Biomedical subjects

M Yamada

Publications and source records attributed to M Yamada.

At least 703 records · Page 39Linked to original sources

PubMed1998

Further characterization of hydrogen peroxide-dependent fatty acid alpha-hydroxylase from Sphingomonas paucimobilis.

Although fatty acid alpha-hydroxylase (FAAH) activity has been detected in various species, FAAH has not been sufficiently characterized. In this report, we describe the properties of FAAH highly purified from Sphingomonas paucimobilis. The FAAH was purified by about 5,200-fold. Blotting analysis with a specific antibody against the FAAH showed that its apparent molecular mass was approximately 43 kDa. FAAH showed alpha-hydroxylation activity in the presence of H2O2, but little if any activity with cumene hydroperoxide, t-butyl hydroperoxide, or t-butyl peroxybenzonate. The Km value for H2O2 was 72 microM. Highly purified FAAH oxidized various non-esterified saturated and unsaturated fatty acids including myristic acid, but not myristoyl-CoA. Potassium cyanide and sodium azide inhibited the FAAH activity in a concentration-dependent manner. Other respiratory chain inhibitors such as rotenone and antimycin A did not inhibit the activity. Among cytochrome P450 inhibitors, SKF-525A markedly inhibited the activity at the concentration of 2 mM, but CO did not. Imidazole, an inhibitor of plant alpha-oxidation, showed no inhibitory effect at 1 mM.

Catalysis↗

PubMed1998

ADP/vanadate mediated photocleavage of myosin light chain kinase at the autoinhibitory region.

The vanadate (Vi)-mediated photocleavage reaction was used to study the interaction between the regulatory segment and the catalytic site of smooth muscle myosin light chain kinase (MLCK). When MLCK was irradiated with long-wave UV (366 nm) in the presence of ADP and Vi, kinase activity was substantially decreased, and the MLCK polypeptide of 130 kDa was cleaved into several smaller fragments with apparent molecular masses of 100, 70, 60, 32, and 28 kDa. Inhibition of kinase activity and photocleavage were both competitively antagonized by the addition of ATP. Inconsistency between the observed maximum levels of UV-induced inhibition of MLCK-mediated phosphorylation (80%) and photocleavage (15-20%) suggested that the photocleavage reaction proceeds as a two-step process. Monoclonal antibodies recognizing the C-terminus of MLCK labeled the 60- and 28-kDa fragments, indicating that MLCK was cleaved at two sites, at 28 and 60 kDa from the C-terminus, within what are believed to be the autoinhibitory region and the catalytic site, respectively. Moreover, Ca2+-calmodulin (Ca2+-CaM) protected against cleavage at the site at 28 kDa from the C-terminus. Analysis of the amino acid composition of the fragment revealed that the cleavage site at 28 kDa from C-terminus occurred at Lys 799 +/- 3 amino acid residues, which is in a region where the CaM-binding and pseudosubstrate regions overlap. These results suggest that the three-dimensional structure of MLCK brings the regulatory segment into direct contact with the ATP-binding site. Moreover, the binding of Ca2+-CaM displaces the regulatory segment away from the catalytic site.

Adenosine Diphosphate↗

PubMed1998

Prophylactic endoscopic ligation of high-risk oesophageal varices in a cirrhotic patient with severe haemophilia A.

One of the main complications in cirrhosis is haemorrhage from oesophageal varices. It is serious and often fatal, especially in cirrhotic patients with haemophilia. We describe the use of endoscopic variceal ligation (EVL) for prophylaxis of oesophageal variceal bleeding in a high-risk patient, a 40-year-old Japanese man, with severe haemophilia A and liver cirrhosis caused by hepatitis C virus. He had large, coil-shaped varices with a red colour sign, predicting the likelihood of haemorrhage. Administration of omeprazole and factor VIII concentrate achieved rapid healing of the post-EVL ulcers and prevented bleeding from them. Four EVL sessions eradicated oesophageal varices completely, and he has had no recurrence of varices for 2 years, indicating that the procedure was of considerable benefit.

Adult↗

PubMed1998

Confocal observation of senile plaques in Alzheimer's disease: senile plaque morphology and relationship between senile plaques and astrocytes.

