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PubMed2006

Contrasting genome-wide distribution of 8-hydroxyguanine and acrolein-modified adenine during oxidative stress-induced renal carcinogenesis.

Oxidative stress is a persistent threat to the genome and is associated with major causes of human mortality, including cancer, atherosclerosis, and aging. Here we established a method to generate libraries of genomic DNA fragments containing oxidatively modified bases by using specific monoclonal antibodies to immunoprecipitate enzyme-digested genome DNA. We applied this technique to two different base modifications, 8-hydroxyguanine and 1,N6-propanoadenine (acrotein-Ade), in a ferric nitrilotriacetate-induced murine renal carcinogenesis model. Renal cortical genomic DNA derived from 10- to 12-week-old male C57BL/6 mice, of untreated control or 6 hours after intraperitoneal injection of 3 mg iron/kg ferric nitrilotriacetate, was enzyme digested, immunoprecipitated, cloned, and mapped to each chromosome. The results revealed that distribution of the two modified bases was not random but differed in terms of chromosomes, gene size, and expression, which could be partially explained by chromosomal territory. In the wild-type mice, low GC content areas were more likely to harbor the two modified bases. Knockout of OGG1, a repair enzyme for genomic 8-hydroxyguanine, increased the amounts of acrolein-Ade as determined by quantitative polymerase chain reaction analyses. This versatile technique would introduce a novel research area as a high-throughput screening method for critical genomic loci under oxidative stress.

Acrolein↗
PubMed1987

[The significance of chemotherapy in oral cancer].

Fifty-three patients with postoperative recurrent epidermoid carcinomas of the head and neck were treated with BEAM, DP, MBD and CAP regimens. Complete response (CR) and partial response (PR) were achieved in 9% and 23% of all evaluable patients, respectively. Especially, in patients with oral cancer (17 cases), the response rate was only 23% (CR 0%, PR 23%). From our results, it was concluded that chemotherapy in any form must be palliative for a patient who has received prior unsuccessful radiotherapy or surgery. For previously untreated patients who present with Stage III or IV advanced carcinoma, combination chemotherapy with CDDP, MTX and PEP should be considered as the initial method of treatment. Adjuvant chemotherapy and intra-arterial chemotherapy for oral cancer should also be considered. Adjuvant chemotherapy combined with UFT and MTX is indicted to be effective for the prevention of local recurrence and micrometastases, but a further randomized study is necessary to confirm these findings.

Adult↗
PubMed1993

[Clinical study of the timed voiding schedule for urinary incontinence in demented elders].

Fourty-one demented elderly patients who had been diapered because of urinary incontinence in Hanwa-Senboku Hospital were treated with the one-week timed voiding schedule. Finally 14 patients (34%) became dry, and free from diapers for more than 1 week after the treatment. With regard to their cystometrogram findings, 10 (67%) of 15 patients having normoactive detrusor function, 3 (14%) of 21 patients having overactive detrusor function and 1 (20%) of 5 patients having underactive detrusor function became free from diapers after the treatment. Therefore a patient with normoactive detrusor function is thought to be a good candidate for the timed voiding schedule. Regardless of their detrusor function, those who showed good responsiveness to the treatment had better activities in daily life (better than D in the Katz index) and small amount of residual urine (less than 80 ml), indicating that not only the detrusor function but also both activities in daily life and the amount of residual urine are the important factors to reestablish voiding behavior in demented elderly patients. The timed voiding schedule is thought to be a useful method to treat incontinence in demented elderly patients, and should be tried first of all, although intermittent catheterization and/or other alternatives would be necessary for those with a large amount of residual urine with poor activities in daily life.

Activities of Daily Living↗
PubMed1998

Improved method for measuring tartrate-resistant acid phosphatase activity in serum.

