[Adaptation of English-written health-related quality of life measure for children].
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Biomedical subjects
Publications and source records attributed to N Yagi.
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The peptide hormone insulin forms a spherical aggregate, called a spherulite, at low pH and high temperature. A spherulite is composed of a core and many fibrils extending from it. These fibrils are thought to be amyloid fibers with a beta-sheet structure. In the present study, spherulites with a diameter of 50-100 microm were examined by X-ray fiber diffraction using a 6 microm beam. When a spherulite was scanned with the microbeam and the observed diffraction patterns were arranged in a two-dimensional array, the direction of the scatter was centrosymmetric, demonstrating a symmetric growth of fibrils. There were diffraction peaks at Bragg spacings of 23 nm, 3.3 nm and 1.2 nm in the direction perpendicular to the fibrils and 0.48 nm along the fibrils. The 0.48 nm reflection shows that the hydrogen bonds between beta-strands are along the fibril. The 23 nm reflection corresponds to the separation between fibrils, the 3.3 nm reflection is due to the arrangement of protofilaments, and the 1.2 nm reflection arises from the arrangement of peptide chains. On the basis of these results, a model of a fibril with an extended insulin molecule is proposed.
High quality real-time imaging of lungs in vivo presents considerable challenges. We demonstrate here that phase contrast x-ray imaging is capable of dynamically imaging the lungs. It retains many of the advantages of simple x-ray imaging, whilst also being able to map weakly absorbing soft tissues based on refractive index differences. Preliminary results reported herein show that this novel imaging technique can identify and locate airway liquid and allows lung aeration in newborn rabbit pups to be dynamically visualized.
In order to evaluate the effectiveness of edge enhancement by refraction in computed tomography, images of a cross section of a euthanized mouse thorax were recorded at low (20 keV) and high (72 keV) x-ray energies at a spatial resolution of about 40 microm. Compared with the images obtained with the detector at 30 cm from an object, when the object was located at 113 cm from the detector, the contrast between tissues and air was improved at both energies. The improvement was more pronounced at 72 keV where the absorption contrast was weaker. This effect was due to refraction at the surfaces of alveolar membranes and small airways which creates areas with apparently high and low linear attenuation coefficients within tissues. The edge enhancement by refraction was also effective in images of a euthanized rabbit thorax at x-ray energies of 40 and 70 keV at a spatial resolution of about 0.15 mm. These results raise the possibility that the refraction contrast may be utilized to obtain a high-resolution tomographic image of human lung and bone with low dose.
The significant degree of X-ray phase contrast created by air-tissue interfaces, coupled with the poor radiographic contrast of conventional chest radiographs, makes the inflated lung an ideal candidate for investigating the potential diagnostic improvement afforded by phase contrast X-ray imaging. In small animals these methods highlight the lung airways and lobe boundaries and reveal the lung tissue as a speckled intensity pattern not seen in other soft tissues. We have compared analyser-based and propagation-based phase contrast imaging modalities, together with conventional radiographic imaging, to ascertain which technique shows the greatest image enhancement for various lung sizes. The conventional radiographic image of a mouse was obtained on a Siemens Nova 3000 mammography system, whilst phase contrast images of mice and rabbit chests were acquired at the medical imaging beamline (20B2) at the SPring-8 synchrotron radiation research facility in Japan. For mice aged 1 day, 1 week and 1 month old it was determined that analyser-based imaging showed the greatest overall image contrast, however, for an adult rabbit both techniques yielded excellent contrast. The success of these methods in creating high quality images for rabbit lungs raises the possibility of improving human lung imaging using phase contrast techniques.
Phase contrast x-ray imaging of small animal lungs reveals a speckled intensity pattern not seen in other tissues, making the lungs highly visible in comparison to other organs. Although bearing a superficial resemblance to alveoli, the cause of this speckle has not been established. With a view to determining the mechanism for the formation of speckle, this paper details the results of propagation-based phase contrast experiments performed on mice lungs, together with packed glass microspheres used to emulate lung tissue. These experimental studies are compared to numerical simulations, based on wave propagation techniques. We find that speckle arises from focusing effects, with multiple alveoli acting as aberrated compound refractive lenses. Both experiments and modelling suggest that this speckle-formation phenomenon may lead to better screening methods for human lungs than conventional radiography.
