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Biomedical subjects

A Nakamura

Publications and source records attributed to A Nakamura.

At least 271 records · Page 15Linked to original sources

Presacral epidermoid cyst: report of a case.

An asymptomatic tumor in the pelvis was incidentally found by ultrasonography in a 67-year-old woman while being examined after presenting with a common cold. Further examinations revealed a presacral cystic tumor, which measured 10 x 12cm in size. The cyst was thus removed in the normal manner for such cases. The pathological diagnosis was an epidermoid cyst. An analysis of 15 other cases previously reported in the literature indicated that large epidermoid cysts should normally be excised through an abdominal approach alone, provided that the tumor is benign.

Aged↗

A novel splicing mutation in the ceruloplasmin gene responsible for hereditary ceruloplasmin deficiency with hemosiderosis.

Hereditary ceruloplasmin deficiency with hemosiderosis (aceruloplasminemia) is a newly recognized autosomal recessive disorder of copper-iron metabolism due to mutations in the ceruloplasmin (Cp) gene. We report here a novel mutation in the Cp gene in a 54-year-old Japanese woman with this disease. She showed clinical triad; diabetes mellitus, retinal degeneration and neurological disorder in her middle age. Laboratory findings were characteristic for no detectable serum ceruloplasmin and increased serum ferritin. Liver biopsy revealed excessive storage of iron in hepatocytes and magnetic resonance imaging of the brain was indicative of increased iron content in the basal ganglia, thalamus and dentate nucleus. The a-->g substitution at the splice acceptor site of the intron 6 (1209-2) caused a 8-bp deletion in Cp mRNA by defective splicing, resulting in a premature termination codon at the amino acid position 388. Truncation of Cp, even if effectively translated, may cause loss of its normal function because of drastic change in its triangular structure.

Alternative Splicing↗

The effect of high altitude and caloric restriction on reactive carbonyl derivatives and activity of glutamine synthetase in rat brain.

Both exposure to high altitude (HA) and caloric restriction (CR) may influence free radical generation. The aim of the present study was to evaluate the effects of 4 wk chronic exposure to 4,000m of HA and CR (60% of CR of control (C) rats) on protein oxidation in brain. Eighteen rats with similar body mass were assigned to CR, HA and C rats. Reactive carbonyl derivatives (RCD), marker of protein oxidation, were measured by Western blot. In addition, the activity and protein content of glutamine synthetase (GS) were determined. The body mass of C rats was significantly higher (P< 0.001) than that of HA and CR groups. The quantified signal intensity of RCD was significantly stronger in C rats than in HA and CR rats. The activity of GS was significantly increased in CR rats, while the protein content of GS was decreased in HA rats compared to C group. The data suggest that both HA and CR decreases the accumulation of RCD in the brain, however the mechanism of the decrease seems to be different during HA and CR.

Altitude↗

Role of beta-adrenoceptor on renal interleukin-6 and tumor necrosis factor in spontaneously hypertensive rats.

We have shown previously in the spontaneously hypertensive rat (SHR) kidney that interleukin-6 (IL-6) and tumor necrosis factor (TNF) mRNA levels were low under conditions of acute anaesthesia and surgical stress. The reasons for the suppression of IL-6 and TNF gene expression in the SHR were investigated by examining the influence of enhanced beta-adrenergic stimulation, high blood pressure, and renal function (renal blood flow, glomerular filtration rate, plasma creatinine levels) on renal IL-6 and TNF mRNAs. The experiments were performed by means of the following three studies; (1) SHR and Wistar rats at 4, 7, 9 week old were injected with lipopolysaccaride (LPS), and then a relationship between blood pressure levels and IL-6 and TNF mRNA levels were estimated, (2) isoproterenol and propranolol were administered into SHR and WKY rats, and the levels of IL-6 and TNF mRNA were compared, (3) under condition of anaesthesia and surgical stress, blood pressure and renal functions in SHR were measured, and then the relationships between these factors and IL-6 or TNF mRNA levels were analyzed. Renal IL-6 and TNF mRNAs in SHR remained low even though blood pressure increased with age and there was no significant correlation between IL-6 or TNF mRNA levels and values of blood pressure or renal function under anaesthesia and surgical stress. However, the inhibition of the IL-6 and TNF mRNAs in SHR was prevented by propranolol treatment. These results suggested that suppression of IL-6 and TNF mRNAs in the SHR kidney could be due to overactivity of beta-adrenergic influences which may importantly contribute to the development of hypertension.

Animals↗

Plant defense-related enzymes as latex antigens.

