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Biomedical subjects

M Matsuda

Publications and source records attributed to M Matsuda.

At least 145 records · Page 8Linked to original sources

Relationship between mitochondrial DNA polymorphism and the individual differences in aerobic performance.

This study focused on the mitochondrial DNA (mtDNA) as the genetic factor most likely to bring about the individual difference in endurance capacity or its trainability. Platelets contain mtDNA but no nuclear DNA, whereas rho(0)-HeLa cells have nuclear DNA but no mtDNA. The oxidative capacity of mitochondria in the cultured cells, which were fused rho(0)-HeLa cell with platelets obtained from individual subjects (the so-called "cybrids"), reflects the individual mtDNA polymorphism in the gene-coding region. The purpose of this study was to investigate the relationship between the oxidative capacity of cybrids and the individual difference in endurance capacity, or its trainability. Forty-one sedentary young males took part in an 8-week endurance training program. They were determined by using their VO(2 max) as an index of endurance capacity on an ergocycle before and after the endurance training program. The relations between VO(2 max) before endurance training or the change of it by endurance training and the oxidative capacity of cybrids were investigated. There was no relation between them, and two groups were drawn from all subjects, based on one standard division of their initial VO(2 max): the higher pre-VO(2 max) group (n = 6) and the lower pre-VO(2 max) group (n = 5) (51.8 +/- 3.5 ml/min/kg vs. 33.3 +/- 3.8 ml/min/kg, p < 0.01). No significant difference was found between the O(2) consumption of the cybrids in the higher initial VO(2 max) group and that in the lower initial VO(2 max) group (16.3 +/- 4.9 vs. 15.9 +/- 2.0 nmol O(2)/min/10(7) cells, NS). Furthermore, neither the cytochrome c oxidase (COX) activity nor the complex I + III activity of cybrids showed a significant difference between the two groups. The oxidative capacity of cybrids between the high trainability group (n = 6) (Delta VO(2 max) 12.1 +/- 1.6 ml/min/kg) and the low trainability group (n = 9) (Delta VO(2 max) 2.3 +/- 0.5 ml/min/kg) was also similar. Thus the mtDNA polymorphism is very unlikely to relate to the individual difference in endurance capacity or its trainability in young sedentary healthy subjects.

Adult↗

PPARgamma ligands increase expression and plasma concentrations of adiponectin, an adipose-derived protein.

Insulin resistance and its dreaded consequence, type 2 diabetes, are major causes of atherosclerosis. Adiponectin is an adipose-specific plasma protein that possesses anti-atherogenic properties, such as the suppression of adhesion molecule expression in vascular endothelial cells and cytokine production from macrophages. Plasma adiponectin concentrations are decreased in obese and type 2 diabetic subjects with insulin resistance. A regimen that normalizes or increases the plasma adiponectin might prevent atherosclerosis in patients with insulin resistance. In this study, we demonstrate the inducing effects of thiazolidinediones (TZDs), which are synthetic PPARgamma ligands, on the expression and secretion of adiponectin in humans and rodents in vivo and in vitro. The administration of TZDs significantly increased the plasma adiponectin concentrations in insulin resistant humans and rodents without affecting their body weight. Adiponectin mRNA expression was normalized or increased by TZDs in the adipose tissues of obese mice. In cultured 3T3-L1 adipocytes, TZD derivatives enhanced the mRNA expression and secretion of adiponectin in a dose- and time-dependent manner. Furthermore, these effects were mediated through the activation of the promoter by the TZDs. On the other hand, TNF-alpha, which is produced more in an insulin-resistant condition, dose-dependently reduced the expression of adiponectin in adipocytes by suppressing its promoter activity. TZDs restored this inhibitory effect by TNF-alpha. TZDs might prevent atherosclerotic vascular disease in insulin-resistant patients by inducing the production of adiponectin through direct effect on its promoter and antagonizing the effect of TNF-alpha on the adiponectin promoter.

3T3 Cells↗

Improved glycemic control and enhanced insulin sensitivity in type 2 diabetic subjects treated with pioglitazone.

