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Biomedical subjects

A Miyazaki

Publications and source records attributed to A Miyazaki.

At least 91 records · Page 5Linked to original sources
PubMed1998

Reduced expression of the macrophage scavenger receptors in macrophage-like cell mutants resistant to brefeldin A.

Brefeldin A (BFA)-resistant mutants, JB15, JB23 and JB33, were isolated from mutagenized murine macrophage-like (J774) cells and their modified low density lipoprotein (LDL) metabolism was studied. When JB23 cells, the most resistant clone, were incubated with acetylated LDL, intracellular accumulation of cholesteryl esters (CE) was reduced by 31% as compared with J774 cells. The cell-association of 125I-acetyl-LDL with, and subsequent endocytic degradation by JB23 cells were reduced by 40-60% compared with J774 cells. Western and Northern blot analyses showed that the protein and mRNA levels of the macrophage scavenger receptors (MSR) were reduced by 68% and 55% respectively in JB23 cells as compared with those in J774 cells. These results indicate that a putative BFA-target molecule(s) might regulate MSR gene expression as well as macrophage-derived foam cell formation.

Animals↗
PubMed1998

Diffuse bilateral thalamic astrocytomas as examined serially by MRI.

We report the case of a 13-year-old girl with diffuse bilateral thalamic astrocytomas. Incoordination was observed at the onset. Cranial computed tomography (CT) showed enlarged thalami, and magnetic resonance imaging (MRI) revealed these lesions to be symmetrically enlarged with high intensity on the T2-weighted image. Owing to these atypical findings in the neuroimaging studies, we had difficulty in making the correct diagnosis of a brain tumor. After the diagnosis of diffuse bilateral thalamic astrocytomas was obtained, we performed hyperfractionated radiotherapy followed by chemotherapy. Radiation therapy was effective for a while, but the girl's condition deteriorated again and she died 8 months after admission. Although diffuse bilateral thalamic astrocytomas are difficult to diagnose because they do not resemble most other neoplasms on neuroimaging studies, pediatricians should keep this entity in mind in order to arrive at a precise and prompt diagnosis.

Adolescent↗
PubMed1998

Effects of teprenone on gastric epithelial restoration in a rabbit cultured cell model.

The mechanism of action of gastrocytoprotective agents is not fully understood. We assessed the effects of an anti-ulcer agent, teprenone, and bile acid on epithelial restoration. Teprenone with or without deoxycholic acid was added to a complete confluent cultured gastric epithelial cell sheet after wounding. Restoration was monitored for 48 h, and the wound size was assessed. Migration velocity was measured, and proliferation was detected by sequential staining with bromodeoxyuridine. The labeling index was calculated. Restoration was completed within 48h in controls, whereas deoxycholic acid retarded repair. The migration velocity was suppressed by deoxycholic acid treatment. The number of proliferative cells peaked at 36 h (labeling index, 1.7%) in controls. In the deoxycholic acid group, the maximal labeling index was 0.5% at 48 h. Teprenone abolished the bile acid-induced retardation. Teprenone showed cytoprotective effects against deoxycholic acid-induced inhibition of epithelial cell migration and proliferation.

Animals↗
PubMed1998

Effect of a prolyl endopeptidase inhibitor, JTP-4819, on radial maze performance in hippocampal-lesioned rats.

The effect of a novel prolyl endopeptidase (PEP) inhibitor, (S)-2-2[[(S)-2-(hydroxyacetyl)-1-pyrrolidinyl]carbonyl]-N-(phenylmethyl) -1-pyrrolidinecarboxamide (JTP-4819), on spatial learning deficits in rats with dorsal hippocampal (DH) lesions was examined using an eight-arm radial maze. The correct performance remained at chance levels even after 18 acquisition trials in rats with DH lesions. JTP-4819 (3.0 mg/kg, p.o.) significantly ameliorated this learning impairment after 34-41 days of treatment. When DH lesions were created in rats after achievement of learning, postoperative performance deteriorated prominently, but gradually recovered with the repetition of trials. JTP-4819 (3.0 mg/kg, p.o.) significantly decreased the number of trials needed to reattain learning criterion. After the behavioral experiment, the choline acetyltransferase (ChAT) activity and [3H]-pirenzepine binding (Kd, Bmax) in the residual hippocampus and cerebral cortex were analyzed. Neither parameter was significantly affected by JTP-4819. In conclusion, JTP-4819 can improve both learning and relearning deficits of spatial memory in DH-lesioned rats, postulating that enhancement of neuropeptide activity via PEP inhibition may be involved in the mechanism of action of JTP-4819.

