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In Japan, due to the increase of old-old, Aging-Associated Cognitive Decline (AACD; Levy, 1994) qualifiers are rapidly increasing. The number of AACD group was 13 (age 74.7 ± 2.8), and the healthy group was 6 (age 71.8 ± 3.7). We had the participants to walk a 9m walking way, of 5m measuring distance made by connecting 2 “Walk Way” (ANIMA Corp.), plus 2m acceleration distance and 2m deceleration distance, 2 trials each of fastest possible gait (Speed up walking; S) and dual task gait of calculating while walking (Dual task walking; DT) in the respective order. Further, intervention task for the DT group was, to chant the answer of calculation starting from 70 minus 7 for the first trial and 50 minus 7 for the second trial, and continue to take away 7. We carried out “Five Cog” for detection of AACD elderly (Yatomi, 2006). The significant differences in gait characteristics between the two groups within the DT task were, gait speed (P=0.037), cadence (P=0.058), stride (P=0.149). On the other hand, there were no differences seen in the S task (n.s.). The S task (Shinkai, 2000), said to be beneficial as a good predictor of vital functions, could not detect AACD elderly. Within the DT task, since there was a gait speed decrease, it is possible to be beneficial as one of screening tasks to detect AACD elderly. It could be possible even for non-specialists, in a short time to detect AACD elderly, by adding a DT task to widely popular gait test.

Epithelial cells are connected to each other around taste pores in rat fungiform papillae. Cytoskeletal components are responsible for the maintenance of intracellular adhesion, and we investigated the identification and localization of actin filaments around taste pores. On the basis of observations made by immunohistochemical transmission electron microscopy comparing with confocal laser scanning microscopy using actin-lectin double staining, actin filaments were found to be localized, encircling the squeezed taste pore cavity, in epithelial cells a few micrometers below the papilla surface. In addition, these observations suggest that the organization of actin filaments around taste pores might be involved in the constriction of taste pores.

To realize the high-voltage vacuum circuit breaker, we investigated the voltage-distribution ratio after current interruption at the double-break vacuum circuit breaker. We used the spiral CuCr (20%) electrodes and the axial magnetic-field CuCr (50%) electrodes as the experimental electrodes. At the double-break vacuum circuit breaker, where two vacuum interruptions that had the same and different characteristics of the electrode structure and contact materials were connected in series, there was a bias in the voltage-distribution ratio of each vacuum interrupter after current interruption. This seems to be because there was difference in the postarc current that passed through each vacuum interrupter after current interruption.

Several Leonid fireballs were successfully photographed by the Japanese Fireball Network and by other observers in Japan on 16 November, 1996. A totals of seven of these were simultaneously observed from two or more stations, from which the orbital and physical data were deduced. The radiant of these fireballs were very small, only 0.1°, similar to that of the 1991 Perseids. The 1996 Leonids showed a lower magnitude distribution index similar to those obtained in the 1961 and 1965 Leonids. All of these showers occurred before perihelion passage of the parent comet. We conclude that we have already encountered the elongated front part of the dust trail of the Leonid parent comet, where the trail is probably composed of larger dust particles.

