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Biomedical subjects

Y Inagaki

Publications and source records attributed to Y Inagaki.

At least 73 records · Page 4Linked to original sources
PubMed1999

Capture of a genomic HMG domain sequence by the En/Spm-related transposable element Tpn1 in the Japanese morning glory.

The non-autonomous transposable element Tpn1 from the Japanese morning glory is an En/Spm-related DNA element found in the second intron of the DFR-B gene for flower pigmentation in the mutable line flecked, which shows variegation for flower color. It carries a genomic DNA segment containing at least four exon sequences encoding part of a HMG-box sequence. Spliced hybrid transcripts containing the DFR-B exon(s) and the HMG exons in Tpn1 were detected in the flower buds of the flecked line, and they were polyadenylated within Tpn1. Thus, Tpn1 can be regarded as a specialized transducing transposon carrying a part of the genomic sequence for a HMG box. The possible implications of the finding for evolution are discussed.

Amino Acid Sequence↗
PubMed1999

A study of anti-carbonic anhydrase II antibodies in rheumatic autoimmune diseases.

Autoantibodies to human carbonic anhydrase II (CAII) were screened by ELISA in 109 sera from Asian Japanese patients with systemic lupus erythematosus (SLE), primary Sjögren's syndrome (Sjs), progressive systemic sclerosis (PSS) and dermatomyositis (DM). Anti-CAII antibodies were positive in 24.1% of SLE, 20.0% of primary Sjs, 16.7% of PSS and 25.0% of DM. On the other hand, sera from atopic dermatitis, bullous pemphigoid and psoriasis patients showed no activity for anti-CAII antibodies. CAII could be a common exonuclear autoantigen in subsets of rheumatic autoimmune diseases.

Antibody Specificity↗
PubMed1999

Absence of R1066X mutation in six Japanese patients with Dubin-Johnson syndrome.

The Dubin-Johnson syndrome (DJS) is a rare autosomal recessive liver disease characterized by chronic conjugated hyperbilirubinemia. The phenotype of this syndrome is thought to be caused by the impaired expression of the canalicular multispecific organic anion transporter (cMOAT), which transports non-bile salt organic anions into the bile. Recently, a mutation from arginine (Arg) to stop-codon at codon 1066 in the cMOAT gene has been reported in one Caucasian patient with DJS. In this study, we investigated whether this mutation is found in Japanese patients with DJS. Genomic DNAs were extracted from the leukocytes of six Japanese patients and the fragments spanning codon 1066 were amplified by polymerase-chain reaction. The digest of the amplified fragments with a restriction enzyme, Taql, demonstrated that all of six patients did not exhibit an R1066X mutation. No mutation at Arg1066 was also confirmed by direct sequencing of the amplified products. These findings suggested that this R1066X mutation was not a major mutation in Japanese patients with DJS. Further investigation will be required in an attempt to search other mutations in cMOAT gene in Japanese patients with DJS.

Adult↗
PubMed1999

Remifentanil administration during monitored anesthesia care: are intermittent boluses an effective alternative to a continuous infusion?

UNLABELLED: This randomized, double-blind study was designed to evaluate the analgesic effectiveness and respiratory stability of remifentanil when administered as intermittent bolus injections, a variable-rate infusion, or a combination of a constant basal infusion supplemented with intermittent boluses during monitored anesthesia care (MAC). Forty-five patients undergoing extracorporeal shock wave lithotripsy (ESWL) procedures were randomly assigned to one of the three modes of remifentanil administration. All patients received midazolam 2 mg i.v., followed by a propofol infusion at 50 microg x kg(-1) x min(-1). Two minutes before administering a series of test shock waves: Group I received a remifentanil infusion of 0.1 microg x kg(-1) x min(-1), and a saline bolus (5 mL); Group II received a saline infusion and a remifentanil bolus (25 microg in 5 mL); and Group III received a remifentanil infusion of 0.05 microg x kg(-1) x min(-1), and a remifentanil bolus (12.5 microg in 5 mL). The average pain intensity was scored on an 11-point scale, with 0 = no pain to 10 = severe pain. During the ESWL procedure, pain was treated by increasing the study drug infusion rate by 25%-50% and administering 5-mL bolus injections of the study medication in Groups I (saline) and II (remifentanil 25 microg). In Group III, intermittent 5-mL boluses (remifentanil 12.5 microg) were administered as needed. Patients in Groups II and III reported lower pain scores in response to the test shocks. Significantly more remifentanil was administered in Group I (379 +/- 207 microg) than in Group II (201 +/- 136 microg). However, more interventions were required for the treatment of intraoperative pain in the intermittent bolus group (Group II). When remifentanil is administered as the analgesic component of a MAC technique, these data support the use of intermittent bolus doses (12.5-25 microg) alone or in combination with a basal infusion (0.05 microg x kg(-1) x min(-1)) as alternatives to a variable-rate continuous infusion. IMPLICATIONS: In this study, three different modes of remifentanil administration were used during monitored anesthesia care for extracorporeal shock wave lithotripsy procedures. These results suggest that using intermittent bolus injections of remifentanil (25 microg) or a continuous infusion (0.05 microg x kg(-1) x min(-1)) supplemented with intermittent bolus (12.5 microg) injections may be more effective than a variable-rate infusion of remifentanil during propofol sedation.

