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Hiroki Shimizu

Publications and source records attributed to Hiroki Shimizu.

At least 19 recordsLinked to original sources
PubMed2006

Construction and structural characterization of versatile lactosaminoglycan-related compound library for the synthesis of complex glycopeptides and glycosphingolipids.

We have established a facile and efficient protocol for the preparative-scale synthesis of various compound libraries related to lactosaminoglycans: cell surface oligosaccharides composed of N-acetyllactosamine as a repeating disaccharide unit, based on chemical and enzymatic approaches. Substrate specificity and feasibility of a bacterial glycosyltransferase, Neisseria meningitidis beta1,3-N-acetylglucosaminyltransferase (LgtA), were investigated in order to synthesize various key intermediates suited for the construction of mammalian O-glycopeptides and glycosphingolipids containing poly-N-acetyllactosamine structures. Recombinant LgtA exhibited the highest glycosyltransferase activity with strongly basic conditions (pH = 10, glycine-NaOH buffer) and a broad range of optimal temperatures from 20 to 30 degrees C. Interestingly, it was found that LgtA discriminates L-serine and L-threonine and functions both as a core-1 beta1,3-N-acetylglucosaminyltransferase and core-2 beta1,3-N-acetylglucosaminyltransferase toward Fmoc-Ser derivatives, while LgtA showed only core-2 beta1,3-N-acetylglucosaminyltransferase activity in the presence of Fmoc-Thr derivatives. Combined use of LgtA with human beta1,4-galactosyltransferase allowed for controlled sugar extension reactions from synthetic sugar amino acids and gave synthetic lactosaminoglycans, such as a decasaccharide derivative, Galbeta(1 --> 4)GlcNAcbeta(1 --> 3)Galbeta(1 --> 4)GlcNAcbeta(1 --> 3)Galbeta(1 --> 4)GlcNAcbeta(1 --> 3)Galbeta(1 --> 4)GlcNAcbeta(1 --> 6)[Galbeta(1 --> 3)]GalNAcalpha1 --> Fmoc-Ser-OH (6), and a dodecasaccharide derivative, Galbeta(1 --> 4)GlcNAcbeta(1 --> 3)Galbeta(1 --> 4)GlcNAcbeta(1 --> 3)Galbeta(1 --> 4)GlcNAcbeta(1 --> 6)[Galbeta(1 --> 4)GlcNAcbeta(1 --> 3)Galbeta(1 --> 4)GlcNAcbeta(1 --> 3)Galbeta(1 --> 3)]GalNAcalpha1 --> Fmoc-Ser-OH (9). A partially protected pentasaccharide intermediate, GlcNAcbeta(1 --> 3)Galbeta(1 --> 4)GlcNAcbeta(1 --> 6)[Galbeta(1 --> 3)]GalNAcalpha1 --> Fmoc-Thr-OH (11), was applied for the microwave-assisted solid-phase synthesis of a MUC1-related glycopeptide 19 (MW = 2610.1). The findings suggest that this sugar extension strategy can be employed for the modification of lactosyl ceramide mimetic polymers to afford convenient precursors for the synthesis of various glycosphingolipids.

Amino Sugars↗
PubMed2006

Predictive value of microvolt T-wave alternans for sudden cardiac death in patients with preserved cardiac function after acute myocardial infarction: results of a collaborative cohort study.

OBJECTIVES: We conducted a collaborative cohort study to evaluate the predictive power of microvolt T-wave alternans (TWA) in patients with preserved left ventricular ejection fraction (LVEF) after myocardial infarction (MI). BACKGROUND: There is little information available about the prognostic value of risk stratification markers in this population. Although these patients have a relatively good prognosis, identifying high-risk patients is important in clinical practice. METHODS: This study enrolled 1,041 post-MI patients with an LVEF > or =40% (average 55 +/- 10%). Microvolt TWA testing was performed 48 +/- 66 days after acute MI, and 10 other risk variables were also evaluated. The end points were prospectively defined as sudden cardiac death or life-threatening arrhythmic events. RESULTS: During a follow-up of 32 +/- 14 months, 38 patients (3.7%) died of nonarrhythmic causes and were not considered for analysis. Of the 1,003 evaluable patients, 18 (1.8%) reached an end point. Microvolt TWA was positive in 169 patients (17%), negative in 747 (74%), and indeterminate in 87 (9%). A positive microvolt TWA test, nonsustained ventricular tachycardia, and ventricular late potentials were predictors of events, and percutaneous coronary intervention decreased the risk rate. On multivariate analysis, a positive microvolt TWA test was the most significant predictor, with a hazard ratio of 19.7 (p < 0.0001). This marker had the highest sensitivity and negative predictive value for events. CONCLUSIONS: In patients with preserved cardiac function, the incidence of indeterminate results of microvolt TWA is low, and a positive test result is associated with arrhythmic events. Microvolt TWA could be used for risk stratification in this low-risk population.

