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Biomedical subjects

T Masuda

Publications and source records attributed to T Masuda.

At least 37 records · Page 2Linked to original sources

A novel plant ferritin subunit from soybean that is related to a mechanism in iron release.

Ferritin is a multimeric iron storage protein composed of 24 subunits. Ferritin purified from dried soybean seed resolves into two peptides of 26.5 and 28 kDa. To date, the 26.5-kDa subunit has been supposed to be generated from the 28-kDa subunit by cleavage of the N-terminal region. We performed amino acid sequence analysis of the 28-kDa subunit and found that it had a different sequence from the 26.5-kDa subunit, thus rendering it novel among known soybean ferritins. We cloned a cDNA encoding this novel subunit from 10-day-old seedlings, each of which contained developed bifoliates, an epicotyl and a terminal bud. The 26.5-kDa subunit was found to be identical to that identified previously lacking the C-terminal 16 residues that correspond to the E helix of mammalian ferritin. However, the corresponding region in the 28-kDa soybean ferritin subunit identified in this study was not susceptible to cleavage. We present evidence that the two different ferritin subunits in soybean dry seeds show differential sensitivity to protease digestions and that the novel, uncleaved 28-kDa ferritin subunit appears to stabilize the ferritin shell by co-existing with the cleaved 26.5-kDa subunit. These data demonstrate that soybean ferritin is composed of at least two different subunits, which have cooperative functional roles in soybean seeds.

Amino Acid Sequence↗

Effects of a kappa agonist, spiradoline mesylate (U62,066E), on activation and vaginocervical-stimulation produced analgesia in rats.

Previous research has demonstrated increased pain threshold during copulation, gestation, and parturition in animals. In the laboratory, mechanostimulation of the vaginocervical region in many animals, as well as humans, can increase responsiveness to noxious but not to innocuous stimuli. This increased pain inhibition to vaginocervical stimulation, which mimics natural parturition, is mediated by spinal and supraspinal neuropeptides, including the opiates. The present research was designed to ascertain the possible effects of a kappa opioid agonist on vaginocervical-stimulated analgesia in rats. Initially, the novel kappa-selective agonist, spiradoline mesylate (U62,066E; 0, 0.1, 1.0, 10.0 mg/kg, i.p.), was injected intraperitoneally and general behavioral arousal in an open field apparatus was recorded. Results from this experiment indicate that stimulation with the kappa-selective drug caused significant decreases in behavioral activity at the high dose as compared to saline and the medium and low doses. Next, the effects of U62,066E (0, 0.1, 1.0, 10.0 mg/kg, i.p.) on the analgesia associated with vaginocervical stimulation were determined in a tail flick apparatus. The kappa drug significantly increased antinociceptive thresholds prior to and during vaginocervical stimulation at the 0.1 and 1.0 mg/kg doses. By contrast, the high dose (10.0 mg/kg) of U62,066E decreased vaginocervical stimulation-produced analgesia. Results are discussed in terms of the potential of nonaddictive kappa-selective opioid compounds being utilized in reproductive pain.

Analgesics↗

The effect of myocardial bridging of the coronary artery on vasoactive agents and atherosclerosis localization.

The relationship between alterations in the immunohistochemical expression of three vasoactive agents [endothelial nitric oxide synthase (eNOS), endothelin-1 (ET-1), and angiotensin-converting enzyme (ACE)] and the occurrence human atherosclerosis was investigated in relation to the myocardial bridge (MB) of the left anterior descending coronary artery (LAD), an anatomical site that experiences increased shear stress. Five millimetre cross-sections of LADs with MB from 22 autopsied cases were taken from the left coronary ostium to the cardiac apex and were immunohistochemically stained with antibodies against eNOS, ET-1, and ACE. The extent of atherosclerosis in each section was calculated using the atherosclerosis ratio (intimal cross-sectional area/medial cross-sectional area) determined by histomorphometry. The results were analysed according to their anatomical location relative to the MB, either proximal, beneath, or distal. The extent of atherosclerosis was significantly lower beneath the MB, compared with proximal and distal segments. The expression of eNOS, ET-1, and ACE was also significantly lower beneath the MB. The expression of these agents correlated significantly with the extent of atherosclerosis. Because nitric oxide, after its production by eNOS, is believed to be degraded by superoxide radicals, the effect of eNOS expression on atherosclerosis remains controversial. However, the present findings clearly indicate that the expression of ET-1 and ACE is directly related to the development of human coronary atherosclerosis in vivo through shear stress.

