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Biomedical subjects

M Kato

Publications and source records attributed to M Kato.

At least 685 records · Page 38Linked to original sources

The effect of magnetic coil orientation on the excitation of the median nerve.

OBJECTIVE: We investigated the effect of magnetic coil orientation on the excitation of the median nerve in healthy subjects. METHODS: An 8-shaped coil, 3.2 cm in outer diameter, was used. The median nerve was stimulated at the elbow while the compound muscle action potentials (CMAPs) of abductor pollicis brevis muscle were recorded at 4 different directions of the induced current: orthodromic, antidromic, medio-lateral and latero-medial. RESULTS: We found that the amplitude of the CMAP was the greatest in a medio-lateral (ML) direction. We also measured the induced electric field in the saline tank that mimicked the forearm. The induced electric field and its spatial gradient were the greatest in the ML direction among 4 directions. CONCLUSION: The fact that the forearm is a restrictive volume conductor may result in the different effects of coil orientation on the excitation of the median nerve at the elbow.

Action Potentials↗

Expression of pancreatic alpha-amylase protein and messenger RNA in hilar primitive bile ducts and hepatocytes during human fetal liver organogenesis: an immunohistochemical and in situ hybridization study.

AIMS/BACKGROUND: This study was conducted to evaluate the expression of pancreatic digestive enzymes in hilar bile ducts and hepatocytes during human fetal liver organogenesis. METHODS: We investigated the expression of pancreatic alpha-amylase protein and messenger RNA (mRNA) in hilar primitive bile ducts and hepatocytes by immunohistochemistry and in situ hybridization techniques, using 11 human fetal livers of various gestational ages. The specificity of the immunohistochemistry and in situ hybridization procedures was confirmed by Western blot analysis and in situ hybridization using sense probes, respectively. RESULTS: Immunoreactivity of pancreatic alpha-amylase protein and expression of pancreatic alpha-amylase mRNA were present not only in the primitive ductal cells of the hilar region including the ductal plate, remodelling bile ducts and remodeled bile ducts but also in primitive hepatocytes of the hilar region, though the immunoreactivity and mRNA signals in the primitive hepatocytes disappeared in the third trimester. There was perfect correlation between immunohistochemistry and in situ hybridization. CONCLUSIONS: These results suggest that primitive biliary cells and hepatocytes of the hilar region in the human fetus do express pancreatic alpha-amylase protein and mRNA, and that the primitive biliary epithelial cells and hepatocytes in the hilar region share a common cell lineage with exocrine pancreatic cells.

Bile Ducts, Intrahepatic↗

[Retropharyngeal abscess in an adult and an elderly woman].

Retropharyngeal abscess is reported to be decreasing in frequency in recent years. We report two cases of retropharyngeal abscess that were diagnosed within four years in Department of Otorhinolaryngology, Gifu University Hospital. Case 1 was a 36-year-old male who was suggested to have an abscess as primary infection and case 2 was a 71-year-old female whose abscess seemed to be a secondary infection following unknown primary infection. Endoscopic or open neck drainage as well as antibacterial chemotherapy mainly with combinations of flomoxef and clindamycin in the case 1 and piperacillin and clindamycin in the case 2 was successfully carried out. They were discharged an 15-day and 24-day after operation in the cases 1 and 2, respectively. A 30 min. culture after sample collection on operation demonstrated aerobe-anaerobe mixed infection in both cases; three aerobes and four anaerobes in the case 1 and three aerobes and two anaerobes in the case 2. These results suggest that retropharyngeal abscess may be a complicated infection involving more bacterial species than has been commonly believed. Measuring susceptibility of isolates to antimicrobial agents, a strain of Capnocytophaga sp. and a strain of anaerobic gram-negative rod were highly resistant to piperacillin, cefroxine and ofloxacin. It is important for adequate antibacterial chemotherapy to grasp the bacteriology of retropharyngeal abscess and analyze susceptibility of antimicrobial agents.

Adult↗

Cloning and molecular analysis of the Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) biosynthesis genes in Pseudomonas sp. strain 61-3.

