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Biomedical subjects

S Sekine

Publications and source records attributed to S Sekine.

At least 19 recordsLinked to original sources

Heterotopic thyroid tissue at the porta hepatis in a fetus with trisomy 18.

Heterotopic thyroid tissue at the porta hepatis in a fetus with trisomy 18 is described. The fetus had an eutopic thyroid gland without any abnormalities. The heterotopic thyroid was found at the porta hepatis and showed histological features similar to the eutopic thyroid. Immunohistochemically, the heterotopic follicles were positive for thyroglobulin, but no calcitonin-positive cells were found. Intra-abdominal heterotopic thyroid is exceedingly rare in locations other than the ovary, and to our knowledge, this is the first report of a fetal case. The present case provides clear evidence that abdominal heterotopic thyroid can occur as a congenital anomaly. Migration abnormality of the median anlage of the thyroid is the most likely histogenesis of heterotopic thyroid at the porta hepatis.

Abnormalities, Multiple↗

Modulation of morphine analgesia in alphaCGRP mutant mice.

A homozygous CGRP-/- mouse line was generated by the targeted disruption of exon 5 in the calcitonin/alphaCGRP gene using homologous recombination. The mutant mice lack alphaCGRP mRNA. Furthermore CGRP immunoreactivity almost completely disappears from the spinal cord and is not at all observed in spinal ganglia and muscle synapses. However, motor end plates were still detected by acetylcholinesterase staining. Antinociceptive behavior tested by the tail flick and hot plate tests did not significantly differ in mutant and wild-type mice, except when challenged by morphine. Paradoxically, morphine analgesia was reduced in mutant mice compared with controls in the tail flick test, but not in the hot plate test. Thus, alphaCGRP differentially modulates opiate pain pathways.

Analgesics, Opioid↗

Selection of cDNAs encoding putative type II membrane proteins on the cell surface from a human full-length cDNA bank.

We have developed a simple method to test whether a hydrophobic segment near the N-terminus of a protein functions as a type II signal anchor (SA) in which the N-terminus faces the cytoplasm. A cDNA fragment containing the putative SA sequence of a target clone was fused in-frame to the 5' end of a cDNA fragment encoding the protease domain of urokinase-type plasminogen activator (u-PA). The resulting fused gene was expressed in COS7 cells. Fibrinolytic activity on the cell surface was measured by placing a fibrin sheet in contact with the transfected COS7 cells after removing the medium. When the cDNA fragment encoded a SA, the fibrin sheet was lysed by the u-PA expressed on the cell surface. The fibrinolytic activity was not detected in the culture medium, suggesting that the u-PA remains on the cell surface anchored via the SA in the membrane without being cleaved by signal peptidase. This fibrin sheet method was successfully applied to select five novel cDNA clones encoding putative type II membrane proteins from a human full-length cDNA bank.

Animals↗

Intrapericardial organized hematoma. A rare complication after open heart surgery.

Intrapericardial organized hematoma, which compresses cardiac chambers late after open heart surgery, is extremely rare. We report a case of intrapericardial organized hematoma in a 63-year-old man who underwent aortic valve replacement 8 years prior, which may have aggravated rheumatic mitral valve regurgitation compressing the mitral valve anulus. Under extracardiopulmonary bypass and cardioplegic heart arrest, we removed the hematoma and replaced the mitral valve with a 27 mm St. Jude Medical valve. There were no bleeding points on the heart and pericardium at operation and no history of blunt chest trauma. The etiology of the hematoma is uncertain.

Aortic Valve Stenosis↗

Severe aspiration pneumonia after surgery for reconstructed gastric tube cancer treated with extracorporeal life support.

A 68-year-old man who had received resection for thoracic esophageal cancer 8 years ago, was operated on for the cancer of the reconstructed gastric tube. On the day of the operation, he accidentally swallowed gastric juice due to an obstruction in the reconstructed gastric tube. He suffered from acute hypoxic respiratory failure which could not be controlled with conventional therapy on postoperative day 1. Therefore, extracorporeal life support was employed at 3.0 L/min. extracorporeal flow for 11 days. Before extracorporeal life support data: PO2/FiO2 = 45, A-aDO2 = 600. During extracorporeal life support, the ventilator setting was pressure control (16 cmH2O) ventilation with a positive end expiratory pressure of 8 cmH2O, respiratory rate of 5 breaths/min., and FiO2 of 0.4. The patient was successfully weaned from extracorporeal life support and extubated on postoperative day 12. After extracorporeal life support data: PO2/FiO2 = 225, A-aDO2 = 465. We report on a successful weaning case from extracorporeal life support and discuss the efficacy these of regarding this patient.

Aged↗

Inhibitory effect of methylene blue-induced photooxidation on intimal thickening of vein graft.

