Biomedical subjects

H Obata

Publications and source records attributed to H Obata.

At least 55 records · Page 3Linked to original sources

PubMed1999

Exposure and sensitization to cat dander: asthma and asthma-like symptoms among adults.

BACKGROUND: Several pets, such as cats, dogs, and rodents, are known to produce allergens. Despite the clinical and laboratory evidence that exposure to pets can cause bronchoconstriction in sensitized subjects, the results of population studies have been contradictory. OBJECTIVES: The purpose of this study was to investigate the relationship between cat ownership and the prevalence of asthma, asthma-like symptoms, and bronchitic symptoms among subjects 20 to 44 years of age in Vancouver, Canada and to determine whether sensitization is responsible for such an association. METHODS: Two thousand nine hundred ninety-nine (88%) randomly selected subjects responded to a mail questionnaire. Of these, 504 participated in laboratory examination, including allergy skin testing. RESULTS: One thousand nineteen study responders (34%) were pet owners at the time of the study (current owners). Current pet owners were found to have a higher prevalence of current asthma, asthma-like symptoms, and bronchitic symptoms compared with those without pets. Cat owners had significantly higher risk of having current asthma and asthma-like symptoms. In the subset who had allergic skin tests, we found that those who were allergic to cat dander had a significantly higher risk of current asthma than those not allergic to cat dander and not owning a cat. CONCLUSION: This study provides evidence that sensitization to cat dander is a more important risk factor for current asthma and asthma-like symptoms than cat ownership itself.

Adult↗

PubMed1999

NF-kappaB involvement in the activation of primary adult T-cell leukemia cells and its clinical implications.

The HTLV-I provirus-encoded Tax protein induces NF-kappaB in Tax-transfected Jurkat T cells or HTLVL-I- infected T cells in vitro. Tax induction of NF-kappaB is presumed to be involved in proliferation and activation of primary leukemia cells in vivo. Recent studies have demonstrated that NF-kappaB activities in human T cells are mediated by at least four c-Rel-related DNA binding proteins - p50, p55, p75 and p85. We examined the significance of NF-kappaB induction in primary adult T cell leukemia cells and the induction kinetics of each of the four NF-kappaB species. Marked NF-kappaB activity was detected using an electrophoretic mobility shift assay (EMSA) in the primary cells of patients with acute disease, but little activity was noted in the cells of chronic patients. NF-kappaB activity was enhanced in a time-dependent manner in acute type cells cultured with mitogen-free medium; there was no induction of activity in chronic type cells. UV crosslinking demonstrated all four species of NFkappaB complex - high levels of p50 and lower levels of p55 and p75, in acute type cells; chronic type cells showed only the p50. As a control, normal resting T cells similarly showed only p50; control cells showed little change in activity when cultured without mitogenic stimulation, analogous to chronic type ATL. Northern blotting revealed enhancement of c-rel (encoding p85) and KBFI (encoding p50 and p55) expression in acute type cells during culture, while there was no significant enhancement of mRNAs in chronic type ATL cells or unstimulated normal T cells. Northern blotting also revealed that Tax is upregulated at the mRNA level in acute- but not chronic-type cells during culture. Expression of c-rel and KBF1 mRNAs in acute type cells appeared to be related to Tax mRNA expression. These results suggest that Tax is capable of inducing nuclear expression of all four NF-kappaB species in primary ATL cells of acute type patients, with marked effects on p55, p75, and p85. Tax induction of NF-kappaB species is regulated, at least in part, at a pretranslational level involving increases in c-rel and KBF1 mRNA.

Acute Disease↗

PubMed1999

Creating cataract in a pig eye.

