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N Kinoshita

Publications and source records attributed to N Kinoshita.

At least 37 records · Page 2Linked to original sources
PubMed2000

Molecular cloning and characterization of Xenopus RGS5.

We identified six genes that encode putative RGS proteins (XRGSI-VI) in developing Xenopus embryos using PCR amplification with degenerate primers corresponding to the conserved region (RGS domain) of known RGS proteins. RT-PCR analysis revealed that mRNAs of these XRGSs are differentially expressed during embryogenesis. At stage 1, only XRGSII mRNA was detected. On the other hand, expression of XRGSVI mRNA increased apparently at stage 14 and expression of three of other XRGS (III, IV, V) elevated between stage 25 and 40. To further characterize XRGS proteins expressed in Xenopus embryos, we isolated a cDNA clone for XRGSIII. Based on determined nucleotide sequence, XRGSIII was considered as a Xenopus homologue of mammalian RGS5 (XRGS5). Genetic analysis using the pheromone response halo assay showed that expression of XRGS5 inhibits yeast response to alpha-factor, suggesting that XRGS5 negatively regulates the G-protein-mediated signaling pathway in developing Xenopus embryos.

Amino Acid Sequence↗
PubMed2000

Identification of 187 single nucleotide polymorphisms (SNPs) among 41 candidate genes for ischemic heart disease in the Japanese population.

To investigate whether common variants in the human genetic background are associated with pathogenesis of ischemic heart diseases, we systematically surveyed 41 possible candidate genes for single-nucleotide polymorphisms (SNPs) by directly sequencing 96 independent alleles at each locus, derived from 48 unrelated Japanese patients with myocardial infarction, including 25.8 kb 5' flanking regions, 56.8 kb exonic and 35.4 kb intronic sequences, and 1.8 kb 3' flanking regions. In this genomic DNA of nearly 120 kb, we identified 187 SNPs: 55 in 5' flanking regions, seven in 5' untranslated regions (UTRs), 52 in coding elements, 64 in introns, eight in 3' UTRs, and one in a 3' flanking region. Among the 52 coding SNPs, 26 were non-synonymous changes. Allelic frequencies of some of the polymorphisms were significantly different from those reported in European populations. For example, the Q506R substitution in the coagulation factor V gene, the so-called "Leiden mutation", has a reported frequency of 2.3% in Europeans, but we detected the Leiden mutation in none of the Japanese genomes that we investigated. The allelic frequencies of the -33A>G SNP in the thrombomodulin gene were also very different; this allele occurred at a 12% frequency in the Japanese patients that we examined, although it had been detected in none of 82 Caucasians reported previously. These data support the hypothesis that some SNPs are specific to particular ethnic groups.

3' Untranslated Regions↗
PubMed2000

Aortic root dilatation among young competitive athletes: echocardiographic screening of 1929 athletes between 15 and 34 years of age.

BACKGROUND: Aortic dilatation can be lethal for young competitive athletes. The prevalence among athletes is not known, however, and thus a reasonable approach to early recognition remains uncertain. METHODS AND RESULTS: Echocardiograms of 1929 normotensive athletes 15 to 34 years of age were analyzed. Five (0.26%) athletes had aortic dilatation; 4 of the 5 played basketball. This made the prevalence of aortic dilatation 0.96% (4 of 415) among basketball and volleyball players, who represented a population of especially tall athletes. Tallness aside, only 2 of the 5 athletes had features of Marfan syndrome. Among the athletes without aortic dilatation, the relation between body surface area and aortic root dimension was nonlinear and best described with a quadratic regression model. Athletes with aortic dilatation fell well outside the 95% confidence interval. CONCLUSION: Because a higher incidence of aortic dilatation is to be anticipated among very tall athletes, inclusion of echocardiography in screening before participation in certain sports should be considered.

Adolescent↗
PubMed2000

Physiological profile of middle-aged and older climbers who ascended Gasherbrum II, an 8035-m Himalayan peak.