Aged↗
PubMed2006

Construction of highly glycosylated mucin-type glycopeptides based on microwave-assisted solid-phase syntheses and enzymatic modifications.

A MUC1-related glycopeptide having five core-2 hexasaccharide branches (C330H527N46O207, MW = 8450.9) was synthesized by a new strategy using a combination of microwave-assisted solid-phase synthesis (MA-SPGS) and enzymatic sugar elongation. Synthesis of a key glycopeptide intermediate was best achieved in a combination of PEGA [poly(ethylene glycol)-poly-(N,N-dimethylacrylamide) copolymer] resin and MA-SPGS using glycosylated amino acid building blocks with high speed and high purity. Deprotection of the glycopeptide intermediate and subsequent glycosyltransferase-catalyzed sugar elongations were performed for generation of the additional diversities with the sugar moieties of glycopeptides using beta1,4-galactosyltransferase (beta1,4-GalT) and two kinds of alpha2,3-sialyltransferases [ST3Gal III; alpha2,3-(N)-SiaT and ST3Gal II; alpha2,3-(O)-SiaT]. These reactions proceeded successfully in the presence of 0.2% Triton X-100 to convert the chemically synthesized trisaccharide glycans to disialylated hexasaccharide.

Amino Acid Sequence↗
PubMed2006

Circadian variation of beat-to-beat QT interval variability in patients with prior myocardial infarction and the effect of beta-blocker therapy.

BACKGROUND: Recent studies have demonstrated that increased QT interval variability (QTV) is associated with a greater susceptibility to ventricular arrhythmias and that patients with prior myocardial infarction (MI) were prone to ventricular arrhythmias during the daytime. The goal of the present study was to investigate the circadian variation of the QTV and to determine whether beta-blocker therapy improves the temporal fluctuation of the ventricular repolarization in patients with MI. METHODS: The study population consisted of 15 MI patients who had not received beta-blocker therapy, 11 MI patients who had received beta-blocker therapy, and 12 healthy subjects. Twenty-four hour Holter monitoring was obtained, and the RR and QT intervals were calculated automatically from 512 consecutive sinus beats for every 2 hours. RESULTS: In the daytime, the QT-SD was significantly greater in the MI group than in the healthy subjects (P<0.01), but there was no difference in the QT-SD when comparing the beta-blocker group to the control group. Moreover, the QT variability index and the QT variance normalized for the mean QT were similar pattern with QT-SD. The heart rate variability did not significantly differ when compared between the three study groups. CONCLUSION: These data indicate that the QTV increases during the daytime in patients with MI and that this circadian effect is prevented by beta-blocker therapy. Thus, beta-blocker therapy may reverse the maladaptation of the ventricular repolarization to the change in the heart rate and may thereby reduce the ventricular arrhythmias and decrease the mortality in patients with MI.

Adrenergic beta-Antagonists↗
PubMed2006

Correlation between beat-to-beat QT interval variability and impaired left ventricular function in patients with previous myocardial infarction.

BACKGROUND: Beat-to-beat QT interval variability (QTV) is associated with sudden cardiac death and New York Heat Association functional class severity. We sought to evaluate the relationship between QTV and left ventricular (LV) function in patients with previous myocardial infarction (MI). METHODS: Fifty-nine patients with previous anterior MI were enrolled. LV ejection fraction (EF), LV end-systolic volume index (LVESVI), and LV end-diastolic volume index (LVEDVI) were measured by LV contrast angiography. QT interval was measured by automated analysis of 512-beat records of 12-lead electrocardiogram. The mean interval, standard deviation and variance in RR and QT intervals, and the QT variability index (QTVI) were calculated for each patient using two leads that corresponded with and without the infarction site. High-frequency power, low-frequency power, total-frequency power, and the ratio of low-frequency to high-frequency power in RR and QT intervals were calculated. RESULTS: While measured indices of RR intervals and indices of QT intervals, which did not correspond with the infarction site, did not correlate with differences in LV function, measured indices of QT intervals, which corresponded with the infarction site, did correlate with differences in LV function. However, there were no correlations between the ratio of low-frequency to high-frequency power in QT intervals and EF or LVEDVI. Correlations between QTVI and LV function were observed, particularly between QTVI and LVESVI (r = 0.712, P < 0.0001). CONCLUSION: In patients with previous anterior MI, there was variability in temporal dispersion of QT interval and a strong correlation between QTV corresponded with the infarcted site and LV function.