Adult↗

Network structure of curdlan in DMSO and mixture of DMSO and water.

The viscoelastic property of curdlan solution in dimethyl sulfoxide (DMSO) was investigated. We discuss the difference in the viscoelastic properties of curdlan solution in DMSO and that in 0.1 N NaOH aqueous solution. The viscoelastic function for curdlan solution in 0.1 N NaOH aqueous solution showed the Newtonian flow at the concentration of curdlan as high as 10 wt %. On the other hand, the Newtonian flow was observed in the concentrations below 7 wt % for curdlan solution in DMSO, and the plateau region appeared beyond this concentration. It was revealed by small angle x-ray diffraction measurements that the difference in the mechanical property would be originated from the difference in the network structure. The overlapping concentration c* was calculated on the basis of the mean field theory, and disagreement between theoretical prediction and experimental result was shown. We clarified that the above disagreement can be explained by the polydispersity of the curdlan sample, assuming adequate distribution functions. The static structure of the gel prepared by adding water to curdlan solution in DMSO was investigated. It was clarified by the dynamic viscoelasticity measurement that the cross-linking density increases with increasing the water content.

Dimethyl Sulfoxide↗

Potential immunosuppressive and antiinflammatory activities of Malaysian medicinal plants characterized by reduced cell surface expression of cell adhesion molecules.

In the search for agents effective against immune-mediated disorders and inflammation, we have screened Malaysian medicinal plants for the ability to inhibit the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on the surface of murine endothelial cells (F-2), and mouse myeloid leukaemia cells (M1), respectively. Of 41 kinds (29 species, 24 genera, 16 families) of Malaysian plants tested, 10 and 19 plant samples significantly downregulated the expression of ICAM-1 and VCAM-1, respectively. Bioassay-directed fractionation of an extract prepared from the bark of Goniothalamus andersonii showed that its ingredients, goniothalamin (1) and goniodiol (2) inhibited the cell surface expression of both ICAM-1 and VCAM-1. The present results suggest that Malaysian medicinal plants may be abundant natural resources for immunosuppressive and antiinflammatory substances.

Animals↗

Complete suppression of insulitis and diabetes in NOD mice lacking interferon regulatory factor-1.

Interferon regulatory factor-1 (IRF-1), a transcriptional factor, regulates type I interferon and interferon-induced genes. It was reported that IRF-1 regulates important molecules required for inflammation and immune reactions. To investigate the role of IRF-1 in the development of autoimmune diabetes, we established IRF-1 deficient (IRF-1(-/-)) non-obese diabetic (NOD) mice. IRF-1-deficient C57BL/6J mice were out-crossed to NOD mice, and F1 were backcrossed to NOD mice. At the N8 generation, the heterozygote for IRF-1 mutation was intercrossed and N8F1 was obtained. Out of three NOD genotypes, IRF-1(+/+) and IRF-1(+/-) developed spontaneous diabetes with an incidence of 47% (9/19) and 50% (10/20) by 30 weeks of age, respectively; whereas IRF-1(-/-) did not develop diabetes (0/18, P< 0.01 vs. (+/+) and (+/-)). Histologically, IRF-1(+/+) and IRF-1(+/-) had various degrees of insulitis, but IRF-1(-/-) had no insulitis. In comparison with IRF-1(+/+), the percentage of CD4(+) and Mac-1(+) splenic cells significantly increased, whereas CD3(+), CD8(+) and B220(+) cells decreased in IRF-1(-/-). Furthermore, spleen cell proliferation in response to Con A or murine GAD65 peptide, a major autoantigen of the pancreatic beta-cell, significantly increased, and the IFN-gamma/IL-10 ratio in the culture supernatant significantly decreased in IRF-1(-/-), suggesting Th2 deviation in cytokine balance. These results indicate that IRF-1 plays a key role in developing insulitis and diabetes in NOD mice.

Animals↗

The effect of allergen-induced airway inflammation on airway remodeling in a murine model of allergic asthma.

