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S Kawabata

Publications and source records attributed to S Kawabata.

At least 55 records · Page 3Linked to original sources

Distribution and molecular characterization of Porphyromonas gingivalis carrying a new type of fimA gene.

Fimbriae of Porphyromonas gingivalis are filamentous appendages on the cell surface and are thought to be one of the virulence factors. The fimA gene encoding the subunit protein of fimbriae, fimbrillin (FimA), was classified into four typeable variants (types I to IV). We previously examined the distribution of P. gingivalis in terms of fimA genotypes in periodontitis patients using a fimA type-specific PCR assay. However, some patients harbored P. gingivalis with untypeable fimA. In this study, we have cloned a new type (type V) of fimA from dental plaque samples. P. gingivalis with type V fimA was isolated from dental plaque of a periodontitis patient, and the isolate was named HNA-99. The deduced amino acid sequences were compared with those of type I P. gingivalis ATCC 33277, type II strain HW24D1, type III strain 6/26, and type IV strain HG564, and the homologies were found to be 45, 44, 43, and 55%, respectively. Southern blot analysis showed that the clinical isolate HNA-99 possessed P. gingivalis-specific genes sod and kgp. However, in terms of serological specificities, type V FimA showed a difference from other types of FimA. In addition, type V P. gingivalis bacteria were detected in 16.4% (12 of 73) of the P. gingivalis-positive patients with periodontitis by PCR assay using specific primers. Thus, a new type of fimA gene is now established, and the fimA genotyping could be useful in determining the disease-associated genotypes of P. gingivalis involved in the development of adult periodontitis.

Adult↗

Effect of l-menthol-ethanol-water system on the systemic absorption of flurbiprofen after repeated topical applications in rabbits.

The effect of the l-menthol-ethanol-water system (MEW system), a skin penetration enhancer, on the systemic absorption of flurbiprofen (FP) after repeated topical applications was investigated. FP (1%) gel containing ethanol (25%) and l-menthol (3%) as penetration enhancers was applied to rabbit dorsal skin and the in vivo absorption rate of FP was compared with the in vitro penetration rate through excised skin. In vivo absorption rate of FP was initially high and decreased with time to a value approximately equal to the in vitro rate. The remaining FP in the gel 6 h after the application was 60% of the initial loading and the systemic bioavailability over the 6 h application was about 10%, suggesting that the rest (30%) had accumulated in the skin tissues. The gel was applied for 6 h on the same site or on a new site after the first 6 h-application to learn the effect of repeated applications on FP absorption. The maximum FP concentration after the second application on the virgin skin was slightly higher than that after the first application, as expected in a typical pharmacokinetic process. In contrast, the same site application induced remarkably lower plasma concentration and area under the curve (AUC). A drug-free gel was also utilized to evaluate the effects of the enhancer system. Pretreatment of the drug-free gel on the same site also decreased the FP absorption, whereas post-treatment increased the plasma level of FP, in spite of the removal of the drug gel. These phenomena could be explained by ethanol in the MEW system acting a local irritant and a drug carrier.

Administration, Topical↗

Surgical strategy for meningioma extension into the optic canal.

Neuroimaging of the extension of meningioma into the optic canal was evaluated for planning the surgical strategy. Intracanalicular extension and localization were retrospectively analyzed in 13 patients with frontal base meningioma near the optic canal, based on the findings of visual field defects, magnetic resonance (MR) imaging, and surgical observations. MR imaging confirmed intracanalicular localization in one of three patients with tumors extending into the optic canals, and indicated the tumor in the others. The visual field defect did not precisely correspond to the tumor localization. Unroofing of the optic canal was performed in four patients and no adverse effects were observed. The interhemispheric approach was employed for tumors localized medially in the canal, and the pterional approach for tumors localized laterally. MR imaging is useful to evaluate the intracanalicular extension, but aggressive confirmation during surgery is essential. Tailored unroofing of the optic canal and removal of the intracanalicular tumor can be performed with few adverse effects and results in good tumor control.

Adult↗

A horseshoe crab receptor structurally related to Drosophila Toll.

