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Biomedical subjects

K Kasai

Publications and source records attributed to K Kasai.

At least 37 records · Page 2Linked to original sources

Brain lateralization for mismatch response to across- and within-category change of vowels.

Differences in hemispheric predominance between across- and within-category change perception of vowels were assessed using a whole-head magnetoencephalography. The magnetic mismatch responses (MMNm) to pure-tone and vowel within-category changes were significantly predominant in the right hemisphere; on the other hand, vowel across-category MMNm did not differ in power between hemispheres. The results suggest that both hemispheres are symmetrically activated in the preattentive across-category change perception of vowels, while the within-category change of a vowel is analyzed as the change in physical features of the stimuli, thus predominantly activating the right hemisphere. Thus, the relative contribution of the left auditory cortex in the preattentive speech processing may occur only at the level of perception of the vowel across-category change.

Acoustic Stimulation↗

Impaired suppression of processing in schizophrenic patients suggested by ERPs obtained in a selective attention task.

In the present study, we focused on the ability of suppression of processing in schizophrenic patients, using event-related potentials (ERPs) recorded during a selective attention task. During the task, subjects were required to focus on one ear, counting deviant stimuli, those deviating in duration from a sequence of standard stimuli. We compared amplitude data of two positive components differing in latency elicited by standard stimuli, which reflect suppression of stimulus processing, between schizophrenic patients and normal controls. Significant between-group differences were obtained specifically in the right ear attended condition, suggesting impaired suppression of processing in schizophrenics mediated in the left hemisphere.

Adult↗

Glycated serum albumin-induced vascular smooth muscle cell proliferation through activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway by protein kinase C.

Proliferation of vascular smooth muscle cells (VSMC) contributes to the pathogenesis of atherosclerosis, and glycated serum albumin (GSA, Amadori adduct of albumin) might be a mitogen for VSMC proliferation, which may further be associated with diabetic vascular complications. In this study, we investigated the involvement of mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK), and protein kinase C (PKC), in GSA-stimulated mitogenesis, as well as the functional relationship between these factors. VSMC stimulation with GSA resulted in a marked activation of ERK. The MAPK kinase (MEK) inhibitor, PD98059, blocked GSA-stimulated MAPK activation and resulted in an inhibition of GSA-stimulated VSMC proliferation. GSA also increased PKC activity in VSMC in a dose-dependent manner. The inhibition of PKC by the PKC inhibitors, GF109203X and Rottlerin (PKCdelta specific inhibitor), as well as PKC downregulation by phorbol 12-myristate 13-acetate (PMA), inhibited GSA-induced cell proliferation and blocked ERK activation. This indicates that phorbol ester-sensitive PKC isoforms including PKCdelta are involved in MAPK activation. Thus, we show that the MAPK cascade is required for GSA-induced proliferation, and that phorbol ester-sensitive PKC isoforms contribute to cell activation and proliferation in GSA-stimulated VSMC.

Animals↗

Prevention of evaporation of small-volume sample solutions for capillary electrophoresis using a mineral-oil overlay.

The suppression of evaporation of water from small volumes of sample solutions or reagents for capillary electrophoresis by the use of a mineral-oil overlay was investigated in affinophoresis applications, in which the affinity constant of a mutant protein of recombinant human galectin-1 to a lactose affinophore, a triply negative charged ion having a lactoside as an affinity ligand, was determined. When an injection was carried out from a minimum of 20 microL of an aqueous solution beneath the oil overlay, no oil contamination inside the capillary was observed, provided the capillary was cleanly cut so that the end was flat, and the polyimide coating had been removed for a distance of about 2 mm from the end. Affinophoresis was carried out using 20 microL of an affinophore solution covered with an oil overlay. The abnormalities in the electropherograms as the result of the evaporation of the water from the solution during storage prior to use in an automatic operation of a capillary electrophoresis instrument were suppressed, with respect to the formation of a base line gap, an increase in the detection time of a marker ion and an increase in the initial current. A solution in a vial could be used repeatedly for a longer period of time when overlaid with mineral oil than in the absence of an overlay. The use of a mineral-oil overlay is a simple but very efficient technique for solving the problem of the evaporation of water from small volumes of aqueous solutions for use in capillary electrophoresis.