Senile plaques in the brains of Alzheimer's disease (AD) were examined by confocal laser scanning microscopy (CLSM) with the following three findings. First, in sections stained with Congo red, the serial CLSM images of optical sections clearly revealed that a classic plaque is composed of a plaque core and a corona. Radially arranged process-like structures, corresponding to bundles of amyloid fibrils, formed amyloid cores and stronger signals were detected in the center of some cores. Second, in sections stained with Congo red and anti-glial fibrillary acidic protein (GFAP), reactive astrocytes were found around the senile plaques and many astrocytic processes surrounded the plaque cores and some processes had penetrated into them. Third, three-dimensional reconstruction on classic plaque revealed that the surface of classic plaque showed a 'coral-like' appearance.

Aged↗

PubMed1998

Benefit of in-phase chest wall vibration on the pulmonary hemodynamics in patients with chronic obstructive pulmonary disease.

OBJECTIVE: Chest wall vibration of contracting intercostal muscles (in-phase vibration, (IPV)) improves O2 saturation in patients with chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate the effect of IPV on the pulmonary hemodynamics in patients with COPD. METHODOLOGY: Twelve patients with COPD (FEV1%, 43.7 +/- 14.4%) underwent pulmonary artery catheterization in the supine position; hemodynamic variables as well as arterial and mixed venous gas tension were measured at baseline and after 15 min of IPV with IPV continued during the measurement. RESULTS: Mean pulmonary arterial pressure (Ppa, 21.3 +/- 5.0-19.1 +/- 4.8 mmHg and pulmonary vascular resistance (PVR, 294.6 +/- 196.0-228.5 +/- 101.7 dyne.sec.cm-5) decreased significantly, but there was no change in heart rate, cardiac index or systemic blood pressure. PaO2 increased (66.5 +/- 10.3-70.0 +/- 9.7 Torr) and PaCO2 decreased (46.6 +/- 8.9-45.0 +/- 10.1 Torr) significantly. Minute ventilation and tidal volume increased significantly in five out of the eight patients in whom ventilation was monitored throughout the study. CONCLUSION: IPV improves gas exchange and pulmonary circulation without affecting systemic circulation.

Aged↗

PubMed1998

Around-the-clock nursing care for the elderly in Japan.

PURPOSE: To describe the first phase of creating a Japanese model of community-based long-term care, called around-the-clock care (ACC), by operationally defining the components of ACC, identifying those eligible for the program, clarifying methods of planning and organization, and exploring outcomes. DESIGN: Exploratory evaluation for the population of frail elderly Japanese citizens living at home. A convenience sample of 44 patients receiving care from four visiting-nurse-service stations during 3 months in 1994 was used. METHODS: Needs assessments; record reviews of patient encounters; evaluations by patients, families, and visiting nurses; and reviews of administrative data. FINDINGS: Nurses perceived that ACC stabilized medical status, reduced the emotional and physical burden of treatment, and improved hygiene. It also reduced family caregiving burdens and the stress of family caregivers' employment responsibilities. The highest rating of ACC by patient and families was for those with the most complex physical needs. Skilled nursing at home permitted early identification and treatment of problems before they became crises or required hospitalization. CONCLUSIONS: Early findings suggest 24-hour nurse-home helper teamwork may be an effective system of community-based long-term care and should be considered a key element of future Japanese health policy.

Aged↗

PubMed1998

ftsE(Ts) affects translocation of K+-pump proteins into the cytoplasmic membrane of Escherichia coli.

The ftsE(Ts) mutation of Escherichia coli causes defects in cell division and cell growth. We expressed alkaline phosphatase (PhoA) fusion proteins of KdpA, Kup, and TrkH, all of which proved functional in vivo as K+ ion pumps, in the mutant cells. During growth at 41 degrees C, these proteins were progressively lost from the membrane fraction. The reduction in the abundance of these proteins inversely correlated with cell growth, but the preformed proteins in the membrane were stable at 41 degrees C, indicating that the molecules synthesized at the permissive temperature were diluted in a growth-dependent manner at a high temperature. Pulse-chase experiments showed that KdpA-PhoA was synthesized, but the synthesized protein did not translocate into the membrane of the ftsE(Ts) cells at 41 degrees C and degraded very rapidly. The loss of KdpA-PhoA from the membrane fractions of ftsE(Ts) cells was suppressed by a multicopy plasmid carrying the ftsE+ gene. While cell growth stopped when the abundance of these proteins decreased 15-fold, the addition of a high concentration of K+ ions specifically alleviated the growth defect of ftsE(Ts) cells but not cell division, and the cells elongated more than 100-fold. We conclude that one of the causes of growth cessation in the ftsE(Ts) mutants is a defect in the translocation of K+-pump proteins into the cytoplasmic membrane.

ATP-Binding Cassette Transporters↗

PubMed1998

Survey of human group C rotaviruses in Japan during the winter of 1992 to 1993.

