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Biomedical subjects

H Uchida

Publications and source records attributed to H Uchida.

At least 19 recordsLinked to original sources

Quantitative evaluation of adhesion of lactobacilli isolated from human intestinal tissues to human colonic mucin using surface plasmon resonance (BIACORE assay).

AIMS: To isolate lactobacilli from the mucus layer of the human intestine and evaluate their adhesion abilities using a BIACORE assay. METHODS AND RESULTS: Thirty strains of lactobacilli were isolated from the mucus layer of normal human intestinal tissues using conventional plate culture. The strains were identified using homology comparisons of the 16S rDNA sequence to databases as Lactobacillus salivarius (26%), Lactobacillus fermentum (13%), Lactobacillus gasseri (10%), Lactobacillus paracasei (7%), Lactobacillus casei (3%), Lactobacillus mucosae (3%) and Lactobacillus plantarum (3%). Lactobacillus plantarum LA 318 shows the highest adhesion to human colonic mucin (HCM) using the BIACORE assay at 115.30 +/- 12.37 resonance unit (RU). The adhesion of cell wall surface proteins from strain LA 318 was significantly higher to HCM than to bovine serum albumin (BSA; P < 0.05). CONCLUSIONS: We isolated 30 strains of lactobacilli. Lactobacillus salivarius was the predominant species of lactobacilli isolated in this study. The adhesion of strain LA 318 isolated from human transverse colon to its mucin was shown. The adhesion could be mediated by lectin-like components on the bacterial cell surface. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study where lactobacilli were isolated from human intestinal tissues and shown to adhere to HCM.

Bacterial Adhesion↗

Purification, characterization, and potential applications of formate oxidase from Debaryomyces vanrijiae MH201.

Formate oxidase was found in cell-free extracts of Debaryomyces vanrijiae MH201, a soil isolate. After purification by column chromatography, the preparation showed a protein band corresponding to a molecular mass (MM) of 64 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The MM, estimated by a gel filtration, was 99 kDa. The preparation showed two and three bands on isoelectric focusing under denaturing and native conditions, respectively. These results suggest that the preparation contained three isoforms, each of which might be composed of alphaalpha, alphabeta, and betabeta subunits with apparently similar MM. The preparation acted on formate with K (m) and V (max) values of 11.7 mM and 262 micromol min(-1) mg(-1), respectively, at pH 4.5 and 25 degrees C, but showed no evidence of activity on the other compounds tested. The optimum pH and temperature were pH 4.0 and 35 degrees C, respectively. The preparation showed activities of 85% of the initial activity after storage at pH 6.0 and 4 degrees C for 8 weeks. When 10 mM formaldehyde was reacted with 2.0 U ml(-1) of the enzyme preparation at pH 5.5 and room temperature in the presence of 2.0 U ml(-1) of a microbial aldehyde oxidase and 100 U ml(-1) of catalase for 180 min, neither of formate nor formaldehyde was detected, suggesting that the reaction involved the quantitative conversion of formaldehyde to carbon dioxide.

Aldehyde Oxidase↗

Genetic correlations among fatty acid compositions in different sites of fat tissues, meat production, and meat quality traits in Duroc pigs.

This study estimated genetic parameters for fatty acids of different sites of fat tissue, meat production, and meat quality traits of Duroc pigs selected during 7 generations for ADG, LM area, backfat thickness (BF), and intramuscular fat (IMF). For this study, 394 barrows and 153 gilts were slaughtered at 105 kg of BW. High heritabilities for C18:0 of outer and inner subcutaneous fat tissue were estimated, respectively, as 0.54 and 0.51; those of intermuscular and intramuscular fat were 0.40 and 0.51, respectively. Genetic and phenotypic correlations of ADG and BF with saturated fatty acids of outer and inner subcutaneous fat were positive, but those with C16:1 and C18:2 were negative, and those with C18:1 were nearly zero. Genetic and phenotypic correlations between LM area and respective fatty acids showed opposite results. Respective genetic and phenotypic correlations of melting points with C18:0 and C18:1 were positive and high, and negative and high, respectively. Genetic correlations between cooking loss and SFA (C14:0, C16:0, and C18:0) of IMF were positive and moderate: 0.56, 0.47, and 0.47, respectively. On the other hand, monosaturated fatty acid of C18:1 was highly and negatively correlated with cooking loss (-0.61). Moreover, high genetic correlation between meat color (pork color standard and lightness) and fatty acid compositions of IMF suggest that the SFA (C14:0, C16:0, and C18:0) were correlated genetically with meat lightness and that unsaturated fatty acid compositions (C18:1 and C18:2) were correlated with meat darkness. Results of this study suggest that the fatty acid composition of adipose tissue is correlated genetically with meat production and meat quality traits.