Electrophoresis, Capillary↗

Glycome project: concept, strategy and preliminary application to Caenorhabditis elegans.

Glycans play a central role as potential mediators between complex cell societies, because all living organisms consist of cells covered with diverse carbohydrate chains reflecting various cell types and states. However, we have no idea how diverse these carbohydrate chains actually are. The main purpose of this article is to persuade life scientists to realize the fundamental importance of taking some action by becoming involved in "glycomics". "Glycome" is a term meaning the whole set of glycans produced by individual organisms, as the third bioinformative macromolecules to be elucidated next to the genome and proteome. Here a basic strategy is presented. The essence of the project includes the following: (a) glycopeptides, but not glycans released from their core proteins, are targeted for linkage to genome databases; (b) Caenorhabditis elegans is used as the first model organism for this project, since its genome project has already been completed; (c) four essential attributes are adopted to characterize each glycopeptide: (i) cosmid identification number (ID), (ii) molecular weight (M(r)), (iii) retention (Rs) of pyridylaminated (PA) oligosaccharides in 2-D mapping, and (iv) dissociation constants (Kd's) of PA-oligosaccharides for a set of lectins. Thus, the obtained ID, M(r), R and Kd's construct the glycome database, which will be open as the previous genome and proteome databases. For the project to proceed the "glyco-catch" method is proposed, where a group of target glycopeptides are captured by means of lectin-affinity chromatography after protease digestion. Already glycopeptides from asialofetuin and ovalbumin were successfully captured by galectin-agarose and Con A-agarose, respectively. Further, to examine the practical validity of the method, we extracted membrane proteins from C. elegans with 1% Triton X-100, and isolated specific glycopeptides by use of the same galectin column. One of the glycopeptides was successfully identified in the C. elegans genome database. Finally, for determination of Kd between glycopeptides and lectins, a recently reinforced frontal affinity chromatography (FAC) is proposed as an alternative to define glycan structures in place of determining every covalent structure.

Animals↗

Correlation between buccolingual inclination and wear of mandibular teeth in ancient and modern Japanese.

This study compared the correlation between the buccolingual inclination and the wear score of the mandibular teeth in Jomon (one of the ancestors of modern Japanese who lived from 5000 to 300 BC) and modern Japanese. Data were obtained from 40 modern male Japanese and 39 male Jomon skulls housed in the University Museum of the University of Tokyo and National Science Museum, Tokyo. For each specimen, three computed tomograms of coronal sections of the mandibular body were available. The second premolar, first molar and second molar of Jomon stood more vertically than those of modern Japanese. There were significant correlations between tooth wear and the buccolingual inclination of mandibular molars. When ancient and modern populations were compared, tooth wear was significantly greater and buccolingual tooth inclination was more vertical in the ancient people, who probably applied stronger masticatory forces than modern people. The results suggest that the mandibular molar orientation is more buccal and there is greater tooth wear in the first and second molars of people with strong masticatory function.

Bicuspid↗

Novel Y283C mutation of the A subunit for coagulation factor XIII: molecular modelling predicts its impaired protein folding and dimer formation.

In an Italian patient with severe factor XIII deficiency, a novel mutation, Y283C (TAT to TGT), was identified heterozygously by nucleotide sequencing analysis in exon VII of the gene for the A subunit. The presence of this mutation was confirmed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in the proband and his brother. Molecular modelling predicts that the mutant molecule would be misfolded. It is probable that the impaired folding of the mutant Y283C A subunit led to its instability, which is at least in part responsible for the factor XIII deficiency of this patient.

Child↗

Expression of endogenous galectin-1 and galectin-3 in intrahepatic cholangiocarcinoma.