The purpose of this study was to create a uniform, reproducible hardened lens nucleus and an anterior capsule in a pig eye that approximate those in the human eye for use in cataract surgery training. The lens nuclei was hardened and anterior capsule elasticity was reduced in pig eyes by injecting the lens with mixtures of formalin and alcohol (ethanol, 2-propanol, or both). A 30 gauge needle was used to pierce the lens 3.0 mm from the limbus. Then, 0.2 mL of the formalin and alcohol mixture was injected. The most effective mixture was formalin, ethanol, and 2-propanol in the ratio 4:3:3. Fifteen minutes after this mixture was injected, the pig eye had nuclear cataract with grade III hardness and an anterior capsule that approximated the capsule in a human cataractous eye. The cataract created by this method will be useful in teaching continuous curvilinear capsulorhexis, divide and conquer and phaco chop phacoemulsification, and nuclear hydrodelineation and rotation.

2-Propanol↗

PubMed1999

Basic subsite theory assumptions may not be applicable to hydrolysis of cellooligosaccharides by almond beta-glucosidase.

Steady-state kinetic parameters for the hydrolysis of cellooligosaccharides by almond beta-glucosidase were evaluated at pH 5.0 and 25 degrees C in relation to the subsite theory (K. Hiromi, Biochem. Biophys. Res. Commun., 40, 1-6, 1970). The value of k0/Km decreased monotonously with increasing degree of polymerization (DP) of the substrates (DP = 2-6). Also, the Km and k0 values for cellotriose were smaller than those for cellobiose. These DP dependencies differ from those of most amylases and glucosidases studied so far, to which the subsite theory has been successfully applied. The subsite parameters could not be consistently obtained, which suggests that one or both of the two basic assumptions of the subsite theory might not be applicable to the hydrolysis of cellooligosaccharides by the enzyme. That is, the intrinsic rate of the hydrolysis may depend on the DP and/or there may be interaction between subsites for binding the glucose residues of a substrate.

Journal Article↗

PubMed1999

Intrathecal co-administration of NMDA antagonist and NK-1 antagonist reduces MAC of isoflurane in rats.

BACKGROUND: Intravenous administration of N-methyl-D-aspartate (NMDA) receptor antagonists and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor antagonists reportedly reduce the minimum alveolar anaesthetic concentration (MAC) for inhalation anaesthetics. If pain perception can be prevented by the intrathecal administration of antinociceptive receptor antagonists, these agents may reduce the requirements for inhalation anaesthetics. We studied the effect of intrathecal administration of an AMPA/kainate receptor antagonist, a metabotropic glutamate (mGlu) receptor antagonist and co-administration of NMDA and a neurokinin-1(NK-1) receptor antagonist drugs at low doses on the MAC. METHODS: After Wistar rats (n=36) were fitted with indwelling intrathecal catheters, the MAC of isoflurane was determined following intrathecal administration of a non-NMDA receptor antagonist (CNQX) at 10 microg, a mGlu receptor antagonist (AP3) at 10 microg, or a combination of NMDA receptor antagonist (APV) at 0.01 microg to 1 microg with NK-1 receptor antagonist (CP96345, CP) at 0.1 microg to 10 microg. Subsequently, a reversal dose of intrathecal NMDA with substance P (SP) was administered, and the MAC of isoflurane was redetermined. Conscious rats (n=15) were also examined for the presence of locomotor dysfunction following the intrathecal co-administration of APV and CP. RESULTS: Neither CNQX nor AP3 reduced the MAC of isoflurane. APV at 0.01 microg plus CP at 1 microg, as well as APV at 0.1 microg plus CP at 10 microg, reduced the MAC of isoflurane, with respective reductions of 7.6% and 14%; (P<0.05). Co-administration of NMDA plus SP reversed the decrease in the MAC of isoflurane. Locomotive activity was not changed. CONCLUSIONS: The NMDA receptor and the NK-1 receptor are important determinants of the MAC of isoflurane, exerting this influence by inhibition of pain transmission in the spinal cord, while mGlu and AMPA receptors have no effect on the MAC of isoflurane.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1999

Expression of transforming growth factor-beta superfamily receptors in rat eyes.

