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Biomedical subjects

Hiroyuki Kataoka

Publications and source records attributed to Hiroyuki Kataoka.

15 recordsLinked to original sources

Detection of aminophenylnorharman, a possible endogenous mutagenic and carcinogenic compound, in human urine samples.

Mutagenic/carcinogenic 9-(4'-aminophenyl)-9H-pyrido[3,4-b]indole [aminophenylnorharman (APNH)] is formed from norharman and aniline in the presence of cytochrome P450 3A4/1A2. Because both precursors are widely distributed in the environment, human exposure is unavoidable. To clarify APNH formation in the human body, amounts of the compound in 24-h human urine collected from smokers and nonsmokers, eating a normal diet, were analyzed by liquid chromatography/electrospray ionization tandem mass spectrometry. In addition, norharman and aniline were also analyzed by high-performance liquid chromatography and gas chromatography, respectively. APNH could be detected in all urine samples at levels 49 to 449 pg for smokers and 21 to 594 pg for nonsmokers per 24-h urine, respectively. The amounts of norharman and aniline were 46 to 185 ng and 0.70 to 8.10 microg for smokers and 52 to 447 ng and 0.49 to 5.72 microg for nonsmokers, respectively, per 24-h urine (none of the levels differing significantly between smokers and nonsmokers). To exclude exogenous exposure to norharman and aniline, we analyzed the levels of APNH, norharman, and aniline in urine samples collected from inpatients receiving parenteral alimentation. Similar to the healthy volunteers, all urine samples contained 12 to 338 pg of APNH, 6 to 75 ng of norharman, and 0.33 to 1.86 microg of aniline per 24-h urine. These results suggest that APNH should be considered as a novel endogenous mutagen/carcinogen; thus, it is very important to determine the biological significance of this carcinogen for human cancer development.

Adult↗

Dynamic deformation and recovery response of red blood cells to a cyclically reversing shear flow: Effects of frequency of cyclically reversing shear flow and shear stress level.

Dynamic deformation and recovery responses of red blood cells (RBCs) to a cyclically reversing shear flow generated in a 30-microm clearance, with the peak shear stress of 53, 108, 161, and 274 Pa at the frequency of 1, 2, 3, and 5 Hz, respectively, were studied. The RBCs' time-varying velocity varied after the glass plate velocity without any time lag, whereas the L/W change, where L and W were the major and minor axes of RBCs' ellipsoidal shape, exhibited a rapid increase and gradual decay during the deformation and recovery phase. The time of minimum L/W occurrence lagged behind the zero-velocity time of the glass plate (zero stress), and the delay time normalized to the one-cycle duration remained constant at 8.0%. The elongation of RBCs at zero stress time became larger with the reversing frequency. A simple mechanical model consisting of an elastic linear element during a rapid elongation period and a parallel combination of elements such as a spring and dashpot during the nonlinear recovery phase was suggested. The dynamic response behavior of RBCs under a cyclically reversing shear flow was different from the conventional shape change where a steplike force was applied to and completely released from the RBCs.

Blood Flow Velocity↗

Computational fluid dynamics analysis of the pediatric tiny centrifugal blood pump (TinyPump).

We have developed a tiny rotary centrifugal blood pump for the purpose of supporting circulation of children and infants. The pump is designed to provide a flow of 0.1-4.0 L/min against a head pressure of 50-120 mm Hg. The diameter of the impeller is 30 mm with six straight vanes. The impeller is supported by a hydrodynamic bearing at its center and rotated with a radial coupled magnetic driver. The bearing that supports rotation of the impeller of the tiny centrifugal blood pump is very critical to achieve durability, and clot-free and antihemolytic performance. In this study, computational fluid dynamics (CFD) analysis was performed to quantify the secondary flow through the hydrodynamic bearing at the center of the impeller and investigated the effects of bearing clearance on shear stress to optimize hemolytic performance of the pump. Two types of bearing clearance (0.1 and 0.2 mm) were studied. The wall shear stress of the 0.1-mm bearing clearance was lower than that of 0.2-mm bearing clearance at 2 L/min and 3000 rpm. This was because the axial component of the shear rate significantly decreased due to the narrower clearance even though the circumferential component of the shear rate increased. Hemolysis tests showed that the normalized index of hemolysis was reduced to 0.0076 g/100 L when the bearing clearance was reduced to 0.1 mm. It was found that the CFD prediction supported the experimental trend. The CFD is a useful tool for optimization of the hydrodynamic bearing design of the centrifugal rotary blood pump to optimize the performance of the pump in terms of mechanical effect on blood cell elements, durability of the bearing, and antithrombogenic performance.