Two types of polyhydroxyalkanoate (PHA) biosynthesis gene loci (phb and pha) of Pseudomonas sp. strain 61-3, which produces a blend of poly(3-hydroxybutyrate) [P(3HB)] homopolymer and a random copolymer (poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) [P(3HB-co-3HA]) consisting of 3HA units of 4 to 12 carbon atoms, were cloned and analyzed at the molecular level. In the phb locus, three open reading frames encoding polyhydroxybutyrate (PHB) synthase (PhbCPs), beta-ketothiolase (PhbAPs), and NADPH-dependent acetoacetyl coenzyme A reductase (PhbBPs) were found. The genetic organization showed a putative promoter region, followed by phbBPs-phbAPs-phbCPs. Upstream from phbBPs was found the phbRPs gene, which exhibits significant similarity to members of the AraC/XylS family of transcriptional activators. The phbRPs gene was found to be transcribed in the opposite direction from the three structural genes. Cloning of phbRPs in a relatively high-copy vector in Pseudomonas sp. strain 61-3 elevated the levels of beta-galactosidase activity from a transcriptional phb promoter-lacZ fusion and also enhanced the 3HB fraction in the polyesters synthesized by this strain, suggesting that PhbRPs is a positive regulatory protein controlling the transcription of phbBACPs in this bacterium. In the pha locus, two genes encoding PHA synthases (PhaC1Ps and PhaC2Ps) were flanked by a PHA depolymerase gene (phaZPs), and two adjacent open reading frames (ORF1 and phaDPs), and the gene order was ORF1, phaC1Ps, phaZPs, phaC2Ps, and phaDPs. Heterologous expression of the cloned fragments in PHA-negative mutants of Pseudomonas putida and Ralstonia eutropha revealed that PHB synthase and two PHA synthases of Pseudomonas sp. strain 61-3 were specific for short chain length and both short and medium chain length 3HA units, respectively.

Acyltransferases↗

Increased cytosolic free Mg2+ and Ca2+ in platelets of patients with vasospastic angina.

This study was designed to test the hypothesis that the cellular metabolism of Ca2+ and Mg2+, which is important in platelet function, is abnormal in the platelets of patients with vasospastic angina. Cytosolic free Mg2+ concentration ([Mg2+]i) and Ca2+ handling were determined in the platelets of 24 patients with vasospastic angina and 24 control subjects by use of mag-fura 2 and fura 2. Platelet aggregation was also examined. Basal [Mg2+]i and cytosolic free Ca2+ concentration ([Ca2+]i) in platelets were significantly higher in patients with vasospastic angina than in control subjects. The amplitude of the [Ca2+]i transient induced by thrombin (0.03-1.0 U/ml) was significantly increased in the presence, but not in the absence, of extracellular Ca2+ in patients with vasospastic angina, as compared with controls. Therefore, the influx of Ca2+ across the plasma membrane may be accelerated in vasospastic angina. Thrombin (0.1-1.0 U/ml)-induced maximum aggregation response was significantly greater in patients with vasospastic angina than in controls. Results suggest that increased [Mg2+]i and altered Ca2+ handling by platelets may be associated with coronary vasospasm.

Adenosine Diphosphate↗

Exercise training restores decreased cellular immune functions in obese Zucker rats.

This study investigated whether exercise training had a beneficial effect on the decreased mitogen response and improved a decreased expression of glucose transporter 1 (GLUT-1) in splenocytes from obese Zucker rats. Experimental groups were lean and sedentary and exercise-trained obese Zucker rats. Exercise training, running on a motor-driven treadmill for 5 days/wk for 40 wk, did not induce a significant decrease in body weight in obese Zucker rats. The plasma insulin concentration, showing a significant increase compared with lean Zucker rats, was unaffected by exercise training. However, the plasma triglyceride concentration in obese Zucker rats was significantly depressed by exercise training, whereas it was still higher than that in lean Zucker rats. In addition, natural killer cell activity and concanavalin A-induced mitogenesis of splenic lymphocytes of obese Zucker rats were significantly restored. In these splenic lymphocytes, glucose uptake was significantly lower compared with that in lean Zucker rats, which was also improved by exercise training. Although the expression of GLUT-1, the major glucose transporter in immune cells, was depressed in splenic lymphocytes of obese Zucker rats, exercise training induced a significant improvement. These results suggest that exercise training has a beneficial effect on the decreased cellular immune functions in obese Zucker rats, which is associated, in part, with the improvement in GLUT-1 expression.

Animals↗

Role of cyclic adenosine monophosphate in reducing superoxide anion generation in guinea pig alveolar macrophages.