Galectins, a family of beta-galactoside-binding animal lectins, might be involved in tumor progression. In this study, the expression patterns of galectin-1 and -3 were examined immunohistochemically in intrahepatic cholangiocarcinoma (ICC), with emphasis on its development and progression as well as its histopathologic features, by use of samples of normal intrahepatic bile duct (n = 20), biliary epithelial dysplasia (n = 15), ICC (n = 40), and a cholangiocarcinoma cell line, CCKS1. In normal intrahepatic bile ducts, galectin-3 was constitutively but weakly expressed, whereas galectin-1 was not expressed. In hepatolithiasis, biliary epithelial dysplasia was strongly positive for galectin-3 but negative for galectin-1. Galectin-3 was frequently and strongly expressed in the cytoplasm of well-differentiated ICCs, and its expression was significantly decreased and less intense or even absent in poorly differentiated ICCs. Galectin-1 was expressed in carcinoma cells in ICC, and its incidence and extent were correlated with histologic dedifferentiation of ICC. Proliferative cell nuclear antigen (PCNA) labeling index (LI) was higher in ICC cases positive for galectin-1 than in those that were negative. Galectin-1 was strongly expressed in cancerous stroma of ICC, and this stromal expression was related to histologic dedifferentiation of ICC. In the carcinoma cell line CCKS1, galectin-1 and -3 were expressed in the cytoplasm of carcinoma cells, and galectin-1 was additionally detected in the culture medium. These results suggest that galectin-1 was newly expressed on carcinoma cells of ICC, and its overexpression seems to be associated with neoplastic progression and proliferative activities, and the expression of galectin-1 in cancerous stroma may also be related to the progression of ICC. Galectin-3 expression in epithelial cells is up-regulated in the preneoplastic and early neoplastic stages of ICC, although galectin-3 tends to disappear at later stages of ICC. HUM PATHOL 32:302-310.

Adult↗

Effects of risperidone on event-related potentials in schizophrenic patients.

In order to examine the effects of risperidone on cognitive impairment in schizophrenia, event-related potentials (ERPs) were recorded before and after switching from conventional neuroleptics to risperidone in schizophrenic patients. ERPs were recorded during two auditory discrimination tasks (an oddball task and a distraction task) in 10 medicated schizophrenic patients during conventional neuroleptic and risperidone treatments. The amplitudes and latencies of N 100 and P300 component were measured in ERPs for target stimuli in the oddball task and in ERPs for target and novel stimuli in the distraction task. Although N 100 amplitude and latency and P 300 amplitude did not change significantly after switching the drug compared to that during conventional neuroleptic treatment, P 300 latency for target stimuli shortened significantly during risperidone treatment in both tasks, accompanied by the shortening of the reaction time in the distraction task. The P 300 latency change did not correlate with the change of the severity of psychopathology. These findings suggest that risperidone may speed the information processing in schizophrenic patients, contributing to the improvement of cognitive functions.

Adult↗

Lymphoepithelioma-like carcinoma of rectum: possible relation with EBV.

Lymphoepithelioma-like carcinoma (LEC) of the colon is very rare. Here we report a case of LEC originating in the rectum that was closely associated with Epstein-Barr virus (EBV) infection. The histologic and immunohistologic features, namely, poorly differentiated adenocarcinoma with lymphoid stroma, showed this tumor to be an LEC. The EBV genome was detected by PCR using DNA obtained from tumor tissue sections. Immunohistochemically, EBV-determined nuclear antigen 2 was detected in the tumor cells, and in situ hybridization using EBV-encoded small RNAs probe showed positive labeling in some tumor cells together with a few stromal lymphoid cells. There are some reports of LEC cases that originated in the colon; however, a relation with EBV was not demonstrated. We report here a case of LEC of the rectum demonstrating a possible relation with EBV.

Aged↗

Relationships among facial type, buccolingual molar inclination, and cortical bone thickness of the mandible.

