Search PubMed⌕ Search

Biomedical subjects

K Furukawa

Publications and source records attributed to K Furukawa.

At least 163 records · Page 9Linked to original sources
PubMed1999

Properties of the peptides liberated from rice protein in sokujo-moto.

In the supernatant of sokujo-moto, a high level of acid carboxypeptidase (ACP) activity and a large amount of peptides were observed, however, the amount of free amino acids liberated was small. In order to determine why these peptides were not hydrolyzed to any significant degree by the ACP, the properties of the peptides in sokujo-moto were investigated in this study. Peptides were fractionated from sokujo-moto by ion exchange column chromatography. ACP purified from rice-koji (rice overgrown with Aspergillus oryzae) was allowed to react with the peptides, and it was found that they were not hydrolyzed to any significant degree by the enzyme. Gel filtration chromatography was performed to ascertain the molecular size distribution of the peptides in sokujo-moto, and it was revealed that they were of low molecular sizes; molecular size: mainly in the range of 200-400, and chain length: 2-3. ACP purified from rice-koji was also allowed to react with various synthetic peptides, and it was found that ACP of rice-koji could not rapidly hydrolyze low-molecular-size peptides, such as dipeptides or tripeptides. Acid protease (AP) purified from rice-koji released peptides of molecular sizes mainly in the range of 300-600 or above from rice protein under acidic conditions (pH 3.6; the pH of sokujomoto). When AP and ACP were allowed to act at the same time on rice protein, mainly low-molecular-size peptides (molecular sizes mainly in the range of 200-400) were produced. From these results, it was estimated that AP released peptides with molecular sizes mainly in the range of 300-600 or above from rice protein and ACP degraded the relatively higher molecular size peptides among them to lower molecular size peptides; consequently only low-molecular-size peptides with molecular sizes mainly in the range of 200-400 were released in the supernatant of sokujo-moto.

Journal Article↗
PubMed1999

Influence of amino acid content in seed mash on peptide uptake by yeast cells in main mash in sake brewing process.

It was found that the peptide content of the main mash in the sake brewing process, seeded with kimoto, was higher than in that seeded with sokujo-moto, although the peptide content in kimoto was lower than in sokujo-moto. We investigated the underlying reasons. As a result, we found that the high concentration of free amino acids originating from kimoto decreased the peptide uptake ability of yeast cells in the main mash seeded with kimoto.

Journal Article↗
PubMed1999

Properties of TCA-insoluble peptides in kimoto (traditional seed mash for sake brewing) and conditions for liberation of the peptides from rice protein.

It was found that a large amount of TCA (trichloroacetic acid)-insoluble peptides were liberated into the supernatant of kimoto on the 7th-10th day of mashing. These TCA-insoluble peptides had five polypeptide groups (12, 21, 31, 38, and 55 kDa) on SDS-PAGE (SDS-polyacrylamide gel electrophoresis), and a large amount of high molecular weight peptides, higher than 10,000, were observed upon gel filtration chromatography using TSKgel G2000swxl (Tosoh Co.). Four of these peptides (12, 21, 31, and 38 kDa on SDS-PAGE) appeared specifically in kimoto, and were not detected at all either in sokujo-moto or in the main mash for sake brewing. These TCA-insoluble peptides were fractionated from the supernatant of kimoto on the 9th day, and it was revealed that free amino acids were produced abundantly from them in the presence of the enzyme of rice-koji. Therefore, it was assumed that the peptides are related to the abundant production of free amino acids in kimoto. For the liberation of these TCA-insoluble peptides from rice protein, the enzyme of rice-koji was indispensable. The enzyme liberating the TCA-insoluble peptides from rice protein was purified from rice-koji, and was presumed to be identical with acid protease (AP) of rice-koji. The presence of a high concentration of glucose (higher than 20%) was also indispensable for the liberation of the TCA-insoluble peptides. Furthermore, it was revealed that the peptides were liberated from rice protein under a limited pH of around 4.5.

Journal Article↗
PubMed1999

Mobile catabolic genes in bacteria.

The recent findings of various mobile catabolic genes have provided some insight into the evolution of microbial degradation systems for xenobiotic compounds. The catabolic genes undergo marked genetic rearrangements due to their presence on transposons or association with mobile genetic elements. Bacterial catabolic transposons fall into three defined structural classes. Class I elements include catabolic genes flanked by two copies of insertion sequences. Class II elements carry short terminal inverted repeats and transpose by the replicative mode in which transposase and resolvase are involved. Conjugative catabolic transposons represent the third class of mobile genetic elements. They carry all the genes required for excision, conjugal transfer to a new host, and integration. This review focuses on the structures, functions and roles of the recently characterized catabolic transposons in bacteria. Also described are the mobile catabolic elements that share structural similarity with the pathogenicity and symbiosis islands.