OBJECTIVE: To elucidate the effects of pioglitazone treatment on glucose and lipid metabolism in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: A total of 23 diabetic patients (age 30-70 years BMI < 36 kg/m2) who being treated with a stable dose of sulfonylurea were randomly assigned to receive either placebo (n = 11) or pioglitazone (45 mg/day) (n = 12) for 16 weeks. Before and after 16 weeks of treatment, all subjects received a 75-g oral glucose tolerance test (OGTT) and hepatic peripheral insulin sensitivity was measured with a two-step euglycemic insulin (40 and 160 mU x min(-1) x m(-2) clamp performed with 3-[3H]glucose and indirect calorimetry HbA1c measured monthly throughout the study period. RESULTS: After 16 weeks of pioglitazone treatment, the fasting plasma glucose (FPG; 184 +/- 15 to 135 +/- 11 mg/dl, P < 0.01), mean plasma glucose during OGTT(293 +/- 12 to 225 +/- 14 mg/dl, P < 0.01), and HbA1c (8.9 +/- 0.3 to 7.2 +/- 0.5%, P < 0.01 ) decreased significantly without change in fasting or glucose-stimulated insulin/C-peptide concentrations. Fasting plasma free fatty acid (FFA; 647 +/- 39 to 478 +/- 49) microEq/l, P < 0.01) and mean plasma FFA during OGTT (485 +/- 30 to 347 +/- 33 microEq/l, P < 0.01) decreased significantly after pioglitazone treatment. Before and after pioglitazone treatment, basal endogenous glucose prodution (EGP) and FPG were strongly correlated (r = 0.67, P < 0.01). EGP during the first insulin clamp step was significantly decreased after pioglitazone treatment (P < 0.05) whereas insulin-stimulated total and nonoxidative glucose disposal during the second insulin clamp was increased (P < 0.01). The change in FPG was related to the change in basal EGP, EGP during the first insulin clamp step, and total glucose disposal during the second insulin clamp step. The change in mean plasma glucose concentration during the OGGTT was strongly related to the change in total body glucose disposl during the second insulin clamp step. CONCLUSIONS: These results suggest that pioglitazone therapy in type 2 diabetic patients decreases lasting and postprandial plasma glucose levels by improving hepatic and peripheral (muscle) tissue sensitivity to insulin.

Adipose Tissue↗

[Trust and cooperation: an experimental study of PD with choice of dependence].

This study examined the relationship between cooperation and trust in interpersonal trust formation. Previous studies of trust and cooperation using prisoner's dilemma (PD) games failed, methodologically as well as conceptually, to distinguish the former from the latter. In response to the criticism on the use of iterated PD games, and for the purpose of investigating dynamic relationship between trust and cooperation, an improvement in research methodology was recently proposed: namely, PD with choice of dependence (PD/D; Kakiuchi & Yamagishi, 1997; Yamagishi & Kakiuchi, 2000). We conducted an experiment to compare formation of trust relations in PD and PD/D. Result indicated a higher level of cooperation in PD/D than in PD. Further analysis of strategies used to build trustful relationship, where two partners trusted each other and reciprocated the other's trusting behavior, revealed that participants in PD/D adopted "cautious and unconditional cooperation strategy" rather than TFT strategy.

Adult↗

PCR cloning of the resuscitation-promoting factor (Rpf) gene from Micrococcus luteus, sequencing and expression in Escherichia coli.

A polymerase chain reaction (PCR) cloning procedure was developed for the resuscitation-promoting factor (Rpf) gene of Micrococcus luteus using strains NCIMB 13267, JCM 1464T, JCM 3347, and JCM 3348. A PCR product of the Rpf gene fragment was ligated into a cloning vector pBluescript II KS (+) with the restriction endonucleases Eco RI and Bam HI. The ligation mixture was used to transform Escherichia coli DH5alpha. The DNA sequence of the Rpf gene cloned from strain JCM 1464T was 84% homologous with that of NCIMB 13267, and from strains JCM 3347 and JCM 3348 it was 100% and 86% homologous, respectively. Recombinant Rpf proteins of M. luteus NCIMB 13267 and JCM 1464T after expression in E. coli BL21 harbouring the pET-19b-Rpf plasmid, and after purification, were approximately 16 kD for both strains.