The purpose of this study was to evaluate the relationships between different facial types, and both the buccolingual molar inclination and cortical bone thickness of the mandible. The material consisted of 31 dry skulls of modern Japanese males from the Museum of the University of Tokyo. They all demonstrated normal occlusion with minimal dental discrepancy, and without crossbite or facial asymmetry. The buccolingual inclination of the second molar (M2) in the long-faced subjects was significantly smaller than the same dimension in the average- and short-faced subjects. It was found that the teeth of long-faced subjects were more lingually inclined than those of the short-faced subjects. The cortical bone thickness of the first molar (M1) and M2 sections was thicker in short-faced subjects than in average- and long-faced subjects. The results of this study provide evidence that a significant, but complex relationship exists between structures of the mandibular body and facial types. The morphological features that relate to masticatory function and facial types are associated with the cortical bone thickness of the mandibular body, and the buccolingual inclination of the first and second molars.

Adolescent↗

Construction of an in vivo nonsense readthrough assay system and functional analysis of ribosomal proteins S12, S4, and S5 in Bacillus subtilis.

To investigate the function of ribosomal proteins and translational factors in Bacillus subtilis, we developed an in vivo assay system to measure the level of nonsense readthrough by utilizing the LacZ-LacI system. Using the in vivo nonsense readthrough assay system which we developed, together with an in vitro poly(U)-directed cell-free translation assay system, we compared the processibility and translational accuracy of mutant ribosomes with those of the wild-type ribosome. Like Escherichia coli mutants, most S12 mutants exhibited lower frequencies of both UGA readthrough and missense error; the only exception was a mutant (in which Lys-56 was changed to Arg) which exhibited a threefold-higher frequency of readthrough than the wild-type strain. We also isolated several ribosomal ambiguity (ram) mutants from an S12 mutant. These ram mutants and the S12 mutant mentioned above (in which Lys-56 was changed to Arg) exhibited higher UGA readthrough levels. Thus, the mutation which altered Lys-56 to Arg resulted in a ram phenotype in B. subtilis. The efficacy of our in vivo nonsense readthrough assay system was demonstrated in our investigation of the function of ribosomal proteins and translational factors.

Amino Acid Substitution↗

4-hydroxynonenal prevents NO production in vascular smooth muscle cells by inhibiting nuclear factor-kappaB-dependent transcriptional activation of inducible NO synthase.

The role of lipid peroxidation products in atherogenesis was studied. We investigated whether 4-hydroxy-2-nonenal (HNE) modulates activation of the nuclear factor (NF)-kappaB system or alters expression of the NF-kappaB target gene product, inducible NO synthase (iNOS), in vascular smooth muscle cells (VSMCs) stimulated by lipopolysaccharide (LPS) in combination with interferon (IFN)-gamma (LPS/IFN). NO production induced by LPS/IFN was dose-dependently inhibited by HNE. NF-kappaB activation by LPS/IFN was inhibited by HNE in a dose-dependent manner. HNE significantly decreased LPS/IFN-stimulated proteolysis of IkappaB-alpha. iNOS promoter activity stimulated by LPS/IFN was also decreased by HNE dose-dependently. The treatment of VSMCs with LPS/IFN strongly stimulated iNOS mRNA and protein expression. The LPS/IFN-induced increases in iNOS mRNA and protein levels were dose-dependently decreased by HNE. Our data suggest that treatment with HNE blocks signaling events required for IkappaB-alpha degradation, thereby preventing NF-kappaB activation. Inhibition of NF-kappaB-regulated gene expression, especially modulation of NO production, may contribute to atherogenesis.

Aldehydes↗

Roles of the cytoplasmic and transmembrane domains of syntaxins in intracellular localization and trafficking.

