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Biomedical subjects

H Masaki

Publications and source records attributed to H Masaki.

At least 91 records · Page 5Linked to original sources

Left ventricular mass regression after implantation of St. Jude Medical cardiac valves in small aortic roots.

In this study, we analyzed the extent of regression of left ventricular hypertrophy in patients who received small St. Jude Medical (SJM) aortic valves and compared the results with those of another group receiving larger valves. Eighty-eight patients received either 19 or 21 mm valves (Group 1, 25 patients) or either 23 or 25 mm valves (Group 2, 53 patients). Echocardiographic studies were done before the operation and 5 years postoperatively. At follow-up a significant reduction in the left ventricular mass was found for both patient groups (p < 0.0001). Doppler echocardiography derived pressure gradients for both groups were obtained during the follow-up period. As expected, the patients in Group 1 had higher peak pressure gradients than did those in Group 2. However, there was no significant difference between the 2 groups or any significant correlations between peak pressure gradients and body surface area (BSA). Actuarial survival was 84.7% at 15 years for Group 1 and 85.9% at 17 years for Group 2. Actuarial freedom from valve related events was 91.4% at 15 years for Group 1 and 82.7% at 17 years for Group 2. There was no significant difference in survival or valve related event free curves between the 2 groups. After implantations of SJM valves in small aortic roots, significant left ventricular mass regression was obtained, and the results were comparable to those for valves of other sizes. The long-term performance of aortic valve replacement with small valves was satisfactory as judged by improvement in the functional class of patients and survival statistics, the durability of the prosthesis, and valve related morbidity comparable to that of valves of other sizes.

Adult↗

Isolation of the gene and characterization of the enzymatic properties of a major exoglucanase of humicola grisea without a cellulose-binding domain.

An exoglucanase gene was cloned from a cellulolytic fungus, Humicola grisea. DNA sequencing of this gene, designated as exo1, revealed that it contained four introns in the coding region. The deduced amino acid sequence of EXO1 was 451 amino acids in length and showed 57.7% identity with that of H. grisea cellobiohydrolase 1 (CBH1), but lacked the typical domain structures of a cellulose-binding domain and a hinge region. Transcriptional analysis of the exo1 and cbh1 genes showed that the expression of these genes was induced by Avicel, and repressed in the presence of glucose. The exo1 gene was expressed in Aspergillus oryzae, and the recombinant EXO1 protein was purified. EXO1 and CBH1 produced by A. oryzae showed relatively higher activity toward Avicel, but showed much lower activity toward carboxymethyl cellulose (CMC) and p-nitrophenyl-beta-D-cellobioside (PNPC), than H. grisea endoglucanase 1 (EGL1). The addition of a cellulose-binding domain and a hinge region to EXO1 caused decreases in its enzymatic activities as well as the deletion of the cellulose-binding domain from CBH1. EXO1 showed relatively weak or no synergistic activity toward Avicel with H. grisea endoglucanases, but showed a significant level of apparent synergism with H. grisea CBH1 and Trichoderma reesei EGLI. CBH1 showed a significant level of apparent endo-exo synergism with H. grisea and T. reesei endoglucanases. H. grisea has at least two different types of major exoglucanase components and shows strong cellulolytic activity through synergism with cellulase components including EXO1 and CBH1.

Amino Acid Sequence↗

Isolation of pathogenic bacteria from induced sputum from hospitalized children with pneumonia in Bangladesh.

A prospective study was carried out on 157 patients admitted to a paediatric hospital in Dhaka, Bangladesh to determine the bacteria present in the induced sputum of paediatric patients with X-ray proven pneumonia. Their ages ranged from 21 days to 11 years; 65 per cent of them were male and 35 per cent were female. The most affected age group was between 6 months and 2 years old. Respiratory secretions produced by induced cough were taken by swab from the oropharynx for culture and smear. The predominant bacteria were Haemophilus influenzae, Streptococcus pneumoniae, Branhamella catarrhalis and Gram-negative bacilli. Serotyping of H. influenzae revealed that 76 per cent were non-typable and 18 per cent were of type b; 23.5 per cent of isolates of H. influenzae were beta-lactamase producing. MIC90 of penicillin against S. pneumoniae and H. influenzae were 0.025 and 3.13 micrograms/ml respectively. Ampicillin, penicillin G (benzylpenicillin), amoxycillin, and gentamicin were administered for the treatment of these patients. All cases were apparently improved, on the basis of clinical evaluation, and discharged from the hospital.