Journal Article↗
PubMed1999

Engineering a hybrid pseudomonad to acquire 3,4-dioxygenase activity for polychlorinated biphenyls.

We constructed a hybrid strain that acquired 3,4-dioxygenase activity for polychlorinated biphenyls (PCBs). This strain, KF707-D34, possessed a chimeric biphenyl dioxygenase gene, of which a portion of bphA1 (coding for a large subunit of biphenyl dioxygenase) of Pseudomonas pseudoalcaligenes KF707 was replaced with that of a PCB-degrader, Burkholderia cepacia LB400 by homologous recombination. KF707-D34 retained the ability to degrade 4,4'-dichlorobiphenyl via 2,3-dioxygenation in a fashion identical to that of KF707 and gained novel capability to degrade 2,5,4'-trichlorobiphenyl and 2,5,2',5'-tetrachlorobiphenyl via 3,4-dioxygenation in a fashion identical to that of LB400. Sequence analysis of bphA1 from KF707-D34 revealed that three nucleotides in the 3'-terminal region of KF707 bphA1 were changed to correspond to those in LB400 bphA1. The resulting BphA1 protein in KF707-D34 was changed at position 376 from threonine (Thr) to asparagine (Asn). The results demonstrate that a minor alteration of the amino acid sequence in BphA1 improved the PCB degradation capability in biphenyl-utilizing bacteria.

Journal Article↗
PubMed1999

Differences in N-acetyllactosamine synthesis between beta-1,4-galactosyltransferases I and V.

Unlike classical beta-1,4-galactosyltransferase (beta-1,4-GalT I), beta-1,4-GalT V (formerly IV**) has little activity towards 1 mM N-acetylglucosamine [Sato et al. (1998) Proc Natl Acad Sci USA 95:472-477]. The human beta-1,4-GalTs I and V were expressed individually in Sf-9 cells by transfection of the full coding sequences, and their N-acetyllactosamine synthetase activities were determined towards different N-acetylglucosamine concentrations. Kinetic studies using the cell homogenates as an enzyme source revealed that beta-1,4-GalTs I and V possess Km values of 0.6 mM and 33 mM towards N-acetylglucosamine, and of 48 microM and 41 microM towards UDP-Gal, respectively. No significant inhibition of N-acetyllactosamine synthesis with alpha-lactalbumin was observed for beta-1,4-GalT V but the significant inhibition with alpha-lactalbumin was observed for beta-1,4-GalT I.

Amino Sugars↗
PubMed1999

Removal of mercury from mercury-contaminated sediments using a combined method of chemical leaching and volatilization of mercury by bacteria.

A method for the removal of mercury sulfide from mercury-contaminated sediments was developed, which consists of chemical leaching and volatilization of mercury by bacteria. More than 85% of the mercury in sediment containing 0.11-37.4 mg/kg of mercury was efficiently extracted with 3 M HCl and 74 mM FeCl3. Subsequent volatilization by bacteria resulted in the removal of 62.9-75.1% of mercury from mercury-contaminated Minamata Bay sediments. Methylmercury was also eliminated from soil at a high efficiency. Thus, this combined method of chemical and microbial treatments could be used for efficient removal of both organic and inorganic mercurials from natural sediments.

Bacteria↗
PubMed1999

Functional analysis of O-linked oligosaccharides in threonine/serine-rich region of Aspergillus glucoamylase by expression in mannosyltransferase-disruptants of yeast.

The glaA gene encoding glucoamylase I (GAI) of Aspergillus awamori var. kawachi was heterologously expressed in mannosyltransferase mutants of Saccharomyces cerevisiae, in which the pmt1 gene and the kre2 gene were disrupted. The GAI enzymes expressed in these yeast mutant cells exhibited a lesser extent of O-glycosylation. Secretion of GAI expressed in the pmt1-disruptant and in the kre2-disruptant, respectively, was almost the same as that of GAI expressed in wild type (wt) strains. The number of O-linked mannose in GAI from wt yeast strain ranged in size from one (Man1) to five (Man5). On the other hand, the O-linked oligosaccharides of GAI from the pmt1-disruptant ranged in size from Man1 to Man4. Man5 was not detected and Man2-Man4 were reduced in proportion to the reduction of Man1. The O-linked oligosaccharides of GAI from the kre2-disruptant ranged from Man1 to Man4, and the molar amount of Man4 was reduced to 27.3%, compared to that of the wt strain. The hydrolyzing abilities for soluble starch and the adsorbing abilities on raw starch were comparable between both disruptants and wt strains. However, the digesting abilities for raw starch of the disruptants were decreased to 70% of those of the wt strains. Stabilities of GAI of the disruptants were reduced toward extreme pH and high temperature, compared to those of the wt strains. These results demonstrate that the O-linked oligosaccharides of GAI are responsible for the enzyme stability and activity toward insoluble substrates but not for secretion.