We describe an improved method for the kinetic measurement of tartrate-resistant acid phosphatase (TrACP; EC 3.1.3.2) activity in serum. Of the TrACP derived from erythrocytes, platelets, and macrophages (osteoclasts and others), that from the first two sources is also resistant to fluoride, whereas skeletal TrACP is sensitive to fluoride. Thus, osteoclast-derived TrACP can be measured specifically by exploiting its sensitivity to fluoride. We measured the activity of tartrate-resistant and fluoride-sensitive acid phosphatase (TrFsACP) by using 2,6-dichloro-4-acetylphenyl phosphate as substrate at pH 6.2. The activity of TrFsACP in serum was increased by adding hexadimethrine bromide (Polybrene) to the reaction mixture. This method was not influenced by hemolysis with hemoglobin concentrations as great as 0.9 g/L. The mean +/- SD values of TrFsACP activity by this method were 20.4 +/- 2.8 and 16.4 +/- 2.3 U/L for young (ages 20-29 years) men (n = 34) and women (n = 50), respectively. The highest mean TrFsACP activity was found among children younger than 15 years, followed by that in elderly subjects (older than 60).

Acid Phosphatase↗
PubMed2006

Diagnosis of dementia using perfusion SPECT imaging at the patient's initial visit to a cognitive disorder clinic.

PURPOSE: This study was performed to evaluate the diagnostic accuracy of brain perfusion SPECT in patients visiting the cognitive disorder clinic for initial evaluation using 3D-SSP compared with using standard transaxial section. MATERIALS AND METHODS: Standard transaxial section displays and 3D-SSP z-score maps obtained after administration of Tc-99m ECD or I-123 IMP were randomly interpreted in 315 patients who visited initially to the cognitive disorder clinic (age 46-88 years; 162 women, 153 men). Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy were calculated and receiver operating characteristic (ROC) analysis was performed. RESULTS: One hundred thirty-seven patients were clinically diagnosed with Alzheimer disease and 178 patients were diagnosed with other disorders and age-associated cognitive decline. Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy calculated using standard transaxial display were 61%, 70%, 61%, 70%, and 66%, respectively, and using the 3D-SSP z-score map were 90%, 74%, 73%, 90%, and 81%, respectively. Particularly, the sensitivity value improved in mild cases compared with severe cases using 3D-SSP. Diagnostic performance with 3D-SSP was superior in both mild dementia (Az = 0.64 [section] vs 0.81 [3D-SSP], P = 0.001) and severe dementia (Az = 0.75 [section] vs 0.90 [3D-SSP], P = 0.002). CONCLUSIONS: Brain perfusion SPECT was useful for diagnosis in patients who come to the cognitive disorder clinic for initial evaluation using 3D-SSP.

Aged↗
PubMed1990

Premature chromosome condensation is induced by a point mutation in the hamster RCC1 gene.

At the nonpermissive temperature, premature chromosome condensation (PCC) occurs in tsBN2 cells derived from the BHK cell line, which can be converted to the Ts+ phenotype by the human RCC1 gene. To prove that the RCC1 gene is the mutant gene in tsBN2 cells, which have RCC1 mRNA and protein of the same sizes as those of BHK cells, RCC1 cDNAs were isolated from BHK and tsBN2 cells and sequenced to search for mutations. The hamster (BHK) RCC1 cDNA encodes a protein of 421 amino acids homologous to the human RCC1 protein. In a comparison of the base sequences of BHK and BN2 RCC1 cDNAs, a single base change, cytosine to thymine (serine to phenylalanine), was found in the 256th codon of BN2 RCC1 cDNA. The same transition was verified in the RCC1 genomic DNA by the polymerase chain reaction method. BHK RCC1 cDNA, but not tsBN2 RCC1 cDNA, complemented the tsBN2 mutation, although both have the same amino acid sequence except for one amino acid at the 256th codon. This amino acid change, serine to phenylalanine, was estimated to cause a profound structural change in the RCC1 protein.

Amino Acid Sequence↗
PubMed2004

Evaluation method of artificial acoustical environment: visualization of sound intensity.

Recent development of sound reproduction systems such as 5.1 surround produces artificial sound field inside the enclosure. The acoustic characteristic of the enclosure is an important factor which determines the quality of the sound field reproduction. This paper provides a useful method for evaluating the characteristics of the sound field in the enclosed space. The proposed method is based on the visualization of the measured sound intensity. In this research, the sound intensity is obtained by measuring the impulse responses from the sound source to the receiving points. The direction and the strength of the intensity can be interpreted as having a strong correlation with the reflected sound from the corresponding direction. The proposed method thus projects that information, i.e., the arriving direction and the strength of the intensity, as the circle having appropriate radius onto the surface of the rectangular wire-frame box. This visualization makes it easy to grasp the fundamental characteristics of the reflections in the enclosure.

