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Biomedical subjects

R Nakamura

Publications and source records attributed to R Nakamura.

At least 145 records · Page 8Linked to original sources
PubMed1996

Rice allergenic protein and molecular-genetic approach for hypoallergenic rice.

Allergenic proteins with a molecular mass of about 14 to 16 kDa were isolated from a rice salt-soluble fraction based on the reactivity with IgE antibodies from patients allergic to rice. cDNA clones encoding these allergenic proteins were isolated from a cDNA library of maturing rice seeds, and the deduced amino acid sequences showed considerable similarity to wheat and barley alpha-amylase/trypsin inhibitors, which have recently been identified as major allergens associated with baker's asthma. An antisense RNA strategy was applied to repress the allergen gene expression in maturing rice seeds. Immunoblotting and ELISA analyses of the seeds using a monoclonal antibody to a 16-kDa allergen showed that allergen content of seeds from several transgenic rice plants was markedly lower than that of the seeds from parental wild type rice.

Allergens↗
PubMed1996

Characterization of the bovine kappa-casein gene promoter.

kappa-Casein gene promoter was localized within a 570-bp fragment (-552/+18) of a 5'-flanking region by the gene transfection assay. Deletion mutation analysis in mammary epithelial cell line, HC11, suggested that there are regulatory element in a region from -439 through -125. Some nuclear proteins from lactating rate mammary gland bind to this region specifically. One of them expressed preferentially during pregnancy bound to a 132-bp fragment (-439/-308) and another expressed preferentially during lactation bound to a 183-bp fragment (-307/-125).

Animals↗
PubMed1995

Carboxy-terminal cytoplasmic domain of mouse butyrophilin specifically associates with a 150-kDa protein of mammary epithelial cells and milk fat globule membrane.

A cDNA encoding mouse butyrophilin was obtained by reverse transcriptase-coupled polymerase chain reaction (RT-PCR) using poly (A)+ RNA from lactating mouse mammary gland as a template and screening a cDNA library with the RT-PCR-amplified fragment as a probe. DNA sequencing and computer analysis revealed that it has a rather long 3'-untranslated sequence and that the carboxy-terminal cytoplasmic domain was well conserved between mouse and bovine butyrophilins. To elucidate the biological function of butyrophilin, the cytoplasmic region expressed as fusion protein with glutathione S-transferase (GST) was purified and incubated with the cell lysate of mouse mammary epithelial cell lines, COMMA-ID and HC11. A 150-kDa protein was shown to specifically associate with the cytoplasmic domain and the protein increased in amount when the cells were treated with basal medium supplemented with lactogenic hormones such as prolactin, insulin and glucocorticoid. N-terminal amino acid sequencing indicated that the protein is xanthine dehydrogenase/oxidase which has been cloned from mouse liver. Further, the cytoplasmic domain also bound xanthine dehydrogenase/oxidase from bovine milk fat globule membrane. These results suggest that butyrophilin might be physiologically associated with xanthine dehydrogenase/oxidase and might function in a complex form in milk fat secretion.

Amino Acid Sequence↗
PubMed1995

Molecular cloning of glycoprotein antigens MGP57/53 recognized by monoclonal antibodies raised against bovine milk fat globule membrane.

A cDNA encoding 57 kDa and 53 kDa antigens (MGP57/53) recognized by monoclonal antibodies raised against bovine milk fat globule membrane (MFGM) (Biochim. Biophys. Acta 1199 (1994) 87-95) was cloned from lactating bovine mammary gland by a combination of reverse transcriptase-coupled polymerase chain reaction (RT-PCR) and 3'-rapid amplification of cDNA ends (3'-RACE). The deduced amino-acid sequence showed that mature MGP57/53 consists of 409 amino-acid residues and the calculated molecular weight and isoelectric point are 45,544 and 6.42, respectively. Computer analysis reveals that it has a significant similarity to mouse mammary epithelial cell surface protein, MFG-E8 and a human breast tumor-associated glycoprotein antigen, BA46-1. An N-terminal cysteine-rich domain and a C-terminal tandemly repeated sequence were highly conserved among them, but bovine MGP57/53 lacks 36 amino-acid residues containing a cluster of 5 prolines found in mouse MFG-E8. Northern blot analysis showed that the cDNA hybridized to about 2.0 kb mRNA of lactating bovine mammary gland. These results strongly support our previous report that the two MFGM antigens originate from a single gene and are isoforms with different N-linked sugar chains.