Age Distribution↗

Tissue-specific expression of human angiotensin II AT1 and AT2 receptors and cellular localization of subtype mRNAs in adult human renal cortex using in situ hybridization.

All studies analyzing the localization of angiotensin II (Ang II) receptors in the human kidney have been performed at the protein level using 125I-Ang II as a probe. In this study, cellular localizations of Ang II type l (AT1-R) and type 2 (AT2-R) receptor mRNAs in the adult human renal cortex were examined for the first time using in situ hybridization, and their expression patterns determined by RNase protection assay were compared with those in other human tissues. In the human renal cortex obtained from tumor-free portions in renal cell carcinoma, AT1-R mRNA levels were about 8- to 10-fold higher than AT2-R mRNA levels. Human liver and aorta predominantly expressed AT1-R mRNA, while human right atrium contained both AT1-R and AT2-R mRNAs. Ligand-binding assays revealed that the total Ang II receptor number in the human renal cortex was 16.0 +/- 3.3 fmol/mg protein, similar to that in liver (17.7 +/- 5. 8) but significantly higher than in right atrium (11.6 +/- 3.2) and aorta (5.6 +/- 2.7). Relative distribution ratios of AT1-R and AT2-R numbers in the renal cortex and right atrium were 82/17 and 56/42%, respectively. In situ hybridization study indicated that strongest AT1-R mRNA signals were located in interlobular arteries and tubulointerstitial fibrous regions surrounding interlobular arteries and glomeruli, followed in decreasing order by glomeruli and cortical tubules. Expression of AT2-R mRNA was highly localized in interlobular arteries. Cells present in tubulointerstitial regions were positive for vimentin and collagen type 1, indicating that the majority of the cells present in the regions are fibroblasts. Presence of strong AT1-R mRNA signals in the tubulointerstitial fibrous regions surrounding arteries and glomeruli and the expression of AT2-R mRNA in the interlobular artery were the first evidence, suggesting a pharmacological framework for the differential effects of Ang II receptor subtype mediated renal function in the adult human kidney.

Adult↗

Role of calcium-sensitive tyrosine kinase Pyk2/CAKbeta/RAFTK in angiotensin II induced Ras/ERK signaling.

In cardiac fibroblasts, angiotensin II (Ang II) induced a rapid increase in extracellular signal regulated kinase (ERK) activity in a pertussis toxin insensitive manner. This ERK activation was abolished by the Gq-associated phospholipase C inhibitor U73122 but was insensitive to protein kinase C (PKC) inhibitors or PKC downregulation by phorbol ester. Intracellular Ca2+ chelation by BAPTA-AM or TMB-8 abolished Ang II induced ERK activation, whereas treatment with EGTA or nifedipine did not affect it. Ca2+ ionophore A23187 also induced a rapid increase in ERK activity to an extent similar to that of Ang II stimulation. Calmodulin inhibitors (W7 and calmidazolium) and tyrosine kinase inhibitors (genistein and ST638) completely blocked ERK activation by Ang II and A23187. Both Ang II and A23187 caused a rapid increase in the binding of GTP to p21(Ras), which was nearly abolished by genistein and calmidazolium. Transfection with the dominant negative mutant of Ras and the Ras inhibitor manumycin completely inhibited Ang II induced ERK activation. It was also found for the first time that cardiac fibroblasts abundantly expressed Ca2+-sensitive tyrosine kinase Pyk2/CAKbeta/RAFTK and that Ang II markedly induced its activation in a Ca2+/calmodulin-sensitive manner. Overexpression of the dominant negative mutant of Pyk2 significantly attenuated Ang II or A23187-induced ERK activities (36% and 38% inhibition compared with that in mock-transfected cells, respectively) and ERK tyrosine phosphorylation levels, as well as an increase in the binding of GTP to p21(Ras). These findings demonstrate that in cardiac fibroblasts, Ang II induced Ras/ERK activation is dominantly regulated by Gq-coupled Ca2+/calmodulin signaling and that Pyk2 plays an important role in the signal transmission for efficient activation of the Ang II induced Ras/ERK pathway.

