Search PubMed⌕ Search

Biomedical subjects

T Aoki

Publications and source records attributed to T Aoki.

At least 487 records · Page 27Linked to original sources

Expressed sequence tags of medaka (Oryzias latipes) liver mRNA.

A medaka liver cDNA library was constructed in lambda ZAPII. The number of clones in this library is approximately 5 x 10(6). Three hundred sixty-one clones were randomly selected and, of these, 33 clones with over 1 kb were sequenced. These sequences were compared with GenBank and the dbEST (Re.96.0). Twenty-five clones of these 33 clones encoded 18 different genes and 2 different expressed sequence tags (ESTs). Sequences of 10 of the 18 clones had not previously been reported in fish. The codon usage of medaka genes is similar to that of Xenopus, mouse, and human genes, but not similar to that of yeast and Escherichia coli genes.

Animals↗

Cloning and characterization of transferrin cDNA and rapid detection of transferrin gene polymorphism in rainbow trout (Oncorhynchus mykiss).

A cDNA clone of rainbow trout (Oncorhynchus mykiss) transferrin was obtained from a liver cDNA library. The 2537-bp cDNA sequence contained an open reading frame encoding 691 amino acids and the 5' and 3' noncoding regions. The amino acid sequences at the iron-binding sites and the two N-linked glycosylation sites, and the cysteine residues were consistent with known, conserved vertebrate transferrin cDNA sequences. Single N-linked glycosylation sites existed on the N- and C-lobe. The deduced amino acid sequence of the rainbow trout transferrin cDNA had 92.9% identities with transferrin of coho salmon (Oncorhynchus kisutch); 85%, Atlantic salmon (Salmo salar); 67.3%, medaka (Oryzias latipes); 61.3% Atlantic cod (Gadus morhua); and 59.7%, Japanese flounder (Paralichthys olivaceus). The long and accurate polymerase chain reaction (LA-PCR) was used to amplify approximately 6.5 kb of the transferrin gene from rainbow trout genomic DNA. Restriction fragment length polymorphisms (RFLPs) of the LA-PCR products revealed three digestion patterns in 22 samples.

Amino Acid Sequence↗

A survey of expressed genes in Japanese flounder (Paralichthys olivaceus) liver and spleen.

Expressed sequence tags (ESTs) were obtained from Japanese flounder (Paralichthys olivaceus). We present the results of single-pass sequencing of 493 ESTs from 350 clones from liver cDNA and 57 ESTs from 41 clones from spleen cDNA. Sequences of the cDNA clones were compared with sequences in the GenBank database. Two hundred and two clones (51.7%) appeared to be completely unknown and are likely to represent newly described genes, whereas 189 clones (48.3%) were identified based on matches to sequences in the databases. Three of the unidentified sequences were isolated from both the liver and spleen cDNA libraries. However, there were no identical sequences between liver and spleen clones.

Animals↗

[Nutritional support in terminal patients--evaluation of 109 cases receiving home parenteral nutrition].

For end-stage patients with malignant disease, home parenteral nutrition (HPN) is an effective and useful treatment in terms of pain control and nutritional support. Therefore, these patients are able to spend their final days at home with their family. In 1990, we started the treatment and have been experienced 109 cases. In this report, we evaluate these cases and which condition or circumstance are necessary to perform HPN for the terminal patients.

Adult↗

[Home enteral nutrition of patients with terminal cancer].

Generally hospitalization is regarded as a prerequisite for patients with cancer who require terminal care. Recently, special attention has been directed to home care for these patients from the standpoints of patients quality of life (QOL) and health economics. In the case of home care for patients with terminal cancer, effective pain control, careful nutrition administration, psychological care for the patient and his/her family members and emergency care are indispensable. Due to the number and extent of these requirements, home care for terminal patients is not yet common. We have been promoting the adoption of home enteral nutrition as a method to meet the nutritional needs of patients with terminal cancer. In this report, we discuss home enteral nutrition focusing on its indication and methodology, as well as related problems and nutrition administration in the future.

Adult↗

[PPI or laparoscopic surgery in the treatment of reflux esophagitis?].