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Effects of 20 mM inorganic phosphate on the cross-bridge behavior after photorelease of ATP from caged ATP was studied by X-ray diffraction in rat skinned psoas muscle fibers at 16 degrees C. In the first 30 ms after the photorelease, tension was similar in the presence and absence of phosphate. The tension development was then suppressed in the presence of phosphate. At 500 ms after the photolysis, it was lower by about 30% in the presence of phosphate. In the presence of phosphate, the intensity of the third meridional reflection from the thick filament at 1/14.4 nm(-1) increased more slowly and was 60% of the level without phosphate at 500 ms after the photolysis. The intensities of the equatorial (1,1) reflection and the actin layer-line at 1/36 nm(-1) decreased to a lower level in the presence of phosphate. These results suggest that phosphate does not affect dissociation of myosin heads from actin, but decreases the number of myosin heads in force-generating conformation and reduces tension. Phosphate may also shift the equilibrium between the detached and attached states towards the former.
Time-resolved dynamical x-ray imaging of individual DNA molecules with picometer-scale precision is demonstrated for the first time. Diffracted x-ray tracking (DXT), a single-molecule experiment with x rays, monitors the rotating motions, rather than the translational motions, of a labeled nanocrystal. DXT can obtain information about the dynamics of single molecules through a quantitative analysis, since the signals from DXT are independent of the chemical conditions.
In mice with acute dextran sulfate sodium colitis, we examined the effect of inducible nitric oxide synthase inhibition by (1S,5S,6R,7R)-7chloro-3-amino-5methyl-2-azabicyclo[4.1.0]heptane hydrochloride (ONO-1714) on colonic biochemistry, injury, and inflammation. Colonic luminal nitrate and nitrite were measured by the Griess reaction; inducible nitric oxide synthase messenger RNA expression by reverse transcription-polymerase chain reaction; and nitrotyrosine by immunohistochemistry. Mice with colitis showed increases in nitrate and nitrite, inducible nitric oxide synthase messenger RNA, and numbers of cells staining for nitrotyrosine. Colonic inflammation was severe. ONO-1714 inhibited increases in nitrate and nitrite and numbers of nitrotyrosine-positive cells; injury and inflammation also were reduced. Dextran sulfate sodium-induced increases in thiobarbituric acid-reactive substances, a lipid peroxidation marker, were blunted by ONO-1714, which also inhibited increases in mucosal inflammatory cytokines. Nitric oxide produced by inducible nitric oxide synthase may contribute to colonic inflammation by nitrosation, oxidative damage, and enhanced inflammatory cytokines.
A mechanical study on skinned rat psoas muscle fibers was performed at about 16 degrees C with X-ray diffraction and caged-ATP photolysis. The amount of photoreleased ATP was set < 0.2 mM for analysis of a 'single turnover' of the cross-bridge ATPase. With regard to the phase of activation, the results under the single turn-over condition were generally consistent with previous results obtained with larger amount of photoreleased ATP. Formation of the ADP-rigor state was mechanically monitored by the 90 degrees out-of-phase component of stiffness at 500 Hz, which was elevated on activation and then decreased to zero with a half-time of 0.2-0.3 s. Intensity changes of the X-ray reflections (e.g. equatorial reflections, actin layer lines and a myosin meridional reflection) indicated that a large number of cross-bridges returned to the rigor structure with a half-time of 0.5-0.7 s. During this phase, tension did not increase but slowly decreased with a half-time of about 1.0 s. The in-phase stiffness increased only 20-30% at the most. These results indicate that, even if the number of cross-bridges formed at any moment during full contraction is small, they can interact with actin and form rigor bonds with a rate of 1 s(-1). The force developed in the rigor formation is probably lost due to the presence of rigor bridges and compliance in the preparation.