Biomedical Engineering↗

Influence of radial clearance and rotor motion to hemolysis in a journal bearing of a centrifugal blood pump.

Hemolysis due to narrow clearance of noncontact bearings is a critical problem for rotary blood pumps. We developed a centrifugal blood pump with a magnetic and hydrodynamic hybrid bearing, and found that the hemolysis in the narrow clearance depends not only on the clearance size, but also on the rotor stability. In this study, we quantified the relation between the hemolysis, radial clearance (c), and rotor stability through the measurement of the rotor motion and hemolysis. As a result, it was confirmed that the rotor of the current pump is stabilized within the oscillation of 20 microm in blood, and the hemolysis decreases with increase in the c, which is the opposite in the unstable rotor motion with the previous pump. In order to theoretically discuss this hemolysis tendency, we implemented hemolysis estimation in the c according to hydrodynamics and hemodynamics. This estimation can represent the measured hemolysis tendency, and revealed that the flow rate has large influence on the hemolysis in the c.

Centrifugation↗

Quantification of the secondary flow in a radial coupled centrifugal blood pump based on particle tracking velocimetry.

Secondary flow in the centrifugal blood pump helps to enhance the washout effect and to minimize thrombus formation. On the other hand, it has an adverse effect on pump efficiency. Excessive secondary flow may induce hemolytic effects. Understanding the secondary flow is thus important to the design of a compact, efficient, biocompatible blood pump. This study examined the secondary flow in a radial coupled centrifugal blood pump based on a simple particle tracking velocimetry (PTV) technique. A radial magnetically coupled centrifugal blood pump has a bell-shaped narrow clearance between the impeller inner radius and the pump casing. In order to vary the flow levels through the clearance area, clearance widths of 0.25 mm and 0.50 mm and impeller washout holes with diameters of 0 mm, 2.5 mm, and 4 mm were prepared. A high-speed video camera (2000 frames per second) was used to capture the particle images from which radial flow components were derived. The flow in the space behind the impeller was assumed to be laminar and Couette type. The larger the inner clearance or diameter of washout hole, the greater was the secondary flow rate. Without washout holes, the flow behind the impeller resulted in convection. The radial flow through the washout holes of the impeller was conserved in the radial as well as in the axial direction behind the impeller. The increase in the secondary flow reduced the net pump efficiency. Simple PTV was successful in quantifying the flow in the space behind the impeller. The results verified the hypothesis that the flow behind the impeller was theoretically Couette along the circumferential direction. The convection flow observed behind the impeller agreed with the reports of other researchers. Simple PTV was effective in understanding the fluid dynamics to help improve the compact, efficient, and biocompatible centrifugal blood pump for safe clinical applications.

Biomedical Engineering↗

Measurement of the rotor motion and corresponding hemolysis of a centrifugal blood pump with a magnetic and hydrodynamic hybrid bearing.

This study proposed a centrifugal blood pump with a novel magnetic and hydrodynamic hybrid passive bearing, which consisted of a plain journal bearing for radial stability and a permanent magnetic bearing for axial and tilting stability. We measured the radial motion of the bearing and performed hemolysis tests for the different radial clearance sizes. In the results, it appeared that the radial motion had two modes: the stable center mode, in which the radius of the radial motion rapidly converged to less than 20 microm; and the unstable circle mode, in which the rotor suspension linearly increased with the rotation speed. It also appeared that the pumps with the radial clearance of 80 microm caused more hemolysis than with the smaller clearance sizes in the circle mode. The circle mode was avoidable by the higher rotation or the asymmetric pump structure, but the mechanism of hemolysis in this mode was still unclear.

Animals↗

Were volatile organic compounds the inducing factors for subjective symptoms of employees working in newly constructed hospitals?

This study demonstrated possible relationships between environmental, personal, and occupational factors and changes in the subjective health symptoms of 214 employees after the relocation of a hospital in a region of Japan. Eight indoor volatile organic compounds (VOCs) were detected in at least one of the 19 rooms investigated, and total VOC (TVOC) concentrations in 8 rooms exceeded the advisable value (400 microg/m(3)) established by the Ministry of Health, Labour and Welfare of Japan. Formaldehyde was detected in all the investigated rooms, but none of the results exceeded the guideline value (100 microg/m(3)). Multiple logistic regression analysis was applied to select variables significantly associated with the subjective symptoms that can be induced by sick building syndrome. The results showed that subjective symptoms of deterioration in the skin, eye, ear, throat, chest, central nervous system, autonomic system, musculoskeletal system, and digestive system among employees were associated mainly with gender difference and high TVOC concentrations (>1200 microg/m(3)). Long work hours (>50 h per week) in females and smoking in males were to be blamed for the deterioration of their symptoms. The present findings suggest that to protect employees from indoor environment-related adverse health effects, it is necessary to reduce the concentration of indoor chemicals in new buildings, to decrease work hours, and to forbid smoking.