BACKGROUND: We have previously speculated that methylene blue-induced photooxidation of adventitial surface for 5 minutes can completely inhibit the intimal and medial growth of surgically prepared saphenous vein in vitro. In this study, inhibitory effect of methylene blue-induced photooxidation on intimal thickening of vein graft in vivo was investigated. METHODS: Jugular vein grafts were photooxidized in 0.01% methylene blue solution for 5 minutes, and interposed into arterial circulation for 4 weeks in rabbits. Vein grafts were studied by morphometry and immunohistochemistry. RESULTS: The intimal thickening of photooxidized vein grafts were suppressed significantly compared with those in the nonphotooxidized group. Proliferated cell nuclear antigen (PCNA) index (total PCNA-positive cells/total cell count x 100%) of vein graft was significantly higher in the nonphotooxidized group than those in the photooxidized group. CONCLUSIONS: Methylene blue-induced photooxidation is effective in the inhibition of intimal thickening of vein graft interposed in the arterial circulation for 4 weeks in vivo.

Animals↗

The identity determinants required for the discrimination between tRNAGlu and tRNAAsp by glutamyl-tRNA synthetase from Escherichia coli.

We previously elucidated the major determinant set for Escherichia coli tRNAGlu identity (U34, U35, C36, A37, G1*C72, U2*A71, U11*A24, U13*G22**Alpha46, and Delta47) and showed that the set is sufficient to switch the identity of tRNAGln to Glu [Sekine, S., Nureki, O., Sakamoto, K., Niimi, T., Tateno, M., Go, M., Kohno, T., Brisson, A., Lapointe, J. & Yokoyama, S. (1996) J. Mol. Biol. 256, 685-700]. In the present study, we attempted to switch the identity of tRNAAsp, which has a sequence similar to that of tRNAGlu, and consequently possesses many nucleotide residues corresponding to the Glu identity determinants (U35, C36, A37, G1*C72, and U11*A24). A simple transplantation of the rest of the major determinants (U34, U2*A71, U13*G22**Alpha46, and Delta47) to the framework of tRNAAsp did not result in a sufficient switch of the tRNAAsp identity to Glu. To confer an optimal glutamate accepting activity to tRNAAsp, two other elements, C4*G69 in the middle of the acceptor stem and C12*G23**C9 in the augmented D helix, were required. Consistently, the two base pairs, C4*G69 and C12*G23, in tRNAGlu had been shown to exist in the interface with glutamyl-tRNA synthetase (GluRS) by phosphate-group footprinting. We also found the two elements in the framework of tRNAGln, and determined that their contributions successfully changed the identity of tRNAGln to Glu in the previous study. By the identity-determinant set (C4*G69 and C12*G23**C9 in addition to U34, U35, C36, A37, G1*C72, U2*A71, U11*A24, U13*G22**Alpha46, and Delta47) the activity of GluRS was optimized and efficient discrimination from the noncognate tRNAs was achieved.

Acylation↗

Effect of modified nucleotides on Escherichia coli tRNAGlu structure and on its aminoacylation by glutamyl-tRNA synthetase. Predominant and distinct roles of the mnm5 and s2 modifications of U34.

Overproducing Escherichia coli tRNAGlu in its homologous host results in the presence of several distinctly modified forms of this molecule that we name modivariants. The predominant tRNAGlu modivariant in wild-type E. coli contains five modified nucleosides: Psi13, mnm5s2U34, m2A37, T54 and Psi55. Four other overproduced modivariants differ from it by, respectively, either the presence of an additional Psi, or the presence of s2U34, or the lack of A37 methylation combined with either s2U34 or U34. Chemical probing reveals that the anticodon loop of the predominant modivariant is less reactive to the probes than that of the four others. Furthermore, the modivariant with neither mnm5s2U34 nor m2A37 has additional perturbations in the D- and T-arms and in the variable region. The lack of a 2-thio group in nucleoside 34, which is mnm5s2U in the predominant tRNAGlu modivariant, decreases by 520-fold the specificity of E. coli glutamyl-tRNA synthetase for tRNAGlu in the aminoacylation reaction, showing that this thio group is the identity element in the modified wobble nucleotide of E. coli tRNAGlu. The modified nucleosides content also influences the recognition of ATP and glutamate by this enzyme, and in this case also, the predominant modivariant is the one that allows the best specificity for these two substrates. These structural and kinetic properties of tRNAGlu modivariants indicate that the modification system of tRNAGlu optimizes the stability of tRNAGlu and its action as cofactor of the glutamyl-tRNA synthetase for the recognition of glutamate and ATP.

Adenosine Triphosphate↗

Chronic sclerosing sialadenitis of the submandibular gland associated with idiopathic retroperitoneal fibrosis.