Fecal specimens from patients with acute diarrhea were collected from 10 prefectures in Japan over a 6-month period (November 1992 to April 1993), and the specimens that were negative for human group A rotaviruses were screened for the presence of human group C rotaviruses (CHRVs) by the reverse passive hemagglutination test. Of 784 specimens examined, 53 samples (6.8%) that were collected in 7 of 10 prefectures were positive for CHRV, indicating that CHRVs are widely distributed across Japan. Most of the CHRV isolates were detected in March and April, and CHRVs mainly prevailed in children ages 3 to 8 years. The genome electropherotypes of eight strains isolated in five individual prefectures were surprisingly similar to each other and were different from those of CHRV strains isolated to date. The outer capsid glycoprotein (VP7) gene homologies of the isolates retrieved in 1993 were subsequently analyzed by the dot blot hybridization method. As a result, the VP7 genes of the isolates revealed very high levels of homology not only with each other but also with the VP7 gene of the OK118 strain isolated in 1988. These results suggest that a large-scale outbreak of CHRV occurred during the winter of 1992 and 1993 in Japan.

Adolescent↗

PubMed1998

Epidemiological study of a food-borne outbreak of enterotoxigenic Escherichia coli O25:NM by pulsed-field gel electrophoresis and randomly amplified polymorphic DNA analysis.

This study investigated the applicability of molecular epidemiological techniques to the identification of the causal agent of an outbreak of diarrhea caused by ingestion of food contaminated with enterotoxigenic Escherichia coli (ETEC). The outbreak occurred at four elementary schools in July 1996 and affected more than 800 people. Illness was most strongly associated with eating tuna paste (relative risk, 1.79; 95% confidence interval = 1.16 to 2.79; P = 0.0001). To evaluate the epidemiological characteristics of the pathogen, the DNAs from numerous isolated ETEC strains were subjected to randomly amplified polymorphic DNA analysis, pulsed-field gel electrophoresis of nuclease S1-treated plasmid DNA, and analysis of genomic DNA restriction fragment length polymorphisms. All ETEC isolates were of the O25:NM (nonmotile) serotype, which carries a heat-stable enterotoxin Ib gene. Genotypic analysis demonstrated that the strains isolated from the patients at all four schools were identical. The isolates of ETEC O25:NM obtained from the tuna paste that had been served for lunch at these schools were genetically indistinguishable from those isolated from the patients. Results suggest that this outbreak was food borne. The molecular biology-based epidemiological techniques used in this study were useful in characterizing the causal agent in this food-borne epidemic.

Animals↗

PubMed1998

Severe form of juvenile corneal stromal dystrophy with homozygous R124H mutation in the keratoepithelin gene in five Japanese patients.

AIM: To confirm the mutation of the keratoepithelin gene in patients with a severe form of superficial juvenile granular corneal dystrophy (GCD). METHODS: Five Japanese probands in whom GCD was diagnosed after histopathological examination and who developed severe manifestations of GCD in their first decade of life were investigated. Other affected family members of two probands were also examined. All probands were the offspring of consanguineous parents. DNA was extracted from their peripheral blood leucocytes and mutational analysis of the gene was performed by the polymerase chain reaction and direct sequencing. RESULTS: Four of the five probands underwent their first keratectomy or keratoplasty in their teens and subsequently underwent a second or third keratoplasty. Each proband had a homozygous G-->A transition at codon 124, replacing Arg-->His, of the keratoepithelin gene. Their moderately affected family members were heterozygous for the mutation. CONCLUSIONS: This finding suggests that the severity of the corneal phenotype depends on the dose effect of the mutant gene.

Adult↗

PubMed1998

Quantitative analysis of lipid deposits from Schnyder's corneal dystrophy.

AIM: To report the quantitation of the lipid composition of a corneal button from a Japanese woman in her 60s with clinically and histopathologically proved Schnyder's corneal dystrophy. METHODS: Total lipids extracted from the corneal button of the patient were analysed by the method of thin layer chromatography flame ionisation detection. Two different solvent systems were used for neutral lipid analysis and phospholipid analysis. Results were compared with three age matched corneal buttons obtained from cadaveric eyes. RESULTS: The lipids that accumulated in the cornea in Schnyder's dystrophy consisted mainly of unesterified cholesterol and phospholipids. The analysis of phospholipids showed sphingomyelin to be the predominant phospholipid in the patient's cornea. CONCLUSION: Findings suggest that this disorder involves a disturbance of the metabolism of cholesterol and/or sphingomyelin metabolism that is limited to the cornea.