Syntaxins are target-soluble N-ethylmaleimide-sensitive factor-attachment protein receptors (t-SNAREs) involved in docking and fusion of vesicles in exocytosis and endocytosis. Many syntaxin isoforms have been isolated, and each one displays a distinct intracellular localization pattern. However, the signals that drive the specific intracellular localization of syntaxins are poorly understood. In this study, we used indirect immunofluorescence analysis to examine the localization of syntaxin chimeras, each containing a syntaxin transmembrane domain fused to a cytoplasmic domain derived from a different syntaxin. We show that the cytoplasmic domains of syntaxins 5, 6, 7 and 8 have important effects on intracellular localization. We also demonstrate that the transmembrane domain of syntaxin 5 is sufficient to localize the chimera to the compartment expected for wild-type syntaxin 5. Additionally, we find that syntaxins 6, 7 and 8, but not syntaxin 5, are present at the plasma membrane, and that these syntaxins cycle through the plasma membrane by virtue of their cytoplasmic domains. Finally, we find that di-leucine-based motifs in the cytoplasmic domains of syntaxins 7 and 8 are necessary for their intracellular localization and trafficking via distinct transport pathways. Combined, these results suggest that both the cytoplasmic and the transmembrane domains play important roles in intracellular localization and trafficking of syntaxins.

Amino Acid Sequence↗

Effects of substitution of conserved amino acid residues on the sugar-binding property of the tandem-repeat 32-kDa galectin of the nematode Caenorhabditis elegans.

The 32-kDa galectin (LEC-1) of the nematode Caenorhabditis elegans (C elegans) is composed of two tandemly repeated homologous sequences, each containing a carbohydrate-recognition domain (CRD). Using the polymerase chain reaction (PCR) with LEC-1 cDNA as a template and "megaprimers", we performed site-directed mutagenesis to substitute conserved amino acid residues in these domains. The resultant mutated LEC-1s were produced in E. coli, and their binding abilities were estimated by affinity chromatography. When one of the conserved amino acid residues in the first lectin domain was substituted, the binding ability of the mutant protein to asialofetuin-agarose was reduced but still remained. The binding ability of such mutants was similar to that of the recombinant half molecule containing the second lectin domain (Ch). However, when mutations were introduced into the second lectin domain, the binding ability of these mutant lectins to asialofetuin-agarose was significantly reduced just like the half recombinant molecule containing the first lectin domain (Nh). The different effects of the substitution of amino acid residues on the two lectin domains suggest that the binding properties of the two sites are different and that LEC-1 acts as a "heterobifunctional crosslinker."

Animals↗

Enhancement of plasminogen activator activity stimulated by LPS in gingival fibroblasts of individuals with Down syndrome.

Individuals with Down syndrome (DS) have a high prevalence of periodontal disease, which develops early and progressed rapidly and extensively, in comparison with healthy controls. The severe periodontal disease in individuals with DS has been considered to result from abnormal factors in their host responses. The mechanisms involved in the periodontal inflammatory processes in individuals with DS are not fully understood. Plasminogen activators (PA) are serine proteases that are well known for their part in the initiation of the fibrinolytic cascade leading to the generation of plasmin in periodontal homeostasis, including fibrinolysis and connective tissue remodeling. The PA-plasmin system affects the progression of periodontal disease. In the present study, we examined the effects of the levels of PA activity stimulated with lipopolysaccharide (LPS) in the gingival fibroblasts from donors with DS (DGF). The levels of PA activity without LPS were low in the DGFs, the same as that in the gingival fibroblasts from donors of healthy controls (NDGF). In contrast, the levels of PA activity with LPS in DGFs were significantly higher than that in the NDGFs. These results suggested that PA plays an important role in inducing extensive and rapid inflammation in the periodontal disease in individuals with DS.

Adolescent↗

A standard of AmpliType PM typing from aged evidentiary samples.

In analyzing aged samples by the AmpliType PM PCR amplification and Typing kit, it was occasionally observed that color developed typing strips had dark allele dots on PM loci but no visible S dot. Since the S dot acts as a minimum dot intensity control to determine positive alleles on the PM loci, it is necessary to apply another control system. To achieve positive PM typing from a degraded DNA sample that is inferred to be derived from a single donor, a standard has been adopted wherein loci from which sufficient PCR products are observed on agarose gel can be typed. The objective determination of sufficient PCR was done by comparison between band peak height of each locus generated from a sample and that of the corresponding locus generated from two nanograms (recommended minimum quantity as template DNA) of the control DNA provided in the kit.

Artifacts↗