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Hiroshi Masumoto

Publications and source records attributed to Hiroshi Masumoto.

At least 19 recordsLinked to original sources
PubMed2007

Angiographic fate of collateral vessels after surgical revascularization of the totally occluded left anterior descending artery.

BACKGROUND: Coronary artery bypass grafting (CABG) is best indicated for chronic total occlusion of the left anterior descending artery (LAD) with collaterals. We investigated angiographic changes in the collateral circulation after CABG. METHODS: Preoperative and postoperative angiograms were reviewed in 42 patients who underwent grafting onto occluded LADs. We described the type, location, and size of collaterals, the Rentrop grading, and collateral frame count (CFC). Regional wall motion of the LAD area was also evaluated with the centerline method. Postoperatively, we measured the lengths of LAD proximal (Lp) and distal (Ld) to the graft anastomotic site. RESULTS: Preoperative collaterals comprised 78 pathways (septal 42%, branch-branch 20%, atrial 19%, bridging 18%). After CABG, residual collaterals were identified, mainly through the septal pathways, in 6 patients (14%), most of whom were diabetic. The residual collaterals were a part of those which had been opacified in earlier phases of the preoperative angiograms (CFC: 17 +/- 3 vs 25 +/- 15, p = 0.01). Also, the Ld was shorter in these patients so that Lp/Ld was greater than in patients without residual collaterals (0.80 +/- 0.24 vs 0.53 +/- 0.28, p = 0.04). We found no association of residual collaterals with the improvement of LAD regional wall motion after CABG. CONCLUSIONS: Even after successful CABG, some collaterals with earlier filling of the LAD remain, mainly through the septum. Although the clinical significance remains to be clarified, complex and diffuse atherosclerosis associated with more distal graft anastomoses may contribute to maintaining collaterals after CABG to the occluded LAD, especially in diabetic patients.

Aged↗
PubMed2006

Histone H3 lysine 56 acetylation: a new twist in the chromosome cycle.

Several recent reports have identified lysine 56 (K56) as a novel site of acetylation in yeast histone H3. K56 acetylation is predicted to disrupt some of the histone-DNA interactions at the entry and exit points of the nucleosome core particle. This modification occurs in virtually all the newly synthesised histones that are deposited into chromatin during S-phase. Cells with mutations that block K56 acetylation show increased genome instability and hypersensitivity to genotoxic agents that interfere with replication. Removal of K56 acetylation takes place in the G2/M phase of the cell cycle and is dependent upon Hst3 and Hst4, two proteins that are related to the NAD+-dependent histone deacetylase Sir2. In response to DNA damage checkpoint activation during S-phase, expression of Hst3/Hst4 is delayed to extend the window of opportunity in which K56 acetylation can act in the DNA damage response. The high abundance of histone H3 K56 acetylation, its regulation and strategic location in the nucleosome core particle raise a number of fascinating issues that we discuss here.

Acetylation↗
PubMed2006

The sirtuins hst3 and Hst4p preserve genome integrity by controlling histone h3 lysine 56 deacetylation.

BACKGROUND: Acetylation of histone H3 lysine 56 (K56Ac) occurs transiently in newly synthesized H3 during passage through S phase and is removed in G2. However, the physiologic roles and effectors of K56Ac turnover are unknown. RESULTS: The sirtuins Hst3p and, to a lesser extent, Hst4p maintain low levels of K56Ac outside of S phase. In hst3 hst4 mutants, K56 hyperacetylation nears 100%. Residues corresponding to the nicotinamide binding pocket of Sir2p are essential for Hst3p function, and H3 K56 deacetylation is inhibited by nicotinamide in vivo. Rapid inactivation of Hst3/Hst4p prior to S phase elevates K56Ac to 50% in G2, suggesting that K56-acetylated nucleosomes are assembled genome-wide during replication. Inducible expression of Hst3p in G1 or G2 triggers deacetylation of mature chromatin. Cells lacking Hst3/Hst4p exhibit many phenotypes: spontaneous DNA damage, chromosome loss, thermosensitivity, and acute sensitivity to genotoxic agents. These phenotypes are suppressed by mutation of histone H3 K56 into a nonacetylatable residue or by loss of K56Ac in cells lacking the histone chaperone Asf1. CONCLUSIONS: Our results underscore the critical importance of Hst3/Hst4p in controlling histone H3 K56Ac and thereby maintaining chromosome integrity.

Acetylation↗
PubMed2006

Preoperative evaluation using asialoscintigraphy in patients undergoing cardiac surgery with noncardiac liver cirrhosis.