Innate immunity against microbial pathogens relies on the pattern recognition of cell wall components on invading microbes. Recent evidence has shown that a mammalian Toll-like receptor (TLR) is activated by bacterial lipopolysaccharides (LPS). The innate immunity in invertebrates is also triggered by LPS, as seen in the hemolymph coagulation in horseshoe crab. We report the cloning of a TLR from the Japanese horseshoe crab Tachypleus tridentatus. A cDNA coding for Tachypleus Toll was isolated from a hemocyte cDNA library and the open reading frame codes for a proprotein including a signal sequence. Like Drosophila Toll, Tachypleus Toll is a type I transmembrane protein with an extracellular domain consisting of two leucine-rich repeats flanked by two cystein-rich clusters and a cytoplasmic domain exhibiting striking similarity with the cytoplasmic domain of interleukin-1 receptor. Tachypleus Toll is most similar to Drosophila Toll in the domain architecture and the overall length.

Animals↗

Purification, characterization, and amino acid sequence of an embryonic lectin in perivitelline fluid of the horseshoe crab.

Hemagglutinating activity in perivitelline fluid of the horseshoe crab embryo dramatically increases during the third and fourth molt of the embryo. A 27-kDa lectin, which we named tachylectin-P (TL-P), was newly identified in perivitelline fluid of the horseshoe crab Tachypleus tridentatus. TL-P preferentially agglutinated human A-type erythrocytes, and the activity was inhibited by N-acetyl group-containing monosaccharides. The amino acid sequence analysis indicated that TL-P is almost structurally the same as a hemocyte-derived lectin with no hemagglutinating activity, tachylectin-1 (TL-1), and that 218 out of 221 amino acid residues in total were conserved between the two lectins. Despite the high sequence similarity, biological and biochemical characteristics of TL-P differed from those of TL-1: (i) unlike TL-1, TL-P agglutinates several animal-derived erythrocytes; (ii) unlike TL-1, TL-P has no significant affinity for bacterial lipopolysaccharides or antibacterial activity; (iii) Based on apparent molecular masses determined by gel filtration, TL-P forms a dimer in solution, while TL-1 is present as a monomer; (iv) and TL-P interacts with endogenous proteins of 13 and 14 kDa present in the perivitelline fluid; however, neither has any affinity for TL-1. We propose that TL-P may have an important role in completing embryonic development by interacting with endogenous glycoproteins or N-acetylhexosamines.

Amino Acid Sequence↗

Horseshoe crab hemocyte-derived antimicrobial polypeptides, tachystatins, with sequence similarity to spider neurotoxins.

Antimicrobial peptides, named tachystatins A, B, and C, were identified from hemocytes of the horseshoe crab Tachypleus tridentatus. Tachystatins exhibited a broad spectrum of antimicrobial activity against Gram-negative and Gram-positive bacteria and fungi. Of these tachystatins, tachystatin C was most effective. Tachystatin A is homologous to tachystatin B, but tachystatin C has no significant sequence similarity to tachystatins A and B. Tachystatins A and B showed sequence similarity to omega-agatoxin-IVA of funnel web spider venom, a potent blocker of voltage-dependent calcium channels. However, they exhibited no blocking activity of the P-type calcium channel in rat Purkinje cells. Tachystatin C also showed sequence similarity to several insecticidal neurotoxins of spider venoms. Tachystatins A, B, and C bound significantly to chitin. A causal relationship was observed between chitin binding activity and antifungal activity. Tachystatins caused morphological changes against a budding yeast, and tachystatin C had a strong cell lysis activity. The septum between mother cell and bud, a chitin-rich region, was stained by fluorescence-labeled tachystatin C, suggesting that the primary recognizing substance on the cell wall is chitin. As horseshoe crab is a close relative of the spider, tachystatins and spider neurotoxins may have evolved from a common ancestral peptide, with adaptive functions.

Amino Acid Sequence↗

Horseshoe crab acetyl group-recognizing lectins involved in innate immunity are structurally related to fibrinogen.

We have characterized and cloned newly isolated lectins from hemolymph plasma of the horseshoe crab Tachypleus tridentatus, which we named tachylectins 5A and 5B (TLs-5). TLs-5 agglutinated all types of human erythrocytes and Gram-positive and Gram-negative bacteria. TLs-5 specifically recognize acetyl group-containing substances including noncarbohydrates; the acetyl group is required and is sufficient for recognition. TLs-5 enhanced the antimicrobial activity of a horseshoe crab-derived big defensin. cDNA sequences of TLs-5 indicated that they consist of a short N-terminal Cys-containing segment and a C-terminal fibrinogen-like domain with the highest sequence identity (51%) to that of mammalian ficolins. TLs-5, however, lack the collagenous domain found in a kind of "bouquet arrangement" of ficolins and collectins. Electron microscopy revealed that TLs-5 form two- to four-bladed propeller structures. The horseshoe crab is equipped with a unique functional homologue of vertebrate fibrinogen, coagulogen, as the target protein of the clotting cascade. Our observations clearly show that the horseshoe crab has fibrinogen-related molecules in hemolymph plasma and that they function as nonself-recognizing lectins. An ancestor of fibrinogen may have functioned as a nonself-recognizing protein.