Proton pump inhibitor (PPI) is very effective in the initial treatment of reflux esophagitis. However, the recurrence ratio is high during maintenance therapy. Ten to 20% patients did not remained healed or the symptom were relapsed in spite of one year maintenance treatment with half or routine dose of PPI. Therefore, prolonged maintenance therapy with PPI is needed for many patients with symptomatic reflux esophagitis. On the other hand, surgical treatment is to repair functional disorder in defensive mechanism at esophagogastric junction which is main pathophysiology of reflux esophagitis. Surgical treatment is radical treatment for reflux esophagitis in this point of view. Nissen fundoplication has become the most commonly used antireflux procedure. Following fundoplication, 90% of the patients become symptom free in the long run. Laparoscopic Nissen fundoplication is as effective as open Nissen fundoplication in the short-term results. The average operation time is 2.5 hours, most of patients were discharged within two days after operation and lead to faster recovery from surgery than open procedure. Satisfaction rates ranged from 87 to 100%. Laparoscopic fundoplication can be performed with less morbidity than open procedure. Therefore, laparoscopic Nissen fundoplication is considered to be radical antireflux procedure. Compared to medical treatment, the life time costs of treatment are less with initial surgical management for men age 48 or below and women age 55 or below. The answer for "PPI or laparoscopic surgery in the treatment of reflux esophagitis" is laparoscopic surgery definitely.

Esophagitis, Peptic↗

[The new criteria for skin prick test of atopic early infants--diagnosis for hypersensitivity of egg white].

To investigate the new criteria for skin prick test (SPT) of seventy-four atopic infants (2-5 months of age at the first visit, Mean 3.8 months, M:F = 54:20) to diagnose for hypersensitivity to egg white. It was classified into three groups by reaction type of SPT in the first visit. Group A were the infants who seemed only late (6 hours) or delayed (48 hours) reaction (n = 26). Group B were seemed immediate (15 minutes) and late or delayed reactions (n = 26), Group C were seemed only immediate reaction (n = 23). Atopic infants and controlled infants without no symptom but have any atopic disease a relative in the third degree, agreed to undergo SPT in the first visit, the prior were undergo 9-12 months of age, too. Serum total IgE (RIST), serum specific IgE antibody of egg white (EWRAST) and peripheral eosinophil counts in the blood (Eo. counts) were determined at the same time of SPT in atopic infants. The best criterion for SPT was the longest diameter of a erythema were greater than 3 mm at late and/or delayed reaction (Sensitivity: 100%, Specificity: 60%) in group A. Two third of infants in group. A were seemed immediate reaction and EWRAST levels were increased to larger than gread two at 9-12 months (p < 0.001). RIST levels and Eo. counts at the first visit were increased compared with the normal levels in the all groups, the prior and EWRAST levels in group B were higher than group A or C (p < 0.05, p < 0.05). RIST and EWRAST levels in group A at 9-12 months were higher than the first visit (p < 0.05, p < 0.01). In conclusion, SPT in atopic early infants were seemed several reactions at the first visit, but all reactions were useful for diagnose for hypersensitivity to egg white.

Dermatitis, Atopic↗

Airway and lung tissue behaviour during endothelin-1 induced constriction in rats: effects of receptor antagonists.

Endothelin (ET) 1, a 21 amino acid constrictor peptide, is one of the most potent agonists of airway smooth muscle and acts on two different receptors, i.e., ETA and ETB receptors. Recently, it has been shown that there are species and organ differences in physiological roles of each ET receptor. In rats, however, the physiological roles of ET receptors remain to be clarified. We questioned whether ET-1 might affect airway and lung tissue via different ET receptor subtypes in rats. To answer this question, we investigated the effects of ET-1 on lung behaviour in anesthetized, open-chested, mechanically ventilated (f = 1 Hz, VT = 9 mL/kg, PEEP = 3 cmH2O (1 cmH2O = 98.1 Pa)) rats in the absence or the presence of ETA and ETB selective antagonists, i.e., BQ-123 and BQ-788, respectively. Using alveolar capsules, we calculated lung elastance (EL), resistance of lung (RL), tissue (Rti), and airway (Raw), and hysteresivity (eta = 2 pifRti/EL) under control conditions and after intravenous administration of ET-1 (10(-8) mol/kg). ET-1 induced significant increases in RL, Rti, Raw, EL, and eta. BQ-123 did not affect ET-1 induced constriction, while BQ-788 significantly reduced delta RL, delta Rti, delta Raw, delta EL during ET-1 induced constriction. The effects of the combination of BQ-123 and BQ-788 were not different compared with BQ-788. Eta was not affected by BQ-123 and BQ-788. These data suggest that ETB, but not ETA, receptors may have significant physiological roles in rat lungs in response to ET-1.

Airway Resistance↗

[A case of interstitial pneumonia induced by intravesical administration of bacillus Calmette-Guerin (BCG)].