Aged↗
PubMed2005

Characterization of Vibrio cholerae neuraminidase by a novel mechanism-based fluorescent labeling reagent.

Vibrio cholerae neuraminidase (VCNA) plays a significant role in the pathogenesis of cholera by removing sialic acid residues from higher-order gangliosides to an unmasked GM1, the essential receptor for cholera toxin. Here we report that a novel mechanism-based fluorescent labeling reagent, 5-acetamido-2-(4-N-5-dimethylaminonaphthalene-1-sulfonyl-2-difluoromethylphenyl)-3,5-dideoxy-d-glycero-alpha-d-galacto-2-nonulopyranosonic acid (1), becomes a unique irreversible inhibitor of VCNA. Characterization of an inactivated VCNA by MALDI-TOF/TOFMS analysis revealed that the Asp-576 and Arg-577 residues, which are located within the (576)DRFF(579) sequence, were specifically labeled with this suicide-type fluorescent substrate. Neither Asp-576 nor Arg-577 has ever been known to contribute to a specific residue in the rigid and highly conserved active site of VCNA investigated by crystallographic analysis, suggesting that a flexible beta-turn structure containing this sequence may have a crucial role in the dynamic nature of substrate recognition and catalytic action by VCNA.

Amino Acid Sequence↗
PubMed2005

Combinatorial synthesis of MUC1 glycopeptides: polymer blotting facilitates chemical and enzymatic synthesis of highly complicated mucin glycopeptides.

The chemoselective polymer blotting method allows for rapid and efficient synthesis of glycopeptides based on a "catch and release" strategy between solid-phase and water-soluble polymer supports. We have developed a heterobifunctional linker sensitive to glutamic acid specific protease (BLase). The general procedure consists of five steps, namely (i) the solid-phase synthesis of glycopeptide containing BLase sensitive linker, (ii) subsequent deprotections and the release of the glycopeptide from the resin, (iii) chemoselective blotting of the glycopeptide intermediates in the presence of water-soluble polymers with oxylamino functional groups, (iv) sugar elongations using glycosyltransferases, and (v) the release of target glycopeptides from the polymer platform by selective BLase promoted hydrolysis. The combined use of the solid-phase chemical syntheses of peptides and the enzymatic syntheses of carbohydrates on water-soluble polymers would greatly contribute to the production of complicated glycopeptide libraries, thereby enhancing applicative research. We report here a high-throughput synthetic system for the various types of MUC1 glycopeptides exhibiting a variety of sugar moieties. It is our belief that this concept will become part of the entrenched repertoire for the synthesis of biologically important glycopeptides on the basis of glycosyltransferase reactions in automated and combinatorial syntheses.

Carbohydrate Sequence↗
PubMed2005

Rapid microwave-assisted solid-phase glycopeptide synthesis.

Coupling of glycosylated Fmoc-Thr or Fmoc-Ser with N-terminal amino acids on a resin proceeded smoothly under microwave irradiation for 20 min with much higher efficiency (98% yield per coupling) than found in more general conditions. Compared with a conventional protocol, the present method greatly reduces the time required for solid-phase glycopeptide synthesis from 4 days to 7 h, as is the case with the synthesis of Muc-1-related 20-residue glycopeptide carrying five core-2 trisaccharide chains. [structure: see text]

Amino Acid Sequence↗
PubMed2005

Discordant repolarization alternans-induced atrial fibrillation is suppressed by verapamil.