Amino Acid Sequence↗

Demonstration of genome DNA diversity of strains of urease-positive thermophilic Campylobacter isolated from the natural environment by pulsed-field gel electrophoresis analysis.

Pulsed-field gel electrophoresis (PFGE) analysis was carried out after separate digestion with Apa I, Sal I and Sma I of the genomic DNA from sixteen isolates of urease-positive thermophilic Campylobacter (UPTC) obtained from the natural environment, namely from oysters and mussels, in Northern Ireland. Five NCTC strains previously isolated in England were used for the analysis. Although the eight isolates of UPTC in Northern Ireland and a strain of UPTC in England showed that one or no fragments appeared after digestion with Apa I around 1,900 to 1,640 kb region of the gel, Apa I was shown to cut the genomic DNA from all of the other twelve strains of UPTC and Sal I and Sma I from all of the 21 strains in a distinctly different and distinguishable manner. Consequently, the present study clearly demonstrated that the sixteen isolates of UPTC in Northern Ireland and the five strains in England gave the diversity of the genomic DNA by using PFGE. Some strains of UPTC examined were shown to carry genomes from 1.6 to 1.9 Mb in length, thus the heterogeneous profiles of PFGE and the length of the genomes are thought to occur among the isolates of UPTC in Northern Ireland, as well as among the five representative strains of UPTC from NCTC.

Animals↗

[Calf flow reserve measured by venous-occlusion near-infrared spectroscopy: relationship to exercise tolerance].

OBJECTIVES: The correlations between calf flow reserve evaluated with venous-occlusion near-infrared spectroscopy, air plethysmography or skin laser Doppler flowmetry were investigated in patients with cardiovascular diseases. The relationships to exercise tolerance during upright bicycle exercise were also examined. METHODS: The calf flow reserve (blood flow after 5 min of arterial occlusion/basal blood flow) evaluated by the above three methods and peak oxygen uptake (peak VO2) on bicycle exercise were measured in 24 male patients (mean age 65.0 +/- 7.7 years, left ventricular ejection fraction 24-86%). RESULTS: There was a good correlation between flow reserve by near-infrared spectroscopy (8.0 +/- 3.8) and air plethysmography (7.9 +/- 3.1; r = 0.90). However, there was a weak correlation between flow reserve by near-infrared spectroscopy and skin laser Doppler flowmetry (3.4 +/- 1.7; r = 0.42). There was a good correlation between flow reserve by near-infrared spectroscopy and peak VO2 (r = 0.69), or flow reserve by air plethysmography and peak VO2 (r = 0.53). However, there was no significant correlation between flow reserve by skin laser Doppler flowmetry and peak VO2 (r = 0.18). CONCLUSIONS: Calf flow reserve evaluated by venous-occlusion near-infrared spectroscopy relates to the flow reserve in the muscle tissue and is minimally affected by the flow reserve in the skin. The flow reserve in the muscle tissue is related to exercise tolerance, but the flow reserve in the skin is not, in patients with cardiovascular diseases.

Aged↗

[Low dose intrathecal morphine and postoperative pain relief in elderly patients].

Patient (ASA PS I-III, mean age 68 +/- 14 yr) who had undergone lower extremity surgery under spinal anesthesia were studied to determine the effect of intrathecal administration of morphine 0.1 mg on intra- and postoperative pain relief and its side effects. They were randomly divided into control (C) and intrathecal morphine (M) groups (n = 25, respectively) and received 10 mg tetracaine in 4 ml of a quarter saline with 7.5 micrograms epinephrine. Incidence of intraoperative tourniquet pain was significantly lower in M group (36.8%) than in C group (64.3%). Postoperative pain was examined in terms of the duration until the first supplemental analgesic within 24 hr. The mean duration was 7.0 +/- 4.3 hr in the control group, but 11 patients in the M group needed it within 24 hr (18.1 +/- 6.8 hr, excluding 6 patients who did not receive analgesic). Although incidences of postoperative nausea, vomiting, and itching were higher in M group than in C group, none required antiemetic or naloxone. Both groups showed no difference in postoperative respiratory depression measured by apnea monitor (Eden Trace II, Mallinkrodt Japan, Tokyo). The results suggest that a low dose of intrathecal morphine is effective on postoperative 24 hr pain relief in elderly patients and that its side effects are negligible.