BACKGROUND: Latex allergy is an increasing hazard to people who frequently come into contact with latex products. Of interest concerning this immediate-type allergy is the cross-reactivity to various vegetable foods and pollen. Despite its high prevalence, no adequate explanation has been provided for the cross-reactive antigens. OBJECTIVE: We have hypothesized that a series of plant defense-related proteins act as latex allergens, as well as vegetable food allergens. To evaluate this hypothesis, hydrolytic enzymes that are very likely to take on defensive roles in rubber trees were examined for their antigenicity. METHODS: By applying chromatographic procedures, defense-related enzymes were separated from nonammoniated latex (NAL). Their antigenicity was examined by immunoblotting and ELISA with sera containing IgE antibodies to crude latex proteins. RESULTS: Three kinds of hydrolytic enzymes (basic beta-1,3-glucanases [35, 36.5, and 38 kd], a basic chitinase/lysozyme [29.5 kd], and an acidic esterase [44 kd]) were separated from NAL. They were recognized by IgE antibodies from a significant number of patients allergic to latex. The basic beta-1,3-glucanases and the acidic esterase were also strongly recognized by IgE antibodies from several atopic subjects who were allergic to various vegetable foods rather than latex products. CONCLUSION: It was ascertained that the three defense-related enzymes separated from NAL constituted part of the latex antigens. Taking together the well-known serologic or immunologic relationships and amino acid sequence similarities of defense-related proteins coming from phylogenetically distant plant species, we can suspect their universal antigenicity and cross-reactivity.

Chitinases↗

Differentially expressed olfactomedin-related glycoproteins (Pancortins) in the brain.

Messenger RNA differential display is conducted to search for genes that are expressed in a region-specific pattern in the rodent brain. Eleven novel gene fragments are isolated. One of these genes which we call pancortin, based on its predominant mRNA expression in the cerebral cortex of the adult, is studied. These pancortin cDNA clones are grouped into four different types of cDNA, designated as pancortin-1 to -4. All pancortin cDNAs share a common sequence in the middle of their structure, having two alternative sequences at both 5'- and 3'-ends, respectively. Deduced amino acid sequence shows that all pancortins have sequences of hydrophobic amino acids at N-terminus and no obvious membrane spanning regions. In situ hybridization histochemistry using oligonucleotide probes specific for 5'- and 3'-end variable parts has revealed that these four pancortin mRNAs are expressed differentially in the adult rodent brain. Robust expression of pancortin-1 and -2 mRNA is observed in the cerebral cortex (including the hippocampus and the olfactory bulb). However, little of pancortin-3 and -4 mRNA is observed there. In the cortex, some neurons are stained by an antibody raised against Pancortin. Immuno-electron microscopic study has revealed that Pancortin-like immunoreactive products are localized mainly in the endoplasmic reticulum and not in the Golgi apparatus indicating that Pancortins are the endoplasmic reticulum-anchored proteins. Our results suggest that each Pancortin is differentially regulated and may perform different functions in the brain.

Amino Acid Sequence↗

Cloning and sequencing of three C-type lectins from body surface mucus of the land slug, Incilaria fruhstorferi.

Three C-type lectins of 15 kDa were isolated from the water-soluble fraction (WSF) of the body surface mucus of the land slug, Incilaria fruhstorferi. Based on their partial amino acid sequences, the nucleotide sequences of cDNAs encoding these lectins, named incilarin A, B and C, were determined. cDNAs of incilarin A, B and C consisted of 673, 663 and 715 bp, and deduced amino acids were 150, 149 and 156 residues, respectively. All three lectins had signal peptides of 17 amino acid residues at their N-termini. They showed 44-55% amino acid sequence identity with each other, and lower but significant homology with the other animal C-type lectins and antifreeze protein. Incilarin A and B seem to possess two intramolecular disulfide bonds in the carbohydrate-binding domain (CRD) conserved among the animal C-type lectins, however, one of these bonds is absent in incilarin C.

Amino Acid Sequence↗

Molecular characterization of epidemic multiresistant Staphylococcus haemolyticus isolates.

Fifty-five Staphylococcus haemolyticus specimens isolated from patients and neonatal intensive care unit staff were tested for susceptibility to 12 antimicrobial agents. There were 34 multidrug-resistant isolates which were resistant to oxacillin, ampicillin, cefazolin, cefmetazole, imipenem, and gentamicin. These isolates had a higher frequency of resistance to tobramicin and ofloxacin, and relatively high MICs (2 to 4 micrograms/mL) for vancomycin, although none of the isolates were vancomycin resistant. To investigate hospital-acquired colonization and infection by multiresistant S. haemolyticus, we examined all isolates by pulsed-field gel electrophoresis (PFGE) after SmaI and SstII digestion, and detected an endemic PFGE pattern in multiresistant isolates. The results suggested that local spread of multiresistant S. haemolyticus was hospital acquired, and that the hospital staffs functioned as a reservoir.