OBJECTIVE AND DESIGN: We examined the effect of airway inflammation on airway remodeling and bronchial responsiveness in a mouse model of allergic asthma. MATERIALS AND METHODS: BALB/c mice were sensitized to ovalbumin (OA), and exposed to aerosolized OA (0.01, 0.1 and 1%). Twenty-four hours after the final antigen challenge, bronchial responsiveness was measured, and bronchoalveolar lavage (BAL) and histological examinations were carried out. RESULTS: Repeated antigen exposure induced airway inflammation, IgE/IgG1 responses, epithelial changes, collagen deposition in the lungs, subepithelial fibrosis associated with increases in the amount of transforming growth factor (TGF)-beta1 in BAL fluid (BALF), and bronchial hyperresponsiveness to acetylcholine. The number of eosinophils in BALF was significantly correlated with TGF-beta1 production in BALF and the amount of hydroxyproline. Furthermore, significant correlations were found between these fibrogenic parameters and the bronchial responsiveness. CONCLUSION: These findings demonstrated that in this murine model airway eosinophilic inflammation is responsible for the development of airway remodeling as well as bronchial hyperresponsiveness in allergic bronchial asthma.

Acetylcholine↗

The distribution and production of cholesteryl ester transfer protein in the human aortic wall.

Cholesteryl ester transfer protein (CETP) has been considered to mediate the transfer of cholesteryl ester from arterial wall, however, the distribution and production of CETP in human arterial wall remains unclear. Present study histopathologically demonstrated the distribution of CETP and CETP mRNA in the human aortic wall by immunohistochemistry and in situ hybridization. While CETP was constantly distributed in the media, the protein was recognized within the intima with fibrocellular thickening and atherosclerotic intima. Double immunostaining methods demonstrated CETP expression in smooth muscle cells in the intima and media. CETP mRNA was detected not only in intimal cells but medial smooth muscle cells. Intimal cells expressing CETP mRNA were considered to be monocyte-derived macrophages and smooth muscle cells by immunohistochemistries using two antibodies against smooth muscle actin and human macrophage on the subserial sections. Our in vivo study provides that CETP is produced by smooth muscle cells in the intima and media of human aorta, and it is suggested that arterial smooth muscle cells positively participate in the removal of excessive cholesteryl ester from the arterial wall by CETP production.

Aorta↗

Immunolocalization of apolipoproteins in aortic atherosclerosis in American youths and young adults: findings from the PDAY study.

The immunohistochemical distribution of apolipoproteins in the abdominal aortas of 142 men, 15-34 years of age, collected in a cooperative multicenter study group (Pathobiological Determinants of Atherosclerosis in Youth) was examined in relationship to serum VLDL+LDL+HDL cholesterol levels. ApoB deposits were limited to the intima of specimens with intimal fibro cellular thickening or atherosclerotic lesions. Apo A-I, E and J were observed in both the intima and media of the aortas with intimal lesions. The pattern of apoJ distribution was similar to that of apoA-I and E. The distribution patterns of these apolipoproteins in these young adults were very similar to those in adults and old men seen in an earlier study. The extent of apolipoprotein distribution in the intima and media increased with age and the stage of atherosclerosis development, but was not correlated significantly with serum VLDL+LDL or HDL cholesterol levels. The infiltration of lipoprotein particles into the aortic wall seems to be more strongly associated with the progression of intimal lesions rather than with serum cholesterol levels.

Adolescent↗

Mechanical properties of artificial tracheas composed of a mesh cylinder and a spiral stent.

Much work has been done on the materials used for mesh-type artificial tracheas, but a precise mechanical evaluation of these structures has not yet been performed. In the present study, we determined the mechanical properties of typical mesh-type artificial tracheas and compared them with those of native trachea. Four types of artificial trachea were made and used for the mechanical tests. The basic frame of all the specimens was composed of a mesh cylinder and a spiral stent. The specimen whose mesh was sealed with collagen sponge showed almost the same behavior in the force-strain curve under compression, suggesting that collagen sealing has little effect on mechanical properties. Agreement between measured and estimated mechanical properties was good, especially in the low strain region, suggesting that artificial tracheas can be designed in terms of mechanical properties by mainly considering the basic frame structure.

Animals↗

A new dysferlin gene mutation in two Japanese families with limb-girdle muscular dystrophy 2B and Miyoshi myopathy.

We found a new dysferlin gene mutation in two Japanese families, one with limb-girdle muscular dystrophy 2B and the other with Miyoshi myopathy. All patients in the limb-girdle muscular dystrophy 2B family showed apparent proximal dominant muscle atrophy and weakness, whereas a patient with Miyoshi myopathy in the second family showed distal muscle involvement at an early stage. The common clinical feature of all patients in both families was preferential involvement of calf muscles rather than the tibialis anterior muscle, which was confirmed by muscle computed tomography scan. All patients in both families shared the same homozygous alleles for chromosome 2p13 markers, and dysferlin gene analysis revealed a novel missense mutation, a G to A transition at nt 5882, which changed aspartic acid to asparagine at codon 1837. Allele-specific polymerase chain reaction analysis was used for confirmation of the mutation and for genotype analysis of the family members.

