Search PubMed⌕ Search

Biomedical subjects

Hiroki Kubota

Publications and source records attributed to Hiroki Kubota.

4 recordsLinked to original sources

Role of mitochondria in the generation of spontaneous activity in detrusor smooth muscles of the Guinea pig bladder.

PURPOSE: The rhythmic electrical activity of gastrointestinal smooth muscles is associated with mitochondrial Ca2+ handling. We examined the role of mitochondria in the generation of spontaneous activity in detrusor smooth muscles. MATERIALS AND METHODS: Changes in the membrane potential and intracellular Ca2+ concentration ([Ca2+]i) were measured in detrusor smooth muscles of the guinea pig using conventional microelectrode techniques and Fura-PE3 (Calbiochem, San Diego, California) fluorescence, respectively. RESULTS: Detrusor smooth muscle cells showed spontaneous action potentials and associated transient increases in [Ca2+]i (Ca transients). The mitochondrial protonophore CCCP (carbonyl cyanide m-chlorophenyl hydrazone) (10 microM) depolarized the membrane, increased [Ca2+]i and caused activation followed by suppression of action potentials and Ca transients. High K solution potassium concentration ([K+]o = 30 mM) depolarized the membrane and increased [Ca2+]i to levels similar to those produced by 10 microM CCCP but this depolarization did not suppress action potentials. Nifedipine (10 microM) decreased the amplitude of CCCP induced increases in [Ca2+]i by about 50%. CCCP induced increases in [Ca2+]i were further reduced by about 70% in Ca2+-free solution and by about 30% in the presence of 10 microM SKF96365, a blocker for store operated Ca entry. In the presence of 10 microM nifedipine and 10 microM cyclopiazonic acid, CCCP induced [Ca2+]i responses were suppressed to about 25% of control values. Under these conditions repetitive applications of 10 microM acetylcholine chloride successively decreased [Ca2+]i responses and finally failed to increase [Ca2+]i. Subsequent CCCP failed to elevate [Ca2+]i. CONCLUSIONS: These results suggest that mitochondria have an important role in Ca2+ buffering in bladder smooth muscles. Mitochondrial Ca2+ is presumably supplied by Ca2+ transport from internal stores and also by capacitative calcium entry through nonselective cation channels. Mitochondrial Ca2+ handling may also be critical for the generation of spontaneous activity in detrusor smooth muscle.

Action Potentials↗

[Improved sensitive determination method for bromate in bread].

An effective clean-up procedure was developed to determine trace levels of bromate in bread by high-performance liquid chromatography with post-column flow reactor detection. Bromate was extracted from bread with deionized pure water. After centrifugation, the supernatant was filtered through a paper filter. The filtrate was filtered through a 0.2 micron nylon filter and chloride ion was removed by an IC-SP M Ag cartridge column or On-Guard Ag cartridge column. The eluate was applied to an Oasis MAX anion exchange cartridge column. The column was washed with 20% acetic acid and water. Bromate was then eluted with 0.5% sodium nitrate solution. The eluate was determined by HPLC with post column flow reactor detection. The method had a quantitation limit of 2 ng/g in bread products. Recoveries of bromate from bread ranged from 68 to 72% at a spiked bromate level of 2-10 ng/g.

Bread↗

Specific determination of bromate in bread by ion chromatography with ICP-MS.

A sensitive method for detecting bromate in bread by ion chromatography with inductively-coupled plasma mass spectrometry (IC/ICP-MS) was developed. Bromate was extracted from bread with water. The clean-up procedure included a 0.2 micron filter, a C18 cartridge for defatting, a silver cartridge to remove halogen anions, a centrifugal ultrafiltration unit to remove proteins, and a cation-exchange cartridge to remove silver ions. A 500 microL sample solution was applied to IC/ICP-MS. The detection limit and the quantitation limit of bromate in the solution were 0.3 ng/mL and 1.0 ng/mL, expressed as HBrO3, respectively, which corresponded to 2 ng/g and 5 ng/g, respectively, in bread. Recovery of bromate was about 90%, and the CV was about 2%. Based on the detection limit in solution and recovery from bread, the detection limit of bromate in bread was estimated to be 2 ng/g.

Bread↗

Estimation of concentrations of antifungal agents allowed as food additives in foods and their daily intake based on official inspection results in Japan in fiscal year 1998.

The mean concentrations and daily intake of four antifungal agents were estimated based on the results of an analysis of 7,005 samples of food obtained in official inspections by Japanese local governments in fiscal year 1998. The mean concentration of diphenyl was 0.0004% of the allowable limit, and those of imazalil, o-phenylphenol, and thiabendazole were 14.0%, 3.5%, and 5.7%, respectively. The daily intakes of these antifungal agents per person, estimated from their concentrations and the daily consumption of the foods, were 0.000326, 1.89, 11.5, and 23.3 micrograms, respectively, and assuming a body weight of 50 kg, the amounts of these antifungal agents consumed were 0.000013%, 0.15%, 0.12%, and 0.47% of the acceptable daily intake, respectively. These values are similar to the values obtained on the basis of the results of the official inspection in fiscal years 1994 and 1996, except that the amount of diphenyl is much lower (1/100).

Antifungal Agents↗