Aveolar macrophages (AM) may contribute to airway inflammation via release of superoxide anion (O2-). Elevation of intracellular cyclic adenosine monophosphate (cAMP) levels has been shown to suppress O2- generation, although the exact mechanism is uncertain. To examine the inhibitory effect of cAMP against different stimuli for O2- generation, we compared the effect of cAMP on O2- generation caused by phorbol myristate acetate (PMA), a direct protein kinase C activator, and N-formyl-methionyl-leucyl-phenylalanine (FMLP), which couples its membrane receptor and stimulates guanosine triphosphate binding protein, in guinea pig AM. Cyclic nucleotide phosphodiesterase (PDE) isoenzyme inhibitors or CPT-cAMP, a cAMP analogue, were used in order to increase intracellular cAMP levels. The O2- generation caused by either PMA or FMLP was reduced by cilostazol (PDE 3 inhibitor) and Ro20-1724 (PDE 4 inhibitor), but not by zaprinast (PDE 5 inhibitor). The degree of reduction in O2- generation was not different between PMA and FMLP. Furthermore, CPT-cAMP also reduced PMA- or FMLP-induced O2- generation to a similar degree, although only high concentrations (10(-5) or 10(-4) mol/l) of this agent were effective in producing significant inhibition. A remarkable elevation of the cAMP level is required to produce the inhibitory effect on O2- generation in guinea pig AM. An elevation of cAMP may suppress O2- generation by inhibiting plural sites of the intracellular signaling pathways.

Animals↗

Signal transduction in activation of human eosinophils: G protein-dependent and -independent pathways.

Degranulation of eosinophils and subsequent release of toxic granule proteins play a key role in allergic diseases such as bronchial asthma. We have previously shown that stimulation of eosinophils with immobilized secretory immunoglobulin A (sIgA) induced the phosphorylation of several proteins including 51-, 65-, 73-, 78-, 100-, 105- and 113-kD proteins. Pervanadate, a protein tyrosine phosphatase inhibitor, also induced at least 7 tyrosine phosphorylated proteins including those observed with immobilized sIgA. Pervanadate also induced inositol phosphate (IP) production and degranulation of eosinophils in a concentration-dependent manner. Eosinophil production of IP and degranulation as well as tyrosine phosphorylation of proteins induced by sIgA were completely inhibited by a tyrosine kinase inhibitor, genistein, and pertussis toxin (PTX), suggesting the involvement of both the PTX-sensitive guanine nucleotide-binding (G) protein and protein tyrosine kinases (PTK) in sIgA-induced activation of eosinophils. In contrast, PTX did not affect tyrosine phosphorylation induced by sIgA or pervanadate. Furthermore, pervanadate-induced IP production was partially inhibited by PTX. Finally, a phospholipase C-gamma2 isoform was tyrosine phosphorylated by pervanadate, but not by sIgA. These findings suggest that at least two different pathways, i.e. PTK-mediated G protein-dependent or -independent PLC activation, are involved in the activation of human eosinophils.

Asthma↗

Cross-linking of the beta2 integrin, CD11b/CD18, on human eosinophils induces protein tyrosine phosphorylation and cellular degranulation.

Adhesion molecules, including integrins, play an important role in the selective recruitment of eosinophils. It has recently been shown that integrins also modulate the functions of eosinophils. Here, we tested the hypothesis that cross-linking of the beta2 integrin, alphaMbeta2Mac-/, leads to intracellular signaling events such as activation of protein tyrosine kinases leading to eosinophil degranulation. Cross-of cell surface CD11b/with anti-tibody and goat anti-G immobilized onto the plate triggered tyrosine phosphorylation of several intracellular proteins, including the one with a 115-kD mass (pp115). The same stimulus also provoked degranulation of eosinophils. These findings suggest that engagement of beta2 integrin on eosinophils triggers the activation of intracellular signaling cascade which leads to cellular degranulation.

Antibodies↗

Effects of pituitary adenylate-cyclase-activating polypeptide on the direct-current electroretinogram of the rabbit eye.

Pituitary adenylate-cyclase-activating polypeptide (PACAP) is a recently discovered neuropeptide present in two different forms, PACAP-27 and PACAP-38. Both peptides stimulate the catalytic enzyme adenylate cyclase in pituitary cells. This enzyme is important also regarding the function of the retinal pigment epithelium (RPE). The purpose of the study was to investigate possible influences of PACAP on the rabbit retina and the RPE as reflected in the direct-current (d.c.) electroretinogram (ERG) and the standing potential of the eye (SP). After unilateral sector vitrectomy, a continuous intraocular perfusion with a reference solution alternated with a test solution was established. The corneal d.c. ERG and the SP were recorded from both eyes with the contralateral eye as a control. Both PACAP-27 (0.1 and 1 microM) and PACAP-38 (1 microM) increased the c-wave amplitude significantly (p = 0.028, p = 0.013 and p = 0.024, respectively, n = 4) while neither PACAP-27 (0.1 and 1 microM, p > 0.05, n = 4) nor PACAP-38 (1 microM, p > 0.05, n = 4) produced any significant effects on the a- and b-wave amplitudes of the d.c. ERG. The SP response to the two substances differed with a significant elevation of the SP level with PACAP-27 (1 microM, p = 0.017, n = 4), while PACAP-38 induced a small, nonsignificant SP elevation (1 microM, p > 0.05, n = 4). Retinal penetrations during PACAP-27 (10 microM) perfusion showed an increase in transepithelial potential (TEP) c-wave (p = 0.003) as well as in slow PIII (p = 0.011, n = 3) amplitude level. The results support the presence of PACAP receptors both on the RPE and in the neural retina.