Cultured quail myocytes were much more resistant to H2O2 toxicity than cultured mouse myocytes. The intracellular concentration of glutathione ([GSH]i) and the activity of gamma-glutamylcysteine synthetase (gamma-GCS) in quail heart cells were about five and three times higher, respectively, than in mouse heart cells, although catalase and glutathione peroxidase (GSHpx) activity was similar in both. Preloading of gamma-glutamylcysteine monoethyl ester (gamma-GCE), a membrane-permeating GSH precursor, increased the H2O2 resistance of cultured mouse myocytes. These observations suggest that the high [GSH]i and the high activity of gamma-GCS in quail myocytes are responsible for their high resistance to H2O2. Both H2O2 sensitivity and [GSH]i of mosaic sheets composed of equal amounts of mouse and quail myocytes approximated those of sheets composed entirely of quail myocytes. From these observations, it is hypothesized that GSH was transferred from quail myocytes to mouse myocytes, probably through gap junctions between them, and that quail myocytes resynthesized GSH by a feedback mechanism, thus maintaining their intracellular GSH levels. When the fluorescent dye lucifer yellow was injected into a beating quail myocyte in a mosaic sheet, it spread to neighboring mouse myocytes but not to neighboring L cells (a cell line derived from mouse connective tissue). These observations indicate that existence of gap junctions in the region of cell contact between mouse and quail myocytes but not between quail myocytes and L cells. When quail myocytes preloaded with [3H]gamma-GCE were cocultured with mouse myocytes and L cells, the radioactivity was transmitted to neighboring mouse myocytes but not L cells. These observations show that GSH and/or its precursors can be transmitted from quail myocytes to mouse myocytes through gap junctions and that this can protect mouse myocytes from H2O2 toxicity. Mouse myocyte sheets composed of 10(4) cells or more showed higher resistance to H2O2 toxicity than single isolated mouse myocytes. Metabolic coupling of GSH between myocytes may contribute at least in part to this high resistance of the cell sheets.

We proposed a unique motion detection method that can be used for motion correction in brain PET imaging. This method uses a specially-designed solid marker that enables position and angle (direction) measurement with one optical movie camera. Therefore, it is applicable to a long, narrow patient port space in which a two-camera system cannot be installed. By refining machining accuracy of the solid marker, angle measurement accuracy was significantly improved. As a result, we obtained sufficient measurement accuracies in the angles as well as in the trans-axial positions. In addition, we have been developing a model to calculate axial positions from measured trans-axial positions and angles. As a result of a multivariate analysis of measured motion tracking data, we found that sufficient accuracy can be obtained under well-controlled experimental conditions. In order to realize head motion correction for jPET-D4, we are also considering marker attaching, camera calibration, device installation, and correction algorithms.

To realize the high voltage vacuum circuit breaker, we investigated the voltage distribution ratio after current interruption at the double-break vacuum circuit breaker. We used the spiral CuCr (20 %) electrodes and the axial magnetic field CuCr (50 %) electrodes as the experimental electrodes. At the double-break vacuum circuit breaker, where two vacuum valves that had same and different characteristics of the electrode structure and contact materials were connected in series, there was a bias in the voltage distribution ratio of each vacuum valve after current interruption. This seems to be because there was difference in the residual current that passed through each vacuum valve after current interruption.

Mastication and swallowing increase the heart rate, and posture change and respiration also modulate the heart rate. To clarify the role of mastication and swallowing in the modulation of the autonomic nervous activity, we investigated how they interact with modulation of the heart rate by changing body positions and respiration in young healthy subjects. R-R intervals of electrocardiogram at rest were significantly changed with different body positions, compared with supine and standing. A net shortening by mastication of a chewing gum base was similar in various postures. Respiration induced a periodic change in the R-R intervals, depending on the body postures, but mastication did not markedly change them in each posture. Dry swallowing at rest and spontaneous swallowing during the mastication in the sitting position induced a similar transient shortening and suppressed the respiration-induced changes after the swallowing. The net transient shortening by dry swallowing at rest was similar in the different postures. These results suggest that signals from mastication and swallowing are summated with those from body positions and respiration for shortening the R-R intervals and that signals from swallowing suppress the respiration-induced periodic changes.