We report a case of a 57-year-old man who developed a fibrosclerosing lesion in the submandibular gland and idiopathic retroperitoneal fibrosis (IRF) involving the unilateral periureteral region within a year. Both lesions were resected surgically because of the suspicion of neoplasm. Pathologic examination revealed similar histologic and immunohistochemical features for both lesions, namely, fibrosclerosis with prominent hyalinizing collagen bundles and proliferation of myofibroblastic cells, and a non-neoplastic reactive nature. There was infiltration by lymphocytes with prominent lymph follicles, plasma cells and macrophages. The histologic and immunohistochemical findings suggest that the two lesions were of a similar pathogenesis, which was possibly mediated by macrophages. We think that the present case may be an unusual form of multifocal fibrosclerosis. Although sialolithiasis is thought to be a major pathogenic factor for chronic sclerosing sialadenitis of the submandibular gland, the present case suggests that certain cases might have an etiology similar to IRF.

Biomarkers↗

Isolation and characterization of a cDNA for human mouse, and rat full-length stem cell growth factor, a new member of C-type lectin superfamily.

cDNA encoding stem cell growth factor (SCGF; 245 aa), a novel human growth factor for primitive hematopoietic progenitor cells, has been previously reported (Hiraoka, A., Sugimura, A., Seki, T., Nagasawa, T., Ohta, N., Shimonishi, M., Hagiya, M. and Shimizu, S. Proc. Natl. Acad. Sci. USA 94, 7577-7582, 1997). Here we report the cloning and characterization of a full-length SCGF cDNA. This protein consists of 323, 328 and 328 aa in the human, murine and rat forms, the latter two of which share 85.1% and 83.3% aa identity, and 90.4% and 90.4% aa similarity to the human protein, respectively. Because the newly identified human clone encodes the protein longer by 78 aa than that previously identified, we term the longer clone as hSCGF-alpha and the shorter one as hSCGF-beta. The computer-assisted homology search reveals that SCGF is a new member of the C-type lectin superfamily, and that SCGF shows the greatest homology to tetranectin among the members of the family (27.2-33.7% aa identity and 46.0-53.6% aa similarity). SCGF transcripts are detected in spleen, thymus, appendix, bone marrow and fetal liver. Fluorescent in situ hybridization mapping indicates that the SCGF gene is located on chromosome 19 at position q13.3 for human form and on chromosome 7 at position B3-B5 for murine form, which are close to flk-2/flt3 ligand and interleukin-11 genes of both human and murine species.

Amino Acid Sequence↗

Identification and characterization of a novel SH3-domain binding protein, Sab, which preferentially associates with Bruton's tyrosine kinase (BtK).

Protein interaction cloning method was used to identify a novel molecule, Sab, which binds to the SH3 domain of Bruton's tyrosine kinase (Btk), the deficient cytoplasmic tyrosine kinase in human X-linked agammaglobulinemia and murine X-linked immunodeficiency. Immunoprecipitation using the anti-Sab antibody identified the protein product of the gene as a 70 kDa molecule. While Sab does not have a proline-rich sequence, it was shown to bind to Btk through the commonly conserved structure among SH3 domains. Remarkably, Sab exhibited a high preference for binding to Btk rather than to other cytoplasmic tyrosine kinases, which suggests a unique role of Sab in the Btk signal transduction pathway.

Adaptor Proteins, Signal Transducing↗

Structure of the mouse klotho gene and its two transcripts encoding membrane and secreted protein.

We previously established a novel mouse model for human aging and identified the genetic foundation responsible for it. A defect in expression of a novel gene, termed klotho (kl), leads to a syndrome resembling human aging in mice. The kl gene encodes a single-pass membrane protein whose extracellular domain carries homology to beta-glucosidases. In this report, we present the entire mouse kl gene organization. The mouse kl gene spans about 50 kilobases and consists of five exons. The promoter region lacks a TATA-box and contains four potential binding sites for SP1. We further show that two kl gene transcripts encoding membrane or secreted protein are generated through alternative transcriptional termination. These findings provide fundamental information for further study of the kl gene which may regulate aging in vivo.

Alternative Splicing↗

Hypoproteinemia caused by tricuspid regurgitation: report of a case.

A case of tricuspid regurgitation (TR) complicated by severe hypoproteinemia is presented herein. A 68-year-old man who had undergone coronary artery bypass grafting (CABG) for postinfarction angina suffered repeated inferior myocardial infarction due to obstruction of the proximal right coronary artery, 3 years after which he developed systemic edema. Investigations revealed TR associated with hypoproteinemia; however, treatment consisting of aggressive diuretic therapy and albumin administration proved ineffective. The hypoproteinemia manifested as protein-losing enteropathy clinically, and the tricuspid valve was replaced to eliminate high venous pressure. The serum protein levels became normalized after the operation. Although TR is generally well tolerated in the absence of pulmonary hypertension, surgical management is recommended for patients with severe protein deficiency resistant to medical treatment.