Adult↗

Analysis of phthalate contamination in infusion solutions by automated on-line in-tube solid-phase microextraction coupled with high-performance liquid chromatography.

Contamination of infusion solutions with phthalates was analyzed, and its origin was determined. Phthalates were determined by on-line in-tube solid-phase microextraction coupled with high-performance liquid chromatography (in-tube SPME-HPLC) with UV detection. In-tube SPME is an extraction technique for organic compounds in aqueous samples, in which analytes are extracted from the sample directly into an open tubular capillary by repeated draw/eject cycles of sample solution. The infusion solutions were used without any pretreatment, and the phthalates in these solutions were automatically analyzed by the on-line in-tube SPME-HPLC system. The limits of detection of phthalates in the infusion solutions were 1-10 ng/mL. With a few exceptions, the recoveries of phthalates added to the infusion solutions were above 80%. Di-n-butyl phthalate (DBP) was detected at a concentration of 7-60 ng/mL in most infusion solutions in plastic containers but was not detected in those in glass bottles. On the other hand, no other phthalates were detected in infusion solutions in either plastic or glass containers. Large amounts of DBP were detected in the adhesive used to affix the paper labels to the plastic bottles and bags, but not in the plastic containers themselves. Furthermore, DBP was shown to be readily eluted from the adhesive into water and alcohol and easily pass through the plastic. These results indicated that the source of the DBP was the adhesive used to affix the paper labels, and DBP contaminated the infusion solutions by passing through the plastic. The in-tube SPME-HPLC method is simple and rapid and provides a useful tool for the screening and determination of phthalate contamination in infusion solutions.

Adhesives↗

Simple and rapid analysis of endocrine disruptors in liquid medicines and intravenous injection solutions by automated in-tube solid-phase microextraction/high performance liquid chromatography.

A simple and rapid method was developed for analyzing contamination of endocrine disruptors in liquid medicines and intravenous injection solutions. Endocrine disrupting compounds such as bisphenol A (BPA), alkylphenols and phthalates were quantitated by on-line in-tube solid-phase microextraction coupled with high performance liquid chromatography (in-tube SPME/HPLC) with UV detection. The liquid medicines and intravenous injection solutions could be used directly without any pretreatment, and the BPA, alkylphenols and phthalates in these solutions were automatically analyzed. The limits of quantification for these compounds were 1-10 ng/ml. Recoveries of these compounds spiked to the intravenous injection solutions was over 80%, except for some phthalates. Di-n-butyl phthalate (DBP) was detected at a concentration of 7-60 ng/ml in most intravenous injection solutions in plastic containers, but it was not detected in solutions in glass bottles. Diethyl phthalate, di-n-propyl phthalate, DBP and di-2-ethylhexyl phthalate (DEHP) were also detected in syrup, lotion and eye drops in plastic containers. On the other hand, BPA and alkylphenols were not detected at all in these solutions. DEHP contamination from an administration set increased when total vitamin formulation was added to the infusion solution. DEHP was easily leached from polyvinyl chloride tubing by polysorbate 80. The in-tube SPME/HPLC method is simple, rapid and automatic, and it provides a useful tool for the screening and determination of endocrine disruptor contamination in liquid medicines and intravenous injection solutions.

Autoanalysis↗

Determination of daidzein and genistein in soybean foods by automated on-line in-tube solid-phase microextraction coupled to high-performance liquid chromatography.

An automated on-line method for the determination of the isoflavones, daidzein and genistein, was developed using in-tube solid-phase microextraction coupled to high-performance liquid chromatography (in-tube SPME-HPLC). In-tube SPME is a new extraction technique for organic compounds in aqueous samples, in which analytes are extracted from the sample directly into an open tubular capillary by repeated draw/eject cycles of sample solution. Daidzein, genistein and their glucosides tested in this study were clearly separated within 8 min by HPLC using an XDB-C8 column with diode array detection. In order to optimize the extraction of these compounds, several in-tube SPME parameters were examined. The glucosides daidzin and genistin were analyzed as aglycones after hydrolysis because the glucosides were not concentrated by in-tube SPME. The optimum extraction conditions for daidzein and genistein were obtained with 20 draw/eject cycles of 40 microl of sample using a Supel-Q porous layer open tubular capillary column. The extracted compounds were easily desorbed from the capillary by mobile phase flow, and carryover was not observed. Using the in-tube SPME-HPLC method, the calibration curves of these compounds were linear in the range 5-200 ng/ml, with a correlation coefficient above 0.9999 (n = 18), and the detection limits (S/N = 3) were 0.4-0.5 ng/ml. This method was successfully applied to the analysis of soybean foods without interference peaks. The recoveries of aglycones and glucosides spiked into food samples were above 97%.