Osmotic shock transiently induces a volume change in the cells, followed by a restoration of the cell volume due to intracellular water regulation. Effect of long-term osmotic shock on the water regulation is not completely understood. Vacuole formation by long-term osmotic shock was investigated to clarify the water exclusion mechanism from cytoplasm into intracellular vacuoles in 3T3-L1 cells. Incubation of cells in hypotonic solution reversibly induced the vacuole formation. Staining of vacuoles with fluorescent dyes revealed that vacuoles were derived from endoplasmic reticulum and Golgi apparatus but not lysosomes. Membrane-impermeable fluorescent dyes were taken up into some vacuoles from cytoplasm and extracellular solution, suggesting that some vacuoles exhibit the dynamic changes for the connection of plasma membrane, and that transporter for membrane-impermeable dyes might be active in some vacuole membranes. External Cl(-), but not Na(+), was required for vacuole formation. DPC suppressed the vacuole formation and increased cell height, and further incubation with DPC increased the number of dead cells. Bumetanide, dimethylamiloride, and HgCl(2) did not suppress the hypotonic stress-induced formation of water vacuoles. These findings suggest that 3T3-L1 cells regulate the intracellular water content through the DPC-sensitive external Cl(-)-dependent vacuole formation during long-term osmotic stress.

In this study, rat primary hepatocytes were cultured on a non-woven polytetrafluoroethylene fabric coated with a copolymer of poly (amino acid) urethane (PAU) to examine the expression of their original functions. In the culture dish, ammonium metabolic activity of hepatocytes cultured using the fabric coated with PAU is higher than that using collagen-coated polystyrene. It is also indicated that it is possible for hepatocytes to maintain the functions for long-term culture in the circulatory culture equipment with the fabric coated with PAU.

To develop liver substitute that restore, maintain and/or improve hepatic function, a variety of hybrid hepatic tissue systems have been studied using isolated porcine hepatocyte because of its easily availability[1]. The growth and function of cultured hepatocytes are known to depend on the mode of cell-cell interaction and cell to substrate interaction[2,3]. So there is currently much interest in designing substrates for enhancing the hepatocytes attachment, proliferation and function. Hepatocytes attach to culture dishes coated with the extracellular matrix (ECM) such as fibronectin and collagen. Hepatocytes exhibit different morphology, growth and differentiated function depending on matrix condition. The natural ECM like collagen and fibronectin usually induce hepatocytes to well spread morphology. In this paper, a new functional polymer of polyamino-acid urethane (PAU)[4], in which a polyurethane segment was combined with poly(γ-methyl-L-glutamate) (PMLG), was investigated as an artificial ECM [5] for enhancing the hepatocytes attachment, proliferation and function.

In this study, porcine primary hepatocytes were cultured with or without ascorbic acid 2-phosphate (Asc2-P) to compare the functional activities with rat primary hepatocytes. Ammonium metabolic activity and albumin secretion activity of porcine hepatocytes were lower than those of rat hepatocytes in early culturing period. However, ammonium metabolic activity of porcine hepatocytes was almost recovered to the maximum level of rat hepatocytes and albumin secretion activities of porcine hepatocytes became higher than that of rat hepatocytes after seventh day culturing. The addition of Asc2-P enhanced remarkably both activities of porcine hepatocytes. For comparison, both activities of rat hepatocytes were not recovered and decreased graduate during the culturing period. Asc2-P did not give an effect on the rat hepatocytes.

The effect on the taste pores of denervation of the chorda tympani nerve in the middle-ear cavity was studied comparing confocal laser microscopy with lingual nerve resection. Taste pore cells were stained for actin with rhodamine-phalloidin and positive fluorescence was observed as a ring shape at the transverse cross sections. Within three days after chorda tympani nerve resection the ring reaction disappeared, although the pore morphology remained intact as seen by scanning electron microscopy. On the other hand, lingual nerve resection did not induce such rapid disappearance of the ring reaction. These results suggest that the chorda tympani nerve plays a predominant role in the maintenance of actin filaments in taste pore cells.