Aged↗

Halofuginone inhibits neointimal formation of cultured rat aorta in a concentration-dependent fashion in vitro.

Halofuginone, an anticoccidial quinoazolinone, can specifically inhibit collagen type alpha1 (I) synthesis and gene expression, and also inhibits cultured smooth muscle cell proliferation. The aim of this study was to investigate the effect of halofuginone on neointimal formation of rat aorta after culture in a concentration-dependent manner in vitro. Thoracic aorta of Wistar rats was removed and manipulated to damage the endothelium under sterile conditions, and culture for 15 days in halofuginone-free or halofuginone-added culture medium (n = 20). Segments of cultured aorta were studied by histologic and immunohistochemical methods. Proliferation of neointimal layers consisting of loose multilayer cellular structure was observed in the halofuginone-free control group after 15 days of rat aorta culture, and neointimal formation was significantly decreased as an increasing concentration of halofuginone was added. As with precultured fresh aorta, no intimal proliferation was observed in the cultured segments of aorta with 500 ng/ml halofuginone added to culture medium. The proliferation of cell nuclear antigen index was significantly higher in the halofuginone-free control group than that in the halofuginone-added groups. The present results suggest that halofuginone can inhibit neointimal formation of rat aorta after culture in a concentration-dependent fashion in vitro.

Animals↗

An effective method for improving hemostasis in aortic root replacement with a composite graft.

Hemostasis during aortic root replacement with a composite graft, consisting of a separate Dacron graft and a valve, was markedly improved by using a supra-annular aortic wall reinforcing suture (SAWR). Between February 1994 and January 1998, 14 consecutive patients (10 men and 4 women) underwent replacement of the aortic valve and ascending aorta with the SAWR technique. No intraoperative bleeding of the aortic proximal suture line or early death occurred in this group. No patient required reoperation for bleeding of anastomoses. Three of 10 patients who underwent elective operations were operated on successfully without blood transfusion. In addition, no clinical complications related to the SAWR technique were observed in the follow-up period. We conclude that the SAWR technique can be performed easily and may be an effective method for improving proximal hemostasis.

Aortic Dissection↗

Cloning of human polyubiquitin cDNAs and a ubiquitin-binding assay involving its in vitro translation product.

During large-scale in vitro translation analysis of a human full-length cDNA bank, we found a clone producing a remarkably smaller translation product than that expected from the open reading frame. The cDNA encodes a polyubiquitin, UbC, composed of nine tandem repeats of the ubiquitin unit. The bank contained twelve UbC cDNAs including four full-length ones. Sequencing analysis of these clones showed that UbC cDNAs can be classified into two types, UbC1 and UbC2, in each of which there are six polymorphic nucleotide variations. The present UbC cDNA was in vitro translated in a rabbit reticulocyte or wheat germ extract to produce a free ubiquitin labeled with [35S]methionine. The labeled ubiquitin could be used as a substrate for thiol ester formation with ubiquitin-activating enzyme E1 or ubiquitin-conjugating enzyme E2.

Amino Acid Sequence↗

Microsatellite instability and frameshift mutations in the Bax gene in hereditary nonpolyposis colorectal carcinoma.

We studied microsatellite instability (MI) and bax gene abnormalities in colorectal carcinomas from 36 patients diagnosed as having hereditary nonpolyposis colorectal cancers (HNPCC) according to the clinical criteria (12 with confirmed HNPCC in group A and 24 at high risk of HNPCC in group B) and from 20 randomly selected patients with other colorectal cancers. MI was examined at 4 dinucleotide microsatellite loci and one mononucleotide locus. Frameshift mutations in the bax gene were detected by polymerase chain reaction-single strand conformation polymorphism analysis. MI was detected in 7 of the 12 patients in group A and 12 of the 24 in group B. Three MI patterns were identified: type 1, MI in both mono- and dinucleotide repeats; type 2, MI only in mononucleotide repeats and type 3, MI only in dinucleotide repeats. Most MI-positive patients in group A showed type 1 MI, whereas in group B, 5 showed type 1, 3 showed type 2 and 4 showed type 3. Frameshift mutations in the bax gene correlated strongly with type 1 and type 2 MI. These results indicate that mutations of different DNA mismatch repair genes may cause several types of MI and result in several different clinical phenotypes of HNPCC. The bax gene may be one of the target genes which play a role in the tumorigenesis of HNPCC.

Colorectal Neoplasms↗