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Biomedical subjects

H Yasuda

Publications and source records attributed to H Yasuda.

At least 55 records · Page 3Linked to original sources

Increase in intracellular Ca(2+) and calcitonin gene-related peptide release through metabotropic P2Y receptors in rat dorsal root ganglion neurons.

We examined the effects of the activation of metabotropic P2Y receptors on the intracellular Ca(2+) concentration and the release of neuropeptide calcitonin gene-related peptide (CGRP) in isolated adult rat dorsal root ganglion neurons. In small-sized dorsal root ganglion neurons (soma diameter<30 microm) loaded with fura-2, a bath application of ATP (100 microM) evoked an increase in intracellular Ca(2+) concentration, while the removal of extracellular Ca(2+) partly depressed the response to ATP, thus suggesting that the ATP-induced increase in intracellular Ca(2+) concentration is due to both the release of Ca(2+) from intracellular stores and the influx of extracellular Ca(2+). Bath application of uridine 5'-triphosphate (UTP; 100 microM) also caused an increase in intracellular Ca(2+) concentration in small-sized dorsal root ganglion neurons and the P2 receptor antagonists suramin (100 microM) and pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS; 10 microM) virtually abolished the response, indicating that the intracellular Ca(2+) elevation in response to UTP is mediated through metabotropic P2Y receptors. This intracellular Ca(2+) increase was abolished by pretreating the neurons with thapsigargin (100 nM), suggesting that the UTP-induced increase in intracellular Ca(2+) is primarily due to the release of Ca(2+) from endoplasmic reticulum Ca(2+) stores. An enzyme-linked immunosorbent assay showed that an application of UTP (100 microM) significantly stimulated the release of CGRP and that suramin (100 microM) totally abolished the response, suggesting that P2Y receptor-mediated increase in intracellular Ca(2+) is accompanied by CGRP release from dorsal root ganglion neurons. These results suggest that metabotropic P2Y receptors contribute to extracellular ATP-induced increase in intracellular Ca(2+) concentration and subsequent release of neuropeptide CGRP in rat dorsal root ganglion neurons.

Adenosine Triphosphate↗

High-energy carbon ion irradiation and the inhibition of negative gravitaxis in Euglena gracilis Z.

PURPOSE: To study the effects of high-energy carbon ion irradiation on negative gravitaxis in the photosynthetic flagellate Euglena gracilis strain Z in a dose-response-dependent manner. MATERIALS AND METHODS: Cells were exposed to 290 MeV amu(-1) carbon ion from the Heavy Ion Medical Accelerator in Chiba (HIMAC) at 12 doses (0, 1, 5, 7.5, 10, 15, 20, 50, 75, 100, 150, 200 Gy for water). r was used to quantify negative gravitaxis observed in a recently developed biomonitoring system. RESULTS: Negative gravitaxis of Euglena was significantly affected by irradiation at a dose >7.5 Gy, 28 h after irradiation. Negative gravitaxis recovered after a few days. The rising phase of r during the first 30 s of orientation was independent of irradiation dose. The recovery mechanism itself was damaged by 290 MeV amu(-1) carbon ions at a dose >50 Gy. CONCLUSIONS: The results indicate that negative gravitaxis in Euglena may be affected by the radiation experienced during a space experiment especially during the occurrence of solar flares. The analyses of time needed to reorient to the gravitational vector suggest that the steering control of the flagellar apparatus is affected by exposure to the 290 MeV amu(-1) carbon ion irradiation, resulting in an altered beating pattern of a flagellum and/or changes of the cell form during reorientation.

Animals↗

Application of solid state integrating dosemeters to the determination of biologically equivalent doses in space.

If the biological responses are well approximated by the efficiencies of solid-state integrating dosemeters (SSID), the biologically equivalent doses can be simply estimated using SSID. For demonstrating the applicability of this method to space radiation dosimetry, biologically equivalent doses for two biological endpoints (enzyme inactivation and cell survival) were evaluated in the 8.8 d Shuttle-Mir mission (STS-89) using three commercial thermoluminescence dosemeters: Mg2SiO4:Tb, BeO:Na and 7LiF:Mg,Ti. The approximate biologically equivalent doses at two positions in the Spacehab module were found to be significantly different for trypsin inactivation, whereas they were almost identical for mammalian cell survival.