Ampicillin↗

Epidemiological analysis of Salmonella enterica Enteritidis isolates in Japan by phage-typing and pulsed-field gel electrophoresis.

Salmonella enterica serotype Enteritidis isolates of phage types (PTs) PT1, PT4, PT13a and PT22 derived from sporadic cases and outbreaks of food poisoning in Japan during 1994 and 1995 were analysed by pulsed-field gel electrophoresis (PFGE). While PT1 strains from 5 different outbreaks showed 14 PFGE patterns, 5 PFGE patterns were observed among PT4 isolates from 5 different outbreaks and 6 independent isolates from imported chicken. Interestingly, 8 out of 9 PT4 strains associated with foreign travel to Southeast Asia were indistinguishable in PFGE pattern from 5 independent isolates of imported chicken from England. Although both PT13a and PT22 were first reported in Japan in 1994, PT22 showed various PFGE patterns compared to PT13a which had the same pattern within an outbreak, unlike PT1. These results could indicate that multiple clonal lines of PT1 and PT22 had already spread while relatively fewer clonal lines of PT4 and PT13a might exist in Japan.

Bacteriophage Typing↗

Gene gun-mediated in vivo analysis of tissue-specific repression of gene transcription driven by the chicken ovalbumin promoter in the liver and oviduct of laying hens.

In order to search tissue-specific elements in the 5'-upstream promoter region, gene gun was used to transfect in vivo plasmid DNAs with varying lengths of truncated ovalbumin promoter fused to the CAT reporter gene to the oviduct and liver of laying hens. The results indicated that in the oviduct, consistently high reporter gene expression was observed irrespective of the length of the truncated ovalbumin gene promoters, whereas in the liver the ovalbumin promoter extending from -3200 to +8 bp suppressed substantially the reporter gene expression compared with consistently high gene expression obtained by the ovalbumin promoters from -2800 to +8 bp or shorter length. It was concluded, therefore, that a tissue-specific silencer-like element might reside most likely in the ovalbumin gene promoter region between -3200 and -2800 bp which represses the ovalbumin gene transcription in the liver, but not in the oviduct of laying hens.

5' Untranslated Regions↗

Positional cloning of the gene for Nijmegen breakage syndrome.

Nijmegen breakage syndrome (NBS), also known as ataxia-telangiectasia (AT) variant, is an autosomal recessive disorder characterized by microcephaly, growth retardation, severe combined immunodeficiency and a high incidence of lymphoid cancers. Cells from NBS patients display chromosome instability, hypersensitivity to ionizing radiation and abnormal cell-cycle regulation after irradiation, all of which are characteristics shared with AT. Recently, the NBS locus was mapped at 8q21 by two independent approaches, complementation studies and linkage analysis. Here, we report the positional cloning of the NBS gene, NBS1, from an 800-kb candidate region. The gene comprises 50 kb and encodes a protein of 754 amino acids. The amino-terminal region of the protein shows weak homology to the yeast XRS2, MEK1, CDS1 and SPK1 proteins. The gene is expressed at high levels in the testes, suggesting that it might be involved in meiotic recombination. We detected the same 5-bp deletion in 13 individuals, and conclude that it is likely to be a founder mutation.

Amino Acid Sequence↗

Disposition of mirosamicin in honeybee hives.

Disposition of mirosamicin, a macrolide antibiotic, to honeybee adults, larvae, honey and royal jelly in the beehive after in-feed administration to adult bees was studied. Treatment was initiated at the end of July when the availability of natural pollen and nectar was poor. The drug was mixed with pollen-substitute paste and administered to honeybee colonies continuously for a week at a dosage of 200 mg/hive/week. High distributions in adult bees, jelly, larvae and a relatively low distribution in honey, of mirosamicin were observed. One day dosing of microsamin in sucrose syrup, a nectar substitute, resulted in a very high and long lasting residue in honey. Both royal and worker jelly, secreted from the jelly glands of adult bees, are acidic, so that a high distribution of a basic drug, such as mirosamicin, in jelly can be expected. This mechanism was considered to be responsible for a high concentration of mirosamicin in honeybee larvae, the host of Paenibacillus-larvae infection (American foulbrood), as primary larval food is jelly.

Animals↗

Immunologic abnormalities exhibited in IL-7 transgenic mice with dermatitis.

