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Biomedical subjects

H Ohya

Publications and source records attributed to H Ohya.

At least 19 recordsLinked to original sources

Oxygen consumption of single bovine embryos probed by scanning electrochemical microscopy.

Oxygen consumption of individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). A probe microelectrode was used to scan near a single embryo surface in a culture medium to monitor the oxygen reduction current at 37 degrees C, under a water-saturated atmosphere of 5% CO2 and 95% air. The oxygen concentration profiles near the embryos were in good agreement with the theoretical spherical diffusion. When an embryo reached the stage of a morula with a 74-microm radius on day 6 after in vitro fertilization, the oxygen concentration difference (deltaC) between the bulk solution and the morula surface was 6.90 +/- 1.35 microM. The oxygen consumption rate (F) of the single morula was estimated to be (1.40 +/- 0.27) x 10(-14) mol s(-1). After the SECM measurement, the embryo was continuously cultured for another 2 days and grew to the stage of a blastocyst with a 100-microm radius. For the blastocyst, the deltaC values for the inner cell mass side and the trophoblast side were 16.40 +/- 1.83 and 9.14 +/- 1.68 microM, respectively. The oxygen consumption rate of the blastocyst was found to be in the range of (2.50 +/- 0.46) x 10(-14) mol s(-1) < F < (4.49 +/- 0.50) x 10(-14) mol s(-1). We have carried out SECM measurements for 19 embryos, and the results were compared in detail with these from an optical microscopic observation. The deltaC values for the morulae on day 6 after in vitro fertilization were strongly related to the morphological embryo quality. The morulae showing a larger deltaC value developed into blastocysts of a larger size, and the deltaC value after the subsequent 2 days of cultivation was found to be increased.

Animals↗

Longitudinally detected ESR magnetometer for wide-range measurements of low fields.

A new magnetometer utilizing a longitudinally detected ESR (LODESR) method was developed. The probe head of the LODESR magnetometer is equipped with a single-turn coil (8 mm in diameter) which has a very wide bandwidth because the reactance of the coil is always smaller than the resistance of the transmission line (50 ohm) at frequencies less than 700 MHz. Thus, an absolute magnetic field could be measured over a wide range (2 to 9 mT) using this magnetometer without changing the probe head.

Journal Article↗

Imaging of electrically detected magnetic resonance of a silicon wafer.

An imaging technique of electrically detected magnetic resonance (EDMR) was newly developed. Because the EDMR signal is obtained from paramagnetic recombination centers, one may expect the image to represent the distribution of defect and/or impurity sites in the sample. We successfully obtained EDMR images of a light-illuminated silicon plate 8 mm in width and 15 mm in length, which was cut from a silicon wafer (n-type, 100 Omega cm), under ESR irradiation at a frequency of 890 MHz (wavelength, 340 mm). The reproducibility of the EDMR image obtained from a sample was amply satisfactory. When the oxidized surface of the silicon was removed, the EDMR signal disappeared. Although the EDMR signal reappeared when the surface of the sample became reoxidized, the EDMR image obtained was slightly different from the earlier one. This finding shows that the EDMR image obtained from the sample shows the distribution of defects at the Si/SiO(2) interface.

Journal Article↗

Pulmonary artery reflection for differentially diagnosing primary pulmonary hypertension and chronic pulmonary thromboembolism.

OBJECTIVES: The purpose of this investigation was to differentiate chronic pulmonary thromboembolism (CPTE) from primary pulmonary hypertension (PPH) by means of the indexes of pulmonary arterial reflection. BACKGROUND: These differences in the primary lesions would make pulmonary artery reflection occur earlier in CPTE than in PPH. Although the analysis of pulsatility of pulmonary arterial pressure is useful in the differential diagnosis of PPH and CPTE, it is not known whether the analysis of pulmonary artery reflection can differentiate CPTE from PPH. METHODS: Since CPTE predominantly involves the proximal arteries, whereas PPH involve the peripheral arteries, we hypothesized that patients with CPTE have a large augmentation index and a short inflection time. For this study, we enrolled 62 patients who had CPTE (31 patients) and PPH (31 patients). We measured pulmonary arterial pressure using a fluid filled system that included a balloon-tipped flow directed catheter. To quantify the pulmonary artery reflection, we used the augmentation index and inflection time. RESULTS: The augmentation index was markedly higher in CPTE than it was in PPH (27.4% +/- 15.2% [SD] and -25.1% +/- 26.9%, respectively, p < 0.001) and was diagnostic in separating the two groups. Inflection time separated the two groups reasonably well (97 +/- 20 ms and 210 +/- 49 ms, respectively, p < 0.001). CONCLUSIONS: The analysis of pulmonary arterial reflection is useful in the differential diagnosis of CPTE and PPH.