Angiotensin II↗

Protective effects of baicalein against cell damage by reactive oxygen species.

Baicalein (5,6,7-trihydroxy-2-phenyl-4H-1-benzopyran-4-one), a naturally occurring flavonoid, was found to prevent human dermal fibroblast cell damage induced by reactive oxygen species such as hydrogen peroxide (H2O2), tert-butyl hydroperoxide (BuOOH) and superoxide anions (.O2-) in a concentration-dependent manner, and was more effective than the iron chelator, deferoxamine, hydroxyl radical (.OH) scavengers such as dimethyl sulfoxide (DMSO) and ethanol (EtOH), the lipid peroxidation chain blocker, alpha-tocopherol (Vit. E) and the xanthine oxidase inhibitor, allopurinol. To probe the mechanism of cell defense, the reaction of baicalein with oxygen free radicals was investigated using electron spin resonance (ESR) spectrometry. Baicalein decreased the signal intensities due to the 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) spin adducts of .OH, .O2- and tert-butyl peroxyl (BuOO.) radicals in a concentration-dependent manner. The IC50 values, which are the 50% inhibition concentrations of baicalein for the free radicals, were 10, 45 and 310 microM, respectively. These results suggested that baicalein possesses free radical scavenging ability which prevents the fibroblast damage induced by these free radical species.

Cyclic N-Oxides↗

Isolation of the creA gene from the cellulolytic fungus Humicola grisea and analysis of CreA binding sites upstream from the cellulase genes.

A carbon catabolite repressor gene, creA, was isolated from the cellulolytic fungus Humicola grisea by using a portion of the Trichoderma reesei cre1 gene as a probe. The deduced amino acid sequence predicts a zinc finger protein of 419 amino acids in length, and its zinc finger regions show high similarity with those of Aspergillus CreAs, T. reesei Cre1, and Saccharomyces cerevisiae MIG1. Northern blot analysis showed that the H. grisea creA gene was highly transcribed when the mycelia were grown on glucose-containing media, but the transcription of the H. grisea endoglucanase 1 gene (egl1) and the exoglucanase 1 gene (exo1) were repressed under these conditions. Results of binding assays with the maltose-binding protein::CreA(1-166) fusion protein and the egl1 and the exo1 upstream regions showed that some 6-bp sites having an identical or similar sequence to the consensus sequence for CreA binding were protected from DNase I digestion.

Amino Acid Sequence↗

Detection of potato virus Y P1 protein in infected cells and analysis of its cleavage site.

The P1 protein is liberated from the N-terminal region of the potyviral polyprotein by cleavage depending on its own autoproteolytic activity. Existence of the 32-kDa P1 protein in tobacco plants infected with potato virus Y ordinary strain (PVY-O) was detected by an antiserum against a recombinant PVY-O P1 protein. In vivo analysis using tobacco protoplasts confirmed that the Phe284-Ser285 was the cleavage site separating the P1 protein from the PVY-O polyprotein. Phe284 was indispensable for proteolysis and Ser285 was needed for optimal cleavage susceptibility.

Amino Acid Sequence↗

[Pathophysiological function of angiotensin II AT1 and AT2 receptors and clinical application of AT1 antagonists].