Animals↗

20-Hz flicker stimulus can isolate the cone function in rat retina.

Cone electroretinograms (ERGs) are typically isolated in humans by flicker stimuli against rod-desensitizing adapting fields. To investigate the manner in which adapting-field luminance affects the cone ERGs, we recorded ERGs in normal albino Sprague-Dawley rats with flicker stimuli presented against adapting fields that ranged in luminance from to 1.75 log cd/m2. A flicker rate of 20 Hz was used to isolate the cone ERGs under all adaptation conditions. We found the amplitudes of cone ERGs to increase with increasing adapting-field luminance. These response characteristics are similar to human ERGs using 30-Hz flicker stimuli, which suggests that flicker stimuli are a useful technique to isolate the cone function in rats.

Adaptation, Ocular↗

Downregulation of angiotensin II type 1 receptor gene transcription by nitric oxide.

Nitric oxide (NO) plays an important role not only in the regulation of blood vessel tone, but also in the growth of vascular smooth muscle cells (VSMC). The precise mechanism involved in the inhibition of VSMC growth by NO is not known. To further explore the effect of NO on VSMC growth, we examined the effect of NO on the expression of angiotensin II type 1 receptor (AT1-R) that is important for hypertrophy and hyperplasia of VSMC. S-nitroso acetyl DL-penicillamine (SNAP; 200 micromol/L), a potent NO donor, suppressed expression level of AT1-R mRNA by 90% and AT1-R number by 60% after 24 hours of stimulation. The suppressive effect was dose-dependent. Actinomycin D, which is an inhibitor of gene transcription, did not affect the decrease of AT1-R mRNA by NO. Cyclic guanosine monophosphate (cGMP) analogue, 8 bromo-cGMP, did not affect AT1-R mRNA level. Deletion mutants of the promoter region of rat AT1a-R gene were fused to luciferase reporter gene and introduced to VSMC. Transfected cells were stimulated with SNAP, and luciferase activity was measured. Inhibitory effect of NO was still observed in the shortest deletion mutant that contained 61 bp upstream from transcription start site. In this DNA segment, two DNA binding protein were observed by gel mobility shift assay, and one of these binding proteins was decreased on stimulation by NO. NO downregulates AT1-R gene expression independently of cGMP. A DNA binding protein that binds to the proximal promoter region of AT1-R gene may be responsible for this inhibitory effect. The inhibition of AT1-R gene expression may be implicated in the anti-atherogenic property of NO.

Angiotensin II↗

Baroreflex control of sympathetic nerve activity and heart rate in obstructive sleep apnea.

-Patients with obstructive sleep apnea are at increased risk for hypertension. The mechanisms underlying this increased risk are not known. We tested the hypothesis that obstructive sleep apnea, independent of factors such as hypertension, obesity, and age, is characterized by impairment of baroreflex sensitivity. We measured muscle sympathetic nerve activity (MSNA) and heart rate responses to activation and deactivation of baroreceptors in newly diagnosed, never treated, normotensive patients with obstructive sleep apnea. These responses were compared with those obtained in healthy control subjects closely matched for age, body mass index, and blood pressure. Heart rate and MSNA changes during infusion of phenylephrine (baroreceptor activation) were similar in the control subjects and patients with sleep apnea. Infusion of nitroprusside (baroreceptor deactivation) elicited similar decreases in mean arterial pressure (MAP) but lesser MSNA increases in patients with sleep apnea than in control subjects. Calculation of DeltaMSNA/DeltaMAP ratio revealed that baroreflex regulation of sympathetic activity for similar blood pressure changes was diminished in patients with sleep apnea in comparison to normal control subjects (P=0.01). However, increases in heart rate during nitroprusside infusion were comparable in both groups. Sympathetic, blood pressure and heart rate responses to the cold pressor test were also similar in the 2 groups. Our results indicate that normotensive patients with sleep apnea have a selective impairment of the sympathetic response to baroreceptor deactivation but not to baroreceptor activation or to the cold pressor test. The impairment of baroreflex sympathetic modulation in patients with sleep apnea is not accompanied by any impairment of baroreflex control of heart rate.