Aerospace Medicine↗

Efficiency of a radiophotoluminescence glass dosemeter for low-earth-orbit space radiation.

Chips of a radiophotoluminescence glass dosemeter (RPLG) were used for measurements of space radiation during a 9.8 d Shuttle-Mir mission (STS-91) at an altitude of 400 km and an inclination of 51.65 degrees. Two of RPLG chips were put into each of 59 positions in or on a life-size human phantom. The RPLG values equivalent to 137Cs gamma ray absorbed doses were found to be systematically lower than those of a Mg2SiO4:Tb thermoluminescence dosemeter (TDMS). In comparison with the organ or tissue absorbed dose and dose equivalent values that were estimated using a combination of TDMS and plastic nuclear track detectors, the efficiencies of the RPLG chips were about 80% for the water absorbed dose and about 40% for the dose equivalent. Whereas the percentage values will change during different missions, such additional information obtained from small RPLG chips is useful for improving the reliability of radiation dosimetry in space.

Aerospace Medicine↗

Increased blood carboxyhaemoglobin concentrations in inflammatory pulmonary diseases.

BACKGROUND: Exhaled carbon monoxide has been reported to increase in inflammatory pulmonary diseases and to be correlated with blood carboxyhaemoglobin (Hb-CO) concentration. A study was undertaken to determine whether arterial blood Hb-CO increases in patients with inflammatory pulmonary diseases. METHODS: The Hb-CO concentration in arterial blood was measured with a spectrophotometer in 34 normal control subjects, 24 patients with bronchial asthma, 52 patients with pneumonia, and 21 patients with idiopathic pulmonary fibrosis (IPF). RESULTS: The mean (SE) Hb-CO concentrations in patients with bronchial asthma during exacerbations (n=24, 1.05 (0.05)%), with pneumonia at the onset of illness (n=52, 1.08 (0.06)%), and with IPF (n=21, 1.03 (0.09)%) were significantly higher than those in control subjects (n=34, 0.60 (0.07)%) (mean difference 0.45% (95% confidence interval (CI) 0.23 to 0.67), p<0.01 in patients with bronchial asthma, mean difference 0.48% (95% CI 0.35 to 0.60), p<0.0001 in patients with pneumonia, and mean difference 0.43% (95% CI 0.26 to 0.61) p<0.001 in patients with IPF). In 20 patients with bronchial asthma the Hb-CO concentration decreased after 3 weeks of treatment with oral glucocorticoids (p<0.001). In 20 patients with pneumonia the Hb-CO concentration had decreased after 3 weeks when patients showed evidence of clinical improvement (p<0.001). The values of C-reactive protein (CRP), an acute phase protein, correlated with Hb-CO concentrations in patients with pneumonia (r=0.74, p<0.0001) and in those with IPF (r=0.46, p<0.01). In patients with bronchial asthma changes in Hb-CO concentrations were significantly correlated with those in forced expiratory volume in 1 second (FEV(1)) after 3 weeks (r=0.67, p<0.01). Exhaled carbon monoxide (CO) concentrations were correlated with Hb-CO concentrations (n=33, r=0.80, p<0.0001). CONCLUSIONS: Hb-CO concentrations are increased in inflammatory pulmonary diseases including bronchial asthma, pneumonia, and IPF. Measurement of arterial Hb-CO may be a useful means of monitoring pulmonary inflammation.

Aged↗

Measurement of des-gamma-carboxy prothrombin levels in hemodialysis patients positive for anti-hepatitis virus C antibody.