Angiotensin II (Ang II) receptors are classified into two subtypes, type 1 (ATF-R) and type 2 (AT2-R) by development of non-peptidic antagonists. Classical Ang II function including vasopressor effect, cardiotropic action and aldosterone production is mainly mediated through AT1-R that present in cardiovascular system, adrenal glands and kidneys. AT1-R is abundantly expressed in whole bodies of fetus and its expression is abruptly decreased after birth, and in the adult AT2-R is expressed in brain nuclei, uterus, adrenal medullary glands and ovary. AT1-R and AT2-R are both G-protein coupled receptors and have 46% similarity in amino acid levels with seventh transmembrane conformation. Signal transduction pathway of AT1-R is mainly CA2+ and activation of protein kinase C, while that of AT2-R is still unknown. Clinical application of AT1-R antagonist started and this causes elevation of plasma Ang II levels, which selectively stimulates AT2-R. Thus, one should realize AT2-R-mediated effect in treatment with AT1-R antagonist. We have shown that AT2-R has anti-AT1-R action, such as inhibitory action against AT1-R-mediated positive chronotropic effect or AT1-R-induced proliferative effect, resulting in the protective effects on Ang II-induced cardiovasucular and renal action. Thus, elucidation of AT2-R function will be important in clinical treatment with AT1-R antagonists.

Angiotensin II↗

[Long-term follow-up of patients with small St. Jude aortic valve prostheses].

Thirty-five patients received small sized 19 or 21 mm valves (group I) and 53 patients received 23 or 25 mm valves (group II). At follow-up a significant reduction in the left ventricular mass was found for both patient groups (p < 0.0001). The patients in group I had higher peak pressure gradients than did those in group II. However, there was no significant difference between the two groups, or any significant correlation between peak pressure gradients and body surface area. Actuarial survival was 84.7% at 15 years for group I and 85.9% at 17 years for group II. Actuarial freedom from valve-related events was 91.4% at 15 years for group I and 82.7% at 17 years for group II. There was no significant difference in survival or valve-related event free curves between the two groups. After implantation of small SIM valves, significant left ventricular mass regression was obtained and the results were comparable to those for valves of other sizes. The long-term performance of aortic valve replacement with small valves was satisfactory as judged by improvement in the functional class of patients, survival statistics, the durability of the prosthesis, and valve related morbidity comparable to that of valves of other sizes.

Aged↗

[Immune complex-mediated glomerulonephritis associated with infective endocarditis with aortic valve vegetation].

A 61-year-old male was referred to our hospital for rapidly progressive azotemia. He was also found to have huge vegetation at the aortic valve causing regurgitation. Biochemical examinations revealed the presence of an immunocomplex associated with decreased circulating complements. In biopsy samples from the kidney, we found the presence of fibrillar crescents, proliferation of mesangial cells, increase in extracellular matrix proteins, atrophy of tubules, infiltration of mononuclear cells in the interstitial regions, high density deposits in the mesangial area and mesangial interposition. Since the patient strongly rejected operative treatment by valvular replacement, we continued non-invasive treatment such as hemodialysis and treatment with penicillin G. This transiently improved the condition of the patient, including biochemical data and cardiac function, but there was no reduction in the size of vegetation at the aortic valve and the bacteria responsible for infective endocarditis were not identified. About three months after admission, overt signs of congestive heart failure emerged and the patients was subjected to intensive care with a respirator and hemodynamic monitoring. Although the cardiac function was improved, concomitant severe pneumonia occurred and the patient died of septic shock. Thus, we report a rare case in whom immune complex-mediated glomerulonephritis was associated with infective endocarditis with aortic valve vegetation.

Antigen-Antibody Complex↗

A clinical analysis of the recovery from sensory disturbance after sagittal splitting ramus osteotomy using a Semmes-Weinstein pressure aesthesiometer.