Animals↗
PubMed1998

A neurotrophic pyrimidine compound, MS-818, enhances EGF-induced restoration of gastric epithelial wounds in vitro.

MS-818 is a novel synthetic pyrimidine compound that stimulates nerve regeneration and promotes synthesis of various growth factors and differentiation of astrocytes. Effects of MS-818 on gastric epithelial cells were assessed using a wound repair model with primary cultured gastric epithelial cells from rabbits. A round wound with a constant cell-free area was created and the process of restoration was monitored by measuring wound size every 12 h. Cell proliferation was monitored by sequential staining with BrdU. As previously reported, EGF (10 ng/ml) accelerated wound repair by promoting cell migration and proliferation. Although MS-818 alone had no effects, MS-818 (10-100 microM) enhanced EGF-induced acceleration of gastric epithelial restoration, including cell migration and proliferation. Although the detailed mechanism of action of this agent is still unclear, MS-818 might have favorable effects on in vivo gastric mucosal repair.

Animals↗
PubMed1998

Liver transplantation in a case of hypoproteinemia and coagulopathy.

A female infant with hypoproteinemia and coagulopathy associated with hypertyrosinemia was successfully treated with living-related liver transplantation (LRLT). On the 12th day of life plasma amino acid analysis revealed a marked elevation of tyrosine, so the patient was fed on a low-tyrosine and low-phenylalanine diet. However, hepatosplenomegaly, hypotonia, alopecia, eczema and psychomotor delay did not improve and recurrent episodes of disseminated intravascular coagulation (DIC) caused her condition to deteriorate. Liver biopsy on the 230th day revealed marked fatty change accompanied by mild to moderate cholestasis. Therefore, LRLT from her father was performed on the 286th day resulting in improvement of all the aforementioned signs and symptoms. Despite a thorough examination, no diagnosis of a known disorder could be established. However, her elder brother had also been born with severe hypoproteinemia and coagulopathy, and died of DIC on the second day of life. Thus, the disorder is designated as a new entity, namely 'congenital hypoproteinemia and coagulopathy associated with hypertyrosinemia'.

Disseminated Intravascular Coagulation↗
PubMed1998

Expression of ACAT-1 protein in human atherosclerotic lesions and cultured human monocytes-macrophages.

The acyl coenzyme A:cholesterol acyltransferase (ACAT) gene was first cloned in 1993 (Chang et al, J Biol Chem. 1993;268:20747-20755; designated ACAT-1). Using affinity-purified antibodies raised against the N-terminal portion of human ACAT-1 protein, we performed immunohistochemical localization studies and showed that the ACAT-1 protein was highly expressed in atherosclerotic lesions of the human aorta. We also performed cell-specific localization studies using double immunostaining and showed that ACAT-1 was predominantly expressed in macrophages but not in smooth muscle cells. We then used a cell culture system in vitro to monitor the ACAT-1 expression in differentiating monocytes-macrophages. The ACAT-1 protein content increased by up to 10-fold when monocytes spontaneously differentiated into macrophages. This increase occurred within the first 2 days of culturing the monocytes and reached a plateau level within 4 days of culturing, indicating that the increase in ACAT-1 protein content is an early event during the monocyte differentiation process. The ACAT-1 protein expressed in the differentiating monocytes-macrophages was shown to be active by enzyme assay in vitro. The high levels of ACAT-1 present in macrophages maintained in culture can explain the high ACAT-1 contents found in atherosclerotic lesions. Our results thus support the idea that ACAT-1 plays an important role in differentiating monocytes and in forming macrophage foam cells during the development of human atherosclerosis.

Adult↗
PubMed1998

Analysis of differentiation-induced expression mechanisms of thyrotropin receptor gene in adipocytes.