OBJECTIVE: Liver cirrhosis is recognized as one of the risk factors for severe complications after cardiac surgery. However, there are no established methods for risk stratification of the patients with liver cirrhosis (LC) regarding cardiac surgery. We present our experience of preoperative evaluation of liver function using asialoscintigraphy. METHODS: Between April 1999 and December 2005, we evaluated preoperative liver function using asialoscintigraphy with technetium-99m galactosyl human serum albumin in four cirrhotic patients undergoing coronary artery bypass grafting (n = 2) and valve replacement (n = 2), whose etiologies of LC were alcohol abuse (n = 1) and hepatitis C virus infection (n = 3). They also underwent other tests for preoperative evaluation of liver function, including the indocyanine green (ICG) test. RESULTS: Asialoscintigraphy revealed that the receptor index and the index of blood clearance in each patient were 0.81/0.73, 0.95/0.5, 0.82/0.62, and 0.97/0.57, respectively. These values closely correlated with the results of the ICG test. All patients were discharged alive from hospital after surgery. However, although one patient who underwent off-pump bypass had an uneventful course, three patients had major complications: pleural effusion (n = 1) and wound infection (n = 2). CONCLUSION: Asialoscintigraphy is a practical, reliable method that can replace the ICG test for estimating hepatic function for risk stratification of cirrhotic patients undergoing cardiac surgery, whose mortality and morbidity are still high.

Aged↗
PubMed2006

An artificially constructed de novo human chromosome behaves almost identically to its natural counterpart during metaphase and anaphase in living cells.

Human artificial chromosomes (HACs) are promising reagents for the analysis of chromosome function. While HACs are maintained stably, the segregation mechanisms of HACs have not been investigated in detail. To analyze HACs in living cells, we integrated 256 copies of the Lac operator into a precursor yeast artificial chromosome (YAC) containing alpha-satellite DNA and generated green fluorescent protein (GFP)-tagged HACs in HT1080 cells expressing a GFP-Lac repressor fusion protein. Time-lapse analyses of GFP-HACs and host centromeres in living mitotic cells indicated that the HAC was properly aligned at the spindle midzone and that sister chromatids of the HAC separated with the same timing as host chromosomes and moved to the spindle poles with mobility similar to that of the host centromeres. These results indicate that a HAC composed of a multimer of input alpha-satellite YACs retains most of the functions of the centromeres on natural chromosomes. The only difference between the HAC and the host chromosome was that the HAC oscillated more frequently, at higher velocity, across the spindle midzone during metaphase. However, this provides important evidence that an individual HAC has the capacity to maintain tensional balance in the pole-to-pole direction, thereby stabilizing its position around the spindle midzone.

Anaphase↗
PubMed2005

Assembly of additional heterochromatin distinct from centromere-kinetochore chromatin is required for de novo formation of human artificial chromosome.

Alpha-satellite (alphoid) DNA is necessary for de novo formation of human artificial chromosomes (HACs) in human cultured cells. To investigate the relationship among centromeric, transcriptionally permissive and non-permissive chromatin assemblies on de novo HAC formation, we constructed bacterial artificial chromosome (BAC)-based linear HAC vectors whose left vector arms are occupied by beta geo coding genes with or without a functional promoter in addition to a common marker gene on the right arm. Although HACs were successfully generated from the vectors with promoter-less constructs on the left arm in HT1080 cells, we failed to generate a stable HAC from the vectors with a functional promoter on the left arm. Despite this failure in HAC formation, centromere components (CENP-A, CENP-B and CENP-C) assembled at the integration sites correlating with a transcriptionally active state of both marker genes on the vector arms. However, on the stable HAC, chromatin immunoprecipitation analysis showed that HP1alpha and trimethyl histone H3-K9 were enriched at the non-transcribing left vector arm. A transcriptionally active state on both vector arms is not compatible with heterochromatin formation on the introduced BAC DNA, suggesting that epigenetic assembly of heterochromatin is distinct from centromere chromatin assembly and is required for the establishment of a stable artificial chromosome.

Cell Line, Tumor↗
PubMed2005

Human centromere protein B induces translational positioning of nucleosomes on alpha-satellite sequences.

The human centromere proteins A (CENP-A) and B (CENP-B) are the fundamental centromere components of chromosomes. CENP-A is the centromere-specific histone H3 variant, and CENP-B specifically binds a 17-base pair sequence (the CENP-B box), which appears within every other alpha-satellite DNA repeat. In the present study, we demonstrated centromere-specific nucleosome formation in vitro with recombinant proteins, including histones H2A, H2B, H4, CENP-A, and the DNA-binding domain of CENP-B. The CENP-A nucleosome wraps 147 base pairs of the alpha-satellite sequence within its nucleosome core particle, like the canonical H3 nucleosome. Surprisingly, CENP-B binds to nucleosomal DNA when the CENP-B box is wrapped within the nucleosome core particle and induces translational positioning of the nucleosome without affecting its rotational setting. This CENP-B-induced translational positioning only occurs when the CENP-B box sequence is settled in the proper rotational setting with respect to the histone octamer surface. Therefore, CENP-B may be a determinant for translational positioning of the centromere-specific nucleosomes through its binding to the nucleosomal CENP-B box.

