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Biomedical subjects

Y Kubo

Publications and source records attributed to Y Kubo.

At least 163 records · Page 9Linked to original sources
PubMed1996

Cloning and characterization of a bifunctional metabotropic receptor activated by both extracellular calcium and glutamate.

A cDNA from salmon brain which induces a consistent activation of Ca2+ dependent Cl- (Ca(2+)-Cl-) current in Xenopus oocytes in the saline solution was isolated by an expression cloning method. The primary structure showed a high homology with the metabotropic glutamate receptors (mGluR), and a limited homology with the Ca2+/polyvalent cation receptors (CaR). The encoded protein was functionally characterized for the sensitivity to extracellular Ca2+ (Cao2+), and the consistent activation of Ca(2+)-Cl- current was determined to be due to Ca2+ contained in the saline solution. It was also confirmed that the encoded protein has a function to sense glutamate. An increase in inositol 1,4,5-triphosphate turnover was observed when either Ca2+ or glutamate was applied. Thus, the encoded protein was identified as a bifunctional metabotropic receptor for both Ca2+ and glutamate. In addition, rat mGluR1 alpha was also shown to be activated by Cao2+. This is the first report of a G-protein coupled receptor which is activated by two totally different physiological agonists.

Amino Acid Sequence↗
PubMed1996

A weakly inward rectifying potassium channel of the salmon brain. Glutamate 179 in the second transmembrane domain is insufficient for strong rectification.

A cDNA encoding for a weakly inward rectifying K+ channel (sWIRK: salmon weakly inward rectifying K+ channel) was isolated from the masu salmon brain by expression cloning. The sWIRK channel exhibited the highest similarity with members of the ROMK1 subfamily, BIR10/KAB-2 (70% amino acid identity) and ROMK1 (46%). An ATP binding motif which is characteristic of this subfamily was also conserved. The sWIRK RNA was detected in the brain, but not in the heart, kidney, skeletal muscle, liver, testis, and ovary. In the brain, the expression was observed in the ependymoglial cells on the surface of the ventricles as well as in the small perineuronal glia-like cells in the midbrain and the medulla. When compared with the strong inward rectifier IRK1 channel, the sWIRK channel showed a much weaker inward rectification property, and the activation kinetics upon hyperpolarization was slower and less voltage-dependent. The slope conductance of the single channel inward current was 37 pS (140 mM K+o), and outward current channel events were also observed. The weak rectification of sWIRK is significant in that it has a negatively charged residue (glutamate) in the M2 region which is reported to cause strong inward rectification. By introducing a point mutation to remove this negative charge (glutamine), the sWIRK E179Q mutant channel lost its inward rectification property completely, and the single channel property (45 pS; 140 mM K+o) was ohmic up to highly depolarized potential, even in the presence of the physiological cytoplasmic blockers such as Mg2+ and polyamines.

Amino Acid Sequence↗
PubMed1996

Early results of a reinforced biosynthetic ovine collagen vascular prosthesis for small arterial reconstruction.

The efficacy of a reinforced biosynthetic ovine collagen (RBOC) vascular prosthesis developed for small arterial reconstruction was assessed by examining 30 grafts in 29 patients with arteriosclerosis obliterans. The operative procedures performed were femorofemoral bypass in 2 patients, above-knee femoropopliteal bypass in 28 patients, and below-knee femoropopliteal bypass in 1 patient. Femoropopliteal bypass was simultaneously performed in two patients undergoing femorofemoral bypass using one or two grafts. The indications for surgery were intermittent claudication in 27 patients and to salvage the limb in 2 patients. The longest follow-up period was 49 months, and there were six graft failures, occurring 1, 1, 9, 17, 17, and 23 months after implantation, respectively; caused by compression of the graft from outside in two, infection in one, anastomotic intimal hyperplasia in one, and unknown factors in two. Thus, the primary cumulative patency rate for above-knee femoropopliteal bypass at 3 years was 83.7%, and the secondary patency rate was 91.2%. No aneurysmal change was observed. Moreover, the RBOC was able to be used without preclotting, and its handling and suturing characteristics were satisfactory. Our findings suggest that this vascular prosthesis may be an acceptable alternative for above-knee femoropopliteal bypass.

Aged↗
PubMed1996

Exogenous lysophosphatidylcholine increases non-selective cation current in guinea-pig ventricular myocytes.