Amino Acid Sequence↗
PubMed1995

Selective degeneration of inhibitory interneurons in the rat spinal cord induced by intrathecal infusion of acromelic acid.

We investigated the characteristics of the neurotoxicity mediated by non-N-methyl-D-aspartic acid (NMDA)-type glutamate receptors in the spinal cord by infusing rats intrathecally with three specific agonists (acromelic acid A (ACRO), kainic acid and 1-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)). When ACRO was infused intrathecally continuously for 2 h, the rats developed dose-dependent flaccid paraplegia during the infusion, followed by pure motor, rigid-spastic, long-lasting paraparesis (ED50: 220 pmol/h). The paraparetic rats showed selective degeneration of interneurons in the spinal cord with about 50% loss of their contents of glycine and aspartic and glutamic acids. The alpha-motoneurons in the ventral horns were largely free from permanent damage. These changes were selectively ameliorated by concomitant administration of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), an antagonist at non-NMDA receptors. Kainate induced long-lasting paraplegia with neuronal damage at doses about 40-times higher than those of ACRO that caused paraplegia. When examined 30 days after kainate infusion, the neurotransmitter markers had decreased markedly in the lumbar cords of the rats with long-lasting paraplegia. Intrathecal infusion of large doses of AMPA caused long-lasting flaccid paraplegia, which were accompanied by extensive necrosis in the caudal spinal cord. It seems that ACRO exerts its unique pharmacological actions by activating a subclass of non-NMDA receptors distinct from those activated by kainate and AMPA, and may become a useful tool for investigating the biological roles of glutamate receptors.

Animals↗
PubMed1995

Classification of rice allergenic protein cDNAs belonging to the alpha-amylase/trypsin inhibitor gene family.

Seven cDNA clones encoding rice allergenic proteins were newly isolated. Comparison of the sequences of ten cDNA clones, including the previously isolated three clones results in their classification into four subfamilies. Homologies in the nucleotide sequences among and within subfamilies are 70-85% and above 95%, respectively. A sequence of twenty five amino-acid residues at the C-terminal proximal region is highly conserved among all clones and resembles that of plant lipid transfer proteins.

Allergens↗
PubMed1995

Immunological characterization and localization of a Porphyromonas gingivalis BApNA-hydrolyzing protease possessing hemagglutinating activity.

A monoclonal antibody (mAb-PC) was produced against a BA pNA-hydrolyzing protease possessing hemagglutinating activity (Pase-C) from Porphyromonas gingivalis. Other P. gingivalis BA pNA-hydrolyzing enzymes (Pase-B and Pase-S) did not react with this antibody. By ELISA or SDS-PAGE and Western immunoblotting analysis, mAb-PC recognized all P. gingivalis and P. endodontalis strains tested but did not recognize other members of the Porphyromonas genus nor other putative periodontopathogenic organisms. Pase-C, extracellular vesicles (ECV) and human strains of P. gingivalis showed two major immunoreactive bands (44 kDa and 40 kDa), whereas a different pattern was obtained with animal strains of P. gingivalis. Biotinylarginyl chloromethane, an irreversible inhibitor of trypsin-like proteases, did not affect the reactivity of Pase-C with mAb-PC on immunoblot. By reversed-phase electronmicroscopy following immunogold labeling, the antibody was shown to bind to the cell surface of P. gingivalis. mAb-PC inhibited the hemagglutinating activity of both P. gingivalis cells and ECV whereas a monoclonal antibody against LPS of P. gingivalis did not. These results suggest that Pase-C is located on the cell surface of P. gingivalis and may participate in erythrocyte binding.