Autoantigens↗
PubMed2005

Rapid generation of long synthetic tandem repeats and its application for analysis in human artificial chromosome formation.

Human artificial chromosomes (HACs) provide a unique opportunity to study kinetochore formation and to develop a new generation of vectors with potential in gene therapy. An investigation into the structural and the functional relationship in centromeric tandem repeats in HACs requires the ability to manipulate repeat substructure efficiently. We describe here a new method to rapidly amplify human alphoid tandem repeats of a few hundred base pairs into long DNA arrays up to 120 kb. The method includes rolling-circle amplification (RCA) of repeats in vitro and assembly of the RCA products by in vivo recombination in yeast. The synthetic arrays are competent in HAC formation when transformed into human cells. As short multimers can be easily modified before amplification, this new technique can identify repeat monomer regions critical for kinetochore seeding. The method may have more general application in elucidating the role of other tandem repeats in chromosome organization and dynamics.

Cell Line, Tumor↗
PubMed2005

A role for cell-cycle-regulated histone H3 lysine 56 acetylation in the DNA damage response.

DNA breaks are extremely harmful lesions that need to be repaired efficiently throughout the genome. However, the packaging of DNA into nucleosomes is a significant barrier to DNA repair, and the mechanisms of repair in the context of chromatin are poorly understood. Here we show that lysine 56 (K56) acetylation is an abundant modification of newly synthesized histone H3 molecules that are incorporated into chromosomes during S phase. Defects in the acetylation of K56 in histone H3 result in sensitivity to genotoxic agents that cause DNA strand breaks during replication. In the absence of DNA damage, the acetylation of histone H3 K56 largely disappears in G2. In contrast, cells with DNA breaks maintain high levels of acetylation, and the persistence of the modification is dependent on DNA damage checkpoint proteins. We suggest that the acetylation of histone H3 K56 creates a favourable chromatin environment for DNA repair and that a key component of the DNA damage response is to preserve this acetylation.

Acetylation↗
PubMed2005

The microcephaly ASPM gene is expressed in proliferating tissues and encodes for a mitotic spindle protein.

The most common cause of primary autosomal recessive microcephaly (MCPH) appears to be mutations in the ASPM gene which is involved in the regulation of neurogenesis. The predicted gene product contains two putative N-terminal calponin-homology (CH) domains and a block of putative calmodulin-binding IQ domains common in actin binding cytoskeletal and signaling proteins. Previous studies in mouse suggest that ASPM is preferentially expressed in the developing brain. Our analyses reveal that ASPM is widely expressed in fetal and adult tissues and upregulated in malignant cells. Several alternatively spliced variants encoding putative ASPM isoforms with different numbers of IQ motifs were identified. The major ASPM transcript contains 81 IQ domains, most of which are organized into a higher order repeat (HOR) structure. Another prominent spliced form contains an in-frame deletion of exon 18 and encodes 14 IQ domains not organized into a HOR. This variant is conserved in mouse. Other spliced variants lacking both CH domains and a part of the IQ motifs were also detected, suggesting the existence of isoforms with potentially different functions. To elucidate the biochemical function of human ASPM, we developed peptide specific antibodies to the N- and C-termini of ASPM. In a western analysis of proteins from cultured human and mouse cells, the antibodies detected bands with mobilities corresponding to the predicted ASPM isoforms. Immunostaining of cultured human cells with antibodies revealed that ASPM is localized in the spindle poles during mitosis. This finding suggests that MCPH is the consequence of an impairment in mitotic spindle regulation in cortical progenitors due to mutations in ASPM.

Adult↗
PubMed2005

Mixed venous-arterial CO2 tension gradient after cardiopulmonary bypass.

Significant venous hypercarbia has been reported in septic shock and circulatory failure. Cardiopulmonary bypass also impairs systemic and pulmonary blood perfusion. The objective of this study was to determine the clinical significance of the increased venous-arterial CO2 tension gradient resulting from venous hypercarbia after cardiopulmonary bypass. On arrival in the intensive care unit, venous and arterial CO2 tensions were measured in the radial and pulmonary arteries in 140 consecutive patients who had undergone coronary (n = 79), valve (n = 34), aortic (n = 20), and other (n = 7) surgery under cardiopulmonary bypass. The mean venous-arterial CO2 tension gradient was 5.0 +/- 3.3 mm Hg (range, 7.7 to 15.7 mm Hg). By linear regression analysis, the factors that significantly correlated with venous-arterial CO2 tension gradient were bypass duration, aortic crossclamp time, initial arterial lactate level, transpulmonary arteriovenous lactate difference, arterial bicarbonate level, base excess, cardiac index, mixed venous O2 saturation, O2 delivery, O2 consumption, and the peak value of creatine kinase. The venous-arterial CO2 tension gradient may reflect impaired perfusion and anaerobic metabolism induced by cardiopulmonary bypass and could be a simple and useful indicator for patient management after surgery under cardiopulmonary bypass.