Acoustics↗
PubMed1985

Ultrastructural evidence of the existence of the surface connected canalicular system in the thrombocyte of the shark (Triakis scyllia).

The existence of the surface-connected canalicular system has been demonstrated ultrastructurally in shark [Triakis scyllia) thrombocytes using the ruthenium red method to fix and stain the glycocalyx. In ultrathin sections fixed routinely in glutaraldehyde followed by osmium tetroxide, numbers of electron-lucent vesicles are distributed throughout the cytoplasm. Since ruthenium red-positive reaction product is deposited on the inner surface of these vesicles, they are defined as the surface-connected canalicular system. Moreover, the semithin sectioned thrombocytes fixed by the ruthenium red method provide evidence that the surface-connected system is not composed of vesicles but also of numerous canaliculi. These results suggest that the surface-connected canalicular system is one of the most basic organelles of the thrombocyte system, common to submammalian species and to mammals.

Animals↗
PubMed1995

[Separate perfusion of upper and lower body under mild hypothermia during operation on the thoracoabdominal aorta].

During last 7 years, we performed 24 operations on the thoracoabdominal aorta. There were 9 true and 15 dissecting aneurysms. There were two cases of ruptured aneurysm and thoracoabdominal replacement was performed as a last stage operation for total aortic replacement in 4 cases. Three cases with aortic dissection died within 30 days after surgery. Femoro-femoral bypass was used in 4 cases (1 case died of brain damage, paraplegia and MOF), left heart bypass in 5 cases and separate perfusion of upper and lower body (SPULB) under deep hypothermia in 7 cases (2 cases died of LOS and cerebrovascular accident occurred at 2 weeks after operation) and SPULB with mild hypothermia in 8 cases for circulatory support. There was one case of renal dysfunction and transient mild liver dysfunction occurred in 7 cases. There was no evidence on relationship between surgical outcome and methods of circulatory supports, but we recently prefer SPULB under mild hypothermia for thoracoabdominal surgery since intraoperative massive bleeding and cardiac arrest can be easily treated and major organs can be protected by introducing hypothermia in this perfusion technique.

Adult↗
PubMed1995

Pharmacokinetic study of trimethadione and its metabolite in blood, liver and brain by microdialysis in conscious, unrestrained rats.

A microdialysis method has been developed in the past two decades to determine levels of drug and endogenous compounds in several organs under physiological conditions. In this study, we determined the pharmacokinetics of the model drug, trimethadione (TMO), and its only metabolite, dimethadione (DMO), in liver, blood and brain by the microdialysis method in freely-moving rats. Sampling times were extended up to 24 hours. The construction of a newly developed microdialysis probe for liver and blood is described. The elimination patterns of TMO in liver, blood and brain dialyzates were almost identical and the calculated t1/2 was approximately 3 hr in each sample. However, in brain, tmax was delayed compared with the others while the relative concentration of DMO (AUC0-24h) was lower in brain compared with liver and blood. These studies suggest that this concurrent and successive microdialysis sampling method will not only be a useful tool for pharmacokinetic and drug metabolism studies in the organs of small animals but will also decrease the number of experimental animals needed for a study.

Animals↗
PubMed2005

The GABA type A receptor alpha5 subunit gene is associated with bipolar I disorder.

Several genetic studies have revealed that bipolar disorders are linked with the chromosomal locus of 15q11-q13, where the gamma-aminobutyric acid (GABA) receptor alpha5 subunit gene (GABRA5) locates. GABA is one of the major neurotransmitters that may be involved in the pathogenesis of bipolar disorder. Five polymorphisms in the GABRA5 gene, -754C>T in the promoter region, IVS1-21G>A, IVS2-26T>A, (*)302C>T in 3'-UTR of exon 5, and a CA repeat polymorphism in the 3' flanking region were examined in a Japanese population. IVS1-21G>A exhibited significant differences in the distribution of the genotype and allele frequency in bipolar I disorder patients but not in bipolar II disorder patients, compared with control subjects. The haplotype analysis showed that IVS1-21G>A/IVS2-26A>T was associated with bipolar I disorder, and the IVS1-21A/IVS2-26T haplotype was a negative risk factor for susceptibility to the disorders (odds ratio: 0.57, 95% confidence interval: 0.44-0.73). These results suggest that the GABRA5 gene may confer susceptibility to bipolar I disorder.

