Biomedical subjects

Akio Kobayashi

Publications and source records attributed to Akio Kobayashi.

70 records · Page 4Linked to original sources

PubMed2002

Human monocyte response to retrieved polymethylmethacrylate particles.

The purpose of this study was to compare retrieved polymethylmethacrylate (PMMA) particles from failed total hip arthroplasties in terms of size, shape, and the response of human monocytes with commercially available particles. PMMA particles were isolated from peri-implant tissues of five failed cemented total hip arthroplasties using tissue digestion and a sucrose density gradient technique. Prepolymerized cement powder and those from which barium sulfate had been removed were examined for comparison. After exposure of peripheral human monocytes to PMMA particles, tumor necrosis factor-alpha and interleukin-6 in medium were measured by using enzyme-linked immunosorbent assays. Image analysis revealed that retrieved particles were larger (retrieved: 1.24 microm; prepolymerized cement powder: 0.83 microm; barium sulfate-free powder: 0.87 microm) and were more irregular in shape and rougher than commercially available particles. Cytokine release was increased by all PMMA particle species. However, commercially available PMMA particles stimulated the release of necrosis factor-alpha and interleukin-6 more strongly than did retrieved particles at very high doses. The observed difference in monocyte response might be due to the volume of the challenged particles. Another possible reason for the difference might be alteration of the surface chemistry of particles in situ and the difference in surface morphology between them.

Humans↗

PubMed2002

Angiotensin I-converting enzyme inhibitory peptides derived from wakame (Undaria pinnatifida) and their antihypertensive effect in spontaneously hypertensive rats.

Seven kinds of angiotensin I-converting enzyme (ACE) inhibitory peptides were isolated from the hydrolysates of wakame (Undaria pinnatifida) by Protease S "Amano" (from Bacillus stearothermophilus) by using three-step high-performance liquid chromatography (HPLC) on a reverse-phase column. These peptides were identified by amino acid composition analysis, sequence analysis, and liquid chromatography-mass spectrometry (LC-MS), as Val-Tyr (IC(50) = 35.2 microM), Ile-Tyr (6.1 microM), Ala-Trp (18.8 microM), Phe-Tyr (42.3 microM), Val-Trp (3.3 microM), Ile-Trp (1.5 microM), and Leu-Trp (23.6 microM). These peptides have resistance against gastrointestinal proteases in vitro. Each peptide was determined to have an antihypertensive effect after a single oral administration in spontaneously hypertensive rats (SHR). Among them, the blood pressure significantly decreased by Val-Tyr, Ile-Tyr, Phe-Tyr, and Ile-Trp in a dose of 1 mg/kg of body weight (BW). The present study showed that antihypertensive effect in the hydrolysates of wakame by Protease S "Amano" was attributed to these peptides.

Amino Acid Sequence↗

PubMed2002

In-situ chemical analyses of trans-polyisoprene by histochemical staining and Fourier transform infrared microspectroscopy in a rubber-producing plant, Eucommia ulmoides Oliver.

The localization of polyisoprene in young stem tissues of Eucommia ulmoides Oliver was investigated by histochemical staining and Fourier transform infrared (FT-IR) microspectroscopy. The fibrous structures were stained with Oil Red O. FT-IR microspectroscopic analysis proved that the fibrous structures were trans-polyisoprene. Granular structures stained with the dye, and characteristic absorptions at 2,960 cm(-1) and 1,430 cm(-1) in FT-IR suggested that trans-polyisoprene accumulated in the vicinity of the cambium layer. We have thus successfully shown for the first time the localization of trans-polyisoprene in plant tissues, and our histological investigation allowed us to presume the main sites of biosynthesis and accumulation of trans-rubber. Furthermore, a new technical approach, the preparation of sections using an electronic freezing unit and the in situ analysis of polyisoprene using FT-IR microspectroscopy, is demonstrated to be a promising method for determining the accumulation of polyisoprene as well as other metabolites.