ABO Blood-Group System↗

Tachylectin-2: crystal structure of a specific GlcNAc/GalNAc-binding lectin involved in the innate immunity host defense of the Japanese horseshoe crab Tachypleus tridentatus.

Tachylectin-2, isolated from large granules of the hemocytes of the Japanese horseshoe crab (Tachypleus tridentatus), is a 236 amino acid protein belonging to the lectins. It binds specifically to N-acetylglucosamine and N-acetylgalactosamine and is a part of the innate immunity host defense system of the horseshoe crab. The X-ray structure of tachylectin-2 was solved at 2.0 A resolution by the multiple isomorphous replacement method and this molecular model was employed to solve the X-ray structure of the complex with N-acetylglucosamine. Tachylectin-2 is the first protein displaying a five-bladed beta-propeller structure. Five four-stranded antiparallel beta-sheets of W-like topology are arranged around a central water-filled tunnel, with the water molecules arranged as a pentagonal dodecahedron. Tachylectin-2 exhibits five virtually identical binding sites, one in each beta-sheet. The binding sites are located between adjacent beta-sheets and are made by a large loop between the outermost strands of the beta-sheets and the connecting segment from the previous beta-sheet. The high number of five binding sites within the single polypeptide chain strongly suggests the recognition of carbohydrate surface structures of pathogens with a fairly high ligand density. Thus, tachylectin-2 employs strict specificity for certain N-acetyl sugars as well as the surface ligand density for self/non-self recognition.

Acetylgalactosamine↗

A newly identified horseshoe crab lectin with specificity for blood group A antigen recognizes specific O-antigens of bacterial lipopolysaccharides.

A 14-kDa lectin, named tachylectin-3, was newly identified from hemocytes of the Japanese horseshoe crab, Tachypleus tridentatus. This lectin exhibited hemagglutinating activity against human A-type erythrocytes, but not against the B- and O-types of erythrocytes and animal erythrocytes, including those of sheep, rabbit, horse, and bovine. The hemagglutinating activity of tachylectin-3 was equivalent to that of a previously identified lectin, named tachylectin-2, with affinity for N-acetyl-D-glucosamine or N-acetyl-D-galactosamine. However, the activity of tachylectin-3 was not inhibited by these two N-acetylhexosamines at 100 mM but was inhibited by a blood group A-pentasaccharide at a minimum inhibitory concentration of 0.16 mM. Furthermore, the hemagglutinating activity was strongly inhibited by bacterial S-type lipopolysaccharides (LPSs) from Gram-negative bacteria but not by R-type LPSs lacking O-antigens. One of the most effective S-type LPSs was from Escherichia coli O111:B4, with a minimum inhibitory concentration of 6 ng/ml. These data suggest that tachylectin-3 specifically recognizes Gram-negative bacteria through the unique structural units of O-antigens. Ultracentrifugation analysis revealed that tachylectin-3 is present in dimer in solution. A cDNA coding for tachylectin-3 was isolated from a hemocyte cDNA library. Tachylectin-3 consisted of two repeating sequences, each with a partial sequence similarity to rinderpest virus neuraminidase. Tachylectin-3 and three previously isolated types of tachylectins were all predominantly expressed in hemocytes and released from hemocytes in response to external stimuli. These lectins present at injured sites suggest that they probably serve synergistically to accomplish an effective host defense against invading microbes.

ABO Blood-Group System↗

Capsular hyaluronic acid of group A streptococci hampers their invasion into human pharyngeal epithelial cells.

Group A streptococci (GAS) cause various diseases, from uncomplicated noninvasive, to severe invasive infections. Capsular hyaluronic acid (HA) is known to resist phagocytosis, however, interaction between HA and epithelial cells have not been clearly understood. In this study, both HA-producing wild strains and HA-nonproducing mutants were employed to examine their invasiveness into confluent cultures of HEp-2, a nonphagocytic human epithelial cell line. Invasion of HEp-2 cells by GAS strains increased over time. The hasA gene encoding hyaluronate synthase of GAS strains was inactivated by allelic replacement. It was found that hasA-inactivated mutants were internalized into HEp-2 cells more efficiently than their parent strains under various conditions in terms of incubation time and inoculum size. Taken together, these findings indicate that GAS can be internalized into HEp-2 cells with considerably high frequencies and that the presence of HA of GAS decreased the invasion efficiency.