Animals↗
PubMed1995

Precursor-product relationship between chicken vitellogenin and the yolk proteins: the 40 kDa yolk plasma glycoprotein is derived from the C-terminal cysteine-rich domain of vitellogenin II.

Chicken vitellogenin, a serum lipoprotein specific for laying hens, has been thought to be proteolytically cleaved into the heavy and light chain lipovitellins and phosvitin, the major yolk granule proteins, during or after transportation into oocyte. In this study, another proteolytic product of vitellogenin has newly been isolated from the 'beta-livetin' fraction of yolk plasma. It is a yolk glycoprotein of 40 kDa (YGP40) with asparagine-linked carbohydrate chain(s) recognized by Concanavalin A and castor bean lectin (RCA-I), and it is identified as a C-terminal cysteine-rich fragment of the major vitellogenin (vitellogenin II), the cysteine-rich domain homologous to D2 region of von Willebrand factor. Another yolk plasma glycoprotein of 42 kDa is suggested to be one of the proteolytic products of the minor vitellogenin (vitellogenin I). Both 40 kDa and 42 kDa glycoproteins were shown to be present in growing oocytes but absent in laying hen's serum. Limited proteolysis of vitellogenin II with cathepsin D produced a 40 kDa protein with reactivity to anti-YGP40 antibody. Gel filtration analysis of vitellogenin II digested with cathepsin D showed that YGP40 dissociated from lipovitellin-phosvitin complex after the proteolytic cleavage. These results suggest that after incorporation from serum via a specific receptor vitellogenin II is cleaved in the oocyte into four fragments, heavy and light chain lipovitellins, phosvitin and YGP40, and that YGP40 is released into the yolk plasma before or during compartmentation of lipovitellin-phosvitin complex into the yolk granule.

Amino Acid Sequence↗
PubMed1995

A bovine IgG heavy chain contains N-acetylgalactosaminylated N-linked sugar chains.

A 56K protein co-purified with bovine milk fat globule membrane (MFGM) proteins bound to Wisteria floribunda agglutinin (WFA) like most MFGM glycoproteins. Treatment with N-glycanase or beta-N-acetylhexosaminidase abolished the lectin binding to the protein. Amino acid sequence and immunoblot analyses revealed that the 56K protein is an IgG heavy chain. Lectin column chromatography of the oligosaccharides released by hydrazinolysis from the purified IgG heavy chains revealed that 0.08% of the total N-linked sugar chains bind to a WFA-agarose column, suggesting that they contain the beta-N-acetylgalactosaminylated structure.

Acetylgalactosamine↗
PubMed1995

Characterization of subspecies of lipoprotein containing apolipoprotein A-I in heterozygotes for familial lecithin:cholesterol acyltransferase deficiency.

We characterized two species of lipoproteins containing apo A-I, one containing only apo A-I (LpA-I) and the other containing both apo A-I and apo A-II (LpA-I/A-II), in three heterozygotes for familial lecithin:cholesterol acyltransferase deficiency (LCAT). In these patients, particle size and the chemical composition of LpA-I differed from those in normal controls. Small particles < 8.8 nm in diameter were predominant, and protein content was higher in patients' LpA-I than that in normal LpA-I. Changes in LpA-I/A-II were mostly quantitative. Percent lipid and protein composition in LpA-I/A-II were similar to those in normal controls. Despite low LCAT mass and activity in the heterozygotes, the molar and fractional rate of cholesterol esterification in their LpA-I and LpA-I/A-II particles were similar to, or higher than, that of normal controls. We conclude that: (i) low LCAT mass and activity is the likely cause of the quantitative and qualitative differences in LpA-I in heterozygotes; and (ii) a deficiency of normal LpA-I particles 11.1 nm in diameter and the existence of small particles < 8.8 nm in diameter may be responsible for the normal, or higher than normal, cholesterol esterification rate of LpA-I and LpA-I/A-II in heterozygotes.