PURPOSE: Transforming growth factor-beta (TGF-beta) superfamily consists of many multifunctional cytokines including TGF-beta, activins, and bone morphogenetic proteins (BMPs). Previous reports show that these factors are expressed in ocular tissue, and exert their effects through type I and type II serine/threonine type receptors. The purpose of this study is to observe the expression of these receptor families in normal rat eyes as the first step in investigating the functions of TGF-beta superfamily members in ocular tissue. METHODS: The expression of receptors was examined immunohistochemically in one eye each of 6 Lewis rats. The type I receptors examined were the TGF-beta type I receptor (TbetaR-I), activin type I receptor (ActR-I), activin type IB receptor (ActR-IB), bone BMP type IA receptor (BMPR-IA), BMP type IB receptor (BMPR-IB), and activin receptor-like kinase-1 (ALK-1). The type II receptors examined were TGF-beta type II receptor (TbetaR-II), activin type II receptor (ActR-II), and BMP type II receptor (BMPR-II). RESULTS: All 6 type I receptors, ActR-II and the alternative spliced long version of BMPR-II were expressed in the following cells: corneal and conjunctival epithelium, corneal keratocytes, corneal endothelium, epithelium of iris and ciliary body, epithelium of lens, sensory retina excluding the outer segments of photoreceptors, retinal pigment epithelium (RPE), and vascular cells. TbetaR-II and the short version of BMPR-II (truncated form) were expressed in the outer segments of the photoreceptors in addition to the above. The results of staining were similar in all the sections examined. CONCLUSIONS: The cornea, ciliary body, iris, lens, retinal cells, RPE and vascular cells expressed the receptors for TGF-beta superfamily at protein level.

Activin Receptors, Type I↗

PubMed1999

The effect of intrathecal magnesium sulphate on nociception in rat acute pain models.

We examined the antinociceptive effect of intrathecally administered magnesium sulphate (MgSO4) in rats, using acute pain models including mechanical pressure, heat and subcutaneous formalin injection. According to the locomotion test 10 microliters of 6.2% MgSO4 did not produce motor paralysis. At the same dose, responses to pressure and heat were intact, compared with controls given saline. MgSO4 produced depression of pain responses only after the first 10 min in the formalin test. Our studies indicated that MgSO4 did not show remarkable antinociceptive effects in acute pain models.

Acute Disease↗

PubMed1999

Guanidine hydrochloride-induced denaturation of Pseudomonas cepacia lipase.

The guanidine hydrochloride-induced denaturation of Pseudomonas cepacia lipase (PCL) was studied at pH 7 by monitoring the changes in the fluorescence and circular dichroism of the enzyme. The denaturation was irreversible as a whole, and the addition of Ca2+ ions decreased the velocity of the denaturation. The denaturation process was well explained consistently by a two-step mechanism, as follows: [see equation in text] where N is the native state of PCL, D(I) an intermediate denatured-state which can be refolded into the native state, and D(F) the final denatured-state that can not be renatured. Ethanol (10%) increased the denaturation velocity by decreasing the refolding step, D(I) + Ca2+ --> N x Ca2+, which would be caused by the stabilization of D(I) by ethanol.

Burkholderia cepacia↗

PubMed1999

[Biochemical and molecular characterization of Salmonella ser. enteritidis phage type 1 isolated from food poisoning outbreaks in Tokyo].

Since the first outbreak in 1990, the incidence of Salmonella ser. Enteritidis (S. Enteritidis) phage type (PT) 1 food poisoning has gradually increased in Tokyo and has reached approximately 30% of the total S. Enteritidis outbreaks reported. To characterise these S. Enteritidis PT1 food poisoning, a total of 198 strains obtained from 44 outbreaks between 1990 and 1996 were examined for antimicrobial resistance, acid producibility from glycols (propylene and ethylene glycol) and plasmid DNA profiles. The 44 PT1 outbreaks analysed were further subdivided into 11 types by epidemiological markers. The most common patterns were type A (plasmid profile carrying only one plasmid (60 kb). SM and TC resistance and non producibility from glycols), and type B (plasmid profile carrying two plasmids (60 and 20 kb), SM resistance and no producibility from glycols) and were responsible for 21 (47.7%) and 15 (34.1%) outbreaks, respectively. In 11 of 44 outbreaks, strains carrying identical epidemiological markers were isolated both from patients and vehicle foods, environments, and/or food-handlers. Similar to PT4 and PT34 outbreaks reported in Japan, egg and egg-related foods were also suspected in 8 of these 11 outbreaks. Of interest, chicken which were not pointed out in PT4 and PT 34 outbreaks was also suspected as a vehicle of transmission in two outbreaks.

