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Biomedical subjects

K Furukawa

Publications and source records attributed to K Furukawa.

At least 73 records · Page 4Linked to original sources
PubMed2001

Differential gene expression of beta-1,4-galactosyltransferases I, II and V during mouse brain development.

Since most brain glycoproteins from beta-1,4-galactosyltransferase (beta-1,4-GalT) I knockout mice were galactosylated without apparent reduction the gene expression of novel beta-1,4-GalTs II and V which are involved in N-linked oligosaccharide biosynthesis in addition to beta-1,4-GalT I was studied during mouse brain development. Isolation and characterization of beta-1,4-GalT II and V cDNAs from mouse brains indicates that they are also functioning in the brain. Northern blot analysis revealed that the beta-1,4-GalT I gene is expressed mainly in mid-embryonic stages, while the expression level of beta-1,4-GalT II transcript remains constant and of beta-1,4-GalT V transcript increases during mouse brain development after birth. In situ hybridization revealed that beta-1,4-GalT II and V signals are present in most neural cells, with a marked difference between them in the hippocampus of adult mouse brain tissue. The differential gene expression of beta-1,4-GalTs I, II and V during mouse brain development could affect the differential galactosylation of brain glycoproteins, as revealed by lectin blot analysis.

Aging↗
PubMed2001

Galactosylation of N-linked oligosaccharides by human beta-1,4-galactosyltransferases I, II, III, IV, V, and VI expressed in Sf-9 cells.

Several studies showed that Sf-9 cells can synthesize the galactosylated N-linked oligosaccharides if beta-1,4-galactosyltransferase (beta-1,4-GalT) is supplied. The full-length human beta-1,4-GalT I, II, III, IV, V, and VI cDNAs were independently transfected into Sf-9 cells, and the galactosylation of endogenous membrane glycoproteins was examined by lectin blot analysis using Ricinus communis agglutinin-I (RCA-I), which preferentially interacts with oligosaccharides terminated with Galbeta1-->4GlcNAc group. Several RCA-I-reactive bands appeared in all of the gene-transfected cells, and disappeared on treatment of blots with beta-1,4-galactosidase or N-glycanase prior to incubation with lectin. Introduction of the antisense beta-1,4-GalT II and V cDNAs separately into human colorectal adenocarcinoma SW480 cells, in which beta-1,4-GalT I, II, and V genes were expressed, resulted in the reduction of RCA-I binding toward N-linked oligosaccharides of the membrane glycoproteins. Differences were found in their K(m) values toward UDP-Gal and GlcNAcbeta-S-pNP and in their acceptor specificities toward oligosaccharides with the GlcNAcbeta1-->4(GlcNAcbeta1-->2)Man branch and with the GlcNAcbeta1-->6(GlcNAcbeta1-->2)Man branch. These results indicate that beta-1,4-GalTs II, III, IV, V, and VI are involved in the N-linked oligosaccharide biosynthesis cooperatively but not in a redundant manner with beta-1,4-GalT I within cells.

Adenocarcinoma↗
PubMed2001

Specificity of carbohydrate structures of gangliosides in the activity to regenerate the rat axotomized hypoglossal nerve.

We previously reported that a ganglioside mixture from bovine brain could prevent neuronal death and promote regeneration in rats with hypoglossal nerve resection. In the present study, we have compared the neurotrophic effects of various glycosphingolipids including lactosyl-ceramide. The findings revealed that GT1b had the activity of neuronal death prevention equivalent to a ganglioside mixture or autograft, while other glycolipids exhibited about 60% activity. However, the capability to promote the regeneration varied among glycolipids, that is, GT1b (86%), GD1b (55%), GD1a (35%), GQ1b (34%), GM1 (20%), lactosyl-ceramide (17%) in the number of horseradish peroxidase-positive neurons as an indicator of regeneration. The experiments with oligosaccharides of GT1b or GD1b and ceramide showed that the carbohydrate moiety mainly exerts neurotrophic effects. These findings suggested that fine structures of carbohydrate moiety in gangliosides are critical in the regenerative activity in this hypoglossal nerve regeneration system.