A 61-year-old man with superficial bladder cancer, which was detected after he complained of hematuria, was treated three times with intravesical BCG administration. Since liver dysfunction was detected thereafter, he was admitted to our hospital. Three days after admission, he complained of dyspnea on exertion associated with severe hypoxemia, as well as abnormal findings on chest X-ray, i.e. extensive bilateral lung densities. We performed bronchoscopic examination and obtained bronchoalveolar lavage fluid (BALF) and lung biopsy specimens (TBLB). In the BALF, a marked increase in the total cell number, particularly lymphocytes with a high CD4/CD8 ratio was noted. TBLB specimens revealed the lesions to be numerous non-caseating granulomas. We failed to obtain definite evidence of BCG in the sputum, urine, blood, and BALF. Instead, we found that a lymphocyte stimulation test for BCG (DLST) was strongly positive. Based on these findings, severe interstitial pneumonia probably induced by hypersensitivity against BCG, was diagnosed. Anti-tuberculous agents, and steroid-pulse therapy followed by oral administration of relatively low dose of steroid ameliorated the abnormal conditions, including chest X-ray film findings and hypoxemia. The population of lymphocytes and CD4/CD8 ratio in the BALF were reduced as well. Serious interstitial pneumonia was induced by the intravesical administration of BCG, which resulted in transitional changes in the BALF cell component.

Administration, Intravesical↗

Genomic organization, expression, and chromosomal mapping of the mouse adrenomedullin gene.

We have isolated and characterized the mouse adrenomedullin (AM) gene (Adm) and determined its chromosomal location. The gene spans approximately 2.1 kb and is organized into four exons separated by three introns. The transcription start site was determined to be the adenine nucleotide at -618. The mouse AM 5'-flanking region contains a TATA box-like sequence and several cis-acting regulatory elements. Analysis of the nucleotide and deduced amino acid sequences revealed that mouse preproAM is a 184-amino-acid polypeptide, from which AM and proAM N-terminal 20 peptide are cleaved. Using restriction fragment length variants on a DNA panel of interspecific backcross mice, we mapped Adm to a distal region of mouse chromosome 7.

Adrenomedullin↗

TRAF5, a novel tumor necrosis factor receptor-associated factor family protein, mediates CD40 signaling.

Signals emanating from CD40 play crucial roles in B-cell function. To identify molecules that transduce CD40 signalings, we have used the yeast two-hybrid system to done cDNAs encoding proteins that bind the cytoplasmic tail of CD40. A cDNA encoding a putative signal transducer protein, designated TRAF5, has been molecularly cloned. TRAF5 has a tumor necrosis factor receptor-associated factor (TRAF) domain in its carboxyl terminus and is most homologous to TRAF3, also known as CRAF1, CD40bp, or LAP-1, a previously identified CD40-associated factor. The amino terminus has a RING finger domain, a cluster of zinc fingers and a coiled-coil domain, which are also present in other members of the TRAF family protein except for TRAF1. In vitro binding assays revealed that TRAF5 associates with the cytoplasmic tail of CD40, but not with the cytoplasmic tail of tumor receptor factor receptor type 2, which associates with TRAF2. Based on analysis of the association between TRAF5 and various CD40 mutants, residues 230-269 of CD40 are required for the association with TRAF5. In contrast to TRAF3, overexpression of TRAF5 activates transcription factor nuclear factor kappa B. Furthermore, amino-terminally truncated forms of TRAF5 suppress the CD40-mediated induction of CD23 expression, as is the case with TRAF3. These results suggest that TRAF5 and TRAF3 could be involved in both common and distinct signaling pathways emanating from CD40.

Amino Acid Sequence↗

Cloning of a cDNA encoding the small subunit of cytochrome b558 (cybS) of mitochondrial fumarate reductase (complex II) from adult Ascaris suum.

Complex II in the mitochondria of the adult parasitic nematode, Ascaris suum, exhibits high fumarate reductase activity in addition to succinate dehydrogenase activity and plays a key role in the anaerobic energy metabolism of the worm. In this study, the amino acid sequence of the small subunit of cytochrome b558 (cybS) in adult complex II was deduced from the cDNA isolated by immunoscreening an A. suum muscle cDNA library. Histidine residues, which are possible heme axial ligands in cytochrome b558, were found in the second transmembrane segment of the subunit. This is the first report of the primary structure of the small subunit in the two-subunit cytochrome b in mitochondrial complex II from a multicellular eukaryote.

Amino Acid Sequence↗

High-performance liquid chromatographic method for the rapid and simultaneous determination of sulfamonomethoxine, miloxacin and oxolinic acid in serum and muscle of cultured fish.

A rapid method for the simultaneous determination of sulfamonomethoxine (SMM), miloxacin (MLX) and oxolinic acid (OA) in serum and muscle of cultured fish by high-performance liquid chromatography has been developed. A Hisep shielded hydrophobic phase column (15 cm x 4.6 mm I.D.) and a mobile phase of 0.05 M citric acid-0.2 M disodium hydrogenphosphate buffer, pH 2.5 in 10 mM tetra-n-butyl ammonium bromide-acetonitrile (85:15) with ultraviolet detection at 265 nm were used. The recoveries of SMM, MLX and OA from serum and muscle samples were 72-101%. The detection limits of the three drugs were 0.05-0.1 microgram/ml or g of sample.

