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Biomedical subjects

M Tamura

Publications and source records attributed to M Tamura.

At least 379 records · Page 21Linked to original sources

Interactions between calcium oxalate monohydrate crystals and Madin-Darby canine kidney cells: endocytosis and cell proliferation.

The present investigation was designed to study the biological responses in cultures of Madin-Darby canine kidney (MDCK) cells exposed to calcium oxalate monohydrate (COM) crystals, the most common type of urinary crystals. The addition of COM crystals significantly accelerated the multiplication of MDCK cells and significantly activated the cell viability. After exposure of MDCK cells to COM crystals, scanning electron microscopy revealed that some crystals adhered to the plasma membrane and others were endocytosed by the cell. This cellular uptake of crystals was time dependent from 1 to 8 h and showed a specificity according to crystal type. However, the endocytosis of aggregated COM crystals was less marked than that of non-aggregated crystals. Pre-treatment with each of the glycosaminoglycans (sodium pentosan polysulphate, heparin, and chondroitin sulphate C) produced a significant reduction of the cellular uptake of COM crystals, suggesting that these glycosaminoglycans may play some critical roles in preventing the cellular uptake of crystals. Although investigation in further detail is necessary, we speculate that these crystal-cell interactions, that is, the cellular uptake of crystals and cell proliferation, may be among the earliest processes in the formation of kidney stones.

Animals↗

Local application of basic fibroblast growth factor into the bone increases bone mass at the applied site in rabbits.

The effect of basic fibroblast growth factor (bFGF) applied locally into the bone under physiological conditions was investigated. An aqueous solution containing 0 microgram (vehicle), 100 micrograms or 400 micrograms recombinant human bFGF was percutaneously applied through a needle into the right ilium in rabbit, and the ilia were harvested 4 weeks after the application. Compared with vehicle-treated animals, bone mineral density measured by dualenergy X-ray increased in the 400 micrograms bFGF group. The width of trabeculae in the bFGF-treated groups was greater than in the vehicle group. These results showed that bFGF applied locally into the bone under physiological conditions affected bone formation, and suggested that such an application might have potential for increasing bone.

Absorptiometry, Photon↗

Near-infrared monitoring of cerebral oxygenation state during carotid endarterectomy.

BACKGROUND: Recent studies have indicated that near-infrared spectroscopy (NIRS) could continuously and noninvasively observe the changes in cerebral oxygenation state during hypoxia and ischemia, using their optical properties. Its validity and usefulness during cerebrovascular surgery, however, still remain to be clarified. METHODS: Using NIRS, we continuously monitored the changes in the concentration of oxyhemoglobin, deoxyhemoglobin, and total hemoglobin ([oxy-HB], [deoxy-Hb], and [total Hb], respectively) and redox state of cytochrome oxidase (cyt ox) during carotid endarterectomy for 22 patients, and we compared the NIRS responses with those of intraoperative somatosensory evoked potentials (SEP) and regional cerebral blood flow (rCBF). RESULTS: In 9 of 22 patients, cross-clamping of the carotid artery caused a continuous decrease [oxy-Hb] and [total Hb], and an increase in [deoxy-Hb]. Cyt ox was partially reduced during the clamping. These NIRS responses demonstrated the occurrence of severe hypoxia in the ipsilateral cerebral tissue. These patients showed a marked decrease in the N20 amplitude of SEP and rCBF. In contrast, the other 13 patients did not show a significant decrease in the cerebral oxygenation state, which showed no remarkable changes in either SEP or in rCBF. CONCLUSIONS: NIRS could successfully jude the cerebral oxygenation state noninvasively during carotid surgery and was more sensitive to ischemic crisis than other indirect methods.

Aged↗

NS-3, a TRH-analog, reverses memory disruption by stimulating cholinergic and noradrenergic systems.

The effects of a TRH-analog, N[[(3R,6R)-6-methyl-5-oxo-3-thiomorpholinyl]carbonyl]-L-histidyl-L - prolinamide tetrahydrate (NS-3, CG3703, montirelin hydrate) were compared with those of physostigmine on learning and memory disruption in the passive avoidance response (PAR) induced by either electrolytic lesion of the nucleus basalis magnocellularis (NBM) or by treatment with the noradrenergic neurotoxin, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) in rats. a) In NBM-lesioned rats, both NS-3 and physostigmine significantly reversed disruption of memory consolidation examined 15 min after the training session when these drugs were injected IP immediately after the training session. In addition, reversal by NS-3 (0.1 mg/kg) of the disruption of memory was observed even in the retention test conducted 24 h after the training session. b) NS-3 (0.5 mg/kg) significantly reversed the disruption of memory retrieval, when the drug was administered 15 min before the test session. c) DSP4 (50 mg/kg IP) caused memory disruption when the retention tests were conducted between 1 and 48 h after the acquisition session. NS-3 (0.1 mg/kg), but not physostigmine, significantly reversed the disruption of memory induced by DSP4 treatment. These findings suggest that the consistent antiamnestic action of NS-3 is due to the enhancement of both central cholinergic and noradrenergic systems, possibly via facilitation of the release of these transmitters.