Adult↗
PubMed1995

Distal radioulnar joint subluxation and dislocation diagnosed by standard roentgenography.

The use of standard lateral roentgenography for diagnosing distal radioulnar joint (DRUJ) subluxation and dislocation was investigated. Using a wrist support, bilateral standard lateral roentgenograms of the wrist were obtained in 42 patients with normal wrists and in 56 patients with a unilateral wrist injury. In normal wrists the difference between the radioulnar distance in the right and the left wrist did not exceed 4 mm when the difference in the pisoscaphoid distance was less than 3 mm. Of the 36 patients with wrist injury whose difference in pisoscaphoid distance was less than 3 mm, 15 had a radioulnar distance of 5 mm or more, and computed tomography (CT) confirmed DRUJ dislocation in 14. Concordance between lateral roentgenograms and CT was present in 33 of 36 patients (92%). These results demonstrate the value of a standardized technique for bilateral lateral roentgenography in diagnosing DRUJ subluxation and dislocation.

Adolescent↗
PubMed1995

Portal and peripheral endotoxins in patients with esophageal varices undergoing surgery.

Under the hypothesis that portal endotoxemia, which has been considered evidence of endogenous endotoxemia, is actually a false-positive reaction of the Limulus test, the conventional method of determination, Toxicolor (TOX) and a new endotoxin-specific method, Endospecy (ES) were investigated, whereby portal endotoxemia was reexamined. Peripheral and portal blood samples were collected from 12 patients at various intervals during surgery for esophageal varices, and then evaluated by TOX and ES, the normal values of which are under 60.0 pg/ml and under 9.8 pg/ml, respectively. The mean peripheral and portal endotoxin (Et) levels by TOX were 23.1 pg/ml and 38.9 pg/ml, 1.5 h after the start of surgery, which continued to increase thereafter, the corresponding levels being 48.1 pg/ml and 58.7 pg/ml 8 h after the start of surgery, respectively. The portal Et levels were significantly higher by ES, indicating portal endotoxemia, the mean peripheral and portal Et levels being 6.9 pg/ml, and 6.5 pg/ml, 1.5 h after the start of surgery, these levels showing changes within a similar range. The levels determined by the conventional TOX method increased with time, to a portal Et level of 5.0 pg/ml 8 h after the start of surgery, whereas those determined by ES changed within the same range suggesting that the onset of portal endotoxemia primarily involves the G-factors and that endogenous endotoxemia does not occur.

Blood Specimen Collection↗
PubMed1995

Acute and late neurotoxicity in the rat spinal cord in vivo induced by glutamate receptor agonists.

We investigated glutamate receptor-mediated neurotoxicity in vivo by means of infusing three specific agonists for non-NMDA receptors (acromelic acid A (ACRO), kainic acid and 1-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)) into the adult rat spinal subarachnoid space. ACRO induced long-lasting pure motor, rigid-spastic paraparesis in a dose-dependent manner (EC50: 220 pmol/h) that was accompanied by selective degeneration of spinal interneurons; leaving large anterior horn cells intact. Kainate and AMPA induced paraplegia but with relatively non-selective neuronal damage when given in doses more than 40-fold larger than those required for ACRO. When AMPA (> 100 nmol/h) was infused continuously using a mini-osmotic pump for more than 2 days, rats displayed progressive changes in motor behavior due to extensive damage in the caudal spinal cord where small neurons in the dorsal horns were the most vulnerable. Co-administration of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) specifically prevented neurotoxicity, suggesting a non-NMDA receptor-mediated mechanism. These results indicate that the non-NMDA receptor is heterogeneous, mediating neuronal damage with different selectivity. It is also suggested that chronic activation of glutamate receptors is capable of inducing slowly progressive neuronal death, which suggests relevance to the pathogenesis of ALS.