Adult↗
PubMed1999

The cost-effectiveness of methohexital versus propofol for sedation during monitored anesthesia care.

UNLABELLED: We designed this study to test the hypothesis that methohexital is a cost-effective alternative to propofol for sedation during local anesthesia. Sixty consenting women undergoing breast biopsy procedures under local anesthesia were randomly assigned to receive an infusion of either propofol (50 microg x kg(-1) x min(-1)) or methohexital (40 microg x kg(-1) x min(-1)). The sedative infusion rate was titrated to maintain an observer's assessment of alertness/sedation (OAA/S) score of 3 (with 1 = awake/alert to 5 = asleep). Fentanyl 25 microg i.v. was administered as a "rescue" analgesic during the operation. We assessed the level of sedation (OAA/S score), vital signs, time to achieve an OAA/S score of 3 at the onset and a score of 1 after discontinuing the infusion, discharge times, perioperative side effects, and patient satisfaction. The direct cost of methohexital was lower than that of propofol, based on the milligram dosage infused during the operation. The sedative onset (to achieve an OAA/S score of 3) and the recovery (to return to an OAA/S score of 1) times, as well as discharge times, did not differ between the two groups. Patients receiving methohexital had a significantly lower incidence of pain on initial injection compared with those receiving propofol (10% vs 23%). Because the use of methohexital (29.4 +/- 2.7 microg x kg(-1) x min(-1)) for sedation during breast biopsy procedures has a similar efficacy and recovery profile to that of propofol (36.8 +/- 15.9 microg x kg(-1) x min(-1)) and is less costly based on the amount infused, it seems to be a cost-effective alternative to propofol for sedation during local anesthesia. However, when the cost of the drug infused and drug wasted was calculated, there was no difference in the overall drug cost. IMPLICATIONS: When administered to maintain a stable level of sedation during local anesthesia, methohexital is an acceptable alternative to propofol. However, the overall drug costs were similar with the two drugs.

Anesthesia, Local↗
PubMed1998

A new low density lipoprotein receptor related protein, LRP5, is expressed in hepatocytes and adrenal cortex, and recognizes apolipoprotein E.

The isolation and characterization of rabbit and human cDNAs revealed a new low density lipoprotein receptor (LDLR)-related protein (LRP) designated as LRP5. Human LRP5 cDNA encodes a 1, 616-amino acid type I membrane-like protein with three ligand binding repeats in its extracellular region. LDLR-deficient cells transduced by recombinant adenovirus containing human LRP5 exhibited increased binding of apolipoprotein E (apoE)-enriched beta-migrating very low density lipoprotein. Northern blotting and in situ hybridization revealed a high level of LRP5 expression in hepatocytes and the adrenal gland cortex. In LDLR-deficient Watanabe heritable hyperlipidemic rabbits, LRP5 mRNA was increased in the liver and accumulated in cholesterol-laden foam cells of atherosclerotic lesions.

Adrenal Cortex↗
PubMed1998

Activation of Proalpha2(I) collagen promoter during hepatic fibrogenesis in transgenic mice.

We previously identified the promoter sequence that is essential for basal and TGF-beta-stimulated transcription of alpha2(I) collagen gene (COL1A2). In the present study, we examined whether the promoter is activated during hepatic fibrogenesis by utilizing transgenic mice harboring the COL1A2 upstream sequence. Intraperitoneal CCl4 administration activated the -17 kb COL1A2 promoter more than 10-fold, whereas partial hepatectomy resulted in no significant change in the promoter activity. The non-parenchymal cell fraction, but not parenchymal hepatocytes, isolated from mice harboring the -313 COL1A2 promoter linked to a beta-galactosidase reporter gene contained large amounts of beta-galactosidase and endogenous COL1A2 mRNAs. beta-galactosidase activity in the cells from CCl4-treated mice was significantly higher than in those from untreated animals. These results indicated that different molecular mechanisms control COL1A2 transcription in CCl4-induced liver injury/fibrosis and physiological regeneration after partial hepatectomy, and that the -313 COL1A2 promoter is activated in a cell type-specific manner during hepatic fibrogenesis.

Animals↗
PubMed1998

Presence of Streptococcus anginosus DNA in esophageal cancer, dysplasia of esophagus, and gastric cancer.