Whole cell, patch-clamp studies were performed to examine the effect of lysophosphatidylcholine (LPC) on the membrane current in guinea-pig ventricular myocytes. The addition of 10 microM LPC to the external solution induced a membrane current which had a reversal potential of 0 mV. When Na+, the main cation in the external solution, was replaced by either K+, N-methyl-D-glucamine (NMG) or 90 mM Ca2+, LPC induced a current with the reversal potential near 0 mV, indicating that the current passed through a Ca2+-permeable non-selective cation channel. The order of the cationic permeability calculated from the reversal potential of the current was Cs+ > K+ > NMG > Na+ > Ca2+. Cl- did not pass through the LPC-induced channel. The LPC-induced current was not blocked by Gd3+ in the external solution, nor by the absence of Ca2+ in the pipette solution. In conclusion, LPC induces a Ca2+-permeable non-selective cation channel in guinea-pig ventricular myocytes.

Animals↗
PubMed1996

Re-endothelialisation in autogenous vein grafts.

OBJECTIVES: To clarify the course of re-endothelialisation (Re-E) in an entire graft and to establish the effect of immersion media for the preservation of endothelial cells. METHODS: Autogenous femoral veins of dogs were immersed in heparinised saline solution (n = 18) or heparinized autogenous blood (n = 18). After immersion, the grafts were implanted into bilateral femoral artery, and were retrieved 1 day to 4-8 weeks after implantation. RESULTS: For the grafts immersed in the heparinised saline solution, the values for % area of endothelial cell coverage before implantation, and at 1 day, 1 week, and 4 weeks after implantation were 44.9%, 6.2%, 14.5%, and 81.3%, respectively. For the grafts immersed in heparinised autogenous blood, the values were 73.5%, 20.6%, 79.2% and 95.5%, respectively. However, such relatively rapid speed of Re-E slowed down considerably after 1 week following implantation in this group. CONCLUSIONS: The use of heparinized autogenous blood is strongly recommended as a preparation media for autogenous vein grafts. Almost all of the endothelial cells fall away in the earlier period after implantation and regenerate multifocally and irregularly. Re-E is incomplete even at 8 weeks after surgery, and we suggest that the area of incomplete Re-E may develop into intimal hyperplasia.

Animals↗
PubMed1996

Hepatocyte proliferation induced in rats by lead nitrate is suppressed by several tumor necrosis factor alpha inhibitors.

Lead nitrate induces liver cell proliferation in rats without accompanying liver cell necrosis. However, the mechanism of this proliferation and its effect on hepatocytes remain unknown. Therefore, we examined the liver and blood level of hepatocyte growth factor and tumor necrosis factor alpha (TNF-alpha) at various intervals to determine whether lead nitrate modifies hepatocyte proliferation by altering the production of these cytokines. We also administered several TNF-alpha inhibitors, dexamethasone, adenosine, (2E)-3-[5-(2,3-dimethoxy-6-methyl-1,4-benzoquinoyl)]-2-nonyl-2- propenoic acid (E 3330), and pentoxifylline, to rats to clarify whether pretreatment with these inhibitors suppresses the increase of TNF-alpha messenger RNA (mRNA) in the liver and prevents the hepatocyte proliferation induced by lead nitrate. Hepatocyte proliferation occurred by 24 hours and reached a peak 48 hours after a single intravenous injection of lead nitrate (100 mumol/kg). TNF-alpha mRNA expression in the liver was increased 1, 6, and 12 hours after the injection, whereas no alteration was observed in liver or blood level of hepatocyte growth factor. Pretreatment with dexamethasone (4.0 mg/kg), E3330 (100 mg/kg) adenosine (0.3 mmol/kg), and pentoxifylline (100 mg/kg), inhibited both TNF-alpha mRNA expression and hepatocyte proliferation 48 hours after the injection. These experimental results strongly support the hypothesis that TNF-alpha positively regulates the hepatocyte proliferation induced in rats by the mitogen, lead nitrate.

Adenosine↗
PubMed1996

Cytotoxic and antioxidative sesquiterpenoids from Heterotheca inuloides.

Four sesquiterpenoids, beta-caryophyllene, beta-caryophyllene 4,5 alpha-oxide, 7-hydroxy-3,4-dihydrocadalin, and 7-hydroxycadalin, isolated from the dried flower of Heterotheca inuloides Cass. (Asteraceae), have been found to exhibit cytotoxic activity against several solid tumor cell lines. Among them, 7-hydroxy-3,4-dihydrocadalin and 7-hydroxycadalin have also been found to inhibit autoxidative and microsomal lipid peroxidation.