Adipose Tissue↗

Direct RT-PCR method for detecting two chrysanthemum viroids using minimal amounts of plant tissue.

A direct reverse transcription-polymerase chain reaction (RT-PCR) method for detecting the chrysanthemum stunt viroid (CSVd) and chrysanthemum chlorotic mottle viroid (CChMVd) to screen for a viroid-free chrysanthemum plant at a small plant size was established and named microtissue direct RT-PCR. A razor or syringe needle was used for RNA template preparations. Under a stereoscopic microscope, a razor or syringe needle was used to pierce, a tissue sample to a depth of 0.1-0.2mm, and the sample was directly transferred to the RT mixtures. Methods using razors or needles for the preparation of templates could detect CSVd and CChMVd with a high sensitivity. The most sensitive method used a razor or syringe needle to acquire template from the shoot tips. Using the microtissue direct RT-PCR method, both viroids could be detected from the high- and low-viroid-concentration plants. The microtissue direct RT-PCR method was more sensitive than a conventional template preparation method. Using the microtissue direct RT-PCR method established in this study, the laborious subculture step could be omitted because detecting viroids and screening for viroid-free plants even at a small plant size before the subculture could be possible. In addition, the microtissue direct RT-PCR method could also be a powerful tool for clarifing the viroid distribution among microtissues, such as shoot apical meristems.

Chrysanthemum↗

Characterization and potential application of purified aldehyde oxidase from Pseudomonas stutzeri IFO12695.

The molecular weight of purified aldehyde oxidase from Pseudomonas stutzeri IFO12695 was estimated to be 160 kDa by a gel filtration method. SDS-PAGE showed that the enzyme consisted of three non-identical subunits with molecular weights of 18, 38, and 83 kDa. The enzyme exhibited an absorption spectrum with maxima at 277, 325, 365, 415, 450, 480, and 550 nm and possessed molybdenum, CMP, iron, sulfur, and FAD as its cofactors, indicating that it belonged to the xanthine oxidase family. A variety of aliphatic and aromatic aldehydes were oxidized; and among them n-hexylaldehyde gave the most rapidly action. When 10 mM formaldehyde was treated with the aldehyde oxidase in the presence of catalase for 240 min, the formaldehyde concentration was reduced to 0.8 mM, suggesting this enzyme might be effective for the removal of formaldehyde contained in wastewater.

Aldehyde Oxidase↗

Impairment of thrombospondin-1 expression during epithelial wound healing in corneas of vitamin A-deficient mice.

The purpose of this study is to investigate the expression of thrombospondin-1 (TSP-1), a multifunctional extracellular matrix protein, during re-epithelialization in wounded corneas of vitamin A-deficient mice. Epithelial defects were created in the corneas of normal and Vitamin A-deficient mice with a microgrinder. Wounded corneas were stained with fluorescein and photographed for evaluation of re-epithelialization. Histological examination and immunohistochemical analysis of TSP-1 expression were also performed on the specimens from wounded corneas. In vitamin A-deficient mice, re-epithelialization of the wounded corneal epithelium was significantly delayed compared with that in normal mice. TSP-1 was detectable neither in the unwounded corneal epithelium of normal mice nor in that of vitamin A-deficient mice. In normal mice, linear staining of TSP-1 was observed on the wounded corneal surface and stroma at 30 min and 8 h to 16 h, respectively, after abrasion, and this TSP-1 expression disappeared at 36 to 48 h, when re-epithelialization was completed. In contrast, no TSP-1 staining was observed in the wounded corneas of vitamin A-deficient mice, except for the endothelial cells, throughout the wound healing process. Histological examination revealed a progressive increase in polymorphonuclear neutrophil infiltration in the stroma of the corneas of vitamin A-deficient mice during the healing process. These findings suggest that vitamin A may modulate the expression of TSP-1 in the corneas to accelerate the re-epithelialization of wounded corneas.