Cholesterol↗

PubMed1998

Presenilin 1 intronic polymorphism is not associated with Alzheimer type neuropathological changes or sporadic Alzheimer's disease.

BACKGROUND: A genetic association between the presenilin 1 (PS-1) intronic polymorphism and sporadic Alzheimer's disease has been a matter of controversy. Recent findings have suggested that the PS-1 polymorphism is not associated with Alzheimer's disease or amyloid beta-protein (Abeta) deposition in brains from patients with Alzheimer's disease. OBJECTIVES: To elucidate the influence of the PS-1 polymorphism on Alzheimer type neuropathological changes and the development of Alzheimer's disease, the relation between the PS-1 polymorphism and quantitative severity of Alzheimer type neuropathological changes in the brains from patients with Alzheimer's disease and non-demented subjects was studied. METHODS: The PS-1 and apolipoprotein E (ApoE) genotypes, were examined, together with the densities of the senile plaques, senile plaques with dystrophic neurites, and neurofibrillary tangles in the brains from 36 postmortem confirmed patients with sporadic Alzheimer's disease and 86 non-demented subjects. Association of the PS-1 polymorphism with sporadic Alzheimer's disease and ages at onset and duration of illness in Alzheimer's disease was also examined. RESULTS: The PS-1 polymorphism was not associated with the senile plaques, senile plaques with dystrophic neurites, or neurofibrillary tangles in Alzheimer's disease or non-demented subjects. There was no association of the PS-1 intronic polymorphism with Alzheimer's disease, ages at onset, or durations of illness in Alzheimer's disease. The results remained nonsignificant even when the PS-1 genotype groups were divided into the subgroups with different ApoE epsilon4 status. CONCLUSIONS: The PS-1 intronic polymorphism does not itself have a direct causal role in the formation of Alzheimer type neuropathological changes or in the development of sporadic Alzheimer's disease.

Age of Onset↗

PubMed1998

Transforming growth factor-beta inhibits proliferation and maturation of cultured guinea pig gastric pit cells.

We studied the effects of transforming growth factor-beta1 (TGF-beta1) on guinea pig gastric mucous cells, cultured in serum-free conditions. Electron microscopy showed that most cells were pre-pit cells, characterized by the presence of a few secretory granules scattered in the cytoplasm. Epidermal growth factor (EGF) stimulated cell growth, [3H]glucosamine uptake, and accumulation of mucus granules positive for galactose oxidase-Schiff reaction. This EGF-induced maturation into pit cells was confirmed morphologically by the appearance of uniformly dense ovoid or spherical mucus granules packed in the ectoplasm. Western blotting with an antiphosphotyrosine antibody showed that TGF-beta1 did not inhibit the EGF-initiated tyrosine phosphorylation of the EGF receptor. Northern blotting with cDNA probes for c-fos and c-myc demonstrated that TGF-beta1 did not affect the EGF-induced expression of the transcripts. However, TGF-beta1-treated cells did not replicate and remained in an immature stage, even in the presence of EGF, suggesting a potential role of TGF-beta1 in the regulation of proliferation and differentiation of a pit cell lineage in vivo.

Animals↗

PubMed1998

Serum hyaluronan as a marker of liver fibrosis in hemophiliacs with hepatitis C virus-associated chronic liver disease.

We measured serum type III procollagen peptide, type IV collagen (IV-C), and hyaluronan (HA) in 36 hemophiliacs with hepatitis C virus (HCV)-related chronic liver disease. We also measured these markers in 19 interferon (IFN)-treated patients before and after a 6-month course of IFN-alpha. The serum concentrations of IV-C and HA were correlated with the stage of fibrosis (IV-C r = 0.38; HA r = 0.78). These markers fell only in complete responders showing sustained clearance of serum HCV RNA by IFN. These results suggest that, in hemophiliacs with chronic liver disease, serum HA measurement might be an alternative to liver biopsy and reflect the response to IFN.

Adolescent↗

PubMed1998

Butyrylcholinesterase K variant and cerebral amyloid angiopathy.

BACKGROUND AND PURPOSE: Cholinesterases are found histochemically in the vessels affected with cerebral amyloid angiopathy (CAA). A gene for the K variant of butyrylcholinesterase (BCHE-K) may be associated with late-onset Alzheimer's disease (AD). In search of genetic risk factors for CAA, we investigated the association of BCHE-K with CAA. METHODS: The association between the severity of CAA and BCHE-K was investigated in 155 autopsy cases of the elderly, including 48 patients with AD. RESULTS: There was no significant association of BCHE-K with the severity of CAA in the total, AD, or non-AD cases. Status of the epsilon4 allele of apolipoprotein E gene did not influence the results. CONCLUSIONS: Our results may suggest that BCHE-K is not a definitive risk factor for CAA in the elderly, although further study with larger samples is necessary to confirm this.

