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Biomedical subjects

T Onouchi

Publications and source records attributed to T Onouchi.

At least 19 recordsLinked to original sources

A microbial chip combined with scanning electrochemical microscopy.

A microbial chip for bioassay was fabricated and its performance was characterized by scanning electrochemical microscopy (SECM). The microbial chip was prepared by spotting a suspension of Escherichia coli on a polystyrene substrate by using a glass capillary pen. The respiration activity of the E. coli spot was imaged with SECM by mapping the oxygen concentration around the spot. The SECM images of the microbial chips clearly showed spots with lower reduction currents, indicating that E. coli in the spots uptake oxygen by respiration. The bactericidal effects of antibiotics (streptomycin and ampicillin) were measured using the E. coli-based microbial chip, and discussed in comparison with the minimum inhibitory concentration (MIC) determined by an agar plate dilution method.

Ampicillin↗

Immunoassay of the MRSA-related toxic protein, leukocidin, with scanning electrochemical microscopy.

Scanning electrochemical microscopy (SECM) was applied to the immunoassay of leukocidin, which is a toxic protein produced by methicillin-resistant Staphylococcus aureus (MRSA), with the intention of developing and early diagnostic for MRSA infection. An antibody-chip for leukocidin was prepared by self-assembling of anti-leukocidin on a protein A-coated glass substrate. A sample solution containing leukocidin was spotted onto the antibody-chip, followed by labeling with horseradish peroxidase (HRP) via a sandwich method. The reduction current of the oxidized form of ferrocenylmethanol generated by the HRP reaction was monitored to view SECM images of the spot of captured leukocidin. The amplitude of reduction current depended on the concentrations of sample solutions used for making spots. This SECM-based immunoassay detects as low as 5.25 pg mL(-1) leukocidin.

Electrochemistry↗

Effect of soy protein on bone metabolism in postmenopausal Japanese women.

We conducted a cross-sectional study of the effects of soybean protein intake on bone mineral density and biochemical markers in 85 postmenopausal Japanese women. Nutrients in the diet of postmenopausal Japanese women visiting the osteoporosis unit, including subjects with normal lumbar spine bone mineral density (L2-4 BMD), were investigated by questionnaire, and the calculated daily energy, protein, soy protein and calcium intake were obtained. L2-4 BMD was measured with dual-energy X-ray absorptiometry, and assays done of serum alkaline phosphatase (ALP) and serum intact osteocalcin (IOC) as bone formation markers and urinary pyridinoline (UPYR) and urinary deoxypyridinoline (UDPYR) as bone resorption markers. Soy protein intake was significantly associated with the Z-score for L2-4 BMD (r = 0.23,p = 0.038) and UDPYR (r = -0.23, p = 0.034). Stepwise multiple regression analyses showed that soy protein intake is significantly associated with the Z-score for L2-4 BMD (beta = 0.225, p = 0.04) and UDPYR (beta = -0.08, p = 0.03) among four nutritional factors. These results suggest that high soy protein intake is associated with a higher bone mineral density and a lower level of bone resorption, but further studies are needed to confirm the causal dynamic mechanisms.

Absorptiometry, Photon↗

Hypercalcemia induced with the plasma levels of parathyroid hormone-related peptide in multiple myeloma.

A 69-year-old man visited our department of neurology with symptoms of paresthesia on the lower extremities and lumbago. Biochemical examination of serum samples showed hypercalcemia (serum concentration 15.6 mg/dl). The levels of intact parathyroid hormone (i-PTH) and 1,25-dihydroxyvitamin D were suppressed, whereas parathyroid hormone-related peptide (PTHrP) was elevated up to 5.4 pM (normal range: below 0.6 pM). Additionally, bone survey revealed a punched-out lesion in radiological examinations of the skull. Bone marrow aspiration demonstrated many atypical plasma cells suggesting multiple myeloma. Nephrogenous cyclic adenosine monophosphate (cAMP), urinary deoxypyridinoline, plasma interleukin 6 (IL-6) and transforming growth factor beta (TGF beta) concentrations were elevated, whereas % of renal tubular reabsorption of phosphate (%TRP) was decreased. The immunohistochemical results demonstrated the expression of PTHrP in atypical plasma cells. These data indicated that hypercalcemia complicating multiple myeloma causes an elevation of renal calcium reabsorption and an increase of bone resorption mediated by PTHrP action.

Aged↗

Lack of correlation between the reduction of sevoflurane MAC and the cerebellar cyclic GMP concentrations in mice treated with 7-nitroindazole.