Animals↗

Preoperative airway evaluation using multi-slice three-dimensional computed tomography for a patient with severe tracheal stenosis.

A 71-yr-old female with a malignant thyroid tumour was to undergo thyroidectomy under general anaesthesia. Preoperative chest x-ray and plain computed tomography (CT) showed severe tracheal stenosis. Three-dimensional figures of the trachea and a virtual bronchoscopic movie were obtained from multi-slice CT to evaluate the stenotic region and to simulate fibroscopic tracheal intubation, respectively. After induction of general anaesthesia with propofol, a tracheal tube was successfully passed through the stenotic region under the guide of a fibroscope as simulated in the virtual movie. We conclude that multi-slice CT is useful for preoperative airway evaluation for patients with stenosis and distortion of the trachea.

Aged↗

Development of porous ceramics with well-controlled porosities and pore sizes from apatite fibers and their evaluations.

Porous hydroxyapatite (HAp) ceramics possessing well-controlled porosities and pore sizes were developed by firing apatite-fiber compacts mixed with carbon beads and agar. The total porosities could be controlled in the range from 40 to 85% by varying compaction pressure (20-40 MPa), firing temperature (1000-1300 degrees C) and carbon/HAp ratio (0/10-10/10 (w/w)). Most of the pores were regarded as open pores. The pore sizes were mainly affected by the carbon-bead diameter (5, 20 or 150 microm) and partly by the compaction pressure and the firing temperature. The pore sizes of the porous HAp ceramics derived from the carbon beads of 150 microm in diameter were distributed in the two separate ranges of several micrometers and more than 100 microm.

Ceramics↗

The effect of pre-operative administration of midazolam on the development of intra-operative hypothermia.

(Midazolam is often used for premedication; it is known to promote vasodilation and may therefore affect redistribution of heat during surgery. We examined the effect of pre-operative administration of midazolam on the development of intra-operative hypothermia. Forty-five patients were randomly allocated to one of three groups to receive no premedication (Group C), IM midazolam 0.04 mg.kg(-1) (Group M1) or 0.08 mg.kg(-1) (Group M2) 30 min prior to anaesthesia. Sedation levels were assessed, and then general anaesthesia was induced and maintained using propofol and fentanyl. During surgery, core temperature, which was similar for the three groups prior to induction of anaesthesia, decreased significantly less in the midazolam groups M1 and M2 compared to the control group C. Patients who were more heavily sedated prior to induction of anaesthesia, had significantly lower core temperatures peri-operatively than those who were less sedated, and core temperatures in unpremedicated patients fell to significantly lower levels during surgery than those who were drowsy. We conclude that pre-operative administration of midazolam produces an effect on the development of peri-operative hypothermia. We found that moderate pre-operative sedation reduces the peri-operative heat loss, possibly by affecting core-to-peripheral heat distribution.

Adolescent↗

Presence of staphylococcal exfoliative toxin A in sera of patients with atopic dermatitis.

BACKGROUND: It has been reported that the toxins that Staphylococcus aureus produces are associated with the exacerbation of atopic dermatitis (AD). It has been shown in many studies that staphylococcal enterotoxin (SE) A and SEB contribute to AD by humoral immunity through IgE production as a superantigen. On the other hand, little attention has been paid to the relationship between AD and exfoliative toxin x (ETx). OBJECTIVE: We investigated the toxins that are frequently detected from the skin of patients and how these toxins affect AD. METHODS: S. aureus, isolated from the skin of 100 patients with mild to severe AD, were examined for the producibility of toxins by polymerase chain reaction. Serum samples were obtained from 21 patients with mild and moderate AD. The levels of SEB, ETA, total IgE, specific IgE, and specific IgG in sera were measured by ELISA. RESULTS: SEB was most frequently detected from S. aureus on the skin of these patients as previously reported. And ETx, to which little attention has been paid so far, was frequently detected next to SEB. Furthermore, ETA was detected from the sera of almost all the AD patients. SEB was not detected at all. Although the level of ETA in the AD group was significantly higher than that of controls, ETA-specific IgE was not detected from their sera. High levels of ETA tended to be detected from infantile patients. Although there were no significant differences in the levels of ETA-IgG between AD and the controls, its prevalence was more than twice as high as the controls in AD. CONCLUSION: These results suggest that many AD patients were exposed to ETx. We conclude that ETx may contribute to exacerbation of AD, particularly in infants, by a mechanism that is not through specific IgE production, unlike SEB.