Aspergillus↗
PubMed1999

Prevention of the death of the rat axotomized hypoglossal nerve and promotion of its regeneration by bovine brain gangliosides.

We have examined the time course of the neuronal death and regeneration of rat axotomized hypoglossal nerve with various conditions of the nerve resection, and established a useful system to measure neurotrophic activities of bioactive substances. In this system, neuronal death can be evaluated by counting surviving neurons in the nucleus of hypoglossal neuron at the brain stem, and the degree of the regeneration can be measured by counting horseradish peroxidase-positive cells at the same region after injection of horseradish peroxidase into tongue. Using this system, the effects of brain gangliosides on rat hypoglossal nerve regeneration following 5 mm transection were examined. The addition of a ganglioside mixture from bovine brain as well as the autograft strongly prevented the death of neurons and promoted the regeneration of the lesioned nerve at 10 weeks after the operation. Further analyses on the dose effects and injection sites of gangliosides were performed. Although the mechanisms of the neurotrophic effects of the gangliosides are unknown, the therapeutic application of gangliosides for neuronal degeneration is a promising approach.

Animals↗
PubMed1999

Localization of 17-kDa myosin light chain isoforms in cultured aortic smooth muscle cells.

Smooth muscle myosin II contains two 17-kDa essential light chain isoforms (LC17gi and LC17nm) of which the relative contents differ among myosins. To understand the roles of LC17 isoforms in the functions of myosin, we performed an immunofluorescence microscopic examination of their localization in primary cultured cells isolated from rat aortic smooth muscle. To identify the isoforms, rabbit polyclonal antibodies were prepared against C-terminal nonapeptides corresponding to either LC17gi or LC17nm from porcine aortic smooth muscle myosin. These isoforms differ in only 5 amino acid residues within the C-terminal 9 residues. These antibodies specifically recognize each LC17 isoform on urea-PAGE of total rat aortic cell lysates. Immediately after plating, the smooth muscle cells stained heterogeneously with each antibody, indicating differing contents of LC17 isoforms among cells. On double staining 1-2 d cultures with both antibodies, LC17nm was detected diffusely throughout the cytoplasm, whereas LC17gi was concentrated in specific regions such as the cell periphery and the base of cytoplasmic processes. These results support the suggestion that myosin containing LC17gi is essential for force-generation by aortic smooth muscle and that myosin containing LC17nm may play an important role in maintaining smooth muscle tension.

Animals↗
PubMed1999

Effects of soybean oil emulsion and eicosapentaenoic acid on stress response and immune function after a severely stressful operation.

OBJECTIVE: To investigate the effects of soybean oil emulsion and oral or enteral administration of eicosapentaenoic acid (EPA) on stress response, cytokine production, protein metabolism, and immune function after surgery for esophageal cancer. SUMMARY BACKGROUND DATA: It has been reported that safflower oil, rich in n-6 polyunsaturated fatty acid (n-6 PUFA), affects the survival rate of septic animals and decreases the immune function. It has also been reported that the administration of fish oil, in contrast, reduces these stress responses and stress-induced immunosuppression. In humans, the effects of soybean oil emulsion and the administration of EPA on stress response and immune function after surgery have not been established. METHODS: Patients who underwent esophagectomy with thoracotomy were divided into three groups. Seven patients were fed by total parenteral nutrition (TPN) with soybean oil emulsion, which accounted for 20% of total calories. Seven patients were given oral or enteral administration of 1.8 g/day EPA, in addition to TPN with soybean oil emulsion. Nine patients served as the control group; these patients received fat-free TPN. Serum interleukin-6 (IL-6), C-reactive protein, concanavalin A (con A)- or phytohemagglutinin (PHA)-stimulated lymphocyte proliferation, natural killer cell activity, and stress hormones were measured. RESULTS: The postoperative level of serum IL-6 was significantly higher in the group receiving soybean oil emulsion than in the fat-free group. Oral or enteral supplementation of EPA with soybean oil emulsion significantly reduced the level of serum IL-6 compared with the patients receiving soybean oil emulsion. Con A- or PHA-stimulated lymphocyte proliferation decreased significantly on postoperative day 7 in all groups of patients. The supplementation of EPA with soybean oil emulsion significantly improved the lymphocyte proliferation and natural killer cell activity on postoperative day 21 compared with the group receiving soybean oil emulsion. CONCLUSIONS: Soybean oil emulsion amplifies, and the supplementation of EPA reduces, the stress response and stress-induced immunosuppression.