BACKGROUND: Although inhibition of nitric oxide synthase (NOS) has been reported to be antinociceptive and to reduce the threshold of general anesthesia, the mechanism of action is largely unknown. Specifically, the relation between the minimum alveolar concentration (MAC)-reducing effects of NOS inhibition and cyclic guanosine monophosphate (cGMP) concentrations in the brain has not been defined. To further characterize the effects of NOS inhibition, the authors studied the relation between the MAC of sevoflurane and the cGMP concentration of the brain after acute and chronic treatment with a neuronally selective NOS inhibitor, 7-nitroindazole (7-NI). METHODS: Sevoflurane MAC and cerebellar cGMP concentrations were determined in mice after acute intraperitoneal administration or after 1, 2, 3, and 4 days of gavage feeding of 7-NI. After acute or chronic treatment with 7-NI, the mice were separated into two groups. Sevoflurane MAC was measured by a tail-clamp method in the first group, and cerebellar cGMP concentrations were measured by enzyme-linked immunosorbent assay in the second group of the mice. RESULTS: In mice, acute intraperitoneal administration of 7-NI dose dependently decreased sevoflurane MAC and cerebellar cGMP; and 4-day-long gavage feeding with 7-NI (500 mg/ kg, every 8 h) time dependently decreased cerebellar cGMP, but sevoflurane MAC was reduced only for the first 2 days and returned to its baseline after 3 days of 7-NI feeding. CONCLUSIONS: Although an acute selective inhibition of neuronal NOS decreases sevoflurane MAC and cerebellar cGMP concentrations in mice, there was a dissociation between the two parameters during long-term neuronal NOS inhibition. There may be cGMP-independent compensatory mechanisms that mediate nociception when NOS is chronically inhibited.

Anesthetics, Inhalation↗

[Salicylic acid].

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Anti-Inflammatory Agents, Non-Steroidal↗

The ontogeny of cytosol and nuclear progestin receptors in male rat brain and its male-female differences.

The fetal and postnatal development of the progestin receptor systems in the intact male rat brain was investigated by means of the in vitro cytosol binding and the nuclear exchange assay using [3H]R5020 [( 17 alpha-methyl-3H]17 alpha, 21-dimethyl-19-nor-4, 9-pregnadiene-3,20-dione). The cortical cytosol receptors, first detectable at day 0, rapidly increased at day 7, reaching a maximum at day 10, then gradually declined thereafter. The receptors in the HPOA appeared clearly at day 1, increased during the first 10 days, then remained constant at days 14-21. The postnatal developmental patterns of cytosol brain progestin receptors in males were essentially similar to those in females, but there were some differences between both sexes. The male HPOA at days 10-14 contained more receptors than the female one. Nuclear progestin binding was low in the neonatal male brain at days 1-3. Despite the low level of serum progesterone, the cortical nuclear binding suddenly increased at days 7-10, then remained high at days 14-21. A similar, though less pronounced, pattern was seen in the HPOA. The male pattern of nuclear binding, thus, essentially resembled the female one. However, lower binding in the cortex and, possibly, HPOA was found in males than in females at days 10-21. After progesterone injection postnatal male rats accumulated a lower concentration of progestin receptors in the cortex and, possibly, HPOA than similarly-treated females. It is concluded from these results that progestin receptors in male rat brain appear immediately after birth and develop differentially in the cortex and HPOA. The sudden onset of increased nuclear translocation of endogenous progestin receptor complexes may occur in the brain at around days 7-10. There is a marked sex difference in the nuclear progestin receptor system in the postnatal brain, particularly the cortex. Moreover, the postnatal male brain has lower capacities of nuclear receptor translocation than does the female one. The progestin receptor system in the cortex and, possibly, HPOA of rats in the early postnatal life might be involved with some processes in the mechanism of sexual differentiation of the brain.

Animals↗

Decreased progestin receptors in the cerebral cortex of hypothyroid postnatal rats.

The effects of neonatal hypothyroidism were examined on the concentrations of cytosol receptors for progestin and estrogen in the cytosols from 10-day old female and male rats. The concentration of cytosol progestin- and estrogen receptors in the 10-day old female and male rats with propylthiouracil-induced hypothyroidism was determined by multiconcentration saturation analysis. Neonatal hypothyroidism markedly depressed the level of progestin receptors in the cortex but not in the hypothalamus-preoptic area (HPOA). There were no differences in the concentration of estrogen receptors in the cortex and HPOA between the control and PTU rats. These results suggest that thyroid hormone may be one of the factors affecting the development of progestin receptors in the cortex. Its significance was discussed in relation to determination of the length of sexual differentiation of the rat brain.