We examined the roles of lipid peroxidation, neutrophil accumulation, and inflammatory cytokines in the protective effect of polaprezinc against aspirin-induced gastric mucosal injury in rats. The intragastric administration of acidified aspirin induced hyperemia and hemorrhagic erosions in rat stomachs. The increase in the total gastric erosive area after aspirin administration was significantly inhibited in a dose-dependent manner by treatment with polaprezinc. The increases in thiobarbituric acid-reactive substances and tissue-associated myeloperoxidase activity 3 hr after aspirin administration were significantly inhibited by pretreatment with polaprezinc. The gastric concentration of TNF-alpha increased after aspirin administration, and the increase was also inhibited in a dose-dependent manner by treatment with polaprezinc. The peak expression of TNF-alpha mRNA 1 hr after aspirin administration was inhibited by 30 mg/kg of polaprezinc. Based on these data, the beneficial effects of polaprezinc on aspirin-induced gastric mucosal injury may be attributed to its antioxidative and antiinflammatory properties.
We examined the effect of daily consumption of dietary diacylglycerol (DG) oil on serum lipid concentrations in patients with diabetes whose serum triacylglycerol (TG) levels were persistently increased despite continuous nutritional counseling at the outpatient clinic. Patients (n = 16) were divided into DG and control groups (n = 8 each). DG was incorporated (target dose 10 g/d) by substituting DG oil (80 g DG/100 g oil) for the ordinary TG cooking oil used at home for 12 wk. The control group continued consuming ordinary TG cooking oil. Dietary records indicated that there were no differences between groups in total energy intake or percentage of energy from fat. In the DG group, TG intake decreased from 26.8 +/- 9.3 to 15.7 +/- 8.9 g/d, whereas DG intake increased from 0.3 +/- 0.1 to 10.6 +/- 3.9 g/d. No differences between groups were observed in body weight, total fat intake or total oil consumption throughout the study period. In the DG group, serum TG levels decreased 39.4% from 2.51 +/- 0.75 mmol/L to 1.52 +/- 0.28 mmol/L. Serum glycohemoglobin A(1c) (HbA(1c)) concentration also decreased 9.7%. In contrast, there were no changes in these variables in the control group. Serum total and HDL cholesterol were not affected in either group. These results indicate that DG oil may be useful as an adjunct to the standard diet therapy of fat restriction in the management of diabetics with hypertriglyceridemia.
X-ray diffraction studies were made using synchrotron radiation on ferret right ventricular papillary muscle under three different thyroid states: euthyroidism, hyperthyroidism, and hypothyroidism. The latter two states were induced by treatment with L-thyroxine and methimazole, respectively. The X-ray equatorial reflections were recorded at a time resolution of 10 ms to study the mass movement of myosin cross-bridges from thick to thin filaments. The myosin isomer content was measured by gel electrophoresis which showed that V3 isomer was predominant in euthyroid muscle and 27% of myosin was V1 isomer in hyperthyroid muscle. The intracellular free Ca concentration was measured by using the aequorin method. The peak Ca concentration was similar in all three states, but in the hypothyroid state the time-to-peak was longer and the decay was slower. The time-to-peak of twitch tension was shorter in hyperthyroidism and longer in hypothyroidism than in euthyroidism. The different time courses of twitch tension in different thyroid states accompanied a cross-bridge movement which closely followed the tension development. In hyperthyroidism, the cross-bridge movement significantly preceded tension development, suggesting that hyperthyroid myosin (V1) has a longer latency period between the shift to the vicinity of the thin filament and force development.