Automation↗

Complementary DNA cloning and characterization of cytochrome P450 2D29 from Japanese monkey liver.

A cDNA was cloned from Japanese monkey liver mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR) using oligonucleotide primers based on the marmoset cytochrome P450 2D19 (CYP2D19) nucleotide sequence. The full-length cDNA encoded a 497 amino acid protein (designated CYP2D29) that is 96, 91, and 88% homologous to human CYP2D6, cynomolgus monkey CYP2D17, and marmoset monkey CYP2D19, respectively. Yeast cells (Saccharomyces cerevisiae AH-22 strain) transfected with pGYR1 vectors containing the CYP2D29 cDNA were cultured, and microsomal fractions were obtained. Reduced carbon monoxide-difference spectra and western blot analysis using polyclonal antibodies raised against rat CYP2D2 demonstrated that in yeast cell microsomal fractions, the level of CYP2D29 holoenzyme was similar to that of CYP2D6 holoenzyme. However, western blot analysis indicated that the level of CYP2D29 in Japanese monkey liver microsomes might be much higher than that of CYP2D6 in human liver microsomes. Japanese monkey liver microsomes exhibited much higher activities than did human liver microsomes, expressed as nmol/min/mg protein, for debrisoquine (DB) 4-hydroxylation and bufuralol (BF) 1"-hydroxylation (typical reactions catalyzed by CYP2D6), whereas recombinant CYP2D29 activity, expressed as nmol/min/nmol CYP, was similar to that of CYP2D6 for DB and BF hydroxylation. In kinetic analyses, the K(m) value of CYP2D29 for DB 4-hydroxylation was much lower than that of Japanese monkey liver microsomes, whereas the K(m) value of CYP2D6 for DB 4-hydroxylation was similar to that of human liver microsomes. In contrast, K(m) values for BF 1"-hydroxylation were similar for Japanese monkey and human liver microsomes and yeast cell microsomal fractions expressing recombinant CYP2D29 or CYP2D6. These results suggest that the properties of Japanese monkey CYP2D29 are similar to those of human CYP2D6, but their populations and/or some other factors in liver microsomes may cause the difference in microsomal DB 4-hydroxylase activities between Japanese monkeys and humans.

Amino Acid Sequence↗

In vitro and in vivo formation of aminophenylnorharman from norharman and aniline.

Norharman is not mutagenic to Salmonella strains, but becomes so to S. typhimurium TA98 and YG1024 with S9 mix in the presence of the non-mutagenic aromatic amine, aniline. The mutagenicity from norharman and aniline in the presence of S9 mix is reported to be due to formation of a mutagenic compound, 9-(4'-aminophenyl)-9H-pyrido[3,4-b]indole (aminophenylnorharman, APNH). In the present study, we examined which enzymes might be involved in in vitro formation of APNH from norharman and aniline. When norharman (5mg) and aniline (2.5mg) were incubated with S9 mix, the S9 fraction of which was prepared from the livers of rats treated with phenobarbital (PB) and beta-naphthoflavone (beta-NF), at 37 degrees C for 20 min, 496 ng of APNH was produced. Formation of APNH was also detected with the microsomal fraction, but not with the cytosol fraction. The addition of a p450 inhibitor, SKF-525A, to the reaction mixture at a dose of 5mM resulted in a decrease of APNH formation, to around 40% of the control level. These results suggest involvement of p450 enzyme(s) in the formation of APNH from norharman and aniline. Moreover, a microsomal fraction from human liver was demonstrated to have the capacity to produce APNH from norharman and aniline, similar to the case with the microsomal fraction from rat liver. When norharman (90 mg/kg) and aniline (90 mg/kg) were administered to rats by gavage, APNH could be detected in the urine, at a rate of 19.6 ng+/-16.9 ng per 24h. The level was increased by treatment of the urine samples with hydrochloric acid, suggesting some APNH was excreted into urine as conjugated forms. Thus, it is likely that APNH may be produced from norharman and aniline in the human body.