BACKGROUND: The purpose of this study was to investigate the physiological characteristics of a group of middle-aged and older Japanese climbers who ascended Gasherbrum II, an 8035-m peak in the Karakoram Range of the Himalayas. METHODS: Body composition, cardiac structure, and respiratory gas exchange during exercise were estimated in eight climbers with differing levels of experience (seven men and one woman, aged 54 to 63 years) 6 months before their expedition. RESULTS: Using supplementary O2, the four experienced climbers ascended beyond Camp 4 (7400 m) without showing any health problems and were able to attempt the summit. In contrast, the others, who had minimal experience at extreme altitude, suffered from altitude sickness on the way to Camp 4. Body mass index values were relatively high, but their low percentage of body fat (14.9%-21.4%) was indicative of the climbers' substantial lean body weight. Cardiac structures were generally normal, although three experienced male climbers had borderline hypertension and eccentric hypertrophy of the left ventricle. Peak VO2 ranged from 30.9 to 45.6 ml/kg/min, and no significant relationship between fitness level and the success or failure of the ascent was evident. CONCLUSIONS: Even sexagenarians are capable of safely climbing 8000-m peaks with supplementary O2. An exceptionally high fitness level, as is seen in elite older athletes, does not appear to be required. What is essential, however, is moderate fitness, good health, and extensive experience.

Aged↗
PubMed2000

Peak aerobic performance and left ventricular morphological characteristics in university students.

OBJECTIVE: To determine whether a relationship exists between left ventricular morphology and aerobic capacity in large numbers of male university students with a physically inactive and active life style. DESIGN: A prospective study. SETTING: Sports medicine research center. PARTICIPANTS: Eleven sedentary normal-weight university students, 17 sedentary overweight university students, and 215 university athletes. MAIN OUTCOME MEASURES: After the echocardiographic examination, an incremental treadmill exercise test until exhaustion was performed to measure peak oxygen uptake (VO2). RESULTS: In sedentary students, absolute peak VO2 in the overweight students was slightly higher than that in normal-weight students (3,024 vs. 2.912 ml/min). Relative peak VO2 (ml/min/kg) was highly negatively correlated with body mass index (kg/m2) in a total of 28 sedentary students. The correlation between absolute peak VO2 and left ventricular dimension was weak in the sedentary overweight students; however, a correlation coefficient of 0.55 was obtained in athletic students. A stepwise multiple regression showed significant determinants of absolute peak VO2 in athletic students for body surface area (45%), left ventricular dimension (7%), and certain sports (6%). CONCLUSIONS: A physically active life style plays a role in increasing both aerobic capacity and left ventricular enlargement. Body size appeared to be a potent stimulus to left ventricular enlargement.

Adult↗
PubMed2000

Can serum carnitine levels distinguish hypertrophic cardiomyopathy from hypertensive hearts?

Although echocardiography is a useful diagnostic tool in hypertrophic cardiomyopathy (HCM), it is sometimes difficult to differentiate it from hypertensive heart disease (HHD): some patients with HCM show symmetrical hypertrophy, whereas patients with HHD sometimes show asymmetrical septal hypertrophy. We used a radioiodinated long-chain fatty acid tracer to visualize the altered myocardial fatty acid metabolism of HCM and HHD. Carnitine is the essential substance for the beta-oxidation of long-chain fatty acids. We recently reported that serum free carnitine levels in HCM were elevated and that they were significantly correlated with the severity of myocardial fatty acid metabolic disorder. Therefore, we investigated serum carnitine levels in patients with HCM and HHD, which can contribute to the differentiation of each other. We studied 56 patients with HCM and 20 patients with essential hypertension. Serum free carnitine levels were significantly higher in patients with HCM than those with HHD (HCM 52.5+/-9.5 nmol/mL, HHD 46.6+/-6.4 nmol/mL, P<0.01), but they showed no statistical difference between patients with HHD and normal subjects. Serum acylcarnitine levels were significantly lower in patients with HCM than those with HHD (HCM 10.1+/-4.0 nmol/mL, HHD 14.5+/-4.9 nmol/mL, P<0.0005), although they did not differ between patients with HHD and normal subjects. Scintigraphic analyses with a long-chain fatty acid analog revealed that myocardial tracer uptake was much reduced in patients with HCM compared with that in patients with HHD (quantitative analysis: HCM 2.11+/-0.12, HHD 2.22+/-0.17, P<0.05; semiquantitative analysis: HCM 13.6+/-6.3, HHD 2.0+/-1.5, P<0.0001). In conclusion, the differences in serum carnitine levels between HCM and HHD reflect altered myocardial fatty acid metabolic impairment, and the levels can help to distinguish these 2 diseases.