We report a case of primary ciliary dyskinesia in which a living-donor lobar lung transplant was performed. A 24-year-old woman with a diagnosis of primary ciliary dyskinesia and bronchiectasis was admitted to Shinshu University Hospital because of persistent dyspnea and pyrexia over a period of 4 months. Although she was given various antibiotics, neutrophilia, elevated plasma C-reactive protein (CRP) levels, and respiratory failure persisted. Chest roentgenograms and computed tomography disclosed severe bronchiectasis and diffuse infiltrative shadows in both lung fields. Pseudomonas aeruginosa was detected in a sputum culture. Although a variety of conventional therapies were administered, the patient's oxygenation progressively deteriorated. She was intubated and assisted by mechanical ventilation. The patient and her family proposed lung transplantation, and we concluded that a living-donor lobar lung transplant would be a suitable treatment for her disease. We transported the patient to Okayama University Hospital by helicopter 10 days after intubation. A living-donor lobar lung transplant was successfully performed with lung tissues donated by the patient's mother and sister for each transplant site.

Effects of cytochalasin D on actin filaments in cells encircling taste pores were examined to clarify the functional role of actin filaments in the maintenance of taste pores in rat fungiform papillae, using a confocal laser microscope and a scanning electron microscope. Fluorescence in the taste pore cells was detected as a ring shape produced by actin staining with rhodamine-phalloidin. Treatment of fungiform papillae with cytochalasin D diminished the positive reactions in the taste pore cells and increased the inner diameter of the ring reactions. However, deformation of the taste pores in fungiform papillae was not detected under a scanning electron microscope after treatment with cytochalasin D. These findings suggest that the organization of actin filaments encircling the taste pores contributes to regulation of the taste pore's size in rat fungiform papillae.

In order to develop a production process for carboxypeptidase Y (CPY, yeast vacuolar protease) secreted by Saccharomyces cerevisiae KS58-2D, medium composition, culture conditions, and expression systems were investigated. We found that the addition of histidine to thiamine-free medium, in which CPY production was almost negligible, raised the intracellular thiamine level, resulting in the increase of CPY production. On the basis of the choice of an expression system that uses an inducible GAL10 promoter, reassessment of histidine concentration in the medium, and optimization of the pH level during cultivation (pH 6.5), active CPY was secreted in a quantity of over 400 mg/l, which was more than tenfold that higher than that previously reported. The process developed could be easily scaled-up to industrial-scale fermentation.

We compared the inhibitory effects of histamine and pentagastrin (PG) on motilin-induced upper gastrointestinal phase III activity in conscious dogs that had surgically prepared Heidenhain pouchs (HP). Contractile activity was measured by means of chronically implanted force transducers, and changes in pH of the perfusate through the HP were monitored simultaneously. Intravenous infusion of PG (4 micrograms/kg-hr) inhibited motilin-induced phase III activity both in the main stomach and in the HP, whereas histamine (40 micrograms/kg-hr) inhibited activity only in the main stomach. Famotidine (0.3 mg/kg, i.v., the dose that completely inhibited gastric acid secretion by PG or histamine) blocked the inhibition of phase III activity induced by histamine but did not affect PG-induced inhibition. L-364,718 (1 mg/kg, i.v.), which had no effect on the PG-induced decrease in the pH of the perfusate lowered by PG, reversed the inhibition of phase III activity by PG in the HP but not in the main stomach. However, L-364,718, when combined with famotidine, potently reversed the PG-induced inhibition of phase III activity both in the main stomach and in the HP. These results show that the inhibitory effect of PG on motilin-induced phase III activity is brought about by two distinctive mechanisms, gastric acid and the cholecystokinin receptors-dependent mechanism, whereas the histamine-induced inhibition is mediated only by gastric acid. In the vagally denervated HP, however, gastric acid is not involved in an inhibitory effect of PG.