Aged↗

[Clock drawing in dementia: its reliability and relation to the neuropsychological measures].

To examine the reliability and validity of clock drawing (CD) for evaluating dementia patients in Japan, we investigated the CD performance and its relation to several neuropsychological tests in 150 demented patients including 105 patients with Alzheimer disease and 30 age- and education-matched non-demented subjects (16 patients with mild cognitive impairment and 14 normals). Patients were also evaluated using the Mini-mental State Exam (MMSE), Wechsler Memory Scale (WMS), Kohs Block Design, and word fluency. CD was scored using the Shulman method. CD scores showed a high interrater reliability (r = 0.97). CD by non-demented subjects was essentially normal. As a screening test for Alzheimer disease (AD, mean MMSE = 18.0), CD had a sensitivity of 57.1% and a specificity of 96.7%. However, four of eight AD patients who showed normal MMSE score (> or = 24) drew abnormal clocks. CD scores in dementia were significantly correlated with performance on the Block Design (r = 0.68), MMSE (r = 0.56), and the Mental Control subtest in the WMS (r = 0.58). Stepwise regression analysis revealed that performance on the MMSE and the Block Design explain 53.8% of the variance in the CD scores. These findings indicate that low CD score by the Shulman method may be reflective of a constructional disability and general severity of dementia. CD is not so sensitive as the result of the original report when the patient group includes milder cases of AD. CD is, however, an efficient screening test for detecting and following the patients with dementia, especially combined with the MMSE.

Aged↗

[A case of mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS), showing temporary improvement during the treatment with eicosapentaenoic acid ethyl ester].

We report a 67-year-old man with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS), successfully treated with eicosapentaenoic acid ethyl ester (EPA-E) for about eight months. He showed bilateral auditory disturbance and slowly progressive gait ataxia at age 50 during treatment of diabetes mellitus (DM) with subcutaneous injection of insulin since age 29. At age 58 he manifested an acute hemiparesis of right extremities for one week with no abnormal findings on neuroradiological examinations. A permanent pacemaker was implanted at age 61 to treat frequent syncopal attacks due to complete atrioventricular block. On admission to our hospital, neurological examinations revealed dementia, auditory disturbance, severe cerebellar ataxia and mild atrophy of proximal muscles with systemic hyporeflexia. Based on a point mutation in position 3243 of mitochondrial DNA, he was diagnosed as having MELAS with severe DM, auditory disturbance and cardiac conduction block. After initiation of treatment with EPA-E at a dose of 2,700 mg/day he showed temporarily an improvement in auditory disturbance, blood glucose control and cerebellar ataxia. In objective evaluations for cerebellar ataxia, we could find significant decreases in times for 20 m walking and heel-knee patting in the ninth month, and in time for tracing of a whirl from the third to the ninth month, compared with those before treatment of EPA-E (p < 0.0001). Because EPA-E is taken into mitochondrial membranes and activates electron transmission enzyme complexes, it might be a candidate for therapy of mitochondrial encephalomyopathy, including MELAS.

Aged↗

End-linked homodimers in fibrinogen Osaka VI with a B beta-chain extension lead to fragile clot structure.