Eicosapentaenoic Acid↗
PubMed1999

Simultaneous assessment of right ventricular function and hypertrophy by Tc-99m MIBI.

PURPOSE: Tc-99m MIBI allows the simultaneous performance of cardiac blood pool scintigraphy and myocardial SPECT. The authors performed studies to determine whether right ventricular function and hypertrophy can be evaluated simultaneously using Tc-99m MIBI in patients with right heart disease. METHODS: Using right heart catheterization, several parameters of pulmonary circulation were measured, including right ventricular ejection fraction, in 23 patients with chronic pulmonary disease and pulmonary vascular disease. Within 1 week, right heart blood pool scintigraphy was performed using the first-pass method using Tc-99m MIBI. The right ventricular ejection fraction calculated from right heart blood pool scintigraphy was compared with that measured using right heart catheterization. Myocardial SPECT was performed 1 hour after right heart blood pool scintigraphy. On the short axis images, which allowed optimal visualization of the right ventricle, the right and left ventricular free walls were established as regions of interest. Myocardial wall counts were determined. The right ventricular uptake: left ventricular uptake ratio was calculated to study the relation of that ratio to mean pulmonary artery pressure and total pulmonary resistance. RESULTS: A significant correlation was observed between the right ventricular ejection fraction calculated on right heart catheterization and that calculated on right heart blood pool scintigraphy. The right ventricular uptake:left ventricular uptake ratio correlated positively with pulmonary artery pressure and total pulmonary resistance. CONCLUSION: These results suggest that Tc-99m MIBI allows simultaneous noninvasive evaluation of cardiac function and structure, which previously was considered impossible in patients with right heart disease.

Female↗
PubMed1999

Left ventricular function at rest and during bicycle exercise in normal subjects: assessment by ECG-gated myocardial perfusion SPET with 99Tcm-tetrofosmin.

99Tcm-labelled myocardial perfusion tracers allow simultaneous assessment of myocardial perfusion and left ventricular function using ECG-gated SPET. The aim of this study was to evaluate left ventricular performance during exercise by means of ECG-gated myocardial perfusion SPET. After the administration of 99Tcm-tetrofosmin (555-740 MBq), eight healthy volunteers aged 27-49 years underwent ECG-gated myocardial perfusion SPET at rest and during supine submaximal exercise (75 and 125 W), for 3 min each. Using ECG-gated SPET data, left ventricular end-diastolic volume (LVEDV) demonstrated a biphasic response during exercise (from 106.4 +/- 17.5 to 119.9 +/- 19.9 to 108.1 +/- 19.2 ml). In contrast, left ventricular end-systolic volume decreased gradually and significantly during exercise (from 47.1 +/- 11.9 to 41.5 +/- 8.9 to 36.1 +/- 10.1 ml; P < 0.05), and left ventricular ejection fraction continued to increase at higher workloads (from 56.1 +/- 6.0 to 63.0 +/- 2.7 to 67.0 +/- 4.3; P < 0.01) despite a fall in LVEDV. There was a progressive increase in cardiac output during exercise, which reached a peak of 7.2 +/- 0.9 l.min-1. We conclude that ECG-gated myocardial perfusion SPET can assess left ventricular function during exercise and may provide useful information for the evaluation of patients with ischaemic heart disease.

Adult↗
PubMed1999

Enhanced expression of interleukin-6 in chronic hepatitis C.

AIMS/BACKGROUND: There is a possibility that proinflammatory cytokines such as interleukin-6 (IL-6) are involved in the inflammatory process of chronic hepatitis C. This study was undertaken to investigate the possible role of IL-6 in the pathophysiology of chronic hepatitis C. METHODS: Serum IL-6 levels in 63 patients with chronic hepatitis C and in 26 normal controls were measured. Light and electron immunostaining studies to localize IL-6 protein as well as in situ hybridization to localize IL-6 messenger RNA were performed on 10 liver biopsy specimens. RESULTS: Serum IL-6 levels were significantly (p<0.01) elevated in chronic hepatitis C compared to those in normal controls. Although no statistically significant correlation was found between serum IL-6 levels and hepatobiliary enzyme levels, a significant correlation (p<0.01) was found between serum IL-6 levels and category II of Knodell's histological activity index score. Non-parenchymal cells in hepatic sinusoids and the cells infiltrating enlarged fibrous portal tracts were definitely positive for IL-6 protein and mRNA by immunohistochemistry and in situ hybridization. In addition, immunoelectron microscopy revealed a weak and occasional positive reaction in the cytoplasm of hepatocytes. The majority of the positive cells in hepatic sinusoids showed CD68 immunoreactivity in consecutive sections indicating that these were Kupffer cells. Sinusoidal endothelial cells and hepatic stellate cells also exhibited a weak reaction. CONCLUSION: These results strongly suggest that Kupffer cells in liver parenchyma and macrophages infiltrating in portal tracts are the main producers of elevated IL-6 in serum. Moreover, there is a possibility that IL-6 produced by hepatocytes could also act as a regenerative stimulus to hepatocytes themselves in an autocrine fashion.