Adult↗
PubMed2000

The risk for second primaries in gastric cancer patients: adjuvant therapy and habitual smoking and drinking.

STUDY PURPOSE: To examine whether adjuvant therapy for gastric cancer increases the risk for second primaries, and whether smoking and drinking increase the risk. SUBJECTS AND METHODS: 1,631 patients who were newly diagnosed with gastric cancer and underwent curative operation at Osaka Medical Center for Cancer and Cardiovascular Diseases during 1978-92. Incidence of second primaries were examined through linkage to Osaka Cancer Registry as of the end of December 1995. Observed number of second primaries (O) was compared with the expected according to the incidence in general population (E). Proportional hazards model was used to estimate hazard rate ratio (HR) for second primaries. RESULTS: 149 second primaries were observed. Seventeen cases, detected within 2 months after diagnosis of gastric cancer, were excluded. O/E ratio was 1.13 for adjuvant chemotherapy, 0.93 for immuno-chemotherapy, and 0.78 for immunotherapy, while 1.14 for operation only (not significant). Age-, sex-, and stage-adjusted HR was 1.02 (95% C.I. 0.69-1.50) for chemotherapy, 0.80 (0.41-1.57) for immuno-chemotherapy, and 0.60 (0.08-4.34) for immunotherapy, as compared with the risk for operation only. Among males, HR for current smokers vs. never smokers was 1.82 (1.02-3.26). CONCLUSIONS: Adjuvant therapy for gastric cancer did not significantly increase the risk for second primaries, while smoking elevated it.

Adolescent↗
PubMed2000

Seasonal variation in mortality from ischemic heart disease and cerebrovascular disease in Okinawa and Osaka: the possible role of air temperature.

Okinawa is located in a subtropical area and is well-known for low mortality due to ischemic heart disease (IH-D) and cerebrovascular disease (CVD). However, the factors that contribute to these low mortality rates remain unclear. We examined the seasonal variation in the mortality due to IHD and CVD among Okinawa and Osaka residents, aged 45 to 84 years, between 1992 and 1996. In addition, we studied if there was a relationship between the monthly mortality rate from IHD or CVD and the monthly mean daily air temperature in Naha City and Osaka City. Data on the monthly mean daily air temperature was obtained from the meteorological stations in Naha City and Osaka City. Our results showed that there were inverse correlations between the monthly mean daily temperature in a city and each of the monthly mortality from IHD in Okinawa (r=-0.794, p<0.01), the monthly mortality from CVD in Okinawa (r=-0.837, p<0.001), and the monthly mortality from CVD in Osaka (r=-0.954, p<0.001). In Osaka, the monthly mortality rate from IHD was at or near its minimum value when the mean daily temperature was approximately 25 degrees C (in September), and it increased in a linear fashion as the mean monthly temperature fell (r=-0.975, p<0.001). The difference between the monthly mortality from IHD or CVD among the Okinawa and the Osaka residents increased in the winter season in comparison with that in the other seasons, with the exception for IHD in July and in August. These findings indicate that the lower mortality from IHD and CVD in Okinawa is affected, at least in part, by Okinawa's warm winter.

Aged↗
PubMed2000

Tubelactomicin A, a novel 16-membered lactone antibiotic, from Nocardia sp. I. Taxonomy, production, isolation and biological properties.

A novel 16-membered lactone antibiotic named tubelactomicin A was isolated from the culture broth of an actinomycete strain. The producing organism, designated MK703-102F1, was identified as a member of Nocardia. Tubelactomicin A was isolated from the culture broth by Diaion HP20 absorption, ethyl acetate extraction, silica gel and Sephadex LH-20 column chromatographies and centrifugal liquid-liquid partition chromatography (CPC). Tubelactomicin A showed strong activity against acid-fast bacteria including the drug-resistant strains.