Bacteriophage Typing↗

PubMed1999

Airway inflammation, exhaled nitric oxide, and severity of asthma in patients with western red cedar asthma.

Examination of induced sputum and measurement of exhaled NO have been advocated as noninvasive methods of assessing the degree of airway inflammation. In this study, we performed follow-up evaluation on 71 subjects with asthma caused by exposure to Western red cedar; 50 subjects had left exposure, whereas the rest continued to work in the same job. Spirometry, methacholine challenge tests, exhaled nitric oxide, and sputum induction were carried out. Of the 50 subjects who left exposure, 12 had no respiratory impairment according to the American Throacic Society guidelines for assessing respiratory impairment in patients with asthma, 17 belonged to Class 1, 12 to Class 2, five to Class 3, and four to Class 4. The percentage of eosinophils in induced sputum showed a significant inverse relationship with FEV1 (r = -0.46, p < 0.001), and a significant positive correlation with levels of exhaled NO (r = 0.42, p < 0.001) and with the class of respiratory impairment (r = 0.52, p < 0.001). Mean percent eosinophils were 1.5 for impairment Class 0, 2.2 for Class 1, 1.7 for Class 2, 6.8 for Class 3, and 16.3 for Class 4. No relationship was found between the levels of exhaled NO and the functional parameters as well as the impairment class. NO levels in ppb were 21 for impairment Class 0, 30 for Class 1, 22 for Class 2, 26 for Class 3, and 49 for Class 4. This study also provides objective evidence that airway inflammation, as indicated by induced sputum, corroborates the rating of respiratory impairment in patients with asthma.

Adult↗

PubMed1999

Sputum eosinophils and exhaled nitric oxide during late asthmatic reaction in patients with western red cedar asthma.

Examination of sputum for eosinophils and measurement of exhaled nitric oxide have been proposed as noninvasive methods of assessing airway inflammation in asthma. The use of these tests in the evaluation of patients with occupational asthma has not been reported. This study investigated the changes in sputum eosinophils and exhaled NO before and at intervals after inhalation challenge with plicatic acid in patients with suspected western red cedar asthma. Of 17 subjects who underwent challenge, nine had a positive bronchoconstrictor reaction (responders) and eight had a negative reaction (nonresponders). At 6 and 24 h after plicatic acid challenge, there was a significant increase in sputum eosinophils among responders, which was inversely related to the fall in forced expiratory volume in one second (FEV1) at 6 h. An increase in sputum eosinophils was also found in three nonresponders. Levels of exhaled NO increased at 24 h after challenge with plicatic acid in both responders and nonresponders, being significant only in nonresponders. No correlation was found between the increase in nitric oxide and the magnitude of the functional changes in the airways. There were significant correlations between the degree of sputum eosinophilia and the level of exhaled NO before and after methacholine and plicatic acid challenge. In conclusion, the late asthmatic reaction induced by plicatic acid in patients with western red cedar asthma is associated with an increase in sputum eosinophils. The usefulness of measuring sputum eosinophils and exhaled nitric oxide in the clinical evaluation of patients with suspected occupational asthma caused by low molecular weight compounds has yet to be determined.

Adult↗

PubMed1999

Steady-state inhibitory kinetic studies on the ligand binding modes of Aspergillus niger glucoamylase.

Inhibitory activities of 1-deoxynojirimycin and gluconolactone on Aspergillus niger glucoamylase were studied in relation to the subsite structure of the enzyme. Although both of these inhibitors are considered to bind at subsite 1 of the enzyme active site, 1-deoxynojirimycin showed competitive type inhibition but gluconolactone was a mixed type (or noncompetitive type) inhibitor for the hydrolysis of p-nitrophenyl alpha-D-glucoside. The former type of inhibition suggested that the main binding mode of the substrate was productive, but the latter, nonproductive. A possible way of explaining these apparent inconsistent results is to assume that the main binding mode of the substrate is productive and gluconolactone forms a nonproductive ternary complex with the enzyme and the substrate.