Animals↗
PubMed1996

Cloning and characterization of a melanin biosynthetic THR1 reductase gene essential for appressorial penetration of Colletotrichum lagenarium.

Melanin biosynthesis of Colletotrichum lagenarium is essential for appressorial penetration of the host plant. A melanin deficient mutant 9141 (Thr-) has a defect in the conversion of 1,3,8-trihydroxynaphthalene to vermelone in the melanin biosynthetic pathway. The mutant formed nonmelanized appressoria and had little infectivity on cucumber leaves. A cosmid clone pCR1 was selected from a cosmid library of wild-type C. lagenarium by means of a heterologous probe BRM2, one of the clustered genes involved in melanin biosynthesis of Alternaria alternata. pCR1 transformed the Thr- mutant 9141 to wild-type phenotype. A DNA fragment (THR1) homologous to BRM2 was subcloned from pCR1 and the nucleotide sequence determined. THR1 contains one open reading frame that encodes a protein of 282 amino acids. A transformant resulting from gene disruption showed a light brown phenotype different from the dark brown phenotype of the wild-type 104-T. The transformant formed nonmelanized appressoria and had little infectivity. The THR1 amino acid sequence contains a region highly similar to the Ver1 gene involved in the conversion of versicolorin A to sterigmatocystin in aflatoxin biosynthesis by Aspergillus parasiticus and to the T4HN reductase gene involved in the conversion of 1,3,6,8-tetrahydroxynaphthalene to scytalone and 1,3,8-trihydroxynaphthalene to vermelone in melanin biosynthesis by Magnaporthe grisea. Expression of the THR1 gene during spore germination of C. lagenarium was detected by RNA blotting. We propose that the C. lagenarium THR1 gene encodes a reductase involved in conversion of 1,3,8-trihydroxynaphthalene to vermelone.

Amino Acid Sequence↗
PubMed1996

Cloning and structural analysis of the melanin biosynthesis gene SCD1 encoding scytalone dehydratase in Colletotrichum lagenarium.

Appressoria of the phytopathogenic fungus Colletotrichum lagenarium contain melanin, which has been implicated as an important factor in the penetration of host plants. A cDNA clone containing the melanin biosynthetic gene encoding scytalone dehydratase (SCD1) from C. lagenarium was identified by hybridization with a heterologous cDNA probe from Magnaporthe grisea. The cDNA clone was used to identify a cosmid containing SCD1 in a genomic library of C. lagenarium, and the nucleotide sequence was determined for both the cDNA and genomic clones. The SCD1 gene contained one open reading frame composed of 188 codons and two deduced introns of 57 and 67 nucleotides. The deduced amino acid sequence of the N-terminal region of SCD1 showed high similarity to the amino acid sequence of scytalone dehydratase from Cochliobolus miyabeanus. A plasmid containing the SCD1 gene transformed the melanin-deficient mutant 9201Y (Scd-) to the wild phenotype but did not complement the conditional scytalone dehydratase-deficient mutant C. lagenarium 8015. Genomic DNA analysis indicated that the SCD1 gene is a single locus in C. lagenarium. Transcripts of the SCD1 gene were detected 2 h after the start of conidial germination.

Amino Acid Sequence↗
PubMed1996

Possible origin of murine AIDS (MAIDS) virus: conversion of an endogenous retroviral p12gag sequence to a MAIDS-inducing sequence by frameshift mutations.

The murine AIDS (MAIDS) virus has a unique sequence in its p12gag region, which is responsible for MAIDS development. A transcript hybridizing with this sequence is expressed in normal C57BL/6 mice. The transcript, designated Edv, has been previously cloned and sequenced (Y. Kubo, Y. Nakagawa, K. Kakimi, H. Matsui, K. Higo, L. Wang, H. Kobayashi, T. Hirama, and A. Ishimoto, J. Gen. Virol. 75:881-888, 1994). Compared with the nucleotide sequence of the helper LP-BM5 ecotropic virus, the pathogenic replication-defective MAIDS virus has a 16-bp deletion and a 1-bp insertion in the 5' and 3' regions of the p12gag sequence, respectively, and the Edv transcript contains only a 3-bp deletion. Therefore, the amino acid sequence of the defective MAIDS virus p12gag region is not homologous to that of the helper virus and the Edv transcript because of the frameshift. To determine whether the amino acid sequence resulting from the frameshift is critical for MAIDS development, we constructed chimeric viruses that contained the p12gag regions of the helper virus and the Edv transcript, respectively, with and without the same frame as the defective MAIDS virus by the artificial frameshift mutations. The mutant viruses with the frameshift mutations induced MAIDS in inoculated mice, but the viruses without the mutations did not. These results suggested that the MAIDS virus was generated by frameshift mutations in the p12gag region of Edv or a related sequence.