Acetylcholine↗

Effect of MDP-Lys(L18) as a mucosal immunoadjuvant on protection of mucosal infections by Sendai virus and rotavirus.

To examine the effect of MDP-Lys(L18), a derivative of muramyl dipeptide (MDP), as a mucosal immunoadjuvant, we investigated its activity to augment host resistance against mucosal infections by Sendai virus and rotavirus in mice. In an experimental infection model using suckling mice (10-day-old) inoculated perorally (p.o.) with 1.5 x 10(6) p.f.u. mouse-1 of rotavirus strain SA11, intrarectal (i.r.) as well as p.o. administration of MDP-Lys(L18) (50 micrograms mouse-1) prior to virus infection markedly reduced rotavirus-induced diarrhea. Furthermore, when MDP-Lys(L18) was administered p.o. (1 mg mouse-1), i.r. (300 micrograms mouse-1) or intranasally (i.n., 100 micrograms mouse-1) various days before Sendai virus infection (2.6 x 10(4) HAD mouse-1), all the mucosal administration of MDP-Lys(L18) significantly protected a lethal infection of Sendai virus, showing a dose-dependent manner. However, the efficacy of MDP-Lys(L18) to induce the prophylactic activity against the viruses somewhat varied according to the administration route and timing. In time course analysis of virus isolation in vivo, the mice administered with MDP-Lys(L18) exhibited a significant reduction of both viruses in the lungs for Sendai virus and in the bowels for rotavirus. These results suggest that MDP-Lys(L18) is a potent mucosal immunoadjuvant to enhance nonspecific host resistance against two mucosal infectious viruses, Sendai virus and rotavirus.

Acetylmuramyl-Alanyl-Isoglutamine↗

A comparison of children with suprasellar germ cell tumors and craniopharyngiomas: final height, weight, endocrine, and visual sequelae after treatment.

BACKGROUND: Although treatment results of craniopharyngiomas and germ cell tumors in children have been reported extensively, the relationship between various posttreatment sequelae and the nature of the tumor is not well understood. METHODS: From 1968-94, 47 children with 23 suprasellar germ cell tumors and 24 craniopharyngiomas were treated at Gunma University Hospital, Japan. Forty survivors were assessed with regard to Karnofsky scores, visual scores, final height, final weight, and other endocrine sequelae as long-term functional prognostic indicators. RESULTS: Kaplan-Meier life-table estimates and Karnofsky scores were not significantly different between the two histologic groups. The visual function of survivors did not differ significantly at diagnosis. However, the final visual function following treatment in the germ cell tumor group was better than in the craniopharyngioma group (p < 0.05). Antidiuretic hormone secretion was significantly impaired at diagnosis in the germ cell tumor group (p < 0.0001). However, there was only a marginally significant difference in posttreatment persistent diabetes insipidus. Craniopharyngioma children were shorter and more obese at diagnosis (p < 0.02), and became much more obese than germinoma children with the same hormone therapy (p < 0.003), although the height difference became insignificant following growth hormone treatment. CONCLUSIONS: Children with craniopharyngiomas tended to have more visual disturbances, increased dwarfism and obesity, and less diabetes insipidus than patients with germ cell tumors despite adequate therapy. The method of quantifying visual function may in itself be useful.

Adrenocorticotropic Hormone↗

Interleukin-1 alpha stimulates interstitial collagenase gene expression in human dental pulp fibroblast.

Interstitial collagenase (matrix metalloprotease-1) is a member of a family of matrix metalloproteases and is thought to play a role in degradation of the extracellular components, such as collagens in normal extracellular matrix remodeling and in many disease processes. We examined interstitial collagenase mRNA expression in human dental pulp fibroblast cultures by Northern blot analysis. These cells did not express interstitial collagenase mRNA in an unstimulated condition. Inflammatory cytokines, such as interleukin-1 alpha, induced interstitial collagenase mRNA expression in these cells. The interstitial collagenase mRNA levels began to increase after 2-h exposure, reaching a maximum after 8 h, then dropping to the unstimulated level at 48 h. These effects were observed in a dose-dependent manner in a dose range of 0.1 to 10 ng/ml. Transforming growth factor-beta reduced the levels of interstitial collagenase mRNA expression that were induced by interleukin-1 alpha. These observations suggest that interstitial collagenase mRNA expression in human dental pulp fibroblasts is regulated by the inflammatory cytokines and that interstitial collagenase may play a role in tissue degradation in inflamed dental pulp.

Adolescent↗

Interleukin-8 gene expression by human dental pulp fibroblast in cultures stimulated with Prevotella intermedia lipopolysaccharide.

