Biomedical subjects

Hiroyuki Kamiya

Publications and source records attributed to Hiroyuki Kamiya.

83 records · Page 5Linked to original sources

PubMed2002

An oxidized nucleotide affects DNA replication through activation of protein kinases in Xenopus egg lysates.

To elucidate the response to oxidative stress in eukaryotic cells, the effect of an oxidized nucleotide, 8-oxo-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP), generated from dGTP with an active oxygen, on DNA synthesis was studied using a cell-free DNA replication system derived from Xenopus egg lysates with a single-stranded DNA template. Amounts of newly synthesized DNA were reduced according to the increasing concentration of 8-oxo-dGTP. Pulse labeling analysis revealed that 8-oxo-dGTP could delay DNA synthesis by reducing the rate of chain elongation. This delay was recovered by addition of a protein kinase inhibitor, staurosporine or bisindolylmaleimide I. These results indicate that a staurosporine- or bisindolylmaleimide I-sensitive protein kinase, such as a protein kinase C family member, may contribute to the delay of DNA synthesis by 8-oxo-dGTP. UV-irradiated single-stranded DNA also caused a delay of DNA synthesis on the undamaged template in the lysates. However, this delay was not recovered by staurosporine or bisindolylmaleimide I. Therefore, the mechanism of delay of DNA synthesis by 8-oxo-dGTP may be different from that by UV lesions. This is the first report that demonstrates an effect of an oxidized nucleotide on DNA replication in eukaryotes.

Animals↗

PubMed2002

Real-time and continuous monitoring of myocardial blood flow using a thermal diffusion method.

OBJECTIVES: There has been no method which can measure regional myocardial blood flow in real-time and continuously. The purpose of this study was to validate myocardial blood flow measurement using a thermal diffusion method. For this purpose, myocardial blood flow measurement was performed using the thermal diffusion method and the electrolytic hydrogen clearance method. METHODS: Seven pigs were used for this study, six were for comparison between the thermal diffusion and electrolytic hydrogen clearance methods, and one was for demonstration of myocardial blood flow measurement using the thermal diffusion method on a beating heart coronary artery bypass model with ischemic preconditioning. RESULTS: A good correlation was found between myocardial blood flow values obtained by the electrolytic hydrogen clearance method and 1/V values obtained by a thermal diffusion probe, the correlation coefficient was 0.841 (P<0.001). During the beating heart coronary artery bypass, the regional myocardial blood flow was recorded in real-time and continuously. CONCLUSIONS: This study demonstrated measurement of regional myocardial blood flow using the thermal diffusion method for the first time and simultaneous measurement using the electrolytic hydrogen clearance method for calibration. It provided a real-time and continuous myocardial blood flow measurement and has a potential to contribute to progress in beating-heart surgery.

Animals↗

PubMed2002

Coronary active perfusion system can maintain myocardial blood flow and tissue oxygenation.

OBJECTIVES: To avoid myocardial ischemia in off-pump coronary artery bypass, we developed a coronary active perfusion system (CAPS). The purposes of this study were to investigate the changes in myocardial blood flow and tissue oxygenation during CAPS perfusion, and to validate the ventricular arrhythmias and hemodynamic deterioration preventing the effect. METHODS: Sixteen pigs were divided into a CAPS perfusion group (group C) and a simple coronary occlusion group (group O). The left anterior descending coronary artery was snared in both groups and 30 min of CAPS perfusion was performed in group C. RESULTS: Ventricular arrhythmias were not observed in group C, but occurred in seven out of eight pigs in group O (P=0.003). None of the hemodynamic variables changed in group C, but they deteriorated in group O. Myocardial blood flow, saturation, and hemoglobin plus myoglobin concentration were maintained with a baseline level in group C, but decreased significantly in group O (P<0.001). CONCLUSION: CAPS is a reliable method to avoid myocardial ischemia during coronary occlusion and it may be useful for off-pump coronary artery bypass

Animals↗

PubMed2002

Long-term follow-up of cardiac valve replacement using bioprosthesis in patients 70 years old and older.