This study evaluated insulin secretion and insulin sensitivity before and after short-term oral administration of doxazosin in patients with essential hypertension. The hypertensive group consisted of 11 nonobese subjects (aged 41.0 +/- 2.5 years (mean +/- SEM), body mass index 24.0 +/- 0.53 kg/m2). The normotensive group consisted of 12 subjects matched to the hypertensive group for age and body mass index. The hypertensive group showed significantly higher concentrations of prestimulated and stimulated plasma insulin and plasma C peptide than normal groups. The insulin-mediated glucose disposal rate during euglycaemic clamp (M-value) was significantly lower in the hypertensive group than in normal controls (7.32 +/- 0.56 vs 8.88 +/- 0.34 mg/kg/min, P0.05). After one month of doxazosin treatment blood pressure was significantly reduced (P0.05). The short-term administration of doxazosin improved the M-value significantly to 8.60 +/- 0.62 mg/kg/min without a significant change in stimulated plasma C-peptide level. These data show that hypertension is associated with increased insulin secretion and impaired insulin sensitivity. Selective alpha 1-adrenergic inhibition with doxazosin improves the decreased glucose disposal rate associated with hypertension.

Fifty three cases of Reinke's edema were classified into 3 groups according to Yonekawa's proposed classification. Of these cases, 14 were Type I, 22 were Type II and 17 were Type III. In each case, psychoacoustic evaluation using the "GRBAS" scale and phonatory function tests (fundamental frequency, air flow rate, sound pressure level and maximum phonation time) using Nagashima PS-77 phonatory function analyzer were performed. Psychoacoustically, the voice quality before surgery was estimated moderately impaired, in general, with high grade Roughness accompanied by Asthenisity and Strainedness. Cases with more severe lesions showed much worse psychoacoustic evaluation results and severely impaired phonatoy function, but they also showed greater improvement after surgery. Phonatory function improved significantly within a month and psychoacoustic evaluation improved significantly from 1 to 3 months after surgery, though neither returned to the normal range. In conclusion, we consider that surgical therapy is appropriate in Type II and III cases, with voice therapy and cessation of smoking also necessary for good recovery.

Sex related differences in drugs metabolizing enzymes was studied using Wistar-rats bearing carrageenan-induced edema. One day after treatment with carrageenan, aminopyrine N-demethylase activity was inhibited in males but not in females. Carrageenan treatment did not alter the activity of aniline hydroxylase in either sex of the rats. These data show that there is a sex-related difference in the ability of carrageenan to alter the hepatic drug metabolism, which is substrate specific, in the rat.

We propose a new method of head motion tracking for positron emission tomography (PET) brain imaging. This method uses one optical camera and one solid marker, differing from the conventional method, which uses two or more cameras and markers (e.g. the POLARIS system). We expect this to be useful for jPET-D4, whose patient port is long and small. We present the principle of the head motion measurement and show the results of preliminary experiments.

By Y. Kubota, K. Hukuda, F. Ishihara, M. Furukawa and Y. ShibaAlmost all the inhabitants of thirty one communities in the Upper Ina District, Nagano Prefecture, were subjected to a systematic survey. There were 4, 891 families of consanguineous marriage among 30, 984, couples, i. e. amounting to 15.79%, of which9.88% were cousin marriages, 1.71% marriages between children of cousins and cousins, 4.19% marriages between second cousins.Eugenic Index of Inbreeding of the population, when expressed on the cousin marriage level, was as follows:0.0988+0.0171/2+0.0419/4=0.1178This index, proposed by the senior author (K.H.), refers to the degree of risk apprehended on the ground of race hygiene, especiany when a simple recessive gene exists in the population. When there are no other types of consanguineous marriage than cousin marriages, their frequency corresponds to the index. When there are consanguineous marriages more remote, their frequency contribute, with due reduction, to the index of inbreeding.Kawashima Village has the highest rate of consanguineous marriage, i. e. 28.4 (= 16.43 +3.62+8.21) %. The Eugeie Index of Inbreceding amounts to 0.2029. The community with the lowest rate of consanguineous marriage is Takatoo Town, which still has 9.5 (=6.65+1.21+1.66) S. The Index of Inbreeding, 0.0767, is still very high.Result of further analysis will follow in separate papers.