Animals↗
PubMed2001

Effects of high sodium intake on cardiovascular aldosterone synthesis in stroke-prone spontaneously hypertensive rats.

OBJECTIVES: Aldosterone is synthesized in extra-adrenal tissues such as the vasculature, heart and brain. The mechanisms underlying the effect of high salt intake on the development and acceleration of vascular injury and cardiac hypertrophy in the stroke-prone spontaneously hypertensive rats (SHRSP) are still not clear. The goal of this study was to determine whether high salt intake increases cardiovascular aldosterone synthesis in SHRSP. METHODS: Four-week-old SHRSP were given tap water or 0.9% NaCl solution for hydration for 4 weeks in addition to a normal salt diet. Isolated rat mesenteric arteries and hearts were perfused for 2 h, and the perfusate was analysed by high-performance liquid chromatography. The concentrations of aldosterone synthase gene (CYP11B2) mRNA and angiotensin II receptor (AT1R) mRNA were determined by competitive polymerase chain reaction. RESULTS: Salt-loaded SHRSP had higher blood pressures than SHRSP with normal salt intake. Plasma aldosterone concentrations and plasma renin activity were decreased by high salt intake. Aldosterone production, the expression of CYP11B2 mRNA and AT1R mRNA in mesenteric arteries and hearts were significantly increased by high salt intake. CONCLUSIONS: These results suggest that high salt intake increases aldosterone production and expression of the AT1R mRNA in the cardiovascular tissue in SHRSP, which may contribute to the development of malignant hypertension in salt-loaded SHRSP.

Aldosterone↗
PubMed2001

Hemadsorption expressed by cloned H genes from subacute sclerosing panencephalitis (SSPE) viruses and their possible progenitor measles viruses isolated in Osaka, Japan.

Most subacute sclerosing panencephalitis (SSPE) viruses, including our Osaka-1, -2, and -3 strains isolated in Osaka, have shown negative hemadsorption (HAD) by African green monkey red blood cells. This property has been thought to be characteristic of SSPE virus as compared to the positive reaction of the standard Edmonston strain of measles virus (MV). However, this assumption has become quite obscure because MV mutates frequently at the genetic level during its multiplication and also because recent field strains isolated by lymphoblastoid cell lines have shown negative HAD. To investigate the above issue, the nucleotide sequences of the hemagglutinin (H) genes from SSPE virus Osaka-1, -2, or -3 strains were compared to those of various MV field strains isolated in Osaka by Vero cells. The H gene sequences of three SSPE strains were relatively conserved without such biased hypermutation as had been observed in the matrix (M) gene of three SSPE strains. However, this analysis of the H gene sequence of the SSPE viruses enabled us to deduce possible progenitor MVs, which are in agreement with the deduction from the M gene analysis we reported previously. The HAD of Vero cells transfected with the cloned H cDNAs from the SSPE strains and their progenitors suggested that negative HAD of the SSPE viruses has been maintained as one of original properties of the progenitor MVs rather than having been acquired as an altered one during long-term persistent infection in the brains of patients with SSPE.

Amino Acid Sequence↗
PubMed2001

Directed evolution of biphenyl dioxygenase: emergence of enhanced degradation capacity for benzene, toluene, and alkylbenzenes.

Biphenyl dioxygenase (Bph Dox) catalyzes the initial oxygenation of biphenyl and related compounds. Bph Dox is a multicomponent enzyme in which a large subunit (encoded by the bphA1 gene) is significantly responsible for substrate specificity. By using the process of DNA shuffling of bphA1 of Pseudomonas pseudoalcaligenes KF707 and Burkholderia cepacia LB400, a number of evolved Bph Dox enzymes were created. Among them, an Escherichia coli clone expressing chimeric Bph Dox exhibited extremely enhanced benzene-, toluene-, and alkylbenzene-degrading abilities. In this evolved BphA1, four amino acids (H255Q, V258I, G268A, and F277Y) were changed from the KF707 enzyme to those of the LB400 enzyme. Subsequent site-directed mutagenesis allowed us to determine the amino acids responsible for the degradation of monocyclic aromatic hydrocarbons.