Aniline Compounds↗

Estimation of dietary HCA intakes in a large-scale population-based prospective study in Japan.

Heterocyclic amines (HCAs) are known mutagens and animal carcinogens formed in cooked meat and fish. Japanese people generally consume more fish than animal meat, and chopped and stir-fried method of meat preparation is favored as much as the grilled cooking method. Therefore, it is supposed that the foods contributing to HCA intake in Japan are different from those in western countries. However, dietary HCA intake levels and their main sources have not been estimated in Japan. The purpose of this study was to assess individual intake levels of specific HCAs in subjects of the Japan Public Health Center (JPHC)-based prospective study on cancer and cardiovascular disease (JPHC study). Individual intake levels of six predominant HCAs (Trp-P-1, MeIQ, MeIQx, 7,8-DiMeIQx, 4,8-DiMeIQx, and PhIP) were calculated for 39035 JPHC study participants (18290 men and 20745 women) from four geographic areas (9412 from Iwate, 12018 from Akita, 9896 from Nagano, and 7709 from Okinawa) based on a self-reported food frequency questionnaire (FFQ). Mean daily HCA intake levels (ng/kg per day) for subjects in the mainland areas (Iwate, Akita, and Nagano) were 1.06 (inter quartile range 0.50-1.35) in men and 1.10 (inter quartile range 0.53-1.40) in women. Mean daily HCA intake levels (ng/kg per day) for subjects in Okinawa Island were lower than those in the mainland; 0.83 (inter quartile range 0.33-1.04) in men and 0.92 (inter quartile range 0.33-1.18) in women. PhIP accounted for more than 60% of total HCA intake, followed by MeIQx and MeIQ. More than 50% of total HCA intake was derived from fish in the mainland, whereas more than 50% was from pork in Okinawa. Our study suggests that grilled fish are the major dietary sources of HCAs in Japanese, except those in Okinawa Island.

Adult↗

Automated sample preparation using in-tube solid-phase microextraction and its application -- a review.

Sample preparation, such as extraction, concentration, and isolation of analytes, greatly influences their reliable and accurate analysis. In-tube solid-phase microextraction (SPME) is a new effective sample preparation technique using an open tubular fused-silica capillary column as an extraction device. Organic compounds in aqueous samples are directly extracted and concentrated into the stationary phase of capillary columns by repeated draw/eject cycles of sample solution, and they can be directly transferred to the liquid chromatographic column. In-tube SPME is an ideal sample preparation technique because it is fast to operate, easy to automate, solvent-free, and inexpensive. On-line in-tube SPME-performed continuous extraction, concentration, desorption, and injection using an autosampler, is usually used in combination with high performance liquid chromatography and liquid chromatography-mass spectrometry. This technique has successfully been applied to the determination of various compounds such as pesticides, drugs, environmental pollutants, and food contaminants. In this review, an overview of the development of in-tube SPME technique and its applications to environmental, clinical, forensic, and food analyses are described.

Automation↗

Stereoselective metabolism of bufuralol racemate and enantiomers in human liver microsomes.

A new HPLC method was developed using a chiral column to efficiently separate four 1"-hydroxybufuralol (1"-OH-BF) diastereomers that are major metabolites of bufuralol (BF). Employing this method, we examined diastereomer selectivity in the formation of 1"-OH-BF from BF racemate or enantiomers in four individual samples of human liver microsomes. Three different human liver microsomes showed a selectivity of 1"R-OH < 1"S-OH for BF enantiomers, which was similar to that of recombinant CYP2D6 expressed in insect cell microsomes, whereas one human liver microsomal fraction yielded a selectivity of 1"R-OH > 1"S-OH for BF enantiomers, which was similar to those of recombinant CYP2C19 expressed in insect cell microsomes. Recombinant CYP1A2 and CYP3A4 showed a selectivity similar to that of CYP2D6, but their BF 1"-hydroxylase activities were much lower than those of CYP2D6. In inhibition studies, quinidine, a known CYP2D6 inhibitor, markedly inhibited BF 1"-hydroxylation in the fractions of human liver microsomes that showed the CYP2D6-type selectivity. Furthermore, omeprazole, a known CYP2C19 inhibitor, efficiently suppressed the formation of 1"-OH-BF diastereomers from BF in the microsomal fraction that showed the CYP2C19-type selectivity. From these results, we concluded that the diastereomer selectivity in the formation of 1"-OH-BF from BF differs between CYP2D6 and CYP2C19, both of which can be determinant enzymes in the diastereoselective 1"-hydroxylation of BF in human liver microsomes.

Adult↗