Rat adipose tissue, as well as differentiated 3T3-L1 cells, has been shown to express TSH receptor (TSHR) mRNA in amounts approaching those in the thyroid. We investigated the molecular mechanisms of TSHR gene expression in adipose cells. Primer extension and cloned cDNA sequences showed that transcription of the TSHR gene in rat adipose tissue was from multiple start sites clustered between -89 to -68 bp and almost identical to those in FRTL-5 thyroid cells. By transient expression analysis, we localized, between -146 and -90 bp, a positive regulatory element, the activity of which was markedly increased after the differentiation of 3T3-L1 cells. Deoxyribonuclease I protection showed that nuclear extracts from differentiated 3T3-L1 cells strongly protected two sequences, from -146 to -127 bp, including a cAMP response element-like sequence and from -112 to -106 bp containing a putative Ets-binding sequence. In differentiated 3T3-L1 cells, disruption or deletion of either sequence was found to result in the loss of enhancer activity, suggesting both elements may synergistically activate the TSHR promoter. Electrophoretic mobility shift analysis revealed the induction of new protein/DNA complexes formed either with the cAMP response element-like site or with putative Ets elements after the differentiation into adipocytes. In contrast, nuclear proteins, whose binding to DNA was diminished after the differentiation of 3T3-L1 cells, were found to interact with the site contiguous to the 5'-end of the putative Ets-binding sequence. Mutations of this binding site, which reduced the protein/DNA complex formation, increased TSHR promoter activity in undifferentiated cells. These observations suggested that differentiation-induced diminution of suppressor interactions may allow the enhancers to synergistically activate the transcription of TSHR gene in adipocytes.

Adipocytes↗
PubMed1998

A novel prolyl endopeptidase inhibitor, JTP-4819--its behavioral and neurochemical properties for the treatment of Alzheimer's disease.

Formation of beta-amyloid and neurofibrillary tangles in the brain due to genetic or other factors is the most frequent cause of Alzheimer's disease. In addition, marked reduction of certain brain neuropeptide levels is a consistent finding in patients with Alzheimer's disease, together with the deterioration of cholinergic neurons. Currently, there is great demand for the development of new drugs to improve memory deficits or to delay the neurodegenerative process in conditions such as Alzheimer's disease. In this report, the pharmacological actions of JTP-4819, a novel specific prolyl endopeptidase (PEP) inhibitor devised for the treatment of Alzheimer's disease, are reviewed with respect to its effects on PEP activity, neuropeptidergic and cholinergic neurons, and memory-related behavior in rats. We also discuss the possible beneficial effect of JTP-4819 on beta-amyloid metabolism and its potential neuroprotective properties.

Alzheimer Disease↗
PubMed1998

Hb Peterborough [beta 111(G13)Val-->Phe] in Japan; rapid identification by ESI/MS using proteolytic digests of oxidized globin.

A variant hemoglobin was found in a Japanese female whose hemoglobin was studied to clarify the cause of a low Hb A1c value, found during a routine medical examination. The detection and identification of the variant was performed by electrospray ionization mass spectrometry. Its structure was revealed to be the same as Hb Peterborough [beta 111(G13)Val-->Phe]. For sequence determination, oxidized globin as well as non-derivatized globin were cleaved by trypsin and lysyl endopeptidase. An abnormal peptide was found in digests of oxidized globin, as shown by electrospray ionization mass spectrometry. Cysteic acid in oxidized peptides enhanced the abundance of fragment ions in tandem mass spectrometry, which helped to quickly and accurately determine the substitution in beta T-12, a peptide in the core region. Electrospray ionization mass spectrometry analysis of the hemolysate also showed a low level of glycated hemoglobin. The patient's hemolysate showed decreased stability in the isopropanol test. An abnormal band was detected on isoelectrofocusing on the cathodic side of normal Hb A. This is the second report of Hb Peterborough and the first of its occurrence in Japan.

Adult↗
PubMed1998

A new hemoglobin variant found during Hb A1c measurement: Hb Hokusetsu [beta52(D3)Asp-->Gly].

A new beta chain variant was accidentally found through the assay of Hb A1c in a diabetic patient. The variant was detected by polyacrylamide gel isoelectrofocusing and electrospray ionization mass spectrometry. For sequence determination, globin was cleaved with combination of trypsin and lysyl endopeptidase and analyzed by high performance liquid chromatography connected to electrospray ionization mass spectrometry. An abnormal betaT-5 peptide was found by reconstructed selected ion monitoring. The collision-induced dissociation spectrum of an ion derived from the abnormal betaT-5 peptide revealed a new substitution, [beta52(D3)Asp-->Gly], named Hb Hokusetsu. The sequence was confirmed with an automatic sequencer using peptides isolated by reversed phase high performance liquid chromatography. Amplification of the beta-globin exon 2 and nucleotide sequencing revealed a GAT-->GGT mutation in codon 52 corresponding to an Asp-->Gly replacement. Electrospray ionization mass spectrometry analysis of the hemolysate showed a reasonable value of 10.4% for glycated globin. The variant migrated as Hb S on isoelectrofocusing. Hematological analysis revealed normal parameters. The patient's hemolysate showed normal stability in the isopropanol test. Oxygen equilibrium studies on the patient's red blood cells and hemolysate showed no significant change in oxygen affinity or cooperativity.