Animals↗

Estrogen receptors and estrogen-inducible progestin receptors in the sexual skin of the monkey.

Estrogen-binding macromolecules were identified in sexual skin cytosol and nuclear fractions from female Japanese monkeys (Macaca fuscata fuscata), by sucrose-glycerol gradient analysis and dextran-coated charcoal adsorption technique. Furthermore, using the highly potent synthetic progestin, R5020 (17,21-dimethyl-19-nor-4,9-pregnadiene-3,20-dione; promegestone), progestin-binding macromolecules were detected in sexual skin cytosols from ovariectomized estrogen-primed monkeys. Both cytosol components sedimented at the 8S region on sucrose-glycerol gradients and had a high affinity for the respective steroids, and competition studies showed a specificity for estrogenic, or progestational compounds. The concentration of cytosol progestin receptors was markedly increased with the elevated level of nuclear estrogen receptors following estrogen priming. The above results demonstrate the existence of cytosol and nuclear estrogen receptor sites, and estrogen-induced progestin receptors in the sexual skin of the female monkeys.

Abdomen↗

Progestin receptors in female rat brain and hypophysis in the development from fetal to postnatal stages.

The fetal and postnatal development of the progestin receptors in the intact female rat brain was investigated by means of the in vitro cytosol binding and the nuclear exchange assay using 3H-R5020 ([17 alpha-methyl-3H] 17 alpha-,21-dimethyl-19-nor-4,9-pregnadiene-3,20-dione). A large 7S peak of progestin binding in the cerebral cortical cytosols was observed at days 7-14 with less evident binding in the hypothalamus-preoptic area (HPOA). The cortical cytosol receptors, first detectable at day 0, rapidly increased at days 1-7, reaching a maximum at day 10, then gradually declined thereafter. The receptors in the HPOA and other brain areas appeared at day 1, increased during the first week, then remained constant at days 10-28. Hypophysial cytosol receptors, first detectable at day 10, increased at day 28. Nuclear progestin binding was low in the HPOA and cortex of the neonates at days 1-3. Interestingly, despite the low level of serum progesterone, the cortical nuclear binding suddenly increased at days 7-10, then remained high at days 14-21. A similar, though less pronounced, pattern was seen in the HPOA. Diethylstilbestrol injection caused an increase in the cytosol binding capacities in the HPOA, but did not in the cortex. These results suggest the appearance of the brain progestin receptor system immediately after birth, and differential patterns of their postnatal development in the intact female rat. The onset of increased nuclear translocation of endogenous progestin-receptor complex may occur in the cortex, and possibly HPOA, at around days 7-10. Progestin receptors in the postnatal rat HPOA are estrogen inducible, but not in the cortex.

Animals↗

Progesterone receptors in the cerebral cortex of neonatal female rats.

Progestin-binding components were detected in cerebral cortical cytosols from female rats at 7 days of age. These components labeled in vitro with a tritiated synthetic progestin. R5020, sediment in the 7S region in sucrose density gradients containing 10% glycerol. The dissociation constants and the number of binding sites of the components were 3.9 X 10(-10)M and 47.5 fmol/mg cytosol protein, respectively, indicating high affinity and low capacity binding. The 7S-binding components were specific for progestational compounds. Estradiol competed weakly. Incubation with pronase abolished the 7S progestin binding. Heat experiments suggested a thermolabile nature of the components. These results suggest that the cytosols from the cerebral cortex of 7-day-old female rats contain progesterone receptors.

Animals↗

The presence of progesterone receptors in the sexual skin of the monkey.

R5020(17,21-dimethyl-19-nor-4,9-pregnadiene-3,20-dione)-binding components with sedimentation coefficient of 8S were detected in sexual skin cytosols from estrogen-primed ovariectomized Japanese monkey (Macaca fuscata fuscata). In contrast, little 8S binding was found in similar preparations from the abdominal skin. The dissociation constants and the number of binding sites of the components were 1.6 x 10(-10)M and 36 fmoles/mg cytosol protein, respectively. The 8S binding components were specific for progestational compounds. Incubation with pronase abolished the 8S binding. Thermal experiments revealed the thermolabile nature of the components. Moreover, the concentration of the R5020-binding components was markedly increased by estradiol-17 beta 3-benzoate injections. We conclude from these results that the cytosols from the sexual skin of estrogen-primed female monkeys contain progesterone receptors.

Animals↗