Aged↗

Influence of contraction force and speed on muscle fiber conduction velocity during dynamic voluntary exercise.

Before using electromyographic (EMG) variables such as muscle fiber conduction velocity (MFCV) and the mean or median frequency (MDF) of an EMG power spectrum as indicators of muscular fatigue during dynamic exercises, it is necessary to determine the influence of a joint angle, contraction force and contraction speed on the EMG variables. If these factors affect the EMG variables, their influence must be removed or compensated for before discussing fatigue. The vastus lateralis of eight normal healthy male adults was studied. EMG signals during non-fatiguing dynamic knee extension exercises were detected with a three-bar active surface electrode array. EMG variables were calculated from the detected signals and compared with the angle of the knee joint, the extension torque and the extension speed. The extension torque was set at four levels with 10% intervals between 40 and 70% of the maximum voluntary contraction. The extension speed was set at five levels with 60 degrees /s intervals between 0 and 240 degrees /s. Because the joint angle unsystematically affected the MFCV, EMG variables at a given joint angle were extracted for comparison. The influence of the extension torque and speed on the extracted EMG variables was clarified with an ANOVA and a regression analysis. The statistical analyses showed that MFCV increased with the extension torque but did not depend on the extension speed. In contrast, MDF was independent of the extension torque but was dependent on the extension speed. MDF thus showed a behavior different from that of MFCV. It became clear that if MFCV is used as an indicator of muscular fatigue during dynamic exercises, it is at least necessary to extract MFCV at a predetermined joint angle and then remove the influence of extension torque on MFCV.

Action Potentials↗

Immunoreactivity to monoclonal antibody, Hep Par 1, in human hepatocellular carcinomas according to histopathological grade and histological pattern.

We evaluated the immunoreactivity to monoclonal antibody for hepatocyte (Hep Par 1) and determined the cellular distribution of the antigen in hepatocellular carcinoma (HCC), based on the histopathological grade and histological pattern. The pathological material included 100 areas selected at random in 61 tissue sections from 12 autopsy livers with HCC. Immunoreactivity was classified into four categories; strongly positive (>90% of the area showed positive staining), moderately positive (5-90% of the area), weakly positive (<5% of the area), and negative (completely negative). All 19 (100%) well-differentiated, trabecular type HCC areas were strongly positive for Hep Par 1. Among 11 well-differentiated, pseudoglandular type HCC areas, 2 (18%) were strongly positive, 5 (46%) were moderately positive, 2 (18%) were weakly positive, and 2 (18%) were negative. Among 36 moderately differentiated, trabecular type HCC areas, 6 (17%) were strongly positive, 17 (47%) were moderately positive, 9 (25%) were weakly positive, and 4 (11%) were negative. None of the four moderately differentiated, pseudoglandular type HCC areas were strongly positive, 3 (75%) were moderately positive, 0 was weakly positive, and 1 (25%) was negative. Among 25 poorly differentiated, compact or trabecular type HCC areas, 15 (60%) were weakly positive and 10 (40%) were negative. All 5 (100%) undifferentiated HCC areas were negative for Hep Par 1. Our results indicate that immunoreactivity to Hep Par 1 in HCC decreases with reduced differentiation of the tumor, suggesting that Hep Par 1 monoclonal antibody is useful as a marker for the diagnosis and differentiation of HCCs.

Journal Article↗

Chemical studies on antioxidant mechanism of curcumin: analysis of oxidative coupling products from curcumin and linoleate.

As a part of a research project on the antioxidant mechanism of natural phenolics in food components, curcumin, a turmeric antioxidant, was investigated in the presence of ethyl linoleate as one of the polyunsaturated lipids. During the antioxidation process, curcumin reacted with four types of linoleate peroxyl radicals. Six reaction products were observed in the reaction and subsequently isolated. Their structures were determined by physical techniques, revealing that they have novel tricyclic structures, including a peroxyl linkage. On the basis of the formation pathway for their chemical structures, an antioxidant mechanism of curcumin in polyunsaturated lipids was proposed, which consisted of an oxidative coupling reaction at the 3'-position of the curcumin with the lipid and a subsequent intramolecular Diels--Alder reaction.