4-Quinolones↗

Membrane-type matrix metalloproteinase 1 is a gelatinolytic enzyme and is secreted in a complex with tissue inhibitor of metalloproteinases 2.

The processing mechanism and gelatinolytic activity of the membrane-type matrix metalloproteinase 1 (MT-MMP-1) were examined by expressing in COS-1 cells a deletion mutant of MT-MMP-1 lacking the trans-membrane domain (delta MT1) and its site-directed mutant with a furin-resistant sequence in the propeptide domain (mutant delta MT1). delta MT1, but not mutant delta MT1, was processed to an active form and exhibited gelatinolytic activity as seen using gelatin zymography. delta MT1 isolated in a complex form with tissue inhibitor of metalloproteinases 2 (TIMP-2) from the stable transfectants demonstrated the NH2-terminal sequence of Ala113-IIe-Gln-Leu, indicating cleavage at one amino acid down-stream from the furin recognition sequence. The delta MT1/TIMP-2 complex formed a ternary complex with proMMP-2 through the COOH termini of TIMP-2 and proMMP-2. A human breast carcinoma cell line (MDA-MB-231 cells) also secreted MT-MMP-1 into culture media, which was purified in a complex form with TIMP-2 and showed gelatinolytic activity as seen using zymography. These results demonstrate for the first time that MT-MMP-1 is a gelatinolytic enzyme and secreted from cells in a complex with TIMP-2, which can form a ternary complex of MT-MMP-1/TIMP2/proMMP-2.

Animals↗

A proteinase inhibitor from egg yolk of hen is an ovoinhibitor analog.

A proteinase inhibitor, tentatively termed vitelloinhibitor, was purified from yolk of hen's ovarian follicles. It resembled egg-white ovoinhibitor not only in inhibitory spectrum (active for bovine trypsin and bovine chymotrypsin) but also in thermal stability, pH stability, antiserum reactivity and amino-acid composition. However, vitelloinhibitor had different molecular weight from that of ovoinhibitor. An alpha 2-proteinase inhibitor preparation, isolated from laying hen's serum in the present study, was found to exhibit two bands, and the larger one of the latter corresponded to vitelloinhibitor in molecular weight. The partial N-terminal amino-acid sequence of vitelloinhibitor was the same as those of the two components of serum inhibitor and all three agreed with that of ovoinhibitor. Vitelloinhibitor is likely to be an ovoinhibitor analog derived from a serum precursor, which might be the larger component of alpha 2-proteinase inhibitor.

Amino Acid Sequence↗

MEK kinase is involved in tumor necrosis factor alpha-induced NF-kappaB activation and degradation of IkappaB-alpha.

Signal-dependent activation of the transcription factor NF-kappaB is dominantly regulated by degradation of IkappaB-alpha protein. However, the signaling pathways that lead to the degradation are not clear. Here we report that mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) kinase, an activator of stress-activated protein kinases/jun kinase-1 (SAPKs/JNK1), is involved in such signaling pathways. The transient overexpression of MEK kinase in NIH3T3 fibroblasts activates kappaB-CAT reporter expression in a synergistic manner with TNFalpha stimulation. In contrast, overexpression of kinase-negative MEK kinase suppresses TNFalpha-induced reporter expression. The overexpression of MEK kinase suppresses the inhibitory activity of co-transfected IkappaB-alpha on the kappaB-CAT or human immunodeficiency virus-long terminal repeat-luciferase reporter expression and causes the simultaneous disappearance of the overexpressed IkappaB-alpha. The disappearance of exogenous IkappaB-alpha by the overexpression of MEK kinase is prevented by calpain inhibitor-I, an inhibitor of IkappaB-alpha degradation. These results suggest that MEK kinase is a signal mediator involved in TNFalpha-induced NF-kappaB activation and that the activation of NF-kappaB by MEK kinase is regulated through the degradation of IkappaB-alpha.

3T3 Cells↗

Construction of a fusion protein between protein A and green fluorescent protein and its application to western blotting.

Aequorea green fluorescent protein (GFP) and protein A were fused and expressed in Escherichia coli. The fluorescent native fusion protein (PA-GFP) migrated at 47 kDa in SDS-PAGE. However, the non-fluorescent denatured PA-GFP migrated at 57 kDa which corresponds to the theoretical molecular mass. Although the reason(s) for this mobility shift between fluorescent and non-fluorescent molecules remains unclear, the small ring structure within the native molecules may affect their mobility. The cell extract, prepared from an E. coli strain producing PA-GFP, was used in Western and dot blots. The sensitivity and specificity of the PA-GFP detection were sufficient for rapid and easy screening.

Amino Acid Sequence↗