Animals↗

Regulated expression of the Shiga toxin B gene induces apoptosis in mammalian fibroblastic cells.

Shiga toxins (Stxs) produced by enterohaemorrhagic Escherichia coli may induce colonic ulceration, bloody diarrhoea and acute renal failure. The A subunit (StxA) is known to inhibit protein synthesis, whereas the B subunits (StxB) bind to Gb3 on the cell surface. However, the mechanisms by which Stxs kill target cells remain unclear. Stx1A or Stx1B genes were introduced into pcDNA3.1 vectors and transfected into NIH3T3 and HeLa cells. The Stx1B gene-transfected cells became apoptotic with accompanying DNA fragmentation, whereas the Stx1A gene-transfected cells were found to be necrotic and no DNA fragmentation occurred. The HeLa/C4 cells integrated with the Stx1B gene with a tetracycline-inducible promoter eventually produced cytoplasmic Stx1B, leading to DNA fragmentation on the addition of doxycycline. These apoptotic changes were abrogated by pretreatment with Z-VAD-fmk. These results suggest that the transfected Stx1B gene induces apoptosis by activating the caspase cascade after Stx1B expression in the cytoplasm.

3T3 Cells↗

Functional and structural diversities of C-reactive proteins present in horseshoe crab hemolymph plasma.

Limulin, a sialic-acid-binding and phosphorylethanolamine-binding hemagglutinin in the hemolymph plasma of the American horseshoe crab (Limulus polyphemus), is a hemolytic C-reactive protein [Armstrong, P.B., Swarnakar, S., Srimal, S., Misquith, S., Hahn, E.A., Aimes, R. T. & Quigley, J.P. (1996) J. Biol. Chem. 271, 14717-14721]. We have now identified three types of C-reactive protein in the plasma of the Japanese horseshoe crab (Tachypleus tridentatus), based on different affinities against fetuin-agarose and phosphorylethanolamine-agarose determined by quantitative precipitin assays using fetuin and an artificial phosphorylethanolamine-protein conjugate. Partial amino acid sequences of the isolated C-reactive proteins identified homologous proteins which were named Tachypleus tridentatus CRP-1 (tCRP-1), tCRP-2 and tCRP-3, each of which possibly constitute isoprotein mixtures. tCRP-2 and tCRP-3, but not tCRP-1, agglutinated mammalian erythrocytes. tCRP-1, the most abundant C-reative protein in the plasma, exhibited the highest affinity to the phosphorylethanolamine-protein conjugate but lacked both sialic-acid-binding and hemolytic activities. tCRP-2 bound to both fetuin-agarose and phosphorylethanolamine-agarose, and exhibited Ca2+-dependent hemolytic and sialic-acid-binding activities, suggestive of limulin-like properties. Furthermore, tCRP-2 exhibited a higher affinity to colominic acid, a bacterial polysialic acid. By contrast, tCRP-3 shows stronger hemolytic, sialic-acid-binding and hemagglutinating activities than tCRP-2. tCRP-3 has no affinity to phosphorylethanolamine-agarose, phosphorylethanolamine-protein conjugate and colominic acid. This suggests tCRP-3 is a novel hemolytic C-reactive protein lacking a common characteristic of phosphorylethanolamine-agarose binding affinity. Twenty-two clones of tCRPs with different deduced amino acid sequences were obtained by PCR using oligonucleotide primers based on the N-terminal and C-terminal sequences of tCRPs and with templates including genomic DNA and cDNA of hemocytes or hepatopancreas derived from one individual. The translation products of the tCRP clones possess high molecular diversity which falls into three related groups, consistent with classification based on their biological activities. Only tCRP-3 contained a unique hydrophobic nonapeptide sequence that appears in the transmembrane domain of a major histocompatibility complex class I heavy chain of rainbow trout, suggesting the importance of the hydrophobic patch to the hemolytic activity of tCRP-3. The structural and functional diversities of tCRPs provide a good model for studying the properties of innate immunity in invertebrates, which survive without the benefit of acquired immunity.