Anti-Bacterial Agents↗
PubMed2000

ABC transporter genes, kasKLM, responsible for self-resistance of a kasugamycin producer strain.

We previously reported that a 7.6-kb DNA fragment from Streptomyces kasugaensis M338-M1, a kasugamycin (KSM) producer, included KSM acetyltransferase gene (kac338) and some other genes possibly involved in KSM biosynthesis. As an extension of that study, a 10-kb SacI-KpnI DNA fragment, located approximately 5-15-kb upstream of kac33, was cloned and a 4.2-kb SacI-EcoRI fragment therefrom was sequenced, revealing one incomplete (designated ORF J) and three complete open reading frames (designated kasK, kasL and kasM). The coding frames of kasK, L and M overlap one another with terminator/initiator ATGA sequence. RT-PCR analysis of a DNA region including kasKLM indicated the presence of one transcript that is long enough to span the three genes. The kasK gene potentially encodes an ATP-binding protein of the ATP-binding cassette (ABC) transporter superfamily. Homology search for the deduced KasK protein shows similarity to other ABC transporters involved in self-resistance of a mithramycin and possibly doxorubicin producer strain. The kasL and kasM genes encode different integral membrane proteins, both having six putative transmembrane helices. An expression plasmid for kasKLM (pTV-KLM) was constructed and these genes were expressed in E. coli JM 109, which had been sensitive to KSM. The transformant acquired resistance to KSM, suggesting that KasK, L and M proteins as a set in S. kasugaensis M338-M1 pump out KSM to protect the producer from its toxic metabolite.

ATP-Binding Cassette Transporters↗
PubMed2000

MK800-62F1, a new inhibitor of apoptotic cell death, from Streptomyces diastatochromogenes MK800-62F1. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological activity.

A new compound, MK800-62F1, was isolated from a cultured broth of Streptomyces diastatochromogenes MK800-62F1. It inhibited H2O2-induced apoptosis in human small cell lung carcinoma Ms-1 cells as well as in human T-cell leukemia Jurkat cells. In addition, MK800-62F1 also inhibited camptothecin-induced apoptosis in Jurkat cells, which was mediated by intracellular H2O2 generation. MK800-62F1 did not exhibit antioxidative activity in vitro, suggesting that inhibition of apoptosis by MK800-62F1 was not due to the scavenging of H2O2, rather it was due to the modulation of the downstream event of H2O2 generation.

Antineoplastic Agents, Phytogenic↗
PubMed2000

[Impairment of septal uptake and washout on 123I-BMIPP myocardial SPECT after pacemaker implantation: report of two cases].

Case 1 involved a 62-year-old woman with complete atrioventricular block and case 2 involved a 70-year-old woman with sick sinus syndrome. In both cases, echocardiography, 123I-15-(p-iodophenyl)-3-R,S-methyl pentadecanoic acid (BMIPP) and 99mTc-tetrofosmin myocardial single photon emission computed tomographic (SPECT) findings were normal. Coronary arteriography and left ventriculography also revealed normal in both cases. After pacemaker implantation (DDD-type in case 1 and VVI-type in case 2), both patient's electrocardiograms revealed left bundle branch block-type conduction disturbance, and echocardiography showed asynchronous hypokinesia in the septal region. One year later, although both patient's ATP loading tetrofosmin SPECT were normal, BMIPP SPECT showed reduced uptake and increased washout in the septal region. These findings suggest that these changes of BMIPP SPECT might be caused by left bundle branch block-type conduction disturbance after pacemaker implantation.

Aged↗
PubMed1999

Effects of Hydrophobizing Methods of Surfaces on the Interaction in Aqueous Solutions.

The hydrophobic interaction between the surfaces hydrophobized by the adsorption of STAC in a STAC solution (system I) and the interaction between the surfaces hydrophobized by the reaction with OTS in water (system II) were investigated using an atomic force microscope, and their difference was compared. Clear differences of the interaction between systems I and II were found with respect to the force curves and the morphology of the surfactants adsorbed on the surfaces. It is postulated that the hydrophobic interaction for system II is attributable to the cavity bridging between the surfaces, but that for system I it is originated from a different mechanism. It is also implied that the existence of the surfactants in the bulk will play an important role for the hydrophobic interaction. Copyright 1999 Academic Press.