Adult↗

In vivo ESR study on hepatic reduction of a nitroxide radical after administration of glucose in rats.

The in vivo reducing ability of a nitroxide radical, 4-hydroxyl-2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPOL), in the liver of rats was estimated by using an electron spin resonance spectrometer equipped with a surface-coil-type resonator following administration of glucose. Both oral and intravenous administration of glucose significantly enhanced the reduction capacity. In vitro studies show that the reduction site of TEMPOL in the hepatic homogenate is located in the mitochondrial respiratory chain and microsomal electron transport system. These findings indicate that the enhancement of activity of these systems caused by the glucose administration prompts reduction of TEMPOL in the liver.

Animals↗

Proteome analysis of Oncorhynchus species during embryogenesis.

To understand the molecular mechanisms underlying normal and abnormal development of two salmonids, masu salmon (Oncorhynchus masou) and rainbow trout (O. mykiss), we used two-dimensional (2-D) electrophoresis to construct a series of 2-D maps during the embryonic period. We identified all visible protein spots on the 2-D map by assigning numbers for masu salmon and rainbow trout, and we determined N-terminal sequences of proteins for one hundred of the spots, that appear at very high concentrations in the whole embryos of masu salmon and rainbow trout. We also characterized embryonic stages according to the periods of appearance of spots. Most of the N-terminal sequences were identical or at least highly similar to partial sequences reported for vitellogenin (Vtg) of O. mykiss. A potential proteolytic processing of Vtg for rainbow trout is discussed in relation to the time of appearance and relative position of Vtg fragments within the complete protein sequence.

Amino Acid Sequence↗

In vivo EPR imaging by using an acyl-protected hydroxylamine to analyze intracerebral oxidative stress in rats after epileptic seizures.

EPR imaging by using an acyl-protected hydroxylamine, 1-acetoxy-3-carbamoyl-2,2,5,5-tetramethylpyrrolidine (ACP), in the head of a living rat after kainic acid (KA)-induced epileptic seizures was performed. ACP is a stable non-radical compound, but is easily deprotected with intracellular esterase to yield a hydroxylamine, which is oxidized by intracellular oxidative stress to yield an EPR-detectable nitroxide radical. From in vivo image data, the average values of EPR signal intensity from the hippocampus, striatum, and cerebral cortex were computed. There was no significant difference in cortical signal intensity between the control and KA-treated rats. The signal intensities from the hippocampus and striatum for the KA-treated rats were significantly higher than those for the control. The in vitro study showed that almost the same quantity of ACP moved into all regions of the brain of the control and KA-treated rats. These findings indicate that following a KA-induced seizure, the oxidative stress in the hippocampus and striatum is enhanced, but not so in the cerebral cortex.

Animals↗

Electrically detected magnetic resonance signal intensity at resonant frequencies from 300 to 900 MHz in a constant microwave field.

A method for electrically detected magnetic resonance (EDMR) measurement at different ESR frequencies under a constant alternating magnetic field has been established wherein the accurate relationship between EDMR signal intensity (from a photoexcited silicon crystal and a silicon diode) and a resonant frequency of 300 to 900 MHz (UHF band) was systematically clarified. EDMR signal intensity from a photoexcited silicon crystal against a resonant frequency fitted the curve of y = a(1 - e(-bx)) well, which approached a constant value at higher frequencies. The increase in the EDMR signal intensity from the silicon diode at higher resonant frequencies was smaller than that from the photoexcited silicon crystal. The difference can be explained by the influence of the skin effect; i.e., the microwaves do not penetrate deep into a highly conductive sample at higher frequencies. EDMR signal intensities of samples vs microwave power were measured at 890 MHz. The EDMR signal intensity from the silicon diode continued to increase as the microwave power was increased, while the signal intensity from the photoexcited silicon crystal saturated within the range. The difference can be similarly explained: due to the skin effect, the microwaves gradually penetrate into the silicon diode as the power increases, so that even when saturation has been reached outside, the microwave field inside the diode does not reach the saturation level.

Electricity↗

New sensitive agents for detecting singlet oxygen by electron spin resonance spectroscopy.

Free radicals are well-established transient intermediates in chemical and biological processes. Singlet oxygen, though not a free radical, is also a fairly common reactive chemical species. It is rare that singlet oxygen is studied with the electron spin resonance (ESR) technique in biological systems, because there are few suitable detecting agents. We have recently researched some semiquinone radicals. Specifically, our focus has been on bipyrazole derivatives, which slowly convert to semiquinone radicals in DMSO solution in the presence of potassium tert-butoxide and oxygen. These bipyrazole derivatives are dimers of 3-methyl-1-phenyl-2-pyrazolin-5-one and have anti-ischemic activities and free radical scavenging properties. In this work, we synthesized a new bipyrazole derivative, 4,4'-bis(1p-carboxyphenyl-3-methyl-5-hydroxyl)-pyrazole, DRD156. The resulting semiquinone radical, formed by reaction with singlet oxygen, was characterized by ESR spectroscopy. DRD156 gave no ESR signals from hydroxyl radical, superoxide, and hydrogen peroxide. DRD156, though, gives an ESR response with hypochlorite. This agent, nevertheless, has a much higher ability to detect singlet oxygen than traditional agents with the ESR technique.