Interleukin (IL)-7 transgenic mice, which we established previously, developed severe dermatitis characterized by massive infiltration of gammadelta T cells in the dermal lesion. To fully understand the pathology of this intriguing skin disease, we examined several immunologic features of dermis infiltrating lymphocytes from the lesional skin of IL-7 transgenic mice. We observed a moderate response to mitogens, a poor response to alloantigens, and the absence of cytotoxic activities to several tumor cell lines and skin derived cells regardless of the presence of IL-2 or IL-7. On the other hand, dermis infiltrating lymphocytes could proliferate with exogenous IL-2 and IL-7. Moreover, reverse transcriptase polymerase chain reaction and fluorescence activated cell sorter analysis revealed that dermis infiltrating lymphocytes expressed various cytokines including IL-4 and IL-7, and several activation markers for T cells (CD44, CD69, IL-2R alpha), in addition to IL-7R alpha. In the sera of the affected mice, hyper epsilon-globulinemia was observed. These findings suggested that dermis infiltrating lymphocytes proliferated in an activated state in the skin lesion in an autocrine and/or paracrine manner and produced Th2 type cytokines that might evoke immunologic abnormalities. This study and previous findings suggest that IL-7 transgenic mouse with dermatitis offer the potential of serving as a useful tool for investigating the immunologic role of cutaneous gammadelta T cells, especially their participation in IgE production in vivo.

Animals↗

Insertional mutation by transposable element, L1, in the DMD gene results in X-linked dilated cardiomyopathy.

X-linked dilated cardiomyopathy (XLDCM) is a clinical phenotype of dystrophinopathy which is characterized by preferential myocardial involvement without any overt clinical signs of skeletal myopathy. To date, several mutations in the Duchenne muscular dystrophy gene, DMD , have been identified in patients with XLDCM, but a pathogenic correlation of these cardiospecific mutations in DMD with the XLDCM phenotype has remained to be elucidated. We report here the identification of a unique de novo L1 insertion in the muscle exon 1 in DMD in three XLDCM patients from two unrelated Japanese families. The insertion was a 5'-truncated form of human L1 inversely integrated in the 5'-untranslated region in the muscle exon 1, which affected the transcription or the stability of the muscle form of dystrophin transcripts but not that of the brain or Purkinje cell form, probably due to its unique site of integration. We speculate that this insertion of an L1 sequence in DMD is responsible for some of the population of Japanese patients with XLDCM.

5' Untranslated Regions↗

Expression and characterization of dog CYP2D15 using baculovirus expression system.

Dog CYP2D15 was expressed in Sf9 cells with a recombinant baculovirus. Infection of Sf9 insect cells with a recombinant dog CYP2D15-virus resulted in the expression of a protein which cross-reacted with a polyclonal antibody against a dog CYP2D15-specific peptide. The difference spectrum of CO-complex of reduced P450 of the infected cell microsomes had a maximal absorbance at 449 nm. The specific content of P450 was calculated to be 0.56 nmol/mg of Sf9 cell microsomal protein. Although the expressed dog CYP2D15 showed high catalytic activity for the hydroxylations of bunitrolol and imipramine at low substrate concentration (10 microM), the catalytic activity for that of debrisoquine (50 microM) was extremely low as compared with that of CYP2D from other species. Dog liver microsomes also showed bunitrolol and imipramine hydroxylase activities, but not debrisoquine hydroxylase activity at the same substrate concentrations. In addition, the expressed CYP2D showed high catalytic activity for imipramine N-demethylation. Thus, our study reveals that the expressed dog CYP2D15 engages in high catalytic activity and has a unique substrate specificity from other CYP2D subfamilies. Western blot analysis suggested that the dog CYP2D15 contents were less than 4% of the total liver P450 content, assuming that 100% of expressed CYP2D15 incorporated heme.

Animals↗

Purification and characterization of a puromycin-hydrolyzing enzyme from blasticidin S-producing Streptomyces morookaensis.

Blasticidin S-producing Streptomyces morookaensis JCM4673 produces an enzyme which inactivates puromycin (PM) by hydrolyzing an amide linkage between its aminonucleoside and O-methyl-L-tyrosine moieties [Nishimura et al. (1995) FEMS Microbiol. Lett. 132, 95-100]. In this study, we purified to homogeneity the enzyme from the cell-free extracts of S. morookaensis. The molecular weight of PM-hydrolyzing enzyme, estimated by SDS-PAGE and gel filtration, was 68 and 66 kDa, respectively, suggesting that this protein is monomeric. The PM-hydrolyzing activity was strongly inhibited by Zn2+, Fe2+, Cu2+, Hg2+, and N-bromosuccinimide, but was stimulated by DTT. The optimum pH and temperature for PM-hydrolyzing activity were 8.0 and 45 degrees C, respectively. Several L-aminoacyl-beta-naphthylamides were good substrates for the enzyme, suggesting that the PM-inactivating enzyme has an aminopeptidase activity. The N-terminal sequence of the first 14 amino acids (Val-Ser-Thr-Ala-Pro-Tyr-Gly-Ala-Trp-Gln-Ser-Pro-Ile-Asp) of the enzyme showed no significant homology with any published hydrolase sequences.

Aminopeptidases↗

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↗