BACKGROUND: Ventricular alternans of repolarization produces serious ventricular arrhythmias in experimental models. The present study investigated the role of alternans of atrial repolarization in patients with atrial fibrillation (AF). METHODS AND RESULTS: Electrophysiological studies were performed in 19 patients without structural heart disease. Monophasic action potentials (MAP) were recorded with 2 Franz catheters during steady state pacing, starting at a cycle length (CL) of 400 ms with subsequent decrements of 10 ms. Duration from the onset of upstroke to 90% repolarization of the MAP were measured. If discordant alternans (DA) was present during pacing, verapamil was administrated, and MAP measurements were repeated. Rapid pacing resulted in concordant alternans to DA in 13 of 19 (68%) patients. AF was initiated after the induction of DA in 8 of 13 patients (p=0.012). Verapamil treatment resulted in a significant decrease in the longest pacing CL at which DA was induced (207+/-19 vs 178+/-17 ms, p<0.0001). CONCLUSIONS: Rapid atrial pacing induced DA and was associated with initiation of AF. Furthermore, induction of DA was suppressed by verapamil. Reducing the spatiotemporal repolarization heterogeneity may be how the calcium-channel blockade prevents initiation of AF.

Action Potentials↗
PubMed2005

Development of highly potent D-glucosamine-based chiral fluorescent labeling reagents and a microwave-assisted beta-selective glycosidation of a methyl glycoside reagent.

We synthesized new chiral fluorescence labeling reagents having a 2,3-anthracenedicarboximide group from D-glucosamine, and it was possible to introduce target alcohols at the anomeric positions of the reagents with beta-selectivity by glycosidations. Especially, it was possible to use methyl glycoside reagent as a glycosyl donor with a Lewis acid and microwave irradiation, and it gave selectively beta-glycoside while the reaction without microwave irradiation gave alpha- and beta-mixed glycosides. Those reagents showed very high chiral discrimination ability, and they made it possible to separate the eight stereoisomers of 4,8,12,16-tetramethylheptadecanol by HPLC after derivatizations.

Carbohydrate Conformation↗
PubMed2004

Thermodynamics of binding of D-galactose and deoxy derivatives thereof to the L-arabinose-binding protein.

We report the thermodynamics of binding of d-galactose and deoxy derivatives thereof to the arabinose binding protein (ABP). The "intrinsic" (solute-solute) free energy of binding DeltaG degrees (int) at 308 K for the 1-, 2-, 3-, and 6-hydroxyl groups of galactose is remarkably constant (approximately -30 kJ/mol), despite the fact that each hydroxyl group subtends different numbers of hydrogen bonds in the complex. The substantially unfavorable enthalpy of binding (approximately 30 kJ/mol) of 1-deoxygalactose, 2-deoxygalactose, and 3-deoxygalactose in comparison with galactose, cannot be readily accounted for by differences in solvation, suggesting that solute-solute hydrogen bonds are enthalpically significantly more favorable than solute-solvent hydrogen bonds. In contrast, the substantially higher affinity for 2-deoxygalactose in comparison with either 1-deoxygalactose or 3-deoxygalactose derives from differences in the solvation free energies of the free ligands.

Calorimetry↗
PubMed2004

Sinus slowing caused by adenosine-5'-triphosphate in patients with and without sick sinus syndrome under various autonomic states.

Adenosine infusion can potentially be used as a diagnostic test for sick sinus syndrome (SSS) based on its negative chronotropic effects. Whether autonomic tone underlies adenosine's negative chronotropic effects remains unknown. This study was to investigate the bradycardiac response of sinus node to ATP in patients with and without clinical SSS by measuring atrial cycle length (ACL) before and after bolus of ATP in different states of autonomic tone. The negative chronotropic effect of ATP was assessed by comparing the mean ACL before ATP administration with the longest ACL after a bolus of ATP infusion (Delta ACL). Our results showed that Delta ACL in patients with SSS were significantly greater than that without SSS (P<.001) in all 4 states, and IHR in patients with SSS were significantly lower than calculated IHR (P<.0001). Moreover, there was no significant difference in Delta ACL between the 4 states in patients with SSS (P = .99). However, Delta ACL was significantly greater during isoproterenol infusion and after propranolol administration in patients without sinus node dysfunction, comparing with baseline state (P<.01), but not after combination of atropine (P = .33). Our results indicate that the negative chronotropic effect of ATP on sinus node is much more dramatic in patients with SSS, in which the intrinsic disease of sinus node is responsible for the abnormal adenosine-mediated sinus arrest, and this effect is influenced by autonomic tone in patients without sinus node dysfunction but not in patients with SSS.

Adenosine Triphosphate↗
PubMed2004

Synthesis of (3R,4S)-3,4,5-trihydroxy-4-methylpentylphosphonic acid as a potential inhibitor of the nonmevalonate pathway.