Dimethyl Sulfoxide↗

[The survey of HIV infection among clients of foreign female prostitutes in Tokyo Metropolitan].

OBJECTIVES: It is important to elucidate the risks to foreign female prostitutes of HIV infection from their clients. We determined the prevalence of HIV among clients of foreign female prostitutes using anonymous testing of condom semen samples. METHODS: A total of 250 client condom semen samples were collected with client age and nationality by 15 Latin American and 37 Thai female prostitutes in Tokyo from March to May 1995. We screened by ELISA (GENELAVIA MIXT, Pasteur Sanofi Diagnostics) for antibody to HIV 1 and 2, and then confirmed by Western Blot (LAV Blot 1, LAV Blot 2, Sanofi Diagnostics Pasteur) and distinguished by Immuno Blot PEPTI-LAV 1, 2, (Sanofi Diagnostics Pasteur). RESULTS: The nationality of customers was reported as Japanese (74.8%), Iranian (7.6%), Pakistan (5.6%), Indian (2.8%), Taiwanese (2.0%), and Chinese (1.2%) with other nationalities less than 1.0%. Estimated age categories of the clients were < 20 (1.2%), 20-29 (41.2%), 30-39 (36.4%), 40-49 (12.4%), > or = 50 (6.4%) and undetermined (2.4%). Of the 250 samples collected, only 219 were sufficient for testing. Two samples (0.90%, 95% CI = 0.4-2.2%) were HIV 1 positive, both from Japanese clients (1.2% of Japanese clients, 95% CI = 0.5-2.9%). These data suggested that the prevalence of HIV in clients of female prostitutes was relatively high, and can be considered a risk group in Japan. The sentinel surveillance by anonymous of client condom semen samples is a very useful method to quantify HIV prevalence in this risk group.

Acquired Immunodeficiency Syndrome↗

Trapping of secreted molecules at single-cell level using an HPLC resin particle under a videomicroscope.

Particles of HPLC resins are used for the trapping of secreted molecules from a single cell. The basic molecules, e.g., histamine, are secreted from a single RBL-2H3 cell by granule exocytosis and are trapped by cation-exchange HPLC resins outside the cell. Since quinacrine is concentrated into the exocytotic and acidic microgranules in RBL-2H3 cells, which are used as a model cell line of mast cells, we measured the change in the fluorescence intensity of the quinacrine released from the cells and that of molecules trapped on the resin using a videomicroscope. By measuring the increase in the fluorescence intensity of the resins, we can estimate the real time course of molecular secretion from a single cell.

Animals↗

Evaluation of in-vivo transdermal absorption of cyclosporin with absorption enhancer using intradermal microdialysis in rats.

The purpose of this study was to evaluate the effect of absorption enhancer on in-vivo transdermal absorption of cyclosporin using intradermal microdialysis in rats. Cyclosporin oily solutions (0.5, 2, 8% w/v) were prepared from Sandimmun (10% w/v oily oral preparation of cyclosporin) by diluting with olive oil. 1-[2-(Decylthio)ethyl] azacyclopentan-2-one (HPE-101) and glycerin were added to the cyclosporin formulation as an absorption enhancer at various concentrations between 1 and 20%. These formulations were applied to the shaved abdomen of rats treated with intradermal microdialysis at a flow rate of 2.5 microL min-1 for 6 h. Cyclosporin was immediately detected and attained a plateau in the dermal dialysate after topical application of cyclosporin oily solution alone. Cyclosporin levels in the dialysate increased with increasing cyclosporin concentrations in the formulation from 0.5 to 8% (w/v). HPE-101 did not influence cyclosporin absorption at concentrations less than 6% (w/v). Addition of 10% (w/v) HPE-101 significantly enhanced an apparent absorption rate of cyclosporin by 4.9 times. However, 20% (w/v) HPE-101 did not show the enhancing activity. On the other hand, addition of glycerin at concentrations of 6, 10, and 20% (v/v) significantly enhanced an apparent absorption rate of cyclosporin by 3.0, 6.4, and 6.9 times, respectively. The time lag for cyclosporin absorption was less than 0.21 h in all tested cases. This microdialysis study shows that glycerin is a suitable enhancer for improving the in-vivo cyclosporin absorption from the skin.

Administration, Cutaneous↗

Serum concentrations of apolipoproteins in patients with thyroid dysfunction.