Adolescent↗

Genetic correlation between serum insulin-like growth factor-1 concentration and performance and meat quality traits in Duroc pigs.

This study was intended to examine whether serum IGF-I concentration is appropriate for use as a physiological predictor for genetic improvement of meat production and meat quality traits in pigs. Heritabilities and genetic correlations were estimated for these traits. The Duroc breed used in this study was selected for seven generations for average daily BW gain (DG) from 30 to 105 kg of BW, loin-eye muscle area (EM), backfat thickness (BF), and intramuscular fat (IMF) content. Serum IGF-I concentration of boars and gilts at the fourth generation of selection and that of boars, gilts, and barrows from the fifth to seventh generations of selection were measured at 8 wk (IGFI-8W) for 832 animals and again at the time they reached 105 kg of BW (IGFI-105KG) for 834 animals. A multivariate REML procedure was used to estimate genetic parameters with a model incorporating generation of selection, sex, common environmental effect of litter, and individual additive genetic effects. Heritability estimates for IGFI-8W and IGFI-105KG were 0.23 +/- 0.02 and 0.26 +/- 0.03, respectively. The estimates of common environmental effect for IGFI-8W and IGFI-105KG were 0.20 +/- 0.02 and 0.03 +/- 0.01, respectively. Positive genetic correlations were estimated between IGFI-8W and DG (0.26 +/- 0.08), EM (0.22 +/- 0.10), and IMF (0.32 +/- 0.10). Moreover, the positive genetic correlation between IGFI-105KG and EM was 0.42 +/- 0.08. These results indicate that serum IGF-I concentration at an early stage of growth was effective for prediction of IMF, but it was not a reliable physiological predictor of genetic merit of meat production traits.

Adipose Tissue↗

Safety of AT-1015, a novel 5-HT2A antagonist, in combination with high-dose aspirin: an open-label study.

OBJECTIVE: To assess the safety of AT-1015 in combination with high-dose aspirin (300 mg daily). Study subjects were 17 healthy male volunteers. METHODS: This was an open-label, single-center study. Subjects received aspirin 300 mg once daily, alone on days 1-4, and together with AT-1015 40 mg twice daily on days 5-11. A follow-up assessment was performed on day 18. The primary outcome measure was bleeding time; secondary outcome measures were vital signs, adverse events, physical examinations, 12-lead electrocardiograms (ECG) and laboratory safety tests. RESULTS: There was a significant increase in bleeding time between screening and the end of the aspirin-only period (mean bleeding time 4.8 vs 7.6 min, p = 0.01), but there were no further significant increases during the combination treatment period. The most common adverse events were dry mouth, epistaxis, gingival bleeding and abdominal pain. All treatment-related adverse events were mild in severity and no major bleeding episodes occurred. There were no clinically significant changes in vital signs, physical examinations, 12-lead ECGs or laboratory safety tests. CONCLUSIONS: AT-1015 was safe and well-tolerated in healthy male volunteers when taken in combination with high-dose aspirin, and did not significantly prolong bleeding time compared with aspirin alone.

Administration, Oral↗

Venous blood flow through vein grafts before harvesting minimizes endothelial cell desquamation immediately after implantation.

AIM: The aim of this paper was to examine effectiveness of maintaining venous blood flow through dissected, but not harvested, grafts on prostaglandin I2 production and endothelial cell regeneration after implantation. METHODS: Using 10 dogs, 8-cm-long bilateral jugular veins were dissected; 1 was harvested, and immersed in heparinized blood for 60 or 120 min. The other was left in situ with retained blood flow until harvest after 60 or 120 min. Two 1-cm-long specimens were obtained from these 4 graft types and subjected to PGI2 assay or silver nitrate staining for measurement of endothelial cell coverage area. The remaining 6-cm portions of the 120-min grafts were autogenously implanted into the abdominal aorta, retrieved after 7 days and assayed and measured. RESULTS: Comparisons between groups suggested beneficial effects of preserving venous blood flow until implantation on endothelial coverage and PGI2 production (p<0.05). CONCLUSION: Venous blood flow through dissected vein grafts effectively helps preserve PGI2 production.