The nonmevalonate pathway is widely found in higher plants and in many eubacteria, including pathogenic ones, but not in mammals. Identifying a nonmevalonate pathway inhibitor would greatly contribute to the search for new herbicides and antibacterial drugs to treat, for example, malaria. We describe here the synthesis of (3R,4S)-3,4,5-trihydroxy-4-methylpentylphosphonic acid as a candidate for inhibiting MEP cytidylyltransferase, which is the third step on the nonmevalonate pathway.

Anti-Bacterial Agents↗
PubMed2003

Optimization of tether length in nonglycosidically linked bivalent ligands that target sites 2 and 1 of a Shiga-like toxin.

A series of bivalent ligands for a Shiga-like toxin have been synthesized, their experimentally determined inhibitory activities were compared with a simplified thermodynamic model, and computer simulations were used to predict the optimal tether length in bivalent ligands. The design of the inhibitors exploits the proximity of the C-2' hydroxyl groups of two P(k)-trisaccharides when bound to two different, neighboring carbohydrate recognizing binding sites located on the surface of Shiga-like toxin. NMR studies of the complex between the toxin and bivalent ligands show that site 2 and site 1 of a single B subunit are simultaneously occupied by a tethered P(k)-trisaccharide dimer. A simplified thermodynamic treatment provides the intrinsic affinities and binding energies for the intermolecular and intramolecular association events and permits the deconvolution of the contributions to the relative binding energies for the set of bivalent ligands. Conformational analysis based on MD simulations for bivalent galabioside dimers containing different tethers demonstrated that the calculated local concentrations of the pendant ligand at the second binding site correlate with the experimentally determined relative affinity values of the respective bivalent ligands, thereby providing a predictive method to optimize tether length.

Cross-Linking Reagents↗
PubMed2003

Impaired left ventricular function is associated with increased recovery time dispersion in patients with previous myocardial infarction.

Left ventricular (LV) dysfunction after myocardial infarction is associated with higher risk of serious ventricular arrhythmias and sudden death. We suspected that heterogeneity in ventricular repolarization contributes to these arrhythmias. To quantify this heterogeneity, we measured the recovery time (the interval between QRS onset and the time of maximum dV/dt in the ST-T segment) using an 87-lead body surface mapping electrocardiogram and estimated recovery time dispersion (the difference between maximum recovery time and minimum recovery time) in each lead. Differences between 110 patients with previous myocardial infarction and 31 healthy controls were compared. Recovery time dispersion [medians (25th, 75th percentiles)] was greatest in patients with a dilated LV [169 ms (154, 201) vs. 155 ms (137, 172), P <.005], impaired ejection fraction [173 ms (155, 202) vs. 152 ms (138, 165), P <.0005] and LV dyskinesis [175 ms (159, 201) vs. 155 ms (137, 161), P <.0005]. This study suggests that LV dysfunction associated with myocardial infarction leads to heterogeneous ventricular repolarization and may provide the electrical substrate for ventricular arrhythmias.

Arrhythmias, Cardiac↗
PubMed2003

Wavelet transform analysis of heart rate variability to assess the autonomic changes associated with spontaneous coronary spasm of variant angina.

We used Wavelet transform (WT) to investigate whether variation in autonomic tone was associated with spontaneous coronary spasm in patients with variant angina by analysis of heart rate variability (HRV). Twenty-one episodes preceding ST-segment elevation were selected under Holter monitoring in 12 men and 3 women with variant angina. HRV indices were calculated at 10 second intervals with the continuous WT, and analyzed within 30 minutes preceding ST-segment elevation. High frequency (HF; 0.15 approximately 2.00 Hz) increased significantly during the 4 minutes prior to ST-segment elevation, low frequency (LF; 0.04 approximately 0.15 Hz) decreased significantly during the period from 10 to 5 minutes and increased significantly during the 2 minutes prior to ST-segment elevation, the LF/HF ratio decreased significantly during the period from 10 to 3 minutes and increased significantly during the 2 minutes prior to ST-segment elevation. The RR interval decreased significantly during the 2 minutes prior to ST-segment elevation. These results suggest that the acute variation in autonomic tone was associated with spontaneous coronary spasm in patients with variant angina. A reduction in sympathetic activity, then enhancement of vagal activity may play a key role in triggering the spontaneous coronary spasm, and the secondary activation of sympathetic activity may worsen the coronary spasm resulting in the attack.

Adult↗