of the population, respectively. In the hindlimb, there were 0.06% (6 cases) and 0.14% (14 cases), respectively. Risks of SDF tendonitis in the forelimb in 3-year-olds or older horses were significantly higher than in 2-year-olds. In contrast, the risk of SL desmitis in the forelimb at age 3 and 4 years was 2.23 and 2.11 times higher, respectively, than in 2-year-olds, but this increased to 5.07 times in those age > or = 5 years. Entire males were at greater risk in comparison to females and geldings. CONCLUSIONS: The results suggest that the prevalence of SDF tendonitis and SL desmitis in the forelimb was associated with the horse's age and sex. The prevalence of SL desmitis increased further with age compared with SDF tendonitis, possibly reflecting a more rapid accumulation of degeneration in this structure. POTENTIAL RELEVANCE: The age-related risk demonstrated in this study provides further support that overstrain injuries are associated with accumulated degeneration. These data provide a valuable resource for further research into the aetiology of tendon injury in the racehorse.

Age Factors↗

PubMed2004

Effects of biotin supplementation on serum biotin levels and physical properties of samples of solar horn of Holstein cows.

Effects of dietary biotin supplementation on serum biotin levels and physical properties of sole horn of 40 Holstein cows were evaluated. The mean serum biotin level in biotin-supplemented cows after 10 mo of biotin supplementation (1163.2 +/- 76.2 pg/mL) was significantly higher (P = 0.007) than that in control cows (382.0 +/- 76.2 pg/mL). The sole horn of biotin-supplemented cows was significantly harder (P = 0.026) and had a significantly lower moisture content (P = 0.021) than that of control cows. No morphologic differences in horn tubules or intertubular horn were found between the biotin-supplemented and control cows. The total lipid content of sole horn was significantly higher (P = 0.030) in the biotin-supplemented cows than in the control cows. These results suggest that dietary biotin supplementation causes increases in serum biotin levels and changes in physical properties and fat content of sole horn.

Animal Nutritional Physiological Phenomena↗

PubMed2003

Physiological changes in the concentrations of biotin in the serum and milk and in the physical properties of the claw horn in Holstein cows.

Physiological changes in the concentrations of biotin in the serum and milk and in the physical properties of the claw horn were examined in Holstein cows. A lower concentration of biotin in the serum and a higher concentration of biotin in milk were found during early and late lactation and during the dry period, and a significant (p<0.05) inverse correlation was found between serum and milk biotin concentrations. A high moisture content and a low level of hardness of the claw horn were found during mid-lactation. Our results indicate that change in the serum biotin concentration probably results from the loss of biotin in the milk of cows during each stage of lactation and also confirm that the moisture content and hardness of the claw horn undergo physiological changes.

Animals↗

PubMed2001

Production of biologically active recombinant bovine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae.

The full-length bovine interferon-gamma (bIFN-gamma) cDNA, including the secretion signal peptide coding region was recloned into baculovirus transfer vectors pAcYM1 and pBm050. These vectors were co-transfected with Autographa californica nuclear polyhedrosis virus (AcNPV) or Bombyx mori nuclear polyhedrosis virus (BmNPV) DNA into Spodoptera frugiperda cells (SF21AE) and Bombyx mori cells (BmN), respectively. The recombinant viruses, named AcBIFN-gamma and BmBIFN-gamma, were then recovered. Recombinant bIFN-gamma (rbIFN-gamma) was accumulated in the culture fluid of AcBIFN-gamma-infected Trichoplusia ni cells and BmBIFN-gamma-infected silkworm larvae. These rbIFN-gamma forms were shown to be glycosylated 20 and 22 kDa proteins as confirmed by SDS-PAGE and tunicamycin treatment. These products were sensitive to cystein proteinase. Both rbIFN-gamma proteins, showed high-level biological activities by plaque reduction assay using vesicular stomatitis virus, and MHC class II antigen induction on bovine macrophage cells.

Animals↗

PubMed2001

Serological diagnosis of equine influenza using the hemagglutinin protein produced in a baculovirus expression system.