The authors have identified a 12-residue carboxyl-terminal extension of Lys-Ser-Pro-Met-Arg-Arg-Phe-Leu-Leu-Phe-Cys-Met in a dysfibrinogen derived from a woman heterozygotic for this abnormality and associated with severe bleeding. This extension is due to a T-to-A mutation that creates AAG encoding Lys at the stop (TAG) codon, thus translating 36 base pairs in the noncoding region of the Bbeta gene. The extra Cys residues appear to be involved in 1 or 2 disulfide bonds between 2 adjacent abnormal fibrinogen molecules, forming a fibrinogen homodimer as indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Indeed, about half of the fibrinogen molecules exist as end-linked dimers oriented in parallel or with an angle, as observed by transmission electron microscopy. These end-linked dimers may well alter the conformations of D and DD regions on fibrin assembly, leading to increased fiber branching at their sites in the growing protofibrils. By scanning electron microscopy, the Osaka VI fibrin network appears to have a lacelike structure composed of highly branched, thinner fibers than the normal fibrin architecture. Such fibrin networks may be easily damaged to form large pores when fluids are allowed to pass through the gels. The fragility of Osaka VI fibrin clots, further confirmed by permeation and compaction studies, may account for the massive bleeding observed in this patient. (Blood. 2000;96:3779-3785)

Adult↗

Effects of mifepristone (RU486) treatment on the development of uterine adenomyosis induced by pituitary grafting in mice.

To evaluate the effects of mifepristone (RU486) on the development of uterine adenomyosis induced by pituitary grafting (PG), 3 groups of mice receiving pituitary grafts at 7 weeks of age were given RU486 in food (20 mg/kg chow) from 3-14 (RU486-3 group) or 10-14 (RU486-10 group) weeks of age, or were given no further treatment (PG control group), respectively. All the mice were killed at 14 weeks of age. The uterine weight was significantly decreased in both RU486-treated groups compared with the PG control group. The incidence of adenomyosis was also decreased significantly in both the RU486-3 group (0/10 mice) and RU486-10 group (2/10 mice) compared with the PG control group (7/9 mice). To look for vascular changes in the uterine tissues, which have been reported to be related to the development of adenomyosis, immunohistochemical staining of von Willebrand factor in the blood vessels was performed. The mean surface area and minor axis of blood vessels in the uterus were thereby found to be significantly decreased in the RU486-10 group compared to the PG control group. The results clearly indicated that RU486, a potent antiprogestin, could inhibit the genesis of uterine adenomyosis in mice, and at the same time caused shrinkage of the vascular system. As in humans, progesterone as well as the vascular system therefore appear to be important factors in the pathogenesis of uterine adenomyosis in this mouse model.

Adenomyoma↗

CalDAG-GEFIII activation of Ras, R-ras, and Rap1.

We characterized a novel guanine nucleotide exchange factor (GEF) for Ras family G proteins that is highly homologous to CalDAG-GEFI, a GEF for Rap1 and R-Ras, and to RasGRP/CalDAG-GEFII, a GEF for Ras and R-Ras. This novel GEF, referred to as CalDAG-GEFIII, increased the GTP/GDP ratio of Ha-Ras, R-Ras, and Rap1 in 293T cells. CalDAG-GEFIII promoted the guanine nucleotide exchange of Ha-Ras, R-Ras, and Rap1 in vitro also, indicating that CalDAG-GEFIII exhibited the widest substrate specificity among the known GEFs for Ras family G proteins. Expression of CalDAG-GEFIII was detected in the glial cells of the brain and the glomerular mesangial cells of the kidney by in situ hybridization. CalDAG-GEFIII activated ERK/MAPK most efficiently, followed by CalDAG-GEFII and CalDAG-GEFI in 293T cells. JNK activation was most prominent in cells expressing CalDAG-GEFII, followed by CalDAG-GEFIII and CalDAG-GEFI. Expression of CalDAG-GEFIII induced neuronal differentiation of PC12 cells and anchorage-independent growth of Rat1A cells less efficiently than did CalDAG-GEFII. Thus, co-activation of Rap1 by CalDAG-GEFIII apparently attenuated Ras-MAPK-dependent neuronal differentiation and cellular transformation. Altogether, CalDAG-GEFIII activated a broad range of Ras family G proteins and exhibited a biological activity different from that of either CalDAG-GEFI or CalDAG-GEFII.

Animals↗

Phosphorylation of Bcl-2 protein by CDC2 kinase during G2/M phases and its role in cell cycle regulation.