Bipolar Disorder↗
PubMed1991

Automated two-dimensional liquid chromatographic system for mapping proteins in highly complex mixtures.

An automated two-dimensional liquid chromatographic system was developed for systematic protein separations which could serve for analytical mapping and preparative separations of proteins. The system applies the principles of the column-switching technique, and consists of two different columns connected in tandem through an electrical column switching valve, two pumping systems to operate each column independently and a system controller to perform sequential chromatography on the two columns. A protein mixture is applied to the first-dimensional anion-exchange column and is separated by stepwise elution with an increasing sodium chloride concentration. The eluent is introduced directly to the second-dimensional reversed-phase column, and is further separated by gradient elution with an increasing acetonitrile concentration. The two elution stages are synchronized by a computer program. By this system, very complex protein mixtures such as crude cerebellar extracts were resolved reproducibly into ca. 200 peaks within 12 h. The method can be used for the total analysis of proteins in various tissues and cells without complicated premanupulation of samples, and allows the simultaneous analysis of a protein isolated by chromatography. The isolated protein is most suitable for use in the strategy of protein and gene sequence analysis.

Animals↗
PubMed1979

Frozen ultrathin-sections for x-ray microanalysis of rat tooth germs.

In order to detect the precise location and pathways of calcium in the ameloblastic layer with EM and EDX, frozen ultrathin-sections, potassium pyroantimonate (PPA) method and 45Ca-autoradiography (A.R) were employed. A population of dense granules containing Ca and P was mainly observed within the nuclei and mitochondria of the frozen-sectioned ameloblasts, whereas PPA-reaction was seen not only within them but in ER, Golgi vesicles and intercellular spaces. No granules were found at the enamel matrix by both methods, but peaks for Ca and P were distinct. Silver grains of 45Ca-A.R were located in nuclei, mitochondria. Golgi complex and intercellular spaces. These results indicate that calcium can transport across the ameloblastic layer through two main pathways, extracellular and transcellular.

Ameloblasts↗
PubMed1997

Pharmacokinetic properties of YM17E, an inhibitor of acyl coenzyme A: cholesterol acyl transferase, and serum cholesterol levels in healthy volunteers.

We conducted a single and repeat oral dose study of YM17E, a novel inhibitor of acyl coenzyme A (CoA): cholesterol acyltransferase, in healthy male volunteers to evaluate the pharmacokinetic profile, tolerability and effect of the drug on serum cholesterol. In the single administration study, YM17E was administered after a meal to two groups of subjects (each containing six subjects taking the drug and three taking placebo) receiving 3, 60 and 300 mg or 15, 60 and 450 mg YM17E, respectively. Plasma concentrations of unchanged drug following single oral administration at 3-300 mg after a meal increased with increasing dose. In contrast, plasma concentrations after administration of 450 mg were almost the same as after 300 mg. Unchanged YM17E was not detected in urine after single administration, suggesting that it was excreted via the bile or urine after metabolism. Five active metabolites (M1, M2-a, M2-b, M3 and M4) were observed in plasma at concentrations comparable to those of unchanged YM17E. Their plasma concentrations increased in a slightly greater than dose-dependent manner from 3 to 300 mg. The effect of food was studied in an open crossover design with a 1-week washout period. Twelve subjects received 150 mg YM17E in both the fasted and post-prandial states. The AUC and Cmax after fasting were closely similar to those after a meal, showing that bioavailability was not affected by food intake. In the repeated oral dose study, the subjects received test drug at 150 mg or 300 mg (n = 6 each) or placebo (n = 3) twice a day (after breakfast and after dinner) for 7 days. On days 1 and 7, the subjects received YM17E once a day (after breakfast) for evaluation of pharmacokinetic properties. After repeated oral administration of 150 mg b.d., plasma concentrations reached steady state by day 5 (mean Cmin 48.6 ng.ml-1). After repeated administration of 300 mg b.d., plasma concentrations prior to each daily morning dose increased up to the 5th day (mean Cmin 166.6 ng.ml-1) and then tended to decrease until the 7th day. No significant signs, symptoms or changes in serum cholesterol levels were observed during the single and repeated oral dose studies at 150 mg b.d. Although statistical analysis was not conducted because of the small number of subjects, all subjects receiving repeated oral administration of 300 mg twice daily showed a 25% decrease in serum cholesterol level on day 7, but also the simultaneous occurrence of diarrhoea.