Good long-term results with the bioprosthetic valves for patients 70 years old and older have been reported. However, because the average lifespan is increasing, we aimed to clarify whether patients older than 70 may still be candidates for valve re-replacement. Seventy-one patients 70 years old and older, who received a total of 81 bioprosthetic valve replacements during 73 procedures between 1988 and 2000, were reviewed. There were 8 hospital and 7 late deaths. Ten-year actual survival after valve replacement was 73.5%, and 82.8% when hospital deaths were excluded. During the follow-up period, 2 patients received mitral valve re-replacement. Ten years of freedom from reoperation were found in 66.7% for all valves and in 50.0% for mitral valves. The average lifespan in Japan is currently 77.64 years for men and 84.62 years for women; therefore, valve degeneration in patients who receive bioprosthesis replacement in their early 70s should be anticipated.

Aged↗

PubMed2002

Substrate recognition by the human MTH1 protein.

A nucleotide pool sanitizing enzyme, the human MTH1 protein, hydrolyzes 2-hydroxy-dATP, 8-hydroxy-dATP, and 8-hydroxyd-GTP. To examine the substrate recognition mechanism of the MTH1 protein, ten nucleotide analogs (8-bromo-dATP, 8-bromod-GTP, deoxyisoinosine triphosphate, 8-hydroxy-dITP, 2-aminopurine-deoxyriboside triphosphate, 2-amino-dATP, deoxyxanthosine triphosphate, deoxyoxanosine triphosphate, dITP, and dUTP) were incubated with the protein. Of these, the former five nucleotides were hydrolyzed with various efficiencies. This results suggests the importance of the anti/syn-conformation and the functional groups on the 2 and 6-positions of the purine ring.

DNA Repair Enzymes↗

PubMed2002

Mutagenicity of 5-formylcytosine, an oxidation product of 5-methylcytosine, in DNA in mammalian cells.

To examine the mutagenicity of 5-formylcytosine (5-fC), an oxidation product of 5-methylcytosine (5-mC), 5-fC was incorporated into predetermined sites of double-stranded shuttle vectors. The nucleotide sequences in which the modified base was incorporated were 5'-AFGCGT-3' and 5'-ACGFGT-3' (F represents 5-fC), the recognition site for the restriction enzyme MluI (5'-ACGCGT-3'). 5-fC was incorporated into the template strand of either the leading or lagging strand of DNA replication. The modified DNAs were transfected into simian COS-7 cells, and the DNAs replicated in the cells were recovered and analyzed after a second transfection into Escherichia coli. 5-fC weakly blocked DNA replication in mammalian cells. The 5-fC residues were mutagenic, with mutation frequencies in double-stranded vectors of 0.03-0.28%. The mutation spectrum of 5-fC was broad, and included targeted (5-fC-->G, 5-fC-->A, and 5-fC-->T) and untargeted mutations. These results suggest that the oxidation of 5-mC results in mutations at and around the modified sites.

5-Methylcytosine↗

PubMed2002

Pharmacokinetics of targeting with liposomes.

The optimization of drug disposition in the body leads to an increase in its therapeutic effect and to a decrease in adverse effects. Liposomes can serve as a potential drug carrier for achieving this. However, the behavior of a drug carrier system under in vivo conditions is complex. Therefore, a more complete understanding of the pharmacokinetics of liposomes themselves, as well as that of the encapsulated drug, is required. The optimization of the pharmacokinetics of liposomes can be performed by linking a pharmacodynamic model of the free drugs that are encapsulated into liposomes. Sensitivity analysis was applied to optimize the delivery system to maximize the antitumor effect of liposomal doxorubicin (DOX). Advanced technology for ligand-mediated selective targeting and intracellular targeting is also introduced for antitumor agents and for gene delivery systems.

Animals↗

PubMedDate not supplied

Effect of simulated microgravity on testosterone and sperm motility in mice.

We examined changes in the serum testosterone level and in sperm in the testis and epididymis by using tail-suspended mice, which are a simulation model of the body fluid shift in space, to evaluate the possibility of spermatogenesis failure in space environment. We also studied pathological disorders of the testis in the tail-suspended mice. Tail suspension was imposed with a tail harness to a degree at which the hindlegs of mice did not touch the floor of the housing unit. In control mice, the tail was similarly fixed with a tail harness to impose the same stress, except that a hindleg remained on the floor. Body weight was not significantly different between the 2 groups during 7 days, and testicular weight was significantly different. The testosterone level was significantly lower in the tail-suspended group (0.71 +/- 1.24 ng/mL) than in the control group (2.38 +/- 3.50 ng/mL; P <.05). Microscopy with hematoxylin and eosin (HE) and periodic acid-Schiff (PAS) staining showed a small proportion of seminiferous tubules with impairment of spermatogenic function in the tail-suspended group, and multinucleated giant cells were occasionally noted. Terminal deoxynucleotidyl tranferase-mediated nick end-labeling staining revealed positive cells even in animals in which impairment was considered to be mild based on HE and PAS staining. Many cells showed intense p53 immunostaining compared to the control group, with more intense staining of the nucleus in the tail-suspended group. The proportion of motile sperm was slightly but not significantly reduced in the tail-suspended group. However, the mean movement velocity of the motile spermatozoa was significantly decreased.