Azo Compounds↗

PubMed2002

Degradation of a peptide in pitcher fluid of the carnivorous plant Nepenthes alata Blanco.

Carnivorous plants acquire substantial amounts of nitrogen from insects. The tropical carnivorous plant Nepenthes produces trapping organs called pitchers at the tips of tendrils elongated from leaf ends. Acidic fluid is secreted at the bottoms of the pitchers. The pitcher fluid includes several hydrolytic enzymes, and some, such as aspartic proteinase, are thought to be involved in nitrogen acquisition from insect proteins. To understand the nitrogen-acquisition process, it is essential to identify the protein-degradation products in the pitcher fluid. To gain insight into protein degradation in pitcher fluid, we used the oxidized B-chain of bovine insulin as a model substrate, and its degradation by the pitcher fluid of N. alata was investigated using liquid chromatography-mass spectrometry (LC-MS). LC-MS analysis of the degradation products revealed that the oxidized B-chain of bovine insulin was initially cleaved at aromatic amino acids such as phenylalanine and tyrosine. These cleavage sites are similar to those of aspartic proteinases from other plants and animals. The presence of a series of peptide fragments as degradation products suggests that exopeptidase(s) is also present in the pitcher fluid. Amino acid analysis and peptide fragment analysis of the degradation products demonstrated that three amino acids plus small peptides were released from the oxidized B-chain of bovine insulin, suggesting that insect proteins are readily degraded to small peptides and amino acids in the pitcher fluid of N. alata.

Amino Acid Sequence↗

PubMed2002

Methylamine treatment changes the allocation of carbohydrate to roots in rice plants.

Methylamine (MA), an ammonia analogue, has been used to investigate ammonia uptake. This compound competes with ammonia to be taken up and also inhibits the nitrate assimilation pathway. However, the effect of MA on plant growth is unknown. In this paper, we describe the responses of the rice plant to MA. The growth of MA-treated plants was inhibited in the aerial parts and stimulated in the roots. MA-treatment also induced a decrease of starch and hexose in shoots whereas hexose, sucrose and starch contents are increased in MA-treated roots. These results indicate that MA can change the mass allocation of biomass to the roots. The properties of MA suggest that a plant alters its growth via a change in the distribution of carbohydrate in resposes to the nitrogen status.

Journal Article↗

PubMed2002

Paenibacillus glycanilyticus sp. nov., a novel species that degrades heteropolysaccharide produced by the cyanobacterium Nostoc commune.

A novel bacterial strain, DS-1T, was isolated that degrades heteropolysaccharide produced by the cyanobacterium Nostoc commune. The isolate was identified by a combination of phenotypic characterization, cellular fatty acid analysis, DNA base composition, DNA-DNA hybridization and 165 rRNA gene sequence analysis. Phylogenetic analysis placed strain DS-1T within the Paenibacillus cluster on a phylogenetic tree and the phenotypic characteristics of this strain appear to be similar to those of Paenibacillus curdlanolyticus IFO 15724T and Paenibacillus kobensis IFO 15729T. The strain was distinguished from P. curdlanolyticus IFO 15724T and P. kobensis IFO 15729T by its ability to degrade the polysaccharide of Nostoc commune, by assimilation of rhamnose, inositol and L-fucose and by its wide range of optimal growth temperature (28-37 degrees C). Like other Paenibacillus species, this strain contains anteiso-C15:0 as a major cellular fatty acid, and it has a DNA G+C content of 50.5 mol %. Based on these results, it is concluded that this isolate should be placed within a novel species of Paenibacillus, Paenibacillus glycanilyticus sp. nov., with the type strain DS-1T (= IFO 16618T = JCM 11221T = NRRL B-23455T).

Bacillus↗

PubMed2002

Antihypertensive effects of hydrolysates of wakame (Undaria pinnatifida) and their angiotensin-I-converting enzyme inhibitory activity.