Aged↗
PubMed2004

Construction of a novel human artificial chromosome vector for gene delivery.

Potential problems of conventional transgenes include insertional disruption of the host genome and unpredictable, irreproducible expression of the transgene by random integration. Alternatively, human artificial chromosomes (HACs) can circumvent some of the problems. Although several HACs were generated and their mitotic stability was assessed, a practical way for introducing exogenous genes by the HACs has yet to be explored. In this study, we developed a novel HAC from sequence-ready human chromosome 21 by telomere-directed chromosome truncation and added a loxP sequence for site-specific insertion of circular DNA by the Cre/loxP system. This 21HAC vector, delivered to a human cell line HT1080 by microcell fusion, bound centromere proteins A, B, and C and was mitotically stable during long-term culture without selection. The EGFP gene inserted in the HAC vector expressed persistently. These results suggest that the HAC vector provides useful system for functional studies of genes in isogenic cell lines.

Animals↗
PubMed2004

Long-term clinical performance of AAI pacing in patients with sick sinus syndrome: a comparison with dual-chamber pacing.

AIMS: In this clinical study, we compared two groups of age-matched patients, AAI and DDD, to evaluate the clinical benefits of AAI pacing in patients with sick sinus syndrome (SSS) and normal atrioventricular (AV) conduction. METHODS AND RESULTS: Ninety-five patients with SSS implanted with AAI pacemakers were compared with 101 SSS patients implanted with DDD pacemakers. Mortality, chronic atrial fibrillation, lead survival rates, and reoperation rates were compared by Kaplan-Meier analysis. Eight AAI devices were switched to DDD due to high-degree (grade 2-3) AV block. The incidence of high-degree AV block was 1.104%/year, with a freedom rate of 88.6% at 10 years. There were no significant differences between the two groups in survival rates (87.8% in AAI vs. 93.4% in DDD at 10 years), freedom from atrial fibrillation (93.6% vs. 90.6%), or freedom from reoperation (71.3% vs. 76.3%). On the other hand, lead failure was twice as frequent in the DDD group than in the AAI group (relative risk=2.045, P=0.0382). CONCLUSION: AAI pacing, a simple system using a single lead and single-chamber pacemaker, can achieve a clinical outcome similar to that of the DDD mode in patients with SSS and normal AV conduction.

Adolescent↗
PubMed2004

The role of CENP-B and alpha-satellite DNA: de novo assembly and epigenetic maintenance of human centromeres.

The centromere is an essential functional domain responsible for the correct inheritance of eukaryotic chromosomes during cell division. Eukaryotic centromeres include the highly conserved centromere-specific histone H3 variant, CENP-A, which has provided a powerful tool for investigating the recruitment of centromere components. However, the trigger that targets CENP-A to a specific genomic locus during centromere assembly remains unknown. Although, on rare occasions, CENP-A chromatin may assemble at non-centromeric DNA, all normal human centromeres are assembled and maintained on alpha-satellite (alphoid) DNA. The importance of alphoid DNA and CENP-B binding sites (CENP-B boxes), typical of normal human centromere DNA configurations, has been demonstrated through their requirement in de novo centromere assembly and Human Artificial Chromosome (HAC) assays. Mechanisms to link the centromere tightly to specific genomic sequences exist in humans and the two yeast species.

Autoantigens↗
PubMed2003

CENP-B interacts with CENP-C domains containing Mif2 regions responsible for centromere localization.

Recently, human artificial chromosomes featuring functional centromeres have been generated efficiently from naked synthetic alphoid DNA containing CENP-B boxes as a de novo mechanism in a human cultured cell line, but not from the synthetic alphoid DNA only containing mutations within CENP-B boxes, indicating that CENP-B has some functions in assembling centromere/kinetochore components on alphoid DNA. To investigate whether any interactions exist between CENP-B and the other centromere proteins, we screened a cDNA library by yeast two-hybrid analysis. An interaction between CENP-B and CENP-C was detected, and the CENP-C domains required were determined to overlap with three Mif2 homologous regions, which were also revealed to be involved in the CENP-C assembly of centromeres by expression of truncated polypeptides in cultured cells. Overproduction of truncated CENP-B containing no CENP-C interaction domains caused abnormal duplication of CENP-C domains at G2 and cell cycle delay at metaphase. These results suggest that the interaction between CENP-B and CENP-C may be involved in the correct assembly of CENP-C on alphoid DNA. In other words, a possible molecular linkage may exist between one of the kinetochore components and human centromere DNA through CENP-B/CENP-B box interaction.

Autoantigens↗