Airway Obstruction↗

Minimized platelet interaction with poly(2-hydroxyethyl methacrylate-block-4-bis(trimethylsilyl)methylstyrene) hydrogel showing anomalously high free water content.

Novel hydrophilic-hydrophobic block copolymers - poly(2-hydroxyethyl methacrylate-block-4-bis(trimethylsilyl)methylstyrene) (poly(HEMA-block-BSMS) or BH polymer) - were prepared as materials potentially useful for blood-contacting devices. Among the BH(X) series including poly(HEMA) itself, platelet adhesion and activation are minimum on the surface of BH(10), the block copolymer composed of 10 mol% of the hydrophobic segment. The unique physicochemical characteristics of BH(10) such as the high free water content and the increased mobility of PHEMA segments seem to play an important role in the prevention of platelet adhesion and activation. The water content of BH(10) was twice that of poly(HEMA) itself, even in the presence of 10 mol% of the hydrophobic moiety [poly(BSMS)]. Further, an anomalous increase in free water content was observed for the BH(10) sample measured by DSC. The glass transition temperature of BH(10) in the dry state was determined to be c. 10 degrees C, which was much lower than that observed for each of the homopolymers (c.f.: Tg [poly(HEMA)] = 70 degrees C; Tg [poly(BSMS)] = 160 degrees C), suggesting an increased mobility of tethered PHEMA segments in the BH(10) film.

Acrylic Resins↗

Ligation of the beta2 integrin triggers activation and degranulation of human eosinophils.

Evidence suggests that cellular adhesion is critical for eosinophil effector functions. Here, we tested the hypothesis that an adhesion molecule, specifically beta2 integrin, participates in intracellular signaling events of eosinophils. Eosinophils stimulated with interleukin (IL)-5 and adherent to protein-coated tissue culture plates via beta2 integrin (CD18) showed tyrosine phosphorylation of a number of proteins. Among these proteins, tyrosine phosphorylation of the 105 kD and 115 kD proteins and the product of the c-cbl protooncogene, Cbl, was specifically inhibited using soluble anti-CD18 monoclonal antibody (mAb) to block eosinophil cell adhesion. Furthermore, phosphoinositide turnover of IL-5-stimulated adherent eosinophils was also inhibited by anti-CD18 mAb, suggesting that cellular adhesion plays important roles in eosinophil signal transduction. alphaM beta2 (Mac-1, CD11b/18) was one of the beta2 integrins involved in eosinophil adhesion to protein-coated plates. We found that direct ligation of eosinophil alphaM beta2 with anti-CD11b mAb coupled to polystyrene microbeads induced tyrosine phosphorylation of a 115 kD protein and Cbl. Furthermore, anti-CD11b mAb microbeads induced increases in both phosphoinositide hydrolysis and the eosinophil degranulation response. Control antibodies, such as mouse myeloma IgG1 and anti-HLA class I antigen mAb, did not induce these cellular responses. These results suggest that engagement of beta2 integrin either by cell adhesion or by anti-CD11b mAb triggers activation of an intracellular signaling cascade, including protein tyrosine phosphorylation and phosphoinositide turnover, and subsequent cellular degranulation in human eosinophils. Tyrosine phosphorylation of a 115 kD protein and Cbl may play important roles in adhesion-dependent cellular functions of eosinophils.

Antibodies, Monoclonal↗

Evidence of potential regulation by interleukin-4 of the soluble intercellular adhesion molecule 1 level in patients with seasonal allergic rhinitis under provocation by a small amount of natural allergen.

Interleukin-4 (IL-4) and intercellular adhesion molecule 1 (ICAM-1) are assumed to be involved in the pathogenesis of allergic disease. In this study, we examined the potential link between IL-4 and soluble ICAM-1 (sICAM-1) levels in patients with allergic rhinitis. The levels of sICAM-1 and IL-4 in sera and in nasal epithelial lining fluids (ELF) from 12 patients with Japanese cedar pollinosis were measured preseason through postseason, and the results were compared with those from 7 healthy subjects. In sera from the allergic subjects, the levels of sICAM- 1 were upregulated during the early part of the season and downregulated during the middle of the season, with upregulation of the IL-4 levels. Moreover, a negative correlation was found between the serum sICAM-1 levels and serum IL-4 levels during the middle (r = -.80) and late (r = -.73) parts of the season. In ELF from allergic subjects, the levels of sICAM-1 were significantly upregulated during the early and middle parts of the season, and began to be downregulated during the late part of the season, with upregulation of the levels of IL-4. In conclusion, IL-4 possibly acts as a potential suppressor of sICAM-1 in the pathogenesis of seasonal allergic rhinitis, at least under provocation by a small amount of natural allergen.

Adult↗