AIM: The angiotensin-I-converting enzyme (ACE) inhibitory and antihypertensive activities of wakame hydrolysates have been investigated in several studies. METHODS: Wakame (Undaria pinnatifida) was hydrolyzed using 17 kinds of proteases and the inhibitory activity of the hydrolysates for ACE was measured. Of these hydrolysates 4 with potent ACE inhibitory activity were administered singly and orally to spontaneously hypertensive rats (SHR). RESULTS: The systolic blood pressure of SHR decreased significantly after single oral administration of protease S 'Amano' and proleather FG-F hydrolysates (10 mg protein/kg body weight). In a long-term feeding experiment, 7-week-old SHR were fed standard chow supplemented with protease S 'Amano'-derived wakame hydrolysates for 10 weeks. In SHR fed the 1 and 0.1% wakame hydrolysates, elevation of systolic blood pressure was still significantly suppressed for 7 weeks. CONCLUSIONS: The hydrolysates derived from wakame by protease S 'Amano' have a powerful ACE-inhibitory activity (IC(50) = 86 microg protein/ml) and were effective in spite of their slight bitterness as 'physiologically functional food' with antihypertensive activity.

Administration, Oral↗

PubMed2002

A novel gene delivery system in plants with calcium alginate micro-beads.

We have produced micrometer-sized calcium alginate beads referred to as "bio-beads" that encapsulate plasmid DNA molecules carrying a reporter gene. In order to evaluate the efficiency of the bio-beads in mediating genetic transfection, protoplasts isolated from cultured tobacco cells (BY-2) were transfected with bio-beads containing a plasmid that carries the modified green fluorescent protein gene CaMV35S-sGFP. With the bio-beads treatment, approximately ten-fold higher GFP expression was observed after 24 h incubation compared to that with the conventional method using a naked plasmid solution. Transfection was up to 0.22% efficient. These results indicate that bio-beads have a possibility for efficient transformation in plants.

Journal Article↗

PubMed2001

Aspartic proteinases are expressed in pitchers of the carnivorous plant Nepenthes alata Blanco.

Carnivorous plants acquire significant amounts of nitrogen from insects. The tropical carnivorous plant Nepenthes accumulates acidic fluid containing aspartic proteinase (AP) in its trapping organs (pitchers), suggesting that the plant utilizes insect protein as a nitrogen source. Aspartic proteinases have been purified and characterized from sterile pitcher fluid of several species of Nepenthes; however, there is, as of yet, no information about sequence and expression of Nepenthes AP genes. To identify the pitcher AP, we cloned plant AP homologs from N. alata and examined their expressions. Five AP homologs ( NaAP1-NaAP5) were obtained by reverse transcription-polymerase chain reaction with degenerate primers designed for the conserved sequences of plant APs. Alignment of deduced amino acid sequences with other plant APs demonstrated that NaAP1-NaAP4 contained a plant-specific insert (PSI), a unique sequence of plant AP. However, NaAP5 did not possess the insert, and had a shorter sequence (by >100 amino acids) than the other APs. Northern analysis using a part of the coding region of NaAP1 as a probe showed that bands of approx. 1.8 kb corresponding to the sizes of NaAP1-NaAP4 mRNA were present in roots, stems, leaves, tendrils, and lower part of the pitchers, but a band of approx. 1.3 kb corresponding to the size of NaAP5 mRNA was not observed in any organs. In pitchers, highest expressions of NaAP1-NaAP4 were seen in the lower part of open pitchers containing natural prey, suggesting that the expressions of NaAP1-NaAP4 are coupled with prey capture. Transcripts of NaAP2 and NaAP4 were detected in the digestive glands, where AP secretion may occur. This result suggests that NaAP2 and NaAP4 are the possible APs secreted into the pitcher of N. alata.

Amino Acid Sequence↗

PubMed1987

Toxoplasma antibodies by indirect latex agglutination tests in zoo animals.