The hemagglutinin (HA) protein of an equine influenza strain, A/equine/La Plata/1/93 (LP/93), was produced using a baculovirus expression system. Silkworm larvae inoculated with recombinant baculovirus expressed high quantities of the HA protein which was then purified to greater than 95% purity by fetuin-affinity chromatography. Purified HA protein was used subsequently in an ELISA for detection of antibodies in horse sera. Two hundred serum samples from vaccinated racehorses were reacted on ELISA plates coated with 40.0 ng/ml of purified HA protein. Subsequent optical density (OD) levels revealed titers which correlated highly with respective hemagglutinin inhibition (HI) antibody titers which ranged from <1:8 to 1:256 (correlation coefficient among them was 0.850). ELISA OD levels and HI titers increased at 5 and 7 days post-inoculation, respectively, in a horse inoculated intranasally with LP/93. Respective antibody levels were observed to change in an essentially parallel manner during a period of 1 month. Similarly, ELISA OD levels correlated with HI titers in horses during a period of 6 weeks following intramuscular inoculation with inactivated single-strain vaccines containing LP/93, A/equine/Kentucky/1/81 (H3N8) or A/equine/Rome/5/91 (H3N8). A similar pattern was also observed in eight horses throughout a 10-week period following inoculation with a commercially available inactivated trivalent vaccine containing A/equine/Newmarket/1/77(H7N7), A/equine/Kentucky/81 and LP/93. From these results, it is suggested that this ELISA system could be used for disease diagnosis and surveillance of HI antibody titers among vaccinated horses.

Animals↗

PubMed2001

Histoplasmosis in the lung of a race horse with yersiniosis.

A 4-year-old female thoroughbred race horse died of acute peritonitis caused by necrotizing granulomatous duodenitis. Yersinia enterocolitica was immunohistochemically demonstrated in macrophages in granulomas developed in the duodenum, lung, liver and abdominal lymph nodes. The yeast-like fungi were found in the cytoplasmic vacuoles of macrophages in the lung that infiltrated into the granulomas and surrounding alveoli with congestive edema. The yeast-like fungi were positively stained by Gomori-Grocott chromic acid methenamine silver stain and immuno-histochemically stained with anti-histoplasma antibody. In this case, it was considered that granulomas formed in the duodenum, lung, liver and abdominal lymph nodes were primarily caused by Yersinia enterocolitica due to idiopathic weakening of the immune system. Yeast-like fungi immunohistochemically identified as histoplasmas secondarily infected the lung. This is the first case regarded as equine histoplasmosis capsulati in Japan.

Animals↗

PubMed2000

Histological and morphometrical studies on the mucosa of the equine guttural pouch (auditory tube diverticulum).

The present study attempted to clarify the characteristics of the guttural pouch mucosa in equines and to evaluate its foreign substance clearance ability. The specimens were collected from nine regions (eight in the guttural pouch mucosa, and one in the nasopharynx mucosa). We first examined the pouch mucosa by light and electron microscopy. We then measured the frequency of goblet cells per 200 epithelial cells, the length of the cilia, the thickness of the epithelial cell layer and lamina propria and statistically analyzed the data. The guttural pouch mucosa consisted of stratified columnar epithelia with brush-like cilia, and there were almost no histological differences between adults and foals. The morphometrical study revealed significant differences in goblet cell frequency (p < 0.001) and the thickness of lamina propria (p < 0.05). By contrast, no statistically significant difference was detected in the length of the cilia or the thickness of the epithelial cell layer. These findings suggest that the guttural pouch mucosa provides foreign substance clearance ability, but that its ability varies among different regions of the epithelium.

Age Factors↗

PubMed1999

A survey of white line disease in Japanese racehorses.

A survey was carried out into white line disease in 1781 Thoroughbred racehorses kept in stables at the Japan Racing Association (JRA) Miho Training Center (MTC) September-October 1996. The survey was conducted while horses were being shod by farriers. The horses that still exhibited damaged white lines after regular trimming were diagnosed as having white line disease. The factors recorded were age, sex, number of diseased horses, number of diseased hooves, number of lesions by region over the bearing border of the hoof and the classified length of such lesions. The percentage of total diseased horses was 11.5% (204 animals), with incidence increasing significantly with age (P< or =0.01). Occurrence was independent of sex (P>0.05) was more frequent in the fore- than in the hindhoof and developed more frequently at the toe than at any other region of the forehoof-bearing border. Most lesions ranged from 20 to 30 mm in length.

Animals↗

PubMed1998

Translocation t(X;21)(q13.3; p11.1) in a girl with Menkes disease.