Animals↗

PubMedDate not supplied

No enhancement of nuclear entry by direct conjugation of a nuclear localization signal peptide to linearized DNA.

Efficient nuclear entry of exogenous DNA is one of the key factors toward gene therapy success with nonviral vectors. To re-address the effects of a nuclear localization signal (NLS) peptide attached directly to DNA, we prepared three dumbbell-shaped, green fluorescent protein (GFP)-encoding DNAs containing one or two NLS peptides. The peptide was conjugated to the loop-forming oligodeoxyribonucleotides by cross-linking reactions between the peptide and a modified uracil base with a dioxaoctylamino linker, and the oligonucleotides were then ligated to the DNA molecules. The NLS-conjugated DNA dumbbells were microinjected into the cytosols and nuclei of simian COS-7 cells. In addition, unconjugated DNA dumbbells, with or without a modified uracil base, were also examined for comparison. The GFP gene was expressed with efficiencies in the order of the unmodified DNA >or= the NLS-conjugated DNA > the unconjugated DNA with the base modification, with both cytosolic and intranuclear microinjections. Thus, we concluded that (i) one or two NLS peptide(s) did not dramatically improve the nuclear entry of DNA and that (ii) chemical modification of DNA reduced the transcription efficiency or stability in the nucleus.

Animals↗

PubMedDate not supplied

A coronary active perfusion system for off-pump coronary artery bypass: advantage over passive perfusion regarding the physiology of the coronary artery.

To avoid myocardial ischemia during off-pump coronary artery bypass, we developed a coronary active perfusion system (CAPS) that perfuses arterial blood to the coronary artery at the diastolic phase of the cardiac cycle by a syringe pump system. We report herein the details and compare CAPS with a passive shunt circuit from the femoral artery (FA shunt), regarding the physiology of the coronary artery. Six pigs were used for this study. After CAPS or FA shunt perfusion was established, coronary flow and coronary pressure were measured. The coronary flows in the native coronary artery, FA shunt perfusion, and CAPS perfusion with syringe pump stroke volume settings ranging from 0.1 to 0.4 ml were 27.2+/-3.0, 4.1+/-1.5, 12.7+/-1.2, 24.8+/-1.9, 33.3+/-1.6, and 42.2+/-1.9 ml/min, respectively. Mean coronary pressures under FA shunt perfusion and CAPS perfusion with stroke settings from 0.1 to 0.4 ml were 23.7+/-4.6, 48.8+/-5.0, 61.3+/-7.5, 70.8+/-5.4, and 82.3+/-5.2 mm Hg, respectively. CAPS had an advantage over the FA shunt regarding coronary flow with safe coronary pressure. Using CAPS, an off-pump coronary artery bypass can be performed safely without myocardial ischemia.

Anastomosis, Surgical↗

PubMedDate not supplied

Surgical treatment of congenital coronary artery fistulas: 27 years' experience and a review of the literature.

BACKGROUND AND AIM OF THE STUDY: A congenital coronary artery fistula (CAF) is a relatively rare congenital anomaly and is defined as an abnormal direct communication between any coronary artery and any of the cardiac chambers. This article reviews our experience over the past 27 years, as well as other literature, and discusses the surgical indications and methods relating to CAF. METHODS: From 1973, 25 patients aged from 2 to 69 years underwent surgical treatment for congenital CAF. Seventeen patients were diagnosed as isolated CAF. All patients under 19 years of age with isolated CAF were asymptomatic. Twenty fistulas originated from the left coronary artery and 9 from the right. The pulmonary artery was the most dominant drainage site. Four patients among the isolated CAF cases were surgically treated without a cardiopulmonary bypass. RESULTS: All patients were discharged from hospital without any perioperative complications. Postoperative coronary angiography was done on all patients with only one slightly residual CAF flow. The average follow-up time was 9.6 years and all patients were asymptomatic and doing well. CONCLUSIONS: Definitive surgical correction is safe and effective, with good results. Therefore, it should be considered even in asymptomatic patients because of the risk of future complications.

Adult↗