1-Deoxynojirimycin↗

PubMed1999

Possibility for discriminating between two representative non two-state thermal unfolding models of proteins by DSC.

Possible differences between two representative non two-state thermal unfolding mechanisms of protein are discussed concerning differential scanning calorimetry. Numerical simulations showed that, by DSC measurement, it is hard to discriminate between the independent model, which assumes independent unfolding domains in a protein, and the sequential model, which assumes intermediate(s) between native and denatured states, especially when values of molecular weight, denaturation enthalpy, and difference in denaturation temperature of each denaturation process are large. DSC curve analysis of Aspergillus niger glucoamylase based on these two models gave essentially the same thermodynamic parameters.

Aspergillus niger↗

PubMed1999

[Labyrinthine fistulas in cholesteatoma].

The purpose of the present study was to investigate the clinical features of cases of cholesteatoma with labyrinthine fistulas, and in particular the pre and post-operative bone-conduction (BC). Cholestatoma patients with bone erosion or a defect found in the first stage operation were analyzed. The operations were conducted between 1992 and 1996. The patients were classified into four types, I, IIa, IIb, and III, according to Dornhoffer and Milewski's classification, which is based on different stages in the bone defect. A type I fistula is an erosion of the bony labyrinth with an intact endosteum. Type IIa is accompanied by an opened perilymphatic space with undisturbed perilymph while type IIb has a disturbed perilymph. A Type III fistula is an opened perilymphatic space with a disturbance of the underlying membranous labyrinth. Only 24 patients with type II and type III fistulas were included in this study. The location of the fistulas was the semicircular canals (SCCs) or/and the vestibula in 21 patients and in the cochlea in 3 patients. We examined the fistula by high-resolution computed tomography scan (CT scan) with 1 mm slice and 1 mm width axial-sections in 14 patients. A bone defect in the labyrinth was detected in 10 cases (71.5%) pre-operatively. Pre-operative BC was worse in the patients with cochlear fistulas than in those with fistulas located in SCCs or the vestibula. Within this latter group there were 13 type IIa (group IIa), patients and 8 type IIb or III (group IIb or III) patients. However, there was no difference in the pre-operative BC between these two sub-groups Tympanoplasty was conducted in all 24 patients. The postoperative BC of group IIa and group IIb or III were compared. Two of the 13 patients in group II a (15.4%) and 3 of 8 in group IIb or III (37.5%) had a deteriorated postoperative BC. Statistical analysis revealed that the postoperative BC was more inclined to become worse in patients with advanced stage IIb or III fistulas.

Adolescent↗

PubMed1998

c-Myc gene single-strand binding protein-1, MSSP-1, suppresses transcription of alpha-smooth muscle actin gene in chicken visceral smooth muscle cells.

The expression of alpha-smooth muscle actin is coordinately regulated by positive and negative cis- elements in the promoter region. Although cis -elements and trans -acting factors involved in the positive regulation of the alpha-smooth muscle (alpha-SM) actin gene have been well characterized, details of negative regulation remain unclear. In functional analyses using cultured gizzard smooth muscle cells, we identified a sequence ranging from -238 to -219 in the promoter region as a novel negative element. Mutation and deletion analyses further revealed that a sequence, TATCTTA (-228 to -222), is essential for negative regulation. Gel shift assay and Southwestern blotting indicated that a nuclear protein factor specifically interacts with single- or double-strand DNA including this sequence, and the protein factor displays a highly potent binding to the sense strand DNA. cDNA cloning and gel shift analysis using anti-MSSP-1 antibodies revealed that this protein factor is a chicken homolog of human MSSP-1 (c- myc gene single-strand binding protein-1). In fact, overexpression of MSSP-1 in cultured smooth muscle cells suppresses the promoter activity. These results suggest a novel function of MSSP-1 regarding the transcriptional regulation of alpha-sm actin gene.