Animals↗
PubMed1996

Two aspects of the inward rectification mechanism. Effects of cytoplasmic blockers and extracellular K+ on the inward rectifier K+ channel.

The inward rectification of the inward rectifier K+ channels has been reported to be caused by both the block of the outward current by cytoplasmic Mg2+ and by intrinsic channel gating. Recently it was uncovered that the apparent intrinsic gating is mostly due to a block by cytoplasmic polyamines which is actually extrinsic to the channel. Furthermore, negatively charged amino-acid residues in the center of the M2 region and in the C-terminal hydrophilic domain were identified to be involved in the binding of these blockers to the channels. The inward rectifier shows consistent inward rectification at various extracellular K+ concentrations ([K+]o). To explain the dependency of the channel activity on Ko+, the hypothesis that Ko+ interacts with the inward rectifier K+ channel and thereby activates it (K(+)-activated K+ channel model) has been postulated. In this manuscript, the history and recent progresses of the study of the inward rectification mechanism, especially the effect of the cytoplasmic blockers and Ko+, is reviewed.

Animals↗
PubMed1996

[Pharmacokinetics of 99mTc-MAG3 and 131I-OIH: comparative study based on 2 compartment model analysis].

We studied 50 patients with mild to moderate renal dysfunction to compare pharmacokinetics of 99mTc-MAG3 with that of 131I-OIH. After simultaneous bolus injection of both 99mTc-MAG3 and 131I-OIH, 8-point venous blood sampling was performed from 2 to 44 min post injection. Aliquoted plasma samples were counted for radioactivity along with the injected standard to obtain % injected dose/ml plasma for each tracer. Using obtained time-concentration data, classical 2 compartment model analysis was performed for both tracers to obtain various pharmacokinetic parameters, including distribution volumes (Vds), inter-compartmental rate constants, and plasma clearance. In these parameters, Vd of central compartment, Vd at steady state, central to peripheral inter-compartmental rate constant, and plasma clearance were significantly larger for 131I-OIH. In all parameters, significant correlation was found between 99mTc-MAG3 and 131I-OIH. The best correlation was seen in plasma clearance (r = 0.891, p < 0.0001). Plasma clearance ratio (99mTc-MAG3/131I-OIH), however, showed weak but significant negative correlation with serum creatinine, although this correlation was not likely to affect the overall correlation of clearance between 131I-OIH and 99mTc-MAG3. From these results, we confirmed that 99mTc-MAG3 clearance could be used as an alternative to 131I-OIH clearance, although pharmacokinetic behavior of 99mTc-MAG3 was not exactly the same as that of 131I-OIH.

Adult↗
PubMed1996

[Expression of major histocompatibility complex of advanced non-small cell lung cancer (NSCLC)].

Expression of major histocompatibility complex of lung cancer was examined to study the malignant potential of the tumor using immunochemical staining (anti-HLA class I monoclonal antibody; w6/32). Nuclear DNA contents and the labeling index of proliferating cell nuclear antigen (PCNA) were measured as well. Forty three advanced (p-stage IIIa) lung cancers resected by lobectomy (33 cases), pneumonectomy (9 cases), and segmentectomy (1 case) entered the study. Mediastinal lymph node dissection was performed in every case during the operation. Of the 43 cases, expression of HLA class I was positive in 17 and negative in 26. The patients with positive HLA expression showed significantly better prognosis than those with negative HLA expression in terms of 5 year survival (p < 0.01). The patients with aneuploid pattern and positive HLA class I expression had the best prognosis on the survival curve (p < 0.01). When the PCNA positive rate was high, the prognosis was poor. And the patients with negative PCNA and positive HLA class I expression had significantly better prognosis than the others (p < 0.01). These findings suggest that the expression of HLA class I on the tumor cells is an important prognostic factor in the patients with NSCLC.

Carcinoma, Non-Small-Cell Lung↗
PubMed1996

[Pulmonary aspergillosis: clinical findings and surgical treatment].