A girl with a 46,X,t(X;21) (q13.3;p11.1) karyotype presented with skin redundancy, especially in the neck, prominent occiput and micrognathia, and later developed hypotonia, hypopigmentation, sparse scalp hair, and profound mental retardation characteristic of Menkes disease. Her serum copper (14 microg/dl) and ceruloplasmin (9 mg/dl) levels were extremely low. Fluorescent in situ hybridization analysis with a 100-kb P1-derived artificial chromosome probe containing the Menkes disease gene demonstrated three twin-signals, one on the normal X chromosome and one each on derivative chromosomes X and 21, indicating that the Xq13.3 breakpoint was located within the gene. Replication pattern analysis showed that the normal X chromosome was late replicating, whereas the derivative X chromosome was selectively early replicating. These results indicated that Menkes disease in our patient resulted from a de novo translocation that disrupts the disease gene.

Adenosine Triphosphatases↗

PubMed1998

Human ubiquitin-protein ligase Nedd4: expression, subcellular localization and selective interaction with ubiquitin-conjugating enzymes.

BACKGROUND: Nedd4 is a ubiquitin-protein ligase containing a calcium/lipid-binding domain, multiple WW domains and a C-terminal Hect domain, which is required for both the ubiquitin transfer and the association with E2 ubiquitin-conjugating enzymes. Nedd4 has been reported to be involved in the selective ubiquitination of some regulatory proteins in transcription and membrane transport. RESULTS: Three mRNA species for human Nedd4 were found to be 6.4-, 7.8- and 9.5-kb in size, and their expression patterns varied among normal tissues and cancer cell lines, indicating the tissue- and cell-specificities of Nedd4 expression. The Nedd4 protein, approximately 120 kDa in weight, was found in the cytoplasm, mainly in the perinuclear region and cytoplasmic periphery, of human cultured cells. Neural differentiation induced not only the down-regulation of Nedd4 but also the localization of the protein to both the cytoplasm and neurites. To identify the ubiquitination pathway that is linked to Nedd4, we demonstrated that specific E2 enzymes, including human Ubc4, UbcH5B, UbcH5C, UbcH6 and UbcH7, could transfer ubiquitin molecules to Nedd4 at the active cysteine residue, whereas E6AP accepted ubiquitins from Ubc4, UbcH5B, UbcH5C and UbcH7. Furthermore, nuclear localization of N-terminal deletion mutant Nedd4 enabled us to investigate the interaction between Nedd4 and E2 enzyme (Ubc4 or UbcH7) in the cell. The simultaneous expression of the full-length Nedd4 and E2 enzyme revealed the both proteins mostly colocalized in the cytoplasmic periphery, while the N-terminal deleted Nedd4 induced the nuclear and perinuclear colocalization with E2 enzyme. CONCLUSION: Our findings suggested that Nedd4 plays an important role in the cell regulation, including neural differentiation through cooperation with specific E2 ubiquitination pathways.

Cell Differentiation↗

PubMed1998

Onychomycosis in white line disease in horses: pathology, mycology and clinical features.

This paper describes onychomycosis in horses and reports the pathological findings, associated fungi and incidence of concurrent white line disease. In addition to these observations, relevance between post mortem and clinical findings of onychomycosis are discussed in 3 necropsied horses. Samples were collected from 100 hooves from a total of 51 Thoroughbreds suffering from white line disease. Of these, 15 hooves from 13 horses were also complicated with severe hoof wall fissure formation. Preparations from the same samples were used both for histopathology and for culture to identify the associated fungi. Onychomycosis was diagnosed when it could be confirmed histologically. Fungal identification was based on morphological characteristics in culture and the associated fungi were determined by comparison with culture morphology. As a result, 10 samples collected from horses were diagnosed as suffering from onychomycosis. Seven of these showed complicating severe inner hoof wall fissures and the soil fungus Scedosporium apiospermum or the teleomorph of this fungus, i.e., Pseudallescheria boydii, was isolated. The fungus was found to be ubiquitous in the fissure cavities, the terminal horn of the white line and the terminal horn-like laminae of the metaplastic white line-like tissue. It can be concluded that onychomycosis frequently causes white line disease and/or makes it worse. Associated with deterioration of the submural condition, the main associated fungus for onychomycosis in this series, was Genus Scedosporium and the most susceptible region was the terminal horn of the hypertrophied white line and/or the terminal horn-like laminae of the metaplastic white line-like tissue.