Although it has been reported that Bcl-2 phosphorylation is associated with certain types of apoptosis, there is much controversy over the functional significance of and the kinases responsible for the phosphorylation. In this study, we examined whether Bcl-2 is phosphorylated by CDC2 kinase, a master regulator of G(2)/M transition in the eukaryotic cell cycle. When CDC2 was activated by okadaic acid in HL-60 cells, Bcl-2 phosphorylation was readily induced. The phosphorylation was correlated with the accumulation of cells in G(2)/M phases, but was not proportional to the level of apoptosis. Furthermore, we found that Bcl-2 was phosphorylated during G(2)/M phases of normal cell cycle. The ability of CDC2 to phosphorylate Bcl-2 was confirmed by in vitro kinase assay with a highly purified CDC2-cyclin B complex. Using synthetic peptides and mutant cell lines, we identified threonine 56, one of two consensus sites for CDC2 within the Bcl-2 sequence, as a residue phosphorylated by CDC2. Mutation at threonine 56 abrogated the cell cycle inhibitory effect of Bcl-2 without affecting anti-apoptotic function. These results suggest that two distinct functions of Bcl-2 (anti-apoptosis and cell cycle inhibition) are differentially regulated by post-translational mechanisms such as phosphorylation. CDC2-mediated phosphorylation of Bcl-2 may play some physiological roles in the negative regulatory events during mitosis.

Amino Acid Substitution↗

Aquaporin adipose, a putative glycerol channel in adipocytes.

Adipose tissue is a major site of glycerol production in response to energy balance. However, molecular basis of glycerol release from adipocytes has not yet been elucidated. We recently cloned a novel member of the aquaporin family, aquaporin adipose (AQPap), which has glycerol permeability. The current study was designed to examine the hypothesis that AQPap serves as a glycerol channel in adipocytes. Adipose tissue expressed AQPap mRNA in high abundance, but not the mRNAs for the other aquaglyceroporins, AQP3 and AQP9, indicating that AQPap is the only known aquaglyceroporin expressed in adipose tissue. Glycerol release from 3T3-L1 cells was increased during differentiation in parallel with AQPap mRNA levels and suppressed by mercury ion, which inhibits the function of AQPs, supporting AQPap functions as a glycerol channel in adipocytes. Fasting increased and refeeding suppressed adipose AQPap mRNA levels in accordance with plasma glycerol levels and oppositely to plasma insulin levels in mice. Insulin dose-dependently suppressed AQPap mRNA expression in 3T3-L1 cells. AQPap mRNA levels and adipose glycerol concentrations measured by the microdialysis technique were increased in obese mice with insulin resistance. Accordingly, negative regulation of AQPap expression by insulin was impaired in the insulin-resistant state. Exposure of epinephrine translocated AQPap protein from perinuclear cytoplasm to the plasma membrane in 3T3-L1 adipocytes. These results strongly suggest that AQPap plays an important role in glycerol release from adipocytes.

3T3 Cells↗

Regulatory proteins of R-Ras, TC21/R-Ras2, and M-Ras/R-Ras3.

We studied the regulation of three closely related members of Ras family G proteins, R-Ras, TC21 (also known as R-Ras2), and M-Ras (R-Ras3). Guanine nucleotide exchange of R-Ras and TC21 was promoted by RasGRF, C3G, CalDAG-GEFI, CalDAG-GEFII (RasGRP), and CalDAG-GEFIII both in 293T cells and in vitro. By contrast, guanine nucleotide exchange of M-Ras was promoted by the guanine nucleotide exchange factors (GEFs) for the classical Ras (Ha-, K-, and N-), including mSos, RasGRF, CalDAG-GEFII, and CalDAG-GEFIII. GTPase-activating proteins (GAPs) for Ras, Gap1(m), p120 GAP, and NF-1 stimulated all of the R-Ras, TC21, and M-Ras proteins, whereas R-Ras GAP stimulated R-Ras and TC21 but not M-Ras. We did not find any remarkable difference in the subcellular localization of R-Ras, TC21, or M-Ras when these were expressed with a green fluorescent protein tag in 293T cells and MDCK cells. In conclusion, TC21 and R-Ras were regulated by the same GEFs and GAPs, whereas M-Ras was regulated as the classical Ras.

Amino Acid Sequence↗