A number of studies of evaluation methods for sensory disturbance after sagittal splitting ramus osteotomy (SSRO) are known. To compare postoperative sensory disturbances among patients in different hospitals, a highly reproducible and standardized sensory test is required. In the present study, we measured the tactile threshold in the region innervated by the mental nerves in 45 patients (90 sides) after SSRO using a Semmes-Weinstein pressure aesthesiometer. The percentage of recovery to the normal level defined by Bell was 72.2% at one week, 82.2% at 4 weeks, and 90.0% at 8 weeks after SSRO. The recovery process was evaluated by classifying the postoperative sensory disturbance into 5 grade levels according to Bell's interpretation scale. The results showed that the SW sensory test is useful for evaluation of the recovery process from sensory disturbance after SSRO. Some improvements of this test were also discussed.

Adult↗

Angiotensin type 2 receptors are reexpressed by cardiac fibroblasts from failing myopathic hamster hearts and inhibit cell growth and fibrillar collagen metabolism.

BACKGROUND: Angiotensin (Ang) II type 1 receptor (AT1-R) induces cardiomyocyte hypertrophy and fibroblast proliferation, whereas the physiological role of AT2-R in cardiac remodeling remains poorly defined. METHODS AND RESULTS: Using Bio14.6 cardiomyopathic (CM) hamsters, we found that AT2-R sites were increased by 153% during heart failure compared with F1B controls. AT1-R numbers were increased by 72% in the hypertrophy stage and then decreased to the control level during heart failure. Such differential regulation of AT2-R and AT1-R during heart failure was consistent with changes in the respective mRNA levels. Autoradiography and immunocytochemistry revealed that both AT2-R and AT1-R are localized at higher densities in fibroblasts present in fibrous regions. Surrounding myocardium predominantly expressed AT1-R, but the level of expression was less than that in fibrous regions. Cardiac fibroblasts isolated from CM hearts during heart failure but not from control hamsters expressed AT2-R (30 fmol/mg protein). Using the cardiac fibroblasts expressing AT2-R, we found that Ang II stimulated net collagenous protein production by 48% and pretreatment with an AT2-R antagonist, PD123319, evoked a further elevation (83%). Ang II-induced synthesis of fibronectin and collagen type I were enhanced by 40% and 53%, respectively, by pretreatment with PD123319. Ang II-induced DNA synthesis (assessed by [3H]thymidine uptake) was significantly increased by PD123319, and the AT2-R agonist CGP42112A reduced the serum-stimulated increase in cell numbers by 23%. Treatment with an AT1-R antagonist, TCV116, for 20 weeks inhibited progression of interstitial fibrosis by 28%, whereas with 44-week PD123319 treatment but not 20-week treatment, the extent of the fibrous region was increased significantly, by 29%. CONCLUSIONS: These findings demonstrate that AT2-R is re-expressed by cardiac fibroblasts present in fibrous regions in failing CM hearts and that the increased AT2-R exerts an anti-AT1-R action on the progression of interstitial fibrosis during cardiac remodeling by inhibiting both fibrillar collagen metabolism and growth of cardiac fibroblasts.

Angiotensin Receptor Antagonists↗

Generation of active oxygen species from advanced glycation end-products (AGE) under ultraviolet light A (UVA) irradiation.

To clarify a possible role of advanced glycation end-products (AGE) on photoaging of human skin, the interaction between AGE and ultraviolet A light (UVA) was examined from both a biological and chemical perspective. Human dermal fibroblasts that were exposed to UVA in the presence of AGE bound with bovine serum albumin (AGE-BSA) exhibited a significant decrease of cell viability as compared to control cells, which were exposed to UVA in the absence of AGE-BSA. Further, UVA-irradiated AGE-BSA reduced nitroblue tetrazolium to its formazan, but the reaction was inhibited by addition of superoxide dismutase in the system. UVA dose-dependent formation of H2O2 in AGE-BSA was also observed. An ESR spin-trapping study revealed the generation of unstable free radicals in AGE-BSA under UVA irradiation. After addition of Fe2+ in the system, an ESR spectrum due to the formation of hydroxyl radicals was observed. On the basis of these results, the authors propose that AGE is an important factor for promoting photoaging in the skin via generation of active oxygen species involving .O2-, H2O2, and .OH.