Administration, Oral↗
PubMed1985

Synthesis of 1-beta-D-ribofuranosyl-1,2-dihydropyrimidin-2-one derivatives and their biological activities.

Several alkyl substituted 1-beta-D-ribofuranosyl-1,2-dihydropyrimidin-2-one derivatives were synthesized by the method of stannic chloride-catalyzed glycosidation method to elucidate their inhibitory activity of cytidine deaminase and also their antitumor activities in vitro and in vivo. Alkyl substitution at position 4 or 6 of the derivatives decreased their inhibitory activity for cytidine deaminase and also decreased antitumor activity against L1210 cells in vitro.

Animals↗
PubMed1998

Pharmacodynamics and pharmacokinetics of a single oral dose of nitrazepam in healthy volunteers: an interethnic comparative study between Japanese and European volunteers.

Potential interethnic differences in drug disposition and effects between Japanese and white subjects hamper the registration in Japan of medications already used in Western countries. This double-blind, placebo-controlled, crossover study was conducted to compare the pharmacodynamics and pharmacokinetics of a single oral dose of nitrazepam (5 mg) in age- and sex-matched Japanese (n = 8) and white (n = 8) healthy volunteers. The study was performed in centers in Japan and the Netherlands using the same methods and study design. Subjects were individually matched for gender, age, and body stature. Drug effects were measured by means of saccadic and smooth pursuit eye movements and visual analog lines obtained from the scales of Bond and Lader. There were no pharmacokinetic differences between the Japanese and white subjects. Clearance of nitrazepam was 0.91 +/- 0.165 mL/min/kg and 1.17 +/- 0.492 mL/min/kg, and half-life (t1/2) was 22.1 +/- 4.96 hours and 21.5 +/- 7.51 hours for the Japanese and European groups, respectively. Pharmacokinetic parameters showed no significant correlation with age, height, or weight. The average time-effect curves for the different parameters were comparable between groups. Compared with placebo, both groups showed similar significant reductions in average peak velocity and increases in saccadic inaccuracy and reaction time. Visual analog scores showed clear sedation in the white subjects, but insignificant effects in the Japanese subjects. Smooth pursuit did not change significantly in either group. Slope and intercept of the concentration-effect relationships for saccadic peak velocity showed considerable intersubject variability, but no clear differences between groups. The pharmacokinetics and pharmacodynamics of nitrazepam were similar in matched healthy Japanese and white subjects. Interethnic comparative studies are feasible, and provide meaningful information about potential racial differences in disposition and action of drugs. Such studies can form a rational basis for comparative clinical trials.

Administration, Oral↗
PubMed1993

[A basic study on a highly sensitive automated method for hypercoagulable state in plasma, fluorogenic prothrombin time (FPT) method].

A highly sensitive automated method, fluorogenic prothrombin time (FPT) method, was developed by a combination of a fluorogenic peptide substrate for thrombin and a centrifugal autoanalyzer (Cobas Bio). Using plasmas from stroke patients, we showed that the second reagent containing fluorogenic peptide substrate should be mixed after the first reagent containing tissue factor and plasma were mixed, that is, two steps method, in order to detect hypercoagulable state. When the first and the second reagent were mixed with plasma at the same time, that is, one step method, FPT was not sensitive to hypercoagulable state. We also showed that patient plasmas should be stored at -80 degrees C and subjected to FPT analysis immediately after thawing, not leaving at 4 degrees C or room temperature. Good correlation was observed on FPT of stroke patients using human tissue factor and bovine tissue factor. Another fluorogenic method was developed by the same principle for the evaluation of factors X and VII concentrations in plasma using deficient plasma of factor X or factor VII. Good correlation was observed on factors X and VII concentrations of stroke patients using human tissue factor and bovine tissue factor.

Blood Coagulation Tests↗