Antioxidants↗

Sweetness and enzymatic activity of lysozyme.

Hen egg lysozyme elicits a sweet taste sensation for human beings. Effects of reduction of disulfide bonds, heat treatment, and chemical modification of hen egg lysozyme on both sweetness and hydrolytic activity were investigated. Both the sweetness and enzymatic activities were lost when the intradisulfide linkage in a lysozyme molecule was reduced and S-3-(trimethylated amino) propylated. The sweetness and enzymatic activity of lysozyme were lost on heating at 95 degrees C for 18 h. These facts suggest that tertiary structures of lysozyme are indispensable for eliciting a sweet taste as well as enzymatic activity. Although the modification of carboxyl residues in a lysozyme by glycine methylester or aminomethansulfonic acid resulted in the loss of enzymatic activity by blocking the catalytic residues, the sweetness was fully retained. These results indicate that the sweetness of lysozyme was independent of its enzymatic activity. The lysozyme purified from goose egg white similarly elicited a sweet taste, although goose (g-type) lysozyme is quite different from hen egg lysozyme (c-type) on the basis of structural, immunological, and enzymatic properties. These findings indicate that a specific protein property of lysozyme is required for sweetness elicitation and that the enzymatic activity and carbohydrates produced by enzymatic reaction are not related to the sweet taste.

Animals↗

Antioxidant mechanism of carnosic acid: structural identification of two oxidation products.

To determine the antioxidant mechanism of food phenolics against the oxidation of food components, the reaction of carnosic acid, an antioxidative constituent of the popular herbs sage and rosemary, was investigated in the presence of ethyl linoleate and the radical oxidation initiator 2,2'-azobis(2,4-dimethylvaleronitrile). During this process, carnosic acid was oxidized to an o-quinone and a hydroxy p-quinone, the chemical structures of which were confirmed by physical and chemical techniques. From a quantitative time course analysis of the production of these quinones, an antioxidant mechanism of carnosic acid is proposed, consisting of the oxidative coupling reaction with the peroxyl radical at the 12- or 14-position of carnosic acid and subsequent degradation reactions.

Abietanes↗

Attending holistically versus analytically: comparing the context sensitivity of Japanese and Americans.

Much research indicates that East Asians, more than Americans, explain events with reference to the context. The authors examined whether East Asians also attend to the context more than Americans do. In Study 1, Japanese and Americans watched animated vignettes of underwater scenes and reported the contents. In a subsequent recognition test, they were shown previously seen objects as well as new objects, either in their original setting or in novel settings, and then were asked to judge whether they had seen the objects. Study 2 replicated the recognition task using photographs of wildlife. The results showed that the Japanese (a) made more statements about contextual information and relationships than Americans did and (b) recognized previously seen objects more accurately when they saw them in their original settings rather than in the novel settings, whereas this manipulation had relatively little effect on Americans.

Attention↗

Disruption of antigen-induced airway inflammation and airway hyper-responsiveness in low affinity neurotrophin receptor p75 gene deficient mice.

1. Recently, much attention has been paid to the relationship between the nervous and immune systems. The present study was conducted to clarify the role of neurotrophin low affinity receptor (p75N) in allergic airway inflammation and hyper-responsiveness (AHR) in mice by employing p75N gene deficient mice. 2. Mice were immunized twice by intraperitoneal injections of ovalbumin (OA) at intervals of 12 days. OA was inhaled 10 days after the secondary immunization and repeated three times at 4 days interval. Twenty-four hours after the last inhalation, airway responsiveness to acetylcholine was measured and bronchoalveolar lavage fluid (BALF) was obtained for examining the number of inflammatory cells and the level of cytokines. Serum immunoglobulin was measured as a marker of systemic immune response before the final inhalation. 3. In wild-type mice, repeated antigen provocation resulted in airway eosinophilia, AHR and elevations in serum IgE and interleukin (IL)-4 and -5 in BALF. In p75N gene deficient mice, none of the above parameters was observed after antigen provocation. The antigen-induced production of interferon (IFN)-gamma and nerve growth factor (NGF) were not altered by depletion of p75N gene. 4. The present findings suggest that p75 gene deficiency disrupt an allergic airway inflammation and AHR in mice by interfering type 2 helper T (Th2) cell responses.

Acetylcholine↗