Animals↗

PubMed1997

Cloning and characterization of NE-dlg: a novel human homolog of the Drosophila discs large (dlg) tumor suppressor protein interacts with the APC protein.

We have cloned a cDNA for a novel human homolog of the Drosophila discs large (dig) tumor suppressor protein, termed NE-dlg (neuronal and endocrine dig). Northern blot analysis revealed that the gene is highly expressed in neuronal and endocrine tissues. Fluorescence in situ hybridization (FISH) and radiation hybrid mapping studies localized the NE-dlg gene to chromosome Xq13. We also found that the NE-dlg gene encoded a 100 kDa protein. Immunolocalization studies using an NE-dlg antibody showed that the protein tended to be expressed in non-proliferating cells, such as neurons, cells in Langerhans islets of the pancreas, myocytes of the heart muscles, and the prickle and functional layer cells of the esophageal epithelium. Proliferative cells, including various cultured cancer cell lines and basal cells in the esophageal epithelium, showed little expression of the NE-dlg protein. In addition, yeast two-hybrid screening and in vitro binding assays revealed that the NE-dlg interacted with the carboxyl-terminal region of the APC tumor suppressor protein. These data suggest that NE-dlg negatively regulates cell proliferation through its interaction with the APC protein.

Adaptor Proteins, Signal Transducing↗

PubMed1997

Effect of transportation on the composition of bronchoalveolar lavage fluid obtained from horses.

OBJECTIVE: To study the effects of extended transportation on the composition of bronchoalveolar lavage fluid (BALF) obtained from horses. ANIMALS: 30 horses (14 males, 16 females: 25 Thoroughbreds and 5 Thoroughbred-Arabian cross-breds; 27 to 30 months old) without a history or clinical signs of respiratory tract disease. Bronchoalveolar lavage was performed on nontransported control horses (groups 1 and 2) and transported horses (group 3). PROCEDURE: 20 horses were used to determine the effect of 41 hours of transportation on the composition of BALF (group 3). Bronchoalveolar lavage fluid was analyzed for recovered volume, number and distribution of nucleated cells, total protein and phospholipid concentrations, and phospholipid composition. RESULTS: Total number of nucleated cells in BALF from group-3 horses increased by approximately four-fold after transportation. Total protein concentration in BALF from group-3 horses also increased by approximately fivefold after transportation. Total phosphorus concentrations in group-3 horses decreased significantly from time 0 to immediately after transportation. In group-3 horses, the most characteristic change in composition of BALF after transport was a significant decrease in the concentration of phosphatidylglycerol. CONCLUSION AND CLINICAL RELEVANCE: The decrease in phosphatidylglycerol concentration in BALF after transportation indicates a reduction in the quantity of surfactant. This change may reflect either a decreased production of surfactant by alveolar type II epithelial cells or an increased removal of surfactant from the alveolar region. It is likely that extended transportation resulted in a decreased concentration of surfactant in BALF. Such a decrease may reduce the pulmonary defence mechanisms in the alveolar region, possibly resulting in infection.

Aging↗

PubMed1996

Mosaicism for del(17)(p11.2p11.2) underlying the Smith-Magenis syndrome.

Smith-Magenis syndrome (SMS) is a multiple congenital anomalies/mental retardation syndrome associated with deletion of band p11.2 of chromosome 17. The deletion is typically detected by high-resolution cytogenetic analysis of chromosomes from peripheral lymphocytes. Fluorescence in situ hybridization (FISH) has been previously used to rule out apparent mosaicism for del(17)(p11.2p11.2) indicated by routine cytogenetics. We now report mosaicism for del(17)(p11.2p11.2) in a child with SMS. The mosaicism had gone undetected during previous routine cytogenetic analysis. FISH analysis of peripheral lymphocytes as well as immortalized lymphoblasts using markers from 17p11.2 revealed that approximately 60% of cells carried the deletion. To our knowledge, this is the first case of SMS associated with mosaicism for del(17)(p11.2p11.2).