Journal Article↗
PubMed1999

Structural change of threonine 89 upon photoisomerization in bacteriorhodopsin as revealed by polarized FTIR spectroscopy.

The all-trans to 13-cis photoisomerization of the retinal chromophore of bacteriorhodopsin occurs selectively, efficiently, and on an ultrafast time scale. The reaction is facilitated by the surrounding protein matrix which undergoes further structural changes during the proton-transporting reaction cycle. Low-temperature polarized Fourier transform infrared difference spectra between bacteriorhodopsin and the K intermediate provide the possibility to investigate such structural changes, by probing O-H and N-H stretching vibrations [Kandori, Kinoshita, Shichida, and Maeda (1998) J. Phys. Chem. B 102, 7899-7905]. The measurements of [3-18O]threonine-labeled bacteriorhodopsin revealed that one of the D2O-sensitive bands (2506 cm(-1) in bacteriorhodopsin and 2466 cm(-1) in the K intermediate, in D2O exhibited 18(O)-induced isotope shift. The O-H stretching vibrations of the threonine side chain correspond to 3378 cm(-1) in bacteriorhodopsin and to 3317 cm(-1) in the K intermediate, indicating that hydrogen bonding becomes stronger after the photoisomerization. The O-H stretch frequency of neat secondary alcohol is 3340-3355 cm(-1). The O-H stretch bands are preserved in the T46V, T90V, T142N, T178N, and T205V mutant proteins, but diminished in T89A and T89C, and slightly shifted in T89S. Thus, the observed O-H stretching vibration originates from Thr89. This is consistent with the atomic structure of this region, and the change of the S-H stretching vibration of the T89C mutant in the K intermediate [Kandori, Kinoshita, Shichida, Maeda, Needleman, and Lanyi (1998) J. Am. Chem. Soc. 120, 5828-5829]. We conclude that all-trans to 13-cis isomerization causes shortening of the hydrogen bond between the OH group of Thr89 and a carboxyl oxygen atom of Asp85.

Aspartic Acid↗
PubMed1999

Phosphorylation of adducin by Rho-kinase plays a crucial role in cell motility.

Adducin is a membrane skeletal protein that binds to actin filaments (F-actin) and thereby promotes the association of spectrin with F-actin to form a spectrin-actin meshwork beneath plasma membranes such as ruffling membranes. Rho-associated kinase (Rho- kinase), which is activated by the small guanosine triphosphatase Rho, phosphorylates alpha-adducin and thereby enhances the F-actin-binding activity of alpha-adducin in vitro. Here we identified the sites of phosphorylation of alpha-adducin by Rho-kinase as Thr445 and Thr480. We prepared antibody that specifically recognized alpha-adducin phosphorylated at Thr445, and found by use of this antibody that Rho-kinase phosphorylated alpha-adducin at Thr445 in COS7 cells in a Rho-dependent manner. Phosphorylated alpha-adducin accumulated in the membrane ruffling area of Madin-Darby canine kidney (MDCK) epithelial cells and the leading edge of scattering cells during the action of tetradecanoylphorbol-13-acetate (TPA) or hepatocyte growth factor (HGF). The microinjection of Botulinum C3 ADP-ribosyl-transferase, dominant negative Rho-kinase, or alpha-adducinT445A,T480A (substitution of Thr445 and Thr480 by Ala) inhibited the TPA-induced membrane ruffling in MDCK cells and wound-induced migration in NRK49F cells. alpha-AdducinT445D,T480D (substitution of Thr445 and Thr480 by Asp), but not alpha-adducinT445A,T480A, counteracted the inhibitory effect of the dominant negative Rho-kinase on the TPA-induced membrane ruffling in MDCK cells. Taken together, these results indicate that Rho-kinase phosphorylates alpha-adducin downstream of Rho in vivo, and that the phosphorylation of adducin by Rho-kinase plays a crucial role in the regulation of membrane ruffling and cell motility.

Amino Acid Sequence↗