We recently reported cloning of Streptococcus anginosus (S. anginosus) DNA fragments containing the 16S ribosomal gene from DNA samples of surgical specimens of gastric cancers. To investigate the specificity of S. anginosus infection, Southern blot analysis with S. anginosus 16S ribosomal DNA probe and PCR analysis with S. anginosus-specific primers were performed in DNA samples prepared from 15 esophageal cancers, 43 gastric cancers, 16 lung cancers, 10 cervical cancers, 14 renal cell carcinomas, 10 colorectal cancers, and 19 bladder cancers. We frequently found S. anginosus DNA sequences in DNA samples from esophageal cancer and gastric cancer tissues, as well as in those from dysplasia of the esophagus of esophageal cancer patients. No S. anginosus DNA bands were detected by Southern blot analysis on DNAs from the noncancerous portions of the esophagus or the stomach. By PCR analysis with 35 cycles, only 7% of the noncancerous portion of the esophagus was shown to contain S. anginosus sequences. No S. anginosus sequences were found in DNAs from cancers in lung, cervix, and kidney, but they were found in 1 of 10 colon cancers.

Base Sequence↗
PubMed1998

Distinctive origins of group I introns found in the COXI genes of three gree algae.

Upon surveying the cytochrome c oxidase subunit I (COXI) gene of green algae, we found group I introns in three species of algae, Chlorella vulgaris (Cv), Scenedesmus quadricauda (Sq) and Protosiphon botryoides (Pb). The comparative analysis of these nucleotide sequences and their secondary structures revealed that the introns of Cv, Sq, and Pb belong to groups IB1, ID, and IB2, respectively. Each of the three introns contained an open reading frame (ORF) that showed a similarity to the sequence of the LAGLIDADG endonuclease family. However, each of the intronic ORFs in Sq and Pb had a discontinuity in the middle of' the sequences coding for the LAGLIDADG endonuclease. Either of the two ORFs could be restored to a sequence homologous to the LAGLIDADG endonuclease by the insertion of a nucleotide in the appropriate position. In Sq, a putative pseudo-knot structure was detected in the intronic ORF This suggests the occurrence of a ribosomal frameshift in the translation of the ORF. because such pseudo-knot structures are common in viral ORFs employing a (-1) ribosomal frameshift. In the phylogenetic tree that was inferred from the amino acid sequences of algal and non-algal intronic ORFs, the three algal ORFs did not make a cluster, but were scattered throughout the tree. In addition. each of the three algal ORFs showed a close relationship to the ORFs of non-algal introns that were inserted at the corresponding site of the COX] gene, suggesting distinctive origins of the three algal introns via independent horizontal transfers.

Amino Acid Sequence↗
PubMed1998

Neutralization of biological activity and inhibition of receptor binding by antibodies against human thrombopoietin.

Thrombopoietin (TPO) is a recently isolated cytokine that primarily regulates megakaryocytopoiesis and thrombopoiesis. We recently reported the development of a variety of antibodies (Abs) to synthetic peptides of human (h)TPO and to recombinant human TPO (rhTPO). In this study, we characterized the Abs and mapped immunologically distinct areas of the molecule. Among the five different antipeptide polyclonal Abs, only one, raised against synthetic peptide D8 to Q28, neutralized the TPO-dependent growth of FDCP-2 cells expressing human Mpl (FDCP-hMpl5 cells). One out of seven anti-rhTPO monoclonal Abs, designated as TN1, also showed neutralizing activity. TN1 was found to be specifically reactive with two proteolytic fragments, residues S1 to R117 and A60 to K122 of hTPO, indicating that the epitope(s) of TN1 is localized in residues A60 to R117 of the molecule. These two neutralizing Abs inhibited the binding of biotinylated rhTPO to FDCP-hMpl5 cells. On the other hand, the other Abs, which reacted with five polypeptides of S47 to D62, L108 to A126, N172 to A190, S262 to T284, and P306 to G332 of hTPO, did not show either the neutralizing activity or the ability to inhibit the binding of biotinylated rhTPO to the cell surface hMpl. These findings indicate that two regions, residues D8 to Q28 and A60 to R117 of hTPO, may contain the domains associated with its receptor, C-Mpl. These Abs characterized here are valuable for studying the structural analysis and the biological function of hTPO mediated by its receptor.

Antibodies↗
PubMed1998

Genetic and immunological analyses of Vls (VMP-like sequences) of Borrelia burgdorferi.