Amino Acid Sequence↗
PubMed1998

Cytotoxic effect of oxidized low density lipoprotein on macrophages.

Macrophage or macrophage-derived foam cell death is one of the characteristic events in the development of cell-poor lipid-rich cores of the advanced atherosclerotic plaques. Although the in vivo mechanism for the death of macrophages is unclear, one possible candidate for the agent which induces macrophage cell death is oxidized low density lipoprotein (Ox-LDL). To investigate the mechanism of Ox-LDL-induced macrophage cell death, we have recently employed macrophage cell genetics and isolated mutant cells resistant to the cytotoxic effect of Ox-LDL from mutagenized populations of murine macrophage-derived J774 cells (Hakamata, H., Miyazaki, A., Sakai, M., Matsuda, H., Suzuki, H., Kodama, T., and Horiuchi, S. (1998) J. Lipid Res. 39, 482-494). The results obtained showed that one mutant form, JO21b cells, was characterized by reduced expression of type I and type II class A macrophage scavenger receptors (MSR-AI/AII) with a concomitant decrease in the uptake of Ox-LDL. Moreover, peritoneal macrophages obtained from MSR-AI/AII-knockout mice showed a higher resistance to the cytotoxic effect of Ox-LDL compared to those of their wild-type littermates. From these results, we have concluded that Ox-LDL cytotoxicity to macrophages is enhanced by effective endocytic uptake of Ox-LDL through MSR-AI/AII. These findings imply a possibility that formation of the cell-poor lipid-rich core is also enhanced by MSR-AI/AII-mediated uptake of Ox-LDL and subsequent macrophage cell death in atherosclerotic lesions.

Animals↗
PubMed1998

A new nonamyloid transthyretin variant, G101S, detected by electrospray ionization/mass spectrometry. Mutations in brief no. 201. Online.

Familial amyloidotic polyneuropathy is caused by transthyretin (TTR) variants. The identification of new variants with and without amyloidosis may help to clarify the mechanism of amyloid fibril formation. We detected several variant TTRs from patients with and without symptoms of amyloidosis using mass spectrometry (MS). TTR was isolated by mixing test serum with anti-transthyretin antiserum, and the generated immunoprecipitate was analyzed by high performanced liquid chromatography/electrospray ionization (HPLC/ESI) MS. Variant TTRs showed extra peaks in addition to normal TTR peaks. A variant found in nonamyloid group was sequenced by HPLC/ESI tandem MS using peptides obtained by protelytic digestion of TTR and by DNA analysis. The structure was new, [G101S], and was found in a 74 years old Japanese male. This mutation results from substitution in a CpG hot spot. The substitution in the surface loop, 98-102, between F and G b-strands may not cause amyloid formation.

Aged↗
PubMed1998

Lipid lowering effects of pravastatin in common marmosets.

In the present study it was examined if the common marmoset was susceptible to pravastatin (CAS 81131-70-6, CS 514, Mevalotin), an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. It has been reported that serum lipid levels in common marmosets resemble those in humans. First serum total cholesterol and lipoprotein cholesterol levels of common marmosets were examined in comparison with those in seven different experimental animals including cynomolgus monkeys, beagle dogs, Watanabe heritable hyperlipidemic rabbits, Japanese White Rabbits, Sprague Dawley rats, Wistar-Imamichi rats and golden hamsters. The animals were fed normal chow diets, and total- and lipoprotein-cholesterol levels were determined under identical conditions. The quantitative and polyacrylamide electrophoresis studies indicated that among animals examined serum lipid and lipoprotein profiles in common marmosets were similar to those in humans. When pravastatin at 1-30 mg/kg was orally administered to common marmosets for 28 days, total, low density lipoprotein (LDL)- and high density lipoprotein-cholesterol levels were decreased in a dose dependent manner, whereas triglycerides and very low density lipoprotein cholesterol did not change. These findings confirm that common marmosets have serum lipid and lipoprotein profiles similar to those in humans suggesting that they are susceptible to lipid lowering effects of HMG-CoA reductase inhibitors.