Actins↗

PubMed1998

Functional involvement of serum response factor in the transcriptional regulation of caldesmon gene.

A 22-bp fragment including the CArG element (CArG1) is essential for the transcription of the caldesmon gene. In this study, we investigated the effects of serum response factor (SRF) on the functional regulation of caldesmon promoter in smooth muscle cells. Gel supershift assay revealed that SRF was one component of the CArG1-protein complex. Dominant-negative mutants of SRF suppressed the promoter activity of caldesmon, whereas wild-type SRF overcame this suppression. These results suggest that SRF functions as a core activating factor of the caldesmon promoter. Furthermore, fractionation of smooth muscle cells' nuclear extracts using DNA affinity paramagnetic particles suggests that SRF transactivates the caldesmon promoter in concert with additional factors in the flow-through fraction recruited to the CArG element.

Animals↗

PubMed1998

Effects of transforming growth factor beta on corneal epithelial and stromal cell function in a rat wound healing model after excimer laser keratectomy.

BACKGROUND: Transforming growth factor beta (TGF-beta) regulates extracellular matrix deposition, cell proliferation, and migration, and is expressed in cornea. TGF-beta is thought to be involved in the corneal wound healing process. METHODS: The central corneal area (3 mm in diameter) of Lewis rats was ablated using PTK mode excimer laser and the wound healing process was observed at 12 and 24 h and 2, 5, 10, and 30 days after treatment. The expression of TGF-beta 1, -beta 2 and -beta 3, TGF-beta type I and type II receptors, alpha 3, alpha 5, beta 4 integrin subunits, laminin and fibronectin was studied immunohistochemically. Antibody neutralizing TGF-beta 1, -beta 2 and -beta 3 was administered intraperitoneally, 50 micrograms daily, for 5 days after the laser treatment to investigate the effects of TGF-beta function blockade. RESULTS: At the leading edge of the regenerating epithelium, no TGF-beta type I and type II receptors and beta 4 integrin subunits were expressed after 24 h. Regenerating epithelium covered the ablated area after 2 days. An abnormal fibrotic layer was formed in the subepithelial area. This layer contained round-shaped cells in the stroma in the early stage (2-5 days after laser ablation) and spindle-shaped fibroblast-like keratocytes after 10 days. Laminin and fibronectin expression increased in the fibrotic layer. The increased stromal cells expressed TGF-beta isoforms and TGF-beta receptors. Neutralizing TGF-beta inhibited the stromal cell increase in the laser ablated area after 5 days. CONCLUSION: TGF-beta may be involved in epithelial cell migration and stromal cell reaction during the corneal wound healing process after excimer laser ablation in rat models.

Activin Receptors, Type I↗

PubMed1998

Three-dimensional organization of collagen fibrils during corneal stromal wound healing after excimer laser keratectomy.

PURPOSE: To investigate the structural changes in corneal stromal collagen fibrils after excimer laser keratectomy in relation to the degree of corneal haze. SETTING: University of Tokyo Hospital, Tokyo, Japan. METHODS: Corneal haze was quantitatively measured by analyzing the light scattering in Scheimpflug images of the corneas of white rabbits after excimer laser keratectomy. Collagen fibril structure was examined using scanning electron microscopy after chemical digestion with sodium hydroxide solution; the same specimens were examined by transmission electron microscopy after re-embedding. RESULTS: Corneal haze reached a peak 4 weeks after excimer laser keratectomy and then gradually decreased. The collagen fibrils of the normal cornea were regularly arranged parallel to the surface of the cornea, with small interfibrillar distances. After excimer laser keratectomy, the arrangement was highly disordered, with increased interfibrillar distances. These structural changes were most prominent 4 weeks after excimer laser keratectomy. CONCLUSION: The structural changes in the collagen fibrils of the corneal stroma, especially the increase in interfibrillar distances and the disordered arrangement, were associated with corneal haze after excimer laser keratectomy.

Animals↗