We measured serum concentrations of apolipoproteins (apo A-I, A-II, B, C-II, C-III and E) in patients with thyroid disease (hyperthyroidism; n = 44, hypothyroidism; n = 15) and in normal subjects (n = 89). We found that apoA-II, B and C-III concentrations revealed significant difference among three groups of the normal (apoA-II; 31.4 +/- 4.9 mg/dl, apoB; 85.8 +/- 16.3 mg/dl, apoCIII; 7.45 +/- 2.99 mg/dl), hyperthyroidism (apoA-II; 29.8 +/- 5.4, apoB; 63.4 +/- 18.9, apoC-III; 6.28 +/- 2.45) and hypothyroidism (apoA-II; 27.5 +/- 5.3, apoB; 108.0 +/- 30.9, apoC-III; 9.43 +/- 2.74). Thyroid hormones showed clear negative correlation to apoB (r = 0.70, p < 0.001). Furthermore, apoC-III was also found to be negatively correlated with thyroid hormone concentrations (r = 0.47, p < 0.001).

Adult↗

Lunar base extension program and closed loop life support systems.

Much of papers describing the Lunar Base Construction have been already published. Lunar Base has been considered to be one of the very useful facilities to conduct the excellent scientific program in near future and also to develop the lunar resources such as 3He and oxygen in order to expand the human activities in space. The scale of lunar base and the construction methods to be adopted should be determined based upon the utilization program to be conducted after the initiation of outpost habitation. In order to construct lunar base, it needs to transport lunar base elements such as habitat modules, experiment modules and so on having 20-30 tons weight and to install, connect and activate on lunar surface. The development of transportation system such as OTV enabling to transport over 30 ton payload weight from earth to moon seems to be very difficult in near future, and it seems reasonable to transport three elements per year as described in many papers already published. Therefore, the initial habitat outpost would not have enough volume to produce foods and has to have ECLS system similar to that of space station already going to be developed. After 2-3 years from the initiation of habitation, the addition of food production facilities could be started to expand the habitability of lunar base and finally the complete closed loop life support system could be installed after spending 6-7 years. This paper describes ECLS technologies to be used in the initial habitat outpost and design philosophy of the closed loop technologies to be utilized in the final stage.

Carbon Dioxide↗

DMPO spin trapping in the presence of Fe ion.

Aminoxyl radical formation from DMPO in the presence of Fe ion was studied to clarify the ambiguous ESR signals obtained by spin trapping with DMPO. It was found that when DMPO was used in a Fenton system, a Fe-DMPO complex was formed immediately. This complex was subsequently attacked by oxidative species originating from H2O2 and thus oxidative degradation of DMPO was induced in the Fenton system. On the other hand, in the case of M4PO, the degradation was found to be very slow, indicating that the 3 position of DMPO was favorably attacked by the oxidative species. Some of the degradation products are probably aminoxyl radicals. This series of the degradation products are probably aminoxyl radicals. This series of reactions may compete with spin trapping and make it difficult to analyze ESR spectra obtained in the presence of Fe ion.

Cyclic N-Oxides↗

Design for a bioreactor with sunlight supply and operations systems for use in the space environment.

An experiment was carried out to determine the characteristics of an operations system that can support fast cultivation of algae at high densities in the weightlessness of space. The experiment was conducted in glass bioreactor tanks, in which light was supplied by radiator rods connected to optical fiber cables. The illumination areas of the tanks were 2600 cm2, 6000 cm2, and 9200 cm2 per liter of solution. The characteristics of O2-CO2 gas exchange, concentration and separation of chlorella in the growth medium, dialysis of ionic salts in the growth medium, etc. were examined. Chloralla ellipsoidea was used in the experiment, yielding the following results: (1) By increasing the ratio of illumination area to volume, growth rates of up to approximately 0.6 g/L h could be obtained in a highly concentrated solution (one that contains 20 g/L or more of algae). (2) The most suitable proportions of carbon dioxide and oxygen gases for growing algae quickly at high concentrations were found to be 10% CO2 and 10% O2 (by volume). (3) There was a high optimum concentration for fast cultivation, and the data obtained resembled the theoretical curve postulated by P. Behrens et al. (4) It was possible to exchange carbon dioxide and oxygen using gas-permeable membrane modules. (5) It was possible to separate the chlorella from the growth medium and recycle the medium.

Bioreactors↗

Sunlight supply and gas exchange systems in the microalgal bioreactor.

The bioreactor with sunlight supply system and gas exchange systems presented here has proved feasible in ground tests and shows much promise for space use as a CELSS device. Our chief conclusions concerning the specification of total system needed for a life support system for a man in a space station are the following. (1) Sunlight supply system: compactness and low electrical consumption. (2) Bioreactor system: high density and growth rate of chlorella. (3) Gas exchange system: enough for O2 production and CO2 assimilation.

Air Conditioning↗