Amino Acid Sequence↗
PubMed2001

Characterization of denitrifying granular sludge treating soft groundwater in an upflow sludge-blanket reactor.

Nitrate removal from soft groundwater using ethanol as a carbon source in an upflow sludge-blanket reactor containing denitrifying granular sludge was investigated. At a hydraulic retention time of 0.83 h, influent nitrate was increased stepwise from 20 to 145 mg N/l (volumetric loading rates (VLRs), 0.60 to 4.2 g N/l/d, respectively) and sludge was periodically wasted to maintain a sludge bed of about 2 3 the liquid volume. Complete nitrate removal was achieved at influent nitrate concentrations up to 75 mg N/l(2.1 g N/l/d). MLSS increased from 20 g/l at a VLR of 0.6 g N/l/d to 51 g/l at a VLR of 1.9 g N/l/d, above which it decreased. VSS increased from 11 g/l to a maximum of 25 g/l at a VLR of 2.1 g N/l/d. Settling velocities showed the same trend with maximum values in a VLR range of 1.5 to 2.1 g N/l/d. However, granule size, calcium and magnesium contents of the granular sludge and protein, carbohydrate and nucleic acid contents of extracellular polymers decreased steadily with an increase in VLR throughout the range of testing. Within the VLR range of 0.6 to 2.1 g N/l/d, corresponding to complete nitrate removal and efficient sludge retention, the granular sludge had a high calcium content of 24 to 22%, magnesium ranged from only 0.7 to 0.1%, proteins from 3.2 to 1.3%, carbohydrates from 4.2 to 1.4%, and nucleic acids from 0.34 to 0.05% of the sludge dry weight. These results suggest an optimum operational VLR in terms of nitrate removal and sludge retention of about 2 g N/l/d.

Journal Article↗
PubMed2001

Efficient dechlorination of tetrachloroethylene in soil slurry by combined use of an anaerobic Desulfitobacterium sp. strain Y-51 and zero-valent iron.

A laboratory test was conducted to examine the combined effect of bioaugmentation of an anaerobic bacterial Desulfitobacterium sp. strain Y-51 and addition of zero-valent iron (Fe0) on the reductive dechlorination of tetrachloroethylene (PCE) in a non-sterile soil slurry. Introduction of a strain Y-51 culture in soil (3 mg vss (volatile suspended solids)/kg soil) containing PCE (at 60 micromol/kg soil) led to complete conversion of PCE to cis-1,2-dichloroethylene (cis-DCE) within 40 d. Treatments of the same soil slurry with Fe0 (0.1-1.0%) resulted in extended PCE dechlorination to ethylene (ETH) and ethane (ETA). The combined use of a strain Y-51 culture and Fe0 showed effective dechlorination of PCE than did the individual use. The cis-DCE produced from biological PCE dechlorination by strain Y-51 was totally converted to non-chlorinated end products by the following chemical reduction by Fe0. Furthermore, anaerobic corrosion of Fe0 was found to stimulate the biological reductive dechlorination of PCE by keeping proper levels of pH and oxidation-reduction potential (ORP) and by producing cathodic hydrogen, which might be used as an electron donor for respiratory PCE dechlorination. These findings suggest that the combined use of bacterial strain Y-51 and Fe0 is effective for practical treatment of PCE and other chlorinated ethylenes in contaminated sites.

Journal Article↗
PubMed2001

Isolation and characterization of Desulfitobacterium sp. strain Y51 capable of efficient dehalogenation of tetrachloroethene and polychloroethanes.