BACKGROUND: The prevalence of anti-hepatitis virus C (HCV) antibody is much higher in hemodialysis (HD) patients than in the normal population. Recently, blood des-gamma-carboxy prothrombin (PIVKA-II) has been demonstrated as a sensitive marker for the early detection of hepatocellular carcinoma (HCC). In this study, we measured blood PIVKA-II in HD patients positive for anti-HCV antibody or hepatitis B virus surface (HBs) antigen to examine if HD therapy may affect the measurement of PIVKA-II. PATIENTS AND METHODS: Ninety-four stable HD patients who had anti-HCV antibodies (n = 86) or HBs antigen (n = 8) without any evidence of HCC were enrolled in the study (age: 60 +/- 11 years, duration of HD: 17 +/- 10 years, male/female = 63/31). Five patients had liver cirrhosis and another 5 patients received warfarin treatment. We simultaneously measured serum PIVKA-II and alpha-fetoprotein (AFP), and compared the association between these markers and HCV RNA titer and laboratory parameters. RESULTS: Serum PIVKA-II became positive (> or = 40 mAU/ml) in only 5.6% (5/89) of patients without warfarin administration, ranging from 47 to 71 mAU/ml. Seventy out of 89 patients (78.7%) were below 20 mAU/ml. Serum PIVKA-II did not correlate with biochemical parameters including HCV RNA, while serum AFP was significantly correlated with serum AST (r = 0.21, p < 0.05), gamma-GTP (r = 0.21, p < 0.01) and platelet counts (r = -0.29, p < 0.01), respectively. In contrast, 5 patients receiving warfarin had an extremely high PIVKA-II value ranging from 1,930 to 19,900 mAU/ml. PIVKA-II was significantly and inversely correlated with the thrombotest value (r = -0.72, p = 0.01). CONCLUSION: The positivity of blood PIVKA-II in HD patients with hepatitis viremia was identical to that in patients without renal failure. Warfarin treatment dramatically increased serum PIVKA-II more than 1,000 mAU/ml. These findings suggested that HD treatment itself did not affect the measurement of PIVKA-II, but vitamin K deficiency can readily influence the PIVKA-II level in dialysis patients.

Adult↗

[Bilateral multiple pulmonary arteriovenous fistulas treated by surgery and embolization].

A 46-year-old man, with an abnormal shadow in left lower area in chest X-ray was admitted to our hospital. Three dimensional computed tomography (3 D-CT) and pulmonary arteriogram revealed bilateral multiple pulmonary arteriovenous fistulas (PAVF). PAVF was simple in the right S8, and were multiple in the right lower lobe. PAVF in left side was treated with percutaneous transcatheter coil embolization. PVAFs in right side were treated by lower lobectomy. This combination therapy seemed to be useful for bilateral multiple PAVFs.

Arteriovenous Fistula↗

Nitrate leakage from deciduous forest soils into streams on Kureha Hill, Japan.

Nitrate leakage from deciduous forest soils into streams was investigated for two adjacent hills. Many of the streams on Kureha Hill, located in Toyama City, Japan, have extremely high nitrate concentrations. The nitrate concentration of Hyakumakidani, one of the streams on Kureha Hill, averaged 158 microeq l(-1) and reached 470 microeq l(-1) during an episodic event. In contrast, the streams on Imizu Hill, adjacent to Kureha Hill, had low concentrations, below 15 microeq l(-1). Even during an episode, the nitrate concentrations increased to no more than 75 microeq l(-1). Both areas have similar blown forest soils, C/N ratios in O horizons, and vegetation consisting primarily of deciduous trees. However, soil incubation experiments, which lasted for 4 weeks, revealed that the nitrification rates in the surface soils of Kureha Hill were much higher than in the soils of Imizu Hill.

Agriculture↗

Yeast Ull1/Siz1 is a novel SUMO1/Smt3 ligase for septin components and functions as an adaptor between conjugating enzyme and substrates.