DNA fragments containing the VMP-like sequence (Vls) were cloned from Borrelia burgdorferi strain 297. Analyses by PCR, PFGE, and Southern hybridization revealed that the Vls sequences existed in multi-copies on the 20-kb borrelial plasmid, but not on chromosomes or other plasmids. One Vls unit of the strain 297 was about 669 bases, and predicted peptides length was 223 amino acids. Homologues of the Vls fragment were detected in three B. burgdorferi strains, a B. garinii strain 20047, and a B. afzelii strain P/Gau. A recombinant VlsII protein prepared in Escherichia coli strain JM109 reacted with antibodies that existed in three of five patients, by immunoblotting. These results suggested that the Vls of B. burgdorferi is expressed in Lyme disease patients.

Amino Acid Sequence↗
PubMed1998

Directionally evolving genetic code: the UGA codon from stop to tryptophan in mitochondria.

For the comprehensive analyses of deviant codes in protistan mitochondria (mt), we sequenced about a 1.1-kb region of a mitochondrial (mt) gene, the cytochrome c oxidase subunit I (coxI) in two chlorarachniophytes, the filose amoeba Euglypha rotunda, the cryptomonad Cryptomonas ovata, the prymnesiophyte (haptophyte) Diacronema vlkianum (Pavlovales), and the diatom Melosira ambigua. As a result of this analysis, we noticed that the UGA codon is assigned to tryptophan (Trp) instead of being a signal for translational termination in two chlorarachniophytes and in E. rotunda. The same type of deviant code was reported previously in animals, fungi, ciliates, kinetoplastids, Chondrus crispus (a red alga), Acanthamoeba castellanii (an amoeboid protozoon), and three of the four prymnesiophyte orders with the exception of the Pavlovales. A phylogenetic analysis based on the COXI sequences of 56 eukaryotes indicated that the organisms bearing the modified code, UGA for Trp, are not monophyletic. Based on these studies, we propose that the ancestral mitochondrion was bearing the universal genetic code and subsequently reassigned the codon to Trp independently, at least in the lineage of ciliates, kinetoplastids, rhodophytes, prymnesiophytes, and fungi. We also discuss how this codon was directionally captured by Trp tRNA.

Amoeba↗
PubMed1997

Sp1 is required for the early response of alpha2(I) collagen to transforming growth factor-beta1.

It is currently debated whether AP1 or Sp1 is the factor that mediates transforming growth factor beta1 (TGF-beta) stimulation of the human alpha2(I) collagen (COL1A2) gene by binding to an upstream promoter element (TbRE). The present study was designed to resolve this controversy by correlating expression of COL1A2, AP1, and Sp1 in the same cell line and under different experimental conditions. The results strongly indicate that Sp1 is required for the immediate early response of COL1A2 to TGF-beta and AP1 is not. The Sp1 inhibitor mithramycin blocked stimulation of alpha2(I) collagen mRNA accumulation by TGF-beta, whereas the AP1 inhibitor curcumin had no effect. Furthermore, antibodies against Jun-B and c-Jun failed to identify immunologically related proteins in the TbRE-bound complex, irrespective of whether they were purified from untreated or TGF-beta-treated cells. AP1 did bind to the TbRE probe in vitro, but only in the absence of the upstream Sp1 recognition sequence. Based on this finding and DNA transfection results, we conclude that the AP1 sequence of the TbRE represents a cryptic site used under experimental conditions that either eliminate the more favorable Sp1 binding site or force the balance toward the less probable. Finally, a combination of cell transfections and DNA-binding assays excluded that COL1A2 transactivation involves the retinoblastoma gene product (pRb), an activator of Sp1, the pRb-related protein p107, an inhibitor of Sp1, or the Sp1-related repressor, Sp3.

Collagen↗
PubMed1997

Thrombin cleaves recombinant human thrombopoietin: one of the proteolytic events that generates truncated forms of thrombopoietin.

A heterogeneity in the molecular weight (Mr) of thrombopoietin (TPO) has been reported. We found several thrombin cleavage sites in human, rat, murine, and canine TPOs, and also found that human TPO undergoes selective proteolysis by thrombin. Recombinant human TPO (rhTPO) was incubated with human platelets in the presence of calcium ions to allow the generation of thrombin, and was cleaved into low Mr peptide fragments. The cleavage was completely inhibited by hirudin, indicating that the proteolysis was mediated by thrombin. In a platelet-free system, analyses of thrombin cleavage by immunoblotting using anti-human TPO peptide antibodies revealed that the four major thrombin-cleaved peptide fragments were selectively generated depending on the digestion time. The amino acid sequences of the thrombin-polypeptides were further analyzed, and two major thrombin cleavage sites were determined. One of them was at AR191-T192 in the C-terminal domain of TPO, and thrombin cleaved first at this site. The other site at GR117-T118 in the N-terminal domain was subsequently cleaved by prolonged thrombin digestion. As a result, the biological activity of TPO was modulated. The generation of truncated forms of TPO by thrombin may be a notable event in view of the platelet-related metabolism of TPO.

Amino Acid Sequence↗