Aged↗

PubMed1998

Stimulation of the preprothyrotropin-releasing hormone gene by epidermal growth factor.

Regulation of the expression of the prepro-TRH (ppTRH) gene by epidermal growth factor (EGF) was investigated. The i.p. injection of EGF significantly stimulated hypothalamic ppTRH messenger RNA levels in rats. To clarify whether this stimulatory effect of EGF could be exerted at the level of gene transcription, the 5'-flanking region (-1893/+127) of the mouse ppTRH gene fused to a luciferase reporter gene was transiently transfected into pituitary GH4C1 cells, and the effect of EGF on gene transcription was measured by a luciferase assay. EGF stimulated ppTRH gene promoter activity in a time- and dose-dependent manner. Deletion analysis revealed that two different regions of the promoter, between -254 and -218 [EGF response element-1 (EGFRE1)] and between -130 and -84 (EGFRE2) were required for full stimulation by EGF. The two EGFREs possessed putative binding sequences for the transcription factor Sp1, and they functioned cooperatively in heterologous promoters. Nuclear extracts from GH4C1 cells specifically bound those two EGFREs in gel retardation assays. Two protein-DNA complexes were found on EGFRE1, whereas four complexes were observed on EGFRE2. Although the binding of nuclear extracts to EGFRE1 was competed for by the consensus Sp1 binding sequence, the complexes on EGFRE1 were not supershifted by an Sp1 antibody. Formation of the slower migrating protein complex on EGFRE1 was prevented by EDTA, suggesting that one of the EGFRE1-binding proteins might be an Sp1-related zinc finger protein. Competition and supershift experiments demonstrated that the EGFRE2-binding protein showing that the slowest migration possessed a characteristic similar to that of Sp1. Selective mutations of the Sp1-binding site in EGFRE2 markedly diminished the EGF-induced stimulation. These results suggest that EGF may function as a positive regulator of ppTRH gene expression, and that the stimulatory effect may be mediated through a cooperative interaction between Sp1 or Sp1-related proteins and additional factors that bind to two separate DNA regions.

Animals↗

PubMed1998

Enhanced miotic response to topical dilute pilocarpine in patients with Alzheimer's disease.

To determine whether miotic response to dilute (0.0625%, two drops) topical pilocarpine could be useful in detecting Alzheimer's disease (AD), we assessed the response in 14 AD patients and 10 control subjects. The miotic response to pilocarpine was significantly greater in AD patients than in control subjects (p < 0.001). In contrast, mydriatic response to the anticholinergic tropicamide (0.01%, one drop) failed to show a difference between the groups (p = 0.54). There was no significant correlation between the miotic and mydriatic responses in individuals. We conclude that dilute pilocarpine could be useful as a diagnostic tool in early AD.

Administration, Topical↗

PubMed1998

Change in erythrocyte shape induced by cyclosporine administration.

In order to estimate the effect of the long term administration of cyclosporine (CsA) on the shape change of erythrocytes, erythrocyte shapes which are observed with a scanning electron microscope were classified according to the nomenclature of Bessis for stomatocyte-echinocyte shape transformation. As a result of observing the erythrocyte shape of fifty-six patients with kidney transplantation treated with CsA, the morphological index of the erythrocytes of patients significantly increased to 0.0835+/-0.0085*** in comparison with 0.0004+/-0.0051 of those from healthy volunteers (control) (***: p<0.001, ANOVA). Such transformations had no relation to the subjects' sex or age. On the other hand, the erythrocytes of patients administered more than 100 ng/ml of CsA and posttransplanted within less than two years were transformed by CsA from the state of discocyte to echinocyte. In rats, the morphological index of erythrocytes of rats treated with 3 mg/kg/d or 5 mg/kg/d of CsA significantly increased in comparison with rats treated with saline (control). Furthermore, the erythrocytes of two patients were observed in terms of shape before the treatment with CsA. In both patients, the echinocyte type of erythrocyte increased by treatment with CsA. In vitro, the morphological index of the erythrocytes incubated with plasma containing CsA significantly increased, to 0.459+/-0.066*** in comparison with 0.064+/-0.029 of the control. It is suggested from these results that CsA treatment induces the echinocyte type of erythrocyte.

Adult↗