We analyzed the clinical course of eight female patients of oculomotor nerve palsy due to unruptured internal artery posterior (IC-PC) communicating artery aneurysm in order to speculate on the mechanism of aneurysmal rupture. Seven of the eight patients had preceding headache or ophthalmalgia, three of them deteriorated oculomotor nerve palsy and one showed worsening of ophthalmalgia. These deteriorations occurred between three to seven days after the first clinical symptom appeared. Neck clipping of aneurysm was performed for all eight patients. These aneurysms were profiled except for one patient whose aneurysm ruptured before operation. None of these seven patients had subarachnoid hemorrhage in the operative field. The patients who showed clinical deterioration had fragile aneurysm wall, wall with bleb, and one aneurysm showed minor blood leakage during operative manipulation. In the four patients who did not show clinical deterioration, aneurysm attached oculomotor nerve, these aneurysm walls were arteriosclerotic and we could not find the fragile portion during operation. Repeated minor bleeding near the oculomotor nerve or direct aneurysmal compression are the main causes of oculomotor nerve palsy with unruptured IC-PC aneurysm. Adhesion or stimulation of aneurysm on the upper border of cavernous sinus cause the headache or ophthalmalgia and these deteriorations are thought to occur gradually. However, these deteriorations occur more rapidly than ever thought and reflect acute aneurysmal growth and its direct affect on the oculomotor nerve. IC-PC aneurysm is the only aneurysm we can observe its aneurysmal growth by its unique clinical presentation. From this study, we conclude that the deterioration of oculomotor nerve palsy, preceding headache or ophthalmalgia reflects rapid growth of unruptured IC-PC aneurysm. We neurosurgeons consider it to be an urgent rupture period and should introduce surgical management to prevent its rupture and the devastating consequence of subarachnoid hemorrhage.
We measured the M-N transition of wild-type bacteriorhodopsin (pH 9, 10 degrees C) by time-resolved x-ray diffraction study at SPring8 BL45XU-A. We confirmed the accumulation of M and N intermediates by absorbance measurements, and we found that the time resolution of x-ray diffraction experiments (244 ms) was sufficient to resolve the M-N transition. From the x-ray diffraction data, three components were decomposed by singular value decomposition analysis. The existence of three components in the M-->N-->BR reaction revealed that BR changes its structure during the M-N transition. Moreover, the difference Fourier maps of reconstituted fast and slow decay components clearly showed that the electron density distributions of the F helix changes in the M-N transition. The observed structural change at the F helix will increase access of the Schiff base and D96 to the cytoplasmic surface and facilitate the proton transfer steps that begin with the decay of the M state.
Two novel lipopeptides, which have the peptide ligands [alpha-melanocyte stimulating hormone (alpha-MSH)] sequence and repeated [Gly-Arg-Gly-Asp-Se (GRGDS) sequence], are designed, synthesized by the solid-phase method, and introduced into liposome membranes by the freeze-thaw method. These liposomes bearing the peptide ligands on their surface are expected to bind to cell membranes. We have confirmed that the lipopeptides are introduced into liposome membranes almost quantitatively, while such a high degree of incorporation has not been accomplished in conventional methods. In this respect, the present method is superior to prepare surface-modified liposomes that are applicable to drug carriers and so on. We have also confirmed by using immunoelectron microscopy that the peptide ligands are actually located in an aqueous phase. It has been shown by flow cytometry that the liposome bearing alpha-MSH peptide ligand binds to B16 cells and the liposome bearing the repeated GRGDS sequence binds to NIH3T3 cells.
Myelin-associated oligodendrocytic basic protein is a member of the proteins constituting the central nervous system myelin. By morphometric analysis, we demonstrated that axons of myelin-associated oligodendrocytic basic protein-deficient mice had larger diameters and more myelin lamellae as compared to those of wild-type mice at the same age. It is known that the number of myelin lamellae increases linearly with axonal diameter, and that the rate of radial axonal growth is the factor controlling the rate of myelin formation. In line with these observations, we found that the regression line for axonal diameter and the number of myelin lamellae in myelin-associated oligodendrocytic basic protein-deficient mice appeared to be identical to that in wild-type mice, indicating that the increase in the number of myelin lamellae was the result of the increase in axonal diameter. Furthermore, we generated myelin basic protein/myelin-associated oligodendrocytic basic protein-double-deficient mice through mating myelin-associated oligodendrocytic basic protein-deficient mice with shiverer mice, an autosomal recessive mutant characterized by a lack of all isoforms of myelin basic protein. With these knock-out mice, we showed that axons of the double-deficient mice had larger diameters and smaller form factor, an index of the deformation of the fiber contour, in ensheathed fibers than those of shiverer mice, although there was no difference in axonal diameter of unmyelinated fibers between them. Taken together, myelin-associated oligodendrocytic basic protein seemed to play a role in controlling axonal diameter and in keeping axons round.