SUMO1/Smt3, a ubiquitin-like protein modifier, is known to conjugate to other proteins and modulate their functions in various important processes. Similar to the ubiquitin conjugation system, SUMO/Smt3 is transferred to substrate lysine residues through the thioester cascade of E1 (activating enzyme) and E2 (conjugating enzyme). In our previous report (Takahashi, Y., Toh-e, A., and Kikuchi, Y. (2001) Gene 275, 223-231), we showed that Siz1/Ull1 (YDR409w) of budding yeast, a member of the human PIAS family containing a RING-like domain, is a strong candidate for SUMO1/Smt3 ligase because the SUMO1/Smt3 modification of septin components was abolished in the ull1 mutant and Ull1 associated with E2 (Ubc9) and the substrates (septin components) in immunoprecipitation experiments. Here we have developed an in vitro Smt3 conjugation system for a septin component (Cdc3) using purified recombinant proteins. In this system, Ull1 is additionally required as well as E1 (Sua1.Uba2 complex), E2 (Ubc9), and ATP. A cysteine residue of the RING-like domain was essential for the conjugation both in vivo and in vitro. Furthermore, a region containing the RING-like domain directly interacted with Ubc9 and Cdc3. Thus, this SUMO/Smt3 ligase functions as an adaptor between E2 and the target proteins.

Amino Acid Sequence↗

Smad7 is induced by norepinephrine and protects rat hepatocytes from activin A-induced growth inhibition.

Activin A induces growth arrest of rat hepatocytes in vitro and in vivo. The alpha(1)-adrenergic agonist, norepinephrine (NE), enhances epidermal growth factor-stimulated DNA synthesis and inhibits activin A-induced growth inhibition, but the mechanisms of these actions are unclear. Smad proteins have recently been identified as intracellular signaling mediators of transforming growth factor-beta family members. In the present study, we explored how NE modulates the Smad signaling pathway in rat cultured hepatocytes. We demonstrate that NE inhibits activin A-induced nuclear accumulation of Smad2/3 and that NE rapidly induces inhibitory Smad7 mRNA expression. Infection of Smad7 adenovirus into rat hepatocytes inhibited activin A-induced nuclear accumulation of Smad2/3, enhanced epidermal growth factor-stimulated DNA synthesis, and abolished the growth inhibitory effect of activin A. We also demonstrated that the induction of Smad7 by NE is dependent on nuclear factor-kappa B (NF-kappa B). The amount of active NF-kappa B complex rapidly increased after NE treatment. Preincubation of the cells with an NF-kappa B pathway inhibitor N-tosyl-l-phenylalanine chloromethyl ketone or infection of the cells with an adenovirus expressing an I kappa B super-repressor (Ad5I kappa B) abolished the NE-induced Smad7 expression. These results indicate a mechanism of transmodulation between the Smad and trimeric G protein signaling pathways in rat hepatocytes.

Activins↗

Real-time observation of a single DNA digestion by lambda exonuclease under a fluorescence microscope field.

A fluorescence microscopy technique has been developed to visualize the behavior of individual DNA and protein molecules. Real-time direct observation of a single DNA molecule can be used to investigate the dynamics of DNA-protein interactions, such as the DNA digestion reaction by lambda exonuclease. In conventional methods it is impossible to analyze the dynamics of an individual lambda exonuclease molecule on a DNA because they can only observe the average behavior of a number of exonuclease molecules. Observation of a single molecule, on the other hand, can reveal processivity and binding rate of an individual exonuclease molecule. To evaluate the dynamics of lambda exonuclease, a stained lambda DNA molecule with one biotinylated terminal was fixed on an avidin-coated coverslip and straightened using a d.c. electric field. Microscopic observation of digestion of a straightened DNA molecule by lambda exonuclease revealed that the DNA digestion rate was approximately 1000 bases/s and also demonstrated high processivity.

Avidin↗

Characterization of a novel mammalian SUMO-1/Smt3-specific isopeptidase, a homologue of rat axam, which is an axin-binding protein promoting beta-catenin degradation.