Animals↗
PubMed1995

Arthroscopic assessment of Kienböck's disease.

Arthroscopic examination was performed on 32 wrists of 32 patients with Kienböck's disease to relate the appearance of the intraarticular structures, particularly the articular cartilage, to the radiographic stage. The articular cartilage showed osteoarthritic changes in stage III, although this was not evident on plain radiographs. Cracking in the distal facet and flapping at the proximal facet of the lunate were identified as features of Kienböck's disease. The incidence of interosseous ligament tears was correlated with radiographically determined stage, whereas changes in triangular fibrocartilage were correlated with age and ulnar variance. Wrist arthroscopy is a useful staging tool for Kienböck's disease, supplying helpful information about the intraarticular pathoanatomy which can be used to guide patient management.

Adolescent↗
PubMed1995

Analysis of proximal fragment sclerosis and surgical outcome of scaphoid non-union by magnetic resonance imaging.

32 patients with scaphoid non-union were examined to clarify the relationship between pre-operative radiographic sclerosis in the proximal fragment and signal intensity on magnetic resonance imaging (MRI). In addition, the correlation between changes in signal intensity in the proximal fragment and surgical outcome after bone grafting and internal fixation was investigated. Proximal fragment sclerosis was observed in seven patients. Comparison of the T1 and T2-weighted images with those of the 25 patients without proximal fragment sclerosis revealed no differences. Therefore, there was no relationship between sclerotic changes on plain radiography and the degree of avascularity of the proximal fragment assessed by MRI. None of the patients who had low signal intensity on both T1 and T2-weighted images achieved union within 5 months after surgery. Thus the detection of signal intensity on both T1 and T2-weighted images can be useful to predict the prognosis after the surgery.

Adolescent↗
PubMed1995

Magnitudes of neurological disorders in a farming town in Kyushu, Japan: frequency of neurological symptoms and signs.

We examined 1,270 elderly residents aged 60 years and over in a cross-sectional study of a standardized clinical examination to determine the frequency of 'abnormal' neurological findings in 27 randomly selected communities in the town of Mifune, Japan. The survey showed one or more neurological symptoms in 89.1%, and one or more signs in 94.4% of the examined elderly. Most of the symptoms and signs increased with age, especially after 70 years, indicating the importance of neurological afflictions in the community health of the elderly.

Age Factors↗
PubMed1995

Different effects of subclasses of HDL containing apoA-I but not apoA-II (LpA-I) on cholesterol esterification in plasma and net cholesterol efflux from foam cells.

We investigated the effects of subclasses of plasma LpA-I (HDL containing apoA-I but not apoA-II) on cholesterol esterification in plasma and net cholesterol efflux from foam cells. LpA-I was composed of particles of three diameters: large (11.1 nm; Lg-LpA-I), medium (8.8 nm; Md-LpA-I), and small (7.7 nm; Sm-LpA-I). Plasma concentrations of LpA-I were positively correlated only with the level of Lg-LpA-I. Plasma concentrations of Lg-LpA-I were inversely correlated with the rate of cholesterol esterification in plasma and VLDL- and LDL-depleted plasma. Plasma concentrations of Md-LpA-I and Sm-LpA-I did not correlate with the rate of cholesterol esterification in plasma or VLDL- and LDL-depleted plasma. When macrophage foam cells were incubated with Md- and Sm-LpA-I, cellular cholesterol mass was reduced by approximately 70%. In contrast, the cellular cholesterol-reducing capacity of Lg-LpA-I was negligible. Lg-LpA-I inhibited net cholesterol removal from foam cells that was mediated by Md- and Sm-LpA-I and cholesteryl ester production with these particles. These results suggest that Md- and Sm-LpA-I may actively participate in cellular cholesterol removal and cholesterol esterification in plasma and HDL, while Lg-LpA-I may regulate these functions of Md- and Sm-LpA-I.

Adolescent↗