Total of 216 animals conserved in Seoul Grand Park were examined on the antibody titers of Toxoplasma by the indirect latex agglutination test, 20 out of 131 cases (15.3%) in mammals, 2 out of 75 cases (2.7%) in birds, and none in reptiles, according to species, 15 out of 68 species (22.1%) in mammals, 2 out of 36 species (5.6%) in birds showed positive antibody titers when the titers of 1:32 or higher were regarded as positive. In mammals, it appeared as positive in 2 out of 6 cases (1 out of 3 species ) in marsupials, 1 out of 15 cases (1 out of 11 species) in primates, 1 out of 1 case in bats, 6 out of 13 cases (5 out of 10 species) in carnivores, 1 in 12 cases (1 species out of 3) in odd-toed ungulates, 9 out of 80 cases (6 species out of 38) in even-toed ungulates, and none in rodents and in whales. In birds, 1 out of 21 cases (1 out of 7 species) in gallinaceous birds and 1 out of 6 (5 species ) in parrots appeared to have the positive antibody titers of Toxoplasma. And, none of reptiles showed positive. Frequencies of positive antibody titers were high in 1: 64, 9 cases in mammals followed by 1: 32, 6 cases, 1: 128, 3 cases, and 1: 256, 2 cases, respectively. Two positive cases in birds appeared to be 1: 64.

Journal Article↗

PubMedDate not supplied

Structural characterization of alpha-terminal group of natural rubber. 1. Decomposition of branch-points by lipase and phosphatase treatments.

Deproteinized natural rubber latex (DPNR-latex) was treated with lipase and phosphatase in order to analyze the structure of the chain-end group (alpha-terminal). The enzymatic treatment decreased the content of long-chain fatty acid ester groups in DPNR from about 6 to 2 mol per rubber molecule. The molecular weight and intrinsic viscosity were reduced to about one-third after treatment with lipase and phosphatase. The Huggins' k' constant of the enzyme-treated DPNR showed the formation of linear rubber molecules. The molecular weight distribution of DPNR changed apparently after treatment with lipase and phosphatase. (1)H NMR spectrum of rubber obtained from DPNR-latex showed small signals due to monophosphate, di-phosphate and phospholipids at the alpha-terminus. Treatment of DPNR-latex with lipase and phosphatase decreased the relative intensity of the (1)H NMR signals corresponding to phospholipids, whereas no change was observed for the signals due to mono- and diphosphates. The residual mono- and diphosphate signals as well as some phospholipid signals after lipase and phosphatase treatments indicate that mono- and diphosphate groups are directly linked at the alpha-terminus with the modified structure, expected by aggregation or linking with phospholipid molecules.

Hevea↗

PubMedDate not supplied

In vivo 15N-enrichment of metabolites in suspension cultured cells and its application to metabolomics.

The incorporation of stable isotopes in suspension cultured cells is very simple and useful as a preliminary experimental method in the experimental scene of plant metabolomics to elucidate the metabolic profiles of mutants and transformants. Stable isotope methods would afford a dynamic explanation of turnover speed that would concern the metabolic flux. Utilization of suspension cultured cells allows genes to be easily induced or suppressed, culture conditions to be controlled, and samples to be easily prepared. Stable isotope tracing allows an index of metabolic flux to be obtained. Here we present an experiment feeding (15)N-labeled inorganic salts to Arabidopsis (cell line T87) and Coptis cultured cells. Results of a comparison of (15)N labeling ratios of amino acids derived from T87 cells cultured under light with those cultured in the dark corresponded to transcriptional expressions revealed by microarray experiments published previously, demonstrating the validity of this procedure. Furthermore, (15)N labeling ratios of Coptis cultured cells revealed arginine and lysine metabolism inhibition, which should result in inhibition of polyamine biosynthesis and cell division. This very simple experiment allowed us to uncover metabolic dynamic features of the plant cell. Therefore this method is very useful for forming working hypotheses and experimental design.