A novel SUMO-1/Smt3-specific isopeptidase, SMT3IP2/Axam2 (Smt3-specific isopeptidase 2), was cloned and characterized. The catalytic domains in the carboxyl-terminal region were very much similar to those of other SUMO-1/Smt3-specific proteases, but the amino-terminal part was quite different. The enzyme specifically bound to Smt3a and Smt3b but not to SUMO-1. The SMT3IP2 expressed by Escherichia coli could cleave SUMO-1, Smt3a, or Smt3b from a SUMO-1/RanGAP1, Smt3a/RanGAP1, or Smt3b/RanGAP1 conjugate, respectively, and had the activity of a carboxyl-terminal hydrolase to produce a glycine residue in the carboxyl terminus of these ubiquitin-like proteins. The sequence data indicated that the amino acid sequence of SMT3IP2 was mostly identical to that of rat Axam, which binds to Axin and promotes the degradation of beta-catenin, although its amino-terminal region was much shorter than that of Axam. Therefore, we designated this isopeptidase SMT3IP2/Axam2. When human SW480 cells were transfected with wild-type SMT3IP2/Axam2, the beta-catenin disappeared. When the cells were transfected with the SMT3IP2/Axam2 C500A mutant, which had neither isopeptidase nor carboxyl-terminal hydrolase activity, or with the 1-352 mutant, which lacked the catalytic domain of the enzyme, again the beta-catenin disappeared, indicating that the enzyme activities were not necessary for the instability of beta-catenin in this transfection assay system and that its competition with Dvl for binding to Axin may be important for the instability of beta-catenin as suggested previously for Axam (Kadoya, T., Kishida, S., Fukui, A., Hinoi, T., Michiue, T., Asashima, M., and Kikuchi, A. (2000) J. Biol. Chem. 275, 37030-37037). The involvement of its enzyme activities in the Wnt signaling pathway remains to be elucidated.

Adaptor Proteins, Signal Transducing↗

[Occupationally induced nitric acid and sulfuric acid burns: an analysis of 2 patients from the aspect of occupational health].

We report two patients who suffered from acid burns while working in chemical factories. Case 1: a 44-year-old man who received burn induced by nitric acid on the face and extremities. Despite his protecting facial mask, he was exposed to nitric acid on his face through a gap between the mask and skin surface. Nitric acid was also sprinkled on his scalp which was not covered by a helmet or a protecting device. In addition, he suffered from acid burn on the right scapular region, the right upper arm, and the lower extremities through the work clothes. Case 2: a 26-year-old man who suffered from sulfuric acid burn on the forearms. Both patients were accidentally exposed to acids while they filled tanks with the acids through a hose. Following the manual of the factories, they washed the exposed skin with water for more than 15 minutes after the exposure. Although they recovered without any serious sequel, there remained partial deep tissue destruction of the skin. We reviewed these two cases from the aspect of industrial medicine, and proposed the following three points for improvement in the workplace to prevent accidental acid burns. 1) re-education or enlightenment activities for the well-experienced workers to avoid negligence to the danger of strong acid. 2) recommendation to take a complete shower to avoid overlooking of unaware acid injury. 3) improvement in the protecting facial mask. In addition, clinicians who examine acid-burn patients should not pass over the presence of deep ulcers lying behind the thick crust on the injured area.

Adult↗

A repeated pancreatectomy in the remnant pancreas 22 months after pylorus-preserving pancreatoduodenectomy for pancreatic adenocarcinoma.

We report a case of a repeated curative pancreatic resection in the remnant distal pancreas 22 months after pylorus-preserving pancreatoduodenectomy (PpPD). The patient was a 52-year-old woman with a past history of PpPD for adenocarcinoma of the head of the pancreas 22 months prior to the present operation. The original tumor was histopathologically diagnosed as a papillary adenocarcinoma with clear surgical margin at the surgical cut end of the pancreas (R0, International Union Against Cancer [UICC] classification). Twenty months after the PpPD, a follow-up computed tomography (CT) scan showed multiple low-density lesions in the body and tail of the pancreas without any other distant metastasis. A second operation, curative resection of the remnant pancreas, with splenectomy and distal gastrectomy, was performed. The second tumor was a papillary adenocarcinoma, the same diagnosis as that of the first tumor, and it also showed similar histopathological findings, including immunohistochemical staining of Ki-67 and p53 protein, and the same pattern of K-ras point mutation. The patient is considered to have shown a rare, unique pancreatic cancer with metachronous carcinogenesis in the remnant pancreas.

Adenocarcinoma, Papillary↗

Synovial cells from a patient with rheumatoid arthritis produce osteoclastogenesis inhibitory factor/osteoprotegerin: reciprocal regulation of the production by inflammatory cytokines and basic fibroblast growth factor.