Amino Acid Sequence↗

Cognitive therapy for a major depressive episode in residual schizophrenia.

The present paper describes cognitive approaches to the treatment of a major depressive episode in a patient with residual schizophrenia. The goal of therapy was to increase and stabilize the patient's physical activity through decreasing dysfunctional cognition pertinent to inertia. A therapeutic strategy of 'scheduling activities' was first selected, but to no effect. The vicious circle of alternating excessive activity and total inertia remained unchanged. Based on a revised cognitive case conceptualization, a second strategy, 'scheduling inertia', was then introduced, in which the patient was asked to stay in bed or take a rest for planned periods of time every day. This intervention helped the patient to counteract her perfectionist beliefs. The results suggest that 'scheduling inertia' may be a useful strategy for improving inactivity in a major depressive episode during the residual phase of schizophrenia.

Activities of Daily Living↗

Simulation of therapeutic parent artery occlusion for basilar head aneurysms. Hemodynamic effect of occlusion sites and diameters of collateral arteries.

We simulated parent artery occlusion therapy for basilar head aneurysms to elucidate the hemodynamic changes induced by different occlusion sites and diameters of the posterior communicating arteries (PCom) as collateral pathways. A vascular model of the vertebrobasilar system with a basilar head aneurysm was constructed. Four types of occlusion were simulated: Basilar artery occlusion distal to (Type A), between (Type B) and proximal to (Type C) the superior cerebellar arteries, and bilateral vertebral artery occlusion (Type D). Glycerol solution was perfused into the model, and the half-life of the dye injected into the aneurysm was calculated and regarded as an index of stagnant flow in the aneurysm. The half-life was increased significantly and nonlinearly after parent artery occlusion, depending on the occlusion site and the ratio of two PCom diameters (diameter ratio). Intra-aneurysmal stagnation developed markedly in Type A and Type B in the diameter ratio higher than 0.70 and considerably in Type C in the ratio higher than 0.80. Additional P1 occlusion of the posterior cerebral artery enhanced the stagnation in Type A and B. Since the results are consistent with the published clinical data, the simulation study will be useful for speculating the efficacy of the therapeutic occlusion.

Basilar Artery↗

Paraclinoid aneurysms of the internal carotid artery: hydraulic simulation study on their locations and shape of the carotid siphon.

Hemodynamics of paraclinoid aneurysms were investigated focusing on the effects of their locations and shape of the carotid siphon. A transparent silicon model of the carotid siphon was constructed and a model aneurysm was attached to the outside of the curvature at three different sites. Glycerol solution was perfused into the model, and the half-life of the dye injected into the aneurysm was calculated as an index of the stagnant flow. Values of half-life changed significantly depending on the aneurysmal location and the siphon angle. When the siphon angle was 0 degree where C2 and C4 segments were parallel to each other, the half-life value was the lowest in the C2-C3 junction aneurysm, highest in the C3 segment aneurysm and intermediate in the C2 segment aneurysm. While the C2-C3 junction aneurysm maintained low values regardless of the angle, the C3 segment aneurysm values decreased and C2 segment aneurysm values increased with increases in the angle. These changes of half-life occur because the point at which the faster moving fluid component strikes the curved wall changes according to the siphon angle. These results are considered useful to determine the surgical indications, treatment modalities and post-surgical management of the aneurysms.

Adult↗

Mucosal Th1- versus Th2-type responses for antibody- or cell-mediated immunity to simian immunodeficiency virus in rhesus macaques.

Simian immunodeficiency virus (SIV)-specific B cell responses and the Th1- or Th2-type profiles of cytokine expression were determined for rhesus macaques immunized with SIV antigens via the iliac lymph nodes (by use of a targeted lymph node [TLN] procedure) or orally with SIV p55gag plus cholera toxin (CT) as a mucosal adjuvant. Analysis of CD4+ T cells purified from SIV-stimulated peripheral blood mononuclear cells of immunized macaques revealed that Th2 cytokine production gradually increased after the second and third TLN immunization. Analysis of SIV-specific B cell responses revealed that peak SIV-specific IgA B cell responses followed the third TLN immunization and occurred during peak Th2-type T cell responses. Oral immunization of macaques with p55gag plus CT induced interferon-gamma-secreting Th1-type and select Th2-type cytokine-producing CD4+ T helper cells, which most likely accounted for the induction of p55-specific systemic IgG and mucosal IgA responses.

Animals↗