A strict anaerobic bacterium, strain Y51, was isolated from soil contaminated with tetrachloroethene (PCE). Strain Y51 is capable of very efficiently dehalogenating PCE via trichloroethene (TCE) to cis-1,2-dichloroethene (cis-1,2-DCE) at concentrations as high as 960 microM and as low as 0.6 microM. Strain Y51 was gram-negative, motile with some lateral flagella, and curved rod-shaped. On the basis of the 16S rDNA sequence, the organism was identified to be a species within the genus Desulfitobacterium. Strain Y51 also had dehalogenation activities toward polychloroethanes such as hexa-, penta-, and tetrachloroethanes, from which dichloroethenes were produced as the final products. The cell extracts mediated the dehalogenation of PCE with reduced methyl viologen as an electron carrier at the specific rate of 5.0 nmol min(-1) mg cell protein(-1) (pH 7.2, 37 degrees C). Dehalogenation was highly susceptible to air oxidation, and to potential alternative electron acceptors such as nitrite or sulfite.

Bacillaceae↗
PubMed2001

Oxygenation reactions of various tricyclic fused aromatic compounds using Escherichia coli and Streptomyces lividans transformants carrying several arene dioxygenase genes.

Bioconversion (biotransformation) experiments on arenes (aromatic compounds), including various tricyclic fused aromatic compounds such as fluorene, dibenzofuran, dibenzothiophene, carbazole, acridene, and phenanthridine, were done using the cells of Escherichia coli transformants expressing several arene dioxygenase genes. E. coli carrying the phenanthrene dioxygenase (phdABCD) genes derived from the marine bacterium Nocardioides sp. strain KP7 converted all of these tricyclic aromatic compounds, while E. coli carrying the Pseudomonas putida F1 toluene dioxygenase (todC1C2BA) genes or the P. pseudoalcaligenes KF707 biphenyl dioxygenase (bphA1A2A3A4) genes was not able to convert these substrates. Surprisingly, E. coli carrying hybrid dioxygenase (todC1::bphA2A3A4) genes with a subunit substitution between the toluene and biphenyl dioxygenases was able to convert fluorene, dibenzofuran, and dibenzothiophene. The cells of a Streptomyces lividans transformant carrying the phenanthrene dioxygenase genes were also evaluated for bioconversion of various tricyclic fused aromatic compounds. The ability of this actinomycete in their conversion was similar to that of E. coli carrying the corresponding genes. Products converted from the aromatic compounds with these recombinant bacterial cells were purified by column chromatography on silica gel, and identified by their MS and 1H and 13C NMR analyses. Several products, e.g., 4-hydroxyfluorene converted from fluorene, and cis-1,2-dihydroxy-1,2-dihydrophenanthridine, cis-9,10-dihydroxy-9,10-dihydrophenanthridine, and 10-hydroxyphenanthridine, which were converted from phenanthridine, were novel compounds.

Actinomycetales↗
PubMed2001

[Lymph node dissection in lung cancer].

The standard surgical treatment for stage I, II, and IIIa non-small cell lung cancer (NSCLC) is lobectomy with mediastinal lymph node dissection. In our series of 214 patients with N2 disease the 5-year survival rate was 23%. Systematic mediastinal lymph node dissection provides further prognostic information on the N factor. Among NSCLC patients with a pathological diagnosis of N2, there are significant differences in the 5-year survival rate based the 3 factors of clinical N factor, N1 station metastasis, and N2 station metastasis.

Adult↗
PubMed2001

[Reevaluation of the surgical indications for lung cancer based on the N factor].

The Standard surgical treatment for stage I, II, and IIIa non-small cell lung cancer (NSCLC) is lobectomy with systemic mediastinal lymph node dissection. More than 50% of our series of 220 patients with cN2 disease were classified as pN0-1. The postoperative 5-year survival rate of patients with cN2 disease was 36%, and that of those with cN2-pN2 disease was 18%. Tumor cell type, surgical technique, or site of tumor had no prognostic significance, although pN, cT, and number of N2 sites were of prognostic significance. We conclude that the indications for surgery are T1-2 N2 disease with a single N2 site.