Abnormalities, Multiple↗

PubMed1996

Precise chromosomal locations of the genes for dentatorubral-pallidoluysian atrophy (DRPLA), von Willebrand factor (F8vWF) and parathyroid hormone-like hormone (PTHLH) in human chromosome 12p by deletion mapping.

The precise chromosomal localization of the gene for dentatorubral-pallidoluysian atrophy (DRPLA) was detected by deletion mapping. Segregation patterns of genotypes of polymerase chain reaction products of DRPLA, von Willebrand factor (F8vWF), antigen CD4(p55) (CD4) and parathyroid hormone-like hormone (PTHLH) loci were studied in patients with del(12)(p13.3p13.3), del(12)(p12.3-p11.2), del(12)(p12.1-p11.2), del(12) (p11.2p11.2) and their parents. The gene for DRPLA was assigned to p13.1-p12.3 of chromosome 12. In addition, genes for F8vWF and PTHLH were mapped to p13.2 and p11.2 of chromosome 12, respectively.

Alleles↗

PubMed1996

A human pancreatic islet inwardly rectifying potassium channel: cDNA cloning, determination of the genomic structure and genetic variations in Japanese NIDDM patients.

Ligand gated potassium channels, such as the ATP-regulated potassium channel, play crucial roles in coupling of stimuli to insulin secretion in pancreatic beta cells. Mutations in the genes might lead to the insulin secretory defects observed in patients with non-insulin-dependent diabetes mellitus (NIDDM). We isolated a cDNA encoding a putative subunit of a ligand gated potassium channel from a human islet cDNA library. The channel, which we designated hiGIRK2, appeared to be an alternative spliced variant and a human homologue of recently reported mbGIRK2, KATP-2/BIR1. Transcripts were detected in human brain and pancreas, but not in other tissues including cardiac muscle. The sizes of transcripts in the pancreas differed from those in the brain, suggesting tissue-specific alternative splicing and possible isoforms. We then isolated human genomic clones, determined the complete genomic structure and localized the gene to chromosome 21 (21q22). The gene was comprised of four exons and the protein was encoded by three exons. The entire coding region of the hiGIRK2 gene was scanned by polymerase chain reaction-single strand conformation polymorphism analysis in 80 Japanese NIDDM patients. We found five nucleotide substitutions; three were silent mutations of the third base of codons, one in the first intron, 9 bases upstream of exon 2, and one in the 3'-untranslated region. We conclude that mutations in the gene encoding hiGIRK2, a (subunit of) ligand gated potassium channel, is not a major determinant of the susceptibility to NIDDM in Japanese.

Alternative Splicing↗

PubMed1996

Evidence that poor metabolizers of (S)-mephenytoin could be identified by haplotypes of CYP2C19 in Japanese.

(S)-Mephenytoin is metabolized by CYP2C19. The purpose of this study was to examine availability of phenotyping of poor metabolizers (PMs) of (S)-mephenytoin by polymerase chain reaction (PCR)/restriction enzyme genotyping of CYP2C19 in a Japanese population. We genotyped 217 unrelated healthy Japanese for functionally defective alleles, CYP2C19m1 and CYP2C19m2. The frequencies of the wild type(wm1) and CYP2C19m1 were 0.726 and 0.274, and the wild type(wm2) and CYP2C19m2 were 0.892 and 0.108 respectively. Although the observed numbers of three genotypes were very similar to those estimated according to the Hardy-Weinberg equilibrium for each defect, CYP2C19m2 was not detected in m1 homozygotes, and CYP2C19m1 was not detected in m2 homozygotes. Two defects were inherited separately in four families indicating CYP2C19m1 and m2 segregate independently at the same gene locus. Based on these data, we calculated the haplotype frequencies of wm1-wm2, CYP2C19m1-wm2 and wm1-CYP2C19m2 to be 0.618, 0.274 and 0.108 respectively. Frequencies of homozygotes for CYP2C19m1 and CYP2C19m2 and compound heterozygotes associated with the PM phenotype, were calculated to be 7.5, 1.2 and 5.9% respectively. In total, 14.6% of Japanese are estimated to be PMs. No significant difference was observed between the frequencies of PMs calculated from our results and that identified by urinary S/R ratio (18%) (p > 0.05, chi 2 = 0.545, fd = 1). Our data indicate that Japanese PMs of (S)-mephenytoin could be identified by PCR-based genotyping of CYP2C19.

Adolescent↗