Cell Survival↗

Regulation of cyclodextrin glucanotransferase synthesis in Bacillus ohbensis.

The production of cyclodextrin glucanotransferase (CGTase) by Bacillus ohbensis is dependent on the presence of starch and inhibited by glucose in the medium. Northern blot analysis revealed that the CGTase gene (cgt) was transcribed to almost the same level irrespective of the presence or absence of starch, but glucose completely repressed the transcription. Furthermore, a relatively high amount of CGTase protein was detected on Western blotting only in the medium with starch, showing the lack of posttranslational control of the CGTase activity. These findings suggest some starch induction mechanism for the cgt gene, possibly at the posttranscriptional level, besides negative transcriptional regulation by glucose.

Bacillus↗

Cloning and sequencing of an alpha-glucosidase gene from Aspergillus niger and its expression in A. nidulans.

We have cloned an extracellular alpha-glucosidase gene from Aspergillus niger with oligonucleotide probes synthesized on the basis of the determined peptide sequences. The nucleotide sequence revealed an open reading frame of 985 amino acids split with three introns, and the deduced amino acid sequence was nearly identical to that of the alpha-glucosidase previously determined. The cloned gene was introduced into Aspergillus nidulans, and its expression in the transformants was shown to be regulated by the carbon sources in the medium, suggesting that a common regulatory expression system is shared by these two species as is the case of other starch-degrading enzymes of Aspergillus species.

Amino Acid Sequence↗

Blood cyst of the pulmonary valve in an adult: report of a case and review of the literature.

We report a case of a blood cyst originating from the pulmonary valve cusp of a 43-year-old Japanese woman with pulmonary stenosis. Cineangiography revealed a pedunculated tumor on the arterial surface of the pulmonary valve. It was successfully removed by a transpulmonary artery approach. The cyst contained old blood and calcified thrombi, and its wall consisted of collagenous fibrous tissue. Immunohistochemically, a monolayer of flat cells lining the cyst was confirmed as endothelium using antibodies to von Willebrand factor and CD34, as well as UEA-I lectin. To the best of our knowledge, blood cysts of the pulmonary valve are rare and only 10 such cases have been reported, including six pediatric cases. This case is the oldest and fifth adult patient with a blood cyst found on the pulmonary valve. A possible histogenesis is discussed.

Adult↗

Increased generation of hydrogen peroxide possibly from mitochondrial respiratory chain after UVB irradiation of murine fibroblasts.

The purpose of this study is to detect the generation of active oxygens in UVB-irradiated murine fibroblasts and to propose new mechanisms. Decreased survival of fibroblasts under UVB irradiation was partially recovered by addition of catalase, DMSO or deferoxamine, suggesting the contribution of several types of active oxygen species. Then we examined the formation of active oxygen species and found that fibroblasts under UVB irradiation generated superoxide anion radicals (.O2-), intracellular H2O2, and hydroxyl radicals as estimated by the ESR-spin trapping method. Addition of thenoyltrifluoroacetone, which is an inhibitor of the mitochondrial respiratory chain, decreased 29% of the intracellular H2O2 levels in UVB-irradiated cells, but allopurinol, which is an inhibitor of xanthine oxidase, had no effect on them. On the basis of these results, we propose a a possible mechanism for damage of murine fibroblasts exposed to UVB in terms of generation of active oxygen species. The mitochondrial respiratory chain reaction stimulated by UVB irradiation enhances the generation of .O2-, which is in turn dismutated to H2O2 and O2 by superoxide dismutase. H2O2 is then converted to hydroxyl radicals, catalyzed by trace elements such as iron, as suggested by Fenton-like reaction. Thus, hydroxyl radicals with higher reaction rate-constants than those of other active oxygen species to biomolecules are indicated to be responsible for the cytotoxicity in cells under UV irradiation.

Allopurinol↗