Adenocarcinoma↗
PubMed2001

In vitro organization of biohybrid rat liver tissue incorporating growth factor- and hormone-releasing biodegradable polymer microcapsules.

To enhance the liver-specific functions of rat hepatocyte aggregates without the addition of exogenous growth factors, polylactic acid-polyglycolic acid (PLGA)/gelatin microcapsules that release insulin, dexamethasone, epidermal growth factors, and glucagon were prepared and incorporated into the hepatocyte aggregates in suspension culture. Precoating the capsules with fibronectin enhanced the incorporation of the microcapsules into the hepatocyte aggregates. In a growth factor- and hormone-free culture medium, these microcapsule-containing aggregates showed a sustained cell number and an ammonium detoxification capacity compared with two types of control culture. One was the culture of microcapsule-free aggregates with albumin-containing control capsules and the other was the culture of capsule-free aggregates that were supplied with the same factors and hormones from the culture medium at concentration levels expected from the release kinetics of the microcapsules. Our new methodology demonstrates that the performance and duration of bioartificial liver systems can be enhanced due to a more efficient maintenance of cell number by using such growth factor- and hormone-releasing microcapsules.

Ammonia↗
PubMed2001

[Tumor ablation with MRI navigation--a novel method of microwave coagulation therapy for hepatic tumor].

Fifty-eight patients with hepatic tumor which consisted of 22 hepatocellular carcinomas and 36 metastatic liver tumors were treated by microwave coagulation therapy with MRI navigation. The tumors were located in all segments of liver except S1. In 24 cases among them, the abdominal approach was difficult, because the tumors were located just below the diaphragm. These cases were selected for thoracoscope-assisted microwave ablation under MR-guidance across the diaphragm. All MR data were collected on a vertically oriented open MRI system (0.5 T SIGNA SP/i system: GE Medical Systems). The microwave electrode was introduced into the liver through a 14G needle via a percutaneous puncture with real-time MR image navigation. Microwave ablations at 60 W for 60 seconds were repeated several times depending on the tumor size. MR imaging may be employed as a reliable guide for percutaneous puncture. Moreover, sufficient safety margin could be obtained for hepatic tumor ablation. MR-guided microwave thermoablation therapy is a feasible method of treatment for hepatic tumors.

Carcinoma, Hepatocellular↗
PubMed2001

The vaso-contractile action of zooxanthellatoxin-B from a marine dinoflagellate is mediated via Ca2+ influx in the rabbit aorta.

We previously showed that zooxanthellatoxin-B, isolated from dinoflagellate, caused a sustained contraction of the aorta in an external Ca2+-dependent manner. To clarify the role of Ca2+ in this action, we examined the effects of zooxanthellatoxin-B as well as a depolarizing stimulus (60 mM KCl), using the simultaneous recording for cytosolic Ca2+ level (fura-2) and developed tension in the rabbit aorta. KCl (60 mM) elicited a rapid cytosolic Ca2+ elevation followed by a pronounced contraction, and time required for half-maximum contraction was 2 min. Zooxanthellatoxin-B caused an increase in cytosolic Ca2+ followed by a gradual contraction, with a time for half-maximum contraction of 5-10 min in a concentration-dependent manner. We found a strong correlation between Ca2+ elevation and the contraction in zooxanthellatoxin-B action. In a Ca2+-free solution, zooxanthellatoxin-B caused neither the contraction nor the increase in cytosolic Ca2+. Furthermore, both pre- and post-treatment with verapamil, a voltage-operated Ca2+-channel blocker, partially suppressed both an increase in cytosolic Ca2+ and the contraction by zooxanthellatoxin-B. Zooxanthellatoxin-B-induced contraction was also inhibited by other voltage-operated Ca2+-channel blockers: nifedipine or diltiazem. These results suggest that zooxanthellatoxin-B-elicited contraction is caused by a Ca2+ influx into the smooth muscle cells, partially via voltage-operated Ca2+ channels.

Animals↗