Animals↗

Activated leucocytes express and secrete macrophage inflammatory protein-1alpha upon interaction with synovial fibroblasts of rheumatoid arthritis via a beta2-integrin/ICAM-1 mechanism.

OBJECTIVE: To examine the expression and regulation of chemotactic factor, macrophage inflammatory protein-1alpha (MIP-1alpha) by fibroblast-like synoviocytes (FLS), monocytes and polymorphonuclear neutrophils (PMN) isolated from the synovial fluid (SF) of rheumatoid arthritis (RA) patients. METHODS: Monocytes or PMN obtained from RA SF were co-cultured with unstimulated semiconfluent RA FLS. Culture supernatants were assayed for MIP-1alpha by enzyme-linked immunosorbent assay. The expression of MIP-1alpha mRNA and protein was also determined by Northern blot analyss and immunohistochemistry respectively. RESULTS: Interaction of activated leucocytes with FLS synergistically increased MIP-1alpha expression and secretion via a mechanism mediated by beta2-integrin/ intercellular adhesion molecule 1. CONCLUSION: MIP-1alpha expression within inflamed joints appears to be regulated not only by inflammatory cytokines but also by the physical interaction of activated leucocytes and FLS, and plays a crucial role in the progression and maintenance of RA synovitis.

Arthritis, Rheumatoid↗

Transfer function analysis of positron-emitting tracer imaging system (PETIS) data.

Quantitative analysis of the two-dimensional image data obtained with the positron-emitting tracer imaging system (PETIS) for plant physiology has been carried out using a transfer function analysis method. While a cut leaf base of Chinese chive (Allium tuberosum Rottler) or a cut stem of soybean (Glycine max L.) was immersed in an aqueous solution containing the [18F] F- ion or [13N]NO3- ion, tracer images of the leaf of Chinese chive and the trifoliate of soybean were recorded with PETIS. From the time sequence of images, the tracer transfer function was estimated from which the speed of tracer transport and the fraction moved between specified image positions were deduced.

Allium↗

Adenovirus-mediated transfection of caspase-8 augments anoikis and inhibits peritoneal dissemination of human gastric carcinoma cells.

Caspase-8 is a member of the cysteine protease family that modulates apoptosis induced by a variety of cell death signals and has recently been found to be activated during the process of anoikis, which is a form of apoptosis caused by loss of anchorage in epithelial cells. We previously demonstrated that the inhibition of anoikis promotes peritoneal dissemination of human gastric carcinoma MKN45 cells, which are anchorage dependent. This suggests that augmentation of anoikis may suppress dissemination of carcinoma cells. To determine whether extrinsic overexpression of caspase-8 can augment anoikis in MKN45 cells, we transfected them with the caspase-8 gene using an adenoviral (Adv) vector (Adv-caspase-8). Here we demonstrate that Adv-caspase-8 infection, at 15 multiplicity of infection (MOI), can augment anoikis in MKN45 cells and suppresses MKN45 peritoneal dissemination in SCID mice. The inhibitory effect on peritoneal dissemination resulted in a prolonged survival compared with that in control mice. In contrast, the Adv-caspase-8 (15 MOI) had no distinct effect on cell viability or growth either of attached MKN45 cells or of s.c. tumor growth in SCID mice. Thus, Adv-mediated overexpression of caspase-8 suppressed peritoneal dissemination mainly through augmentation of anoikis. In addition, Adv-caspase-8-mediated augmentation of anoikis was similarly observed in another gastric carcinoma MKN74 cell line. In contrast, Adv-p53 could not augment anoikis in MKN45 cells. These results imply that Adv-mediated gene transfer of caspase-8 can selectively induce apoptosis in detached carcinoma cells and, thus, shows potential as a novel cancer therapy against dissemination of gastric and probably other carcinoma cells originating from epithelial tissues.

Adenocarcinoma↗