Between 1980 and 1995, 10 patients underwent thoracotomy for pulmonary aspergillosis. In six patients, hemoptysis and bloody sputum were the chief complaints. The other complaints were nonspecific. Six patients had a history of pulmonary tuberculosis, and two of those patients underwent upper lobectomy. Aspergillosis had developed in the residual space. A fungus ball was observed on the preoperative chest X-ray and CT scan films in seven patients. Lobectomy was done in three patients, segmentectomy in two, and partial pulmonary resection in four. The patients with lesions that had grown in the residual space underwent curettage with muscle plombage. Three patients underwent thoracoplasty. An additional operation was done in two patients because of poor residual lung expansion. No patient had recurrence. We conclude that surgical treatment should be based on symptoms and on pathological findings.

Adult↗
PubMed1996

[Repair of diaphragmatic hernia through the thoracoabdominal spiral incision].

Traumatic diaphragmatic hernia requires urgent surgical treatment. A 51-year-old female was injured in a speeding car. She had dyspnea, and resistance on the abdominal wall on physical examination. Intra-thoracic and abdominal visceral injury was suspected. A chest roentogenogram and CT scan revealed an obvious diaphragmatic herniation on the left which necessitated emergency operation. On the right semi-lateral position, left thoracoabdominal spiral incision was made through the 7th intercostal space. Good exposure of the chest and abdomen was easily obtained. Herniated organs and the abdominal cavity were thoroughly examined with ease. The diaphragm was repaired directly. She had an uneventful postoperative recovery, and was discharged in 17 days. Thoracoabdominal spiral incision offered an excellent operative exposure for the patient with a possible combined thoracic and abdominal visceral injury.

Female↗
PubMed1996

Free thyroxine concentrations in serum measured by equilibrium dialysis in chronic renal failure.

The serum concentration of free thyroxine (FT4) is often low in patients with chronic renal failure (CRF) with low serum concentrations of triiodothyronine (T3). We evaluated the serum FT4 concentration by using both an equilibrium dialysis RIA kit (D-FT4) and a labeled-antibody kit (M-FT4) in two different groups of CRF patients, undergoing chronic hemodialysis (HD, n = 145) or not (non-HD, n = 30), and in a group of normal healthy subjects (n = 58). Thyroid peroxidase antibodies and thyroglobulin antibodies were not detected in any patient. Serum FT4 concentrations (mean +/- SD, pmol/L) by the D- and M-FT4 assays were, respectively, 21.5 +/- 4.6 and 16.6 +/- 2.0 in the healthy subjects, 17.8 +/- 4.3 and 13.9 +/- 3.6 in the non-HD patients, and 16.9 +/- 4.9 and 10.7 +/- 1.9 in the HD patients. By the D-FT4 assay, results for both CRF groups were significantly different from those for the healthy group (P <0.01), as were the results for each pair of groups by the M-FT4 assay (P <0.01). FT4 values were reported as being within the healthy reference range by D-FT4 in 73 of 113 HD subjects who had low T3 and low M-FT4 values. Serum FT4 concentrations measured by both assay kits showed a significant inverse correlation with the serum concentration of creatinine (P <0.01), but the serum concentrations of sex-hormone-binding globulin did not differ significantly among the three groups. Our results indicate that the low FT4 concentration measured by D-FT4 in patients with CRF, particularly those on HD, probably reflects the actual, mild nonthyroidal illness of renal failure.

Adult↗
PubMed1996

Effects of extracellular cations and mutations in the pore region on the inward rectifier K+ channel IRK1.

It is known that the rectification property of the inward rectifier K+ channel, IRK1, is caused by a block of outward current by cytoplasmic Mg2+ and polyamines, and that the voltage dependence of rectification shifts according to the equilibrium potential of K+. Here it is shown that extracellular K+ (K+ o) but not intracellular K+ (K+ i) affects the activation kinetics of IRK1. A mutant in which the conserved positively charged residue arginine was replaced by a tyrosine (R148Y) exhibited slower activation and a negative shift of the conductance-voltage relationship. In addition, the conductance did not saturate at negative potentials as was observed for the wild type. When using T1+o instead of K+ o as permeant ion, the differences between the wild type and the mutant were qualitatively similar but less prominent. These results suggest that extracellular cations (e.g., K+ o or T1+ o) play a role in the activation of IRK1. Since the effects of K+ o or T1+ o were altered in the mutant, the site R148 presumably is involved in channel regulation by extracellular cations.

Amino Acid Sequence↗