Amino Acids↗

PubMedDate not supplied

Purification and properties of an enzyme capable of degrading the polysaccharide of the cyanobacterium, Nostoc commune.

A novel Nostoc commune-polysaccharide (NPS)-degrading enzyme with a molecular mass of 128.5 kDa was purified from Paenibacillus glycanilyticus DS-1. The optimum pH and temperature of the enzyme activity were 5.5 and 35 degrees C, respectively. The enzyme completely degraded NPS to oligosaccharides, ranging from tetra to hexasaccharides and could degrade the xylan weakly whereas xanthan, gellan, cellulose, curdlan and p-nitrophenyl-beta-D-xylopyranoside were not degraded. Homology analysis of the N-terminal amino acid sequence of the NPS-degrading enzyme against the PIR and SWISS-PROT databases indicated that the sequence was not homologous to any other polysaccharide-degrading enzyme.

Amino Acid Sequence↗

PubMedDate not supplied

Trichosetin, a novel tetramic acid antibiotic produced in dual culture of Trichoderma harzianum and Catharanthus roseus Callus.

The dual culture of Trichoderma harzianum and Catharanthus roseus callus produced an antimicrobial compound with a remarkable activity against the Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis. Structural elucidation revealed that this compound, which we have named trichosetin, is a novel tetramic acid (2,4-pyrrolidinedione) antibiotic and a homolog of the fungal metabolite equisetin. This compound however, was not produced in the individual culture of T. harzianum or C. roseus callus.

Anti-Bacterial Agents↗

PubMedDate not supplied

Characterization and subcellular localization of chlorophyllase from Ginkgo biloba.

Chlorophyllase (Chlase) catalyzes the initial step of chlorophyll (Chl)-degradation, but the physiological significance of this reaction is still ambiguous. Common understanding of its role is that Chlase is involved in de-greening processes such as fruit ripening, leaf senescence, and flowering. But there is a possibility that Chlase is also involved in turnover and homeostasis of Chls. Among the de-greening processes, autumnal coloration is one of the most striking natural phenomena, but the involvement of Chlase during autumnal coloration is not clear. Previously, it was shown that Chlase activity and expression level of the Chlase gene were not increased during autumnal coloration in Ginkgo biloba, indicating that Chlase does not work specially in the de-greening processes in G. biloba. In this study, we characterized the recombinant Chlase and analyzed its subcellular localization to understand the role of the cloned Chlase of G. biloba (GbCLH). GbCLH exhibited its highest activity at pH 7.5, 40 degrees C. Kinetic analysis revealed that GbCLH hydrolyzes pheophytin (Pheo) a and Chl a more rapidly than Pheo b and Chl b. Transient expression analysis of 40 N-terminus amino acids of GbCLH fused with GFP (green fluorescent protein) and subcellular fractionation showed that GbCLH localizes within chloroplasts. Together with our previous results, property of GbCLH and its location within the chloroplasts suggest that GbCLH plays a role in the turnover and homeostasis of Chls in green leaves of G. biloba.

Amino Acid Sequence↗

PubMedDate not supplied

Metabolic fingerprinting and profiling of Arabidopsis thaliana leaf and its cultured cells T87 by GC/MS.

Cell suspension cultures are now recognized as important model materials for plant bioscience and biotechnology. Very few studies of metabolic comparisons between cell cultures and original plants have been reported, even though the biological identity of cultured cells with the normally grown plant is of great importance. In this study, a comparison of the metabolome for primary metabolites extracted from the leaves of Arabidopsis thaliana and cultured cells from an Arabidopsis suspension culture (cell line T87) was performed. The results suggest that although cell suspension cultures and Arabidopsis leaves showed similarities in the common primary metabolite profile, nonetheless, moderate differences in quantitative profile were revealed.

Amino Acids↗