DNA Primers↗
PubMed1999

[Recurrent respiratory tract infection caused by Moraxella catarrhalis in patients with pneumoconiosis].

Moraxella catarrhalis is recognised as a major pathogen in chronic respiratory diseases. Patients with pneumoconiosis have repeated respiratory infections, and the control is very important for good prognosis. From 1988 to 1993, fifty strains of Moraxella catarrhalis were obtained from 9 patients with pneumoconiosis attended at Nagasaki Rosai Hospital. Restriction enzyme analysis of chromosomal DNA by Hind III. Hae III, Cla I was performed on 50 strains of M. catarrhalis. Twelve strains (56%) out of 23 isolates with interval of less than 5 months had identical bacterial restriction endonuclease digestion patterns with three different enzymes, and all isolates with interval more than 6 months had different patterns. Tweny out of 13 episodes by the same strains occurred during 3 months. The acquisition and clearance of M. catarrhalis from the respiratory tract is a dynamic process.

Aged↗
PubMed1999

Expression of DCC protein in colorectal tumors and its relationship to tumor progression and metastasis.

A loss of heterozygosity (LOH) at the DCC gene locus was detected in colorectal tumors, and this LOH might be related to metastasis. The aim of this study was to determine DCC protein expression in colorectal cancer and to evaluate its prognostic value. Allelic loss of the DCC locus was observed in 16 of the 23 patients (66.7%). In all 16 patients with LOH, DCC expression was decreased in the cancer tissue compared with the adjacent normal mucosa. All 23 colorectal tumors had decreased expression of this protein relative to the adjacent normal colonic mucosa in Western blot analysis. The levels of DCC protein were significantly lower in cancer tissues than in adenoma tissues. Decreased DCC protein expression was also observed by immunohistochemistry in the colorectal cancer cases. There were significant correlations between DCC protein expression and histologic type, venous invasion, and hematogenous metastasis. Patients with DCC-protein-negative tumors had a greater relative risk of recurrence compared with those whose tumors were DCC protein-positive. The 5-year survival rate was 91.0% in patients with DCC-protein-positive tumors, and 58.8% in those with DCC-protein-negative tumors; these differences between the two groups of patients were significant (p < 0.01). In multivariate analysis using the Cox regression model, DCC protein expression emerged as an independent prognostic indicator. These findings suggested that a decrease in DCC expression may have an important role in the progression of colorectal cancers and may be a biologic marker of prognostic significance.

Adenoma↗
PubMed1999

Mechanisms of FK 506-induced hypertension in the rat.

-Tacrolimus (FK 506) is a powerful, widely used immunosuppressant. The clinical utility of FK 506 is complicated by substantial hypertension and nephrotoxicity. To clarify the mechanisms of FK 506-induced hypertension, we studied the chronic effects of FK 506 on the synthesis of endothelin-1 (ET-1), the expression of mRNA of ET-1 and endothelin-converting enzyme-1 (ECE-1), the endothelial nitric oxide synthase (eNOS) activity, and the expression of mRNA of eNOS and C-type natriuretic peptide (CNP) in rat blood vessels. In addition, the effect of the specific endothelin type A receptor antagonist FR 139317 on FK 506-induced hypertension in rats was studied. FK 506, 5 mg. kg-1. d-1 given for 4 weeks, elevated blood pressure from 102+/-13 to 152+/-15 mm Hg and increased the synthesis of ET-1 and the levels of ET-1 mRNA in the mesenteric artery (240% and 230%, respectively). Little change was observed in the expression of ECE-1 mRNA and CNP mRNA. FK 506 decreased eNOS activity and the levels of eNOS mRNA in the aorta (48% and 55%, respectively). The administration of FR 139317 (10 mg. kg-1. d-1) prevented FK 506-induced hypertension in rats. These results indicate that FK 506 may increase blood pressure not only by increasing ET-1 production but also by decreasing NO synthesis in the vasculature.

Animals↗