Animals↗
PubMed1998

Isolation of macrophage-like cell mutants resistant to the cytotoxic effect of oxidized low density lipoprotein.

A high concentration of oxidized low density lipoprotein (Ox-LDL) showed a cytotoxic effect on mouse macrophage-derived J774 cells. Mutant cells were selected from these cells that were resistant to the cytotoxic effect of Ox-LDL. One mutant form, named JO21b cells, was characterized in the present study. In spite of a marked resistance to the cytotoxic effect of Ox-LDL, JO21b cells were apparently as sensitive as the parent cells not only to toxic moieties of Ox-LDL, such as 7-ketocholesterol and lysophosphatidylcholine, but also to t-butyl hydroperoxide, an artificial lipid hydroperoxide analog. However, the cellular association of 125I-labeled Ox-LDL with, and subsequent endocytic degradation by JO21b cells was reduced by 70-80% compared with J774 cells. Similarly, accumulation of cholesteryl esters in JO21b cell by Ox-LDL was also reduced by 70%. Northern blot analyses of type I and type II macrophage scavenger receptors (type I and type II MSR) demonstrated that the mRNA levels of JO21b cells were lower than those of J774 cells. Moreover, peritoneal macrophages obtained from MSR-knockout mice showed a higher resistance to the cytotoxic effect of Ox-LDL than those from their wild-type littermates. Our results suggest, therefore, that macrophage scavenger receptor-mediated endocytic uptake of oxidized low density lipoproteins (Ox-LDL) may play an enhancing role in Ox-LDL cytotoxicity to macrophages or macrophage-derived cells.

Animals↗
PubMed1998

[Detection and characterization of aberrant blood proteins and quantification of glycated hemoglobin by mass spectrometry].

Electrospray Ionization Mass Spectrometry can be applied to detect aberrant proteins using intact molecules. Direct examination of hemolysate might well facilitate rapid ascertainment of a variant hemoglobin (Hb) provided that the mass difference between normal and abnormal chains is larger than the resolution power of standard instruments (i.e. = 10 Da). We propose immunoprecipitation as a preparation method of plasma and cell proteins other than Hb prior to MS. Amino acid sequences of various variants detected by MS were determined by MS/MS. Some of these variants were new. These new variants were; 1: Hb Sagami[beta 139(H17)Asn-->Thr]. 2: Hb Hokusetsu[beta 52(D3)Asp-->Gly]. 3: a variant transthyretin, amyloidogenic, [38Asp-->Ala]. 4: a variant transthyretin, non-amyloidogenic, [101Gly-->Ser]. The abundance of ion peaks showed the approximate ratio of each component, which was in agreement with the ratio obtained by chromatography and by ESIMS in the analyses of glycated hemoglobin. Samples with low kidney function (BUN > 50 mg/dl, creatinine > 2.5 mg/dl) showed higher values of glycated Hb on routine HPLC than the MS method. Samples containing high carbamylated Hb might cause this discrepancy.

Glycated Hemoglobin↗
PubMed1998

[Evaluation of anomalous bile ducts using helical CT cholangiography].

PURPOSE: To evaluate the usefulness of helical CT cholangiography (DIC-CT) for laparoscopic cholecystectomy. MATERIAL AND METHODS: Prior to laparoscopic cholecystectomy, 151 patients were examined by DIC-CT. DIC-CT was performed 30 minutes after drip infusion of 100 ml of Iotroxic acid meglumine. Two-dimensional axial (2D) images with 2.5 mm or 1.5 mm slice thickness were reformatted from the raw data, and three-dimensional (3D) images were reconstructed using the surface reconstruction method. Two- and three-dimensional images were compared with the operative cholangiograms in all subjects. RESULTS: DIC-CT images of 16 cases of aberrant bile duct (10.6%) and 4 cases of anomalous junction of the cystic duct (2.6%) corresponded to the operative cholangiogram. In one case of an aberrant posterior bile duct, which drained into the common bile duct more proximally to the papilla Vater than the cystic duct did, it was difficult to diagnose correctly the anomalous junction of the bile duct on 2D images but easy on 3D images. Although another case was misdiagnosed to have double cystic ducts on 3D images, the 2D images and operative cholangiogram showed normal junction of a cystic duct. CONCLUSION: Both 2D and 3D images were useful and necessary in the evaluation of anomalous bile ducts before laparoscopic cholecystectomy.

Adult↗