To understand the involvement of osteoclastogenesis inhibitory factor (OCIF), also called osteoprotegerin (OPG), in the pathogenesis of bone destruction in rheumatoid arthritis (RA), we investigated the cytokine network involved in the production of OCIF by human fibroblast-like synovial (HFLS) cells from a patient with RA. Inflammatory cytokines, such as interleukin (IL)-1beta, IL-6 plus soluble IL-6 receptor (sIL-6R), IL-17, and tumor necrosis factor (TNF)-alpha, which are elevated in synovial fluid in RA, upregulated the production of OCIF to a level (5-20 ng/ml) sufficient to inhibit osteoclastogenesis in vitro. These inflammatory cytokines (except for IL-6 plus sIL-6R) stimulate OCIF production directly or indirectly through stimulation of prostaglandin E2 (PGE2) synthesis. In contrast to the findings with inflammatory cytokines, basic fibroblast growth factor (bFGF) inhibited the production of OCIF by the cells in a dose-dependent manner. While bFGF enhanced both the inflammatory cytokine-mediated release of PGE2 and the PGE2-mediated OCIF production, it significantly suppressed OCIF production by negating the direct stimulatory effect of the inflammatory cytokines. These findings suggest that bFGF in the synovial fluid of patients with RA may lead to severe joint destruction by suppressing the production of OCIF by HFLS cells.

Arthritis, Rheumatoid↗

Expression of R-cadherin in the regenerating chick sciatic nerve.

The ultrastructural localization of R-cadherin in normal and regenerating chick sciatic nerves was investigated immunocytochemically, and was compared with that of N-cadherin. R-cadherin was found on the plasmalemmae of axons and Schwann cells where cell-cell contacts were made in the normal unmyelinated fibers. It was also noted that R-cadherin was expressed on the axolemmae where regenerating axons contacted with each other, and formed fasciculations. The normal myelinated fibers displayed no immunoreactivity except at the mesaxon. These findings of R-cadherin were almost the same as those of N-cadherin of our previous study. N- and R-cadherin seemed to be co-expressed at the cell-cell contact points as mentioned above in the double labeling study. It is probable that cadherins contribute to the pathfinding of regenerating axons by causing them to form fasciculation. However, it seemed that there was no selective sorting of axons by N- and R-cadherin during regeneration as far as the present study was concerned.

Animals↗

High-angle annular dark-field STEM observation of Xe nanocrystals embedded in Al.

High-angle annular dark-field scanning transmission electron microscope (HAADF-STEM) observation of Xe precipitates embedded in crystalline membranes has been made using electron probes of atomic dimensions and HAADF-STEM images of Xe precipitates qualitatively different from conventional TEM observation results have been obtained. Multislice-based HAADF-STEM simulation has been made and it has been revealed that the intensity of images of Xe atoms at positions displaced from Al matrix columns decreases rapidly as the thickness increases. Even in a thin specimen, the off-site Xe atoms of the precipitate at deep locations, were not observable. Therefore, different images are expected for specimens of different thicknesses or depths of these precipitates. These results indicate that the observation of precipitates in crystalline membranes requires some care.

Journal Article↗

Involvement of PIAS1 in the sumoylation of tumor suppressor p53.

Sumoylation of p53 by the ubiquitin-like protein, SUMO-1/sentrin/PIC1, has been shown to stimulate its transcriptional activation activity. The SUMO E3 ligase, a key enzyme in the recognition of substrates to be sumoylated, has not yet been identified. We isolated PIAS1 (protein inhibitor of activated STAT1) as a SUMO-1 binding protein by yeast two-hybrid screening. In addition, PIAS1 bound p53 and Ubc9, the E2 for SUMO. PIAS1 that was mutated in the RING finger-like domain bound p53 and SUMO-1, but not Ubc9. PIAS1 catalyzed the sumoylation of p53 both in U2OS cells and in vitro in a domain-dependent manner. These data suggest that PIAS1 functions as a SUMO ligase, or possibly as a tightly bound regulator of it, toward p53.

Amino Acid Sequence↗