Interleukin (IL)-8 mRNA expression was investigated in human dental pulp fibroblast cultures after stimulation with lipopolysaccharide (LPS) prepared from Prevotella intermedia and inflammatory cytokines. The expression of IL-8 mRNA and the release of IL-8 induced by P. intermedia LPS in pulpal fibroblast cultures were detected by Northern blot analysis and ELISA, respectively. The sufficient concentration of P. intermedia LPS on the IL-8 mRNA expression was 0.1 microgram/ml in pulpal fibroblast cultures. IL-8 mRNA levels began to increase after 2 h of exposure, reached a maximum at 4 to 8 h, and declined after 48 h, reaching the unstimulated level by 60 h. IL-8 production by the pulpal fibroblasts began to increase after 8 h of exposure upon stimulation with 10 microgram/ml of P. intermedia LPS. By contrast Salmonella LPS and synthetic lipid A did not increase IL-8 mRNA concentrations in pulpal fibroblast cultures. Recombinant human IL-1 alpha, beta, and tumor necrosis factor-alpha were capable of stimulating these cells to express IL-8 mRNA but natural human interferon-beta, gamma, and recombinant human IL-6 were incapable in our assay. These results suggest that pulpal fibroblasts are immunoresponsive cells and can elaborate IL-8 upon stimulation with P. intermedia LPS.

Adolescent↗

Scanning electron microscopic characteristics of small-incision intraocular lenses.

PURPOSE: To evaluate the surface characteristics of commonly used, small-incision, intraocular lenses (IOLs). METHODS: Representative samples of five groups of foldable IOLs (4 silicone and 1 acrylic) underwent surface and edge-finish examination using a slit lamp. The IOLs were folded using a folding block and forceps. All the IOLs then were examined using a scanning electron microscope. A one-piece polymethylmethacrylate IOL was used for comparing surface-finish characteristics. The IOLs were examined for optic surface quality, edge finish, haptic, haptic/optic junction, and possible post-folding modifications. RESULTS: Slit-lamp evaluation of the surface quality of all of the silicone lenses demonstrated a smooth finish of the optic surface, edge, and haptics. Scanning electron microscopic analysis of the IOLs demonstrated adequately finished haptics or footplates and optics. Excess molding flash was seen on the edges of the some of the silicone IOLs, and no molding flash was observed on others. The acrylic IOL had a somewhat sharper optic edge. Irregular finish of the haptic/optic junctions of some of the IOLs (both silicone and acrylic) was noted. CONCLUSIONS: Currently available foldable IOLs have demonstrated adequate lens finish. However, irregularities of the haptic/optic junctions and molding flash are present on most IOLs evaluated, indicating room for improvement in the finish of foldable IOLs. Phacoemulsification with capsular bag IOL placement may decrease the clinical significance of these relatively subtle lens finish irregularities.

Acrylamide↗

A simple and novel algorithm for time-resolved multiwavelength oximetry.

Using time-resolved spectroscopy, we have developed an experimental approach to obtain the absolute changes in concentration in the scattering medium of living tissues. The time-resolved Beer-Lambert equation can be applied to living tissue due to the fact that the optical attenuation by absorption can be separated from that by scattering, and the intensity of the light along the non-linear scattered optical path is exponentially attenuated by the absorption. Based on the above, the absolute concentration of haemoglobin as well as oxygen saturation in the rat head can be determined in situ under various respiratory conditions where multiwavelength measurements were performed. The optically assessed values agree with those determined directly by the gas analysis of our previous report. The present method is very simple and therefore opens up wide applications for time-resolved spectrophotometry in clinical medicine as a technique for quantitative near-infrared oxygen monitoring.

Algorithms↗

Characterization of the nucleocapsid protein of Hantaan virus strain 76-118 using monoclonal antibodies.

We characterized the antigenic sites on the nucleocapsid protein (NP) of Hantaan virus (HTN) using 10 monoclonal antibodies (MAbs). At least seven antigenic sites were revealed by a competitive binding assay and divided into three partially overlapping antigenic regions (I, II and III). Regions I [amino acids (aa) 1-103], II (aa 104-204) and III (aa 205-402) were mapped on NP by examining the reactivity of truncated gene products. Those that corresponded to region I reacted with immune mouse serum, indicating that the region contained major linear epitopes as reported with Four corners virus (FCV) and Puumala virus (PUU) NP. At least one MAb to each region inhibited viral growth when they were introduced into cells by scrape-loading. In addition, they conferred protection from a lethal HTN challenge to newborn mice. A PEPSCAN assay localized the epitope of MAb E5/G6 between aa 166-175. Since E5/G6, which had the highest inhibitory effect both in cells and in mice, showed no virus neutralization activity by ordinary neutralization test, this region is suggested to be important for the virus growth after entry into the cells.

Amino Acid Sequence↗

Study of the changes of serum hyaluronic acid during porcine liver transplantation: influence of warm ischemia.

Twelve porcine liver transplantations were performed to investigate whether serum hyaluronic acid (HA) serves as a marker of warm ischemic injury. Group 1 was a control without warm ischemia (n = 7), and pigs in Group 2 were sacrificed by intracardiac KCl injection 60 min before harvesting (n = 5). All pigs survived more than 4 days in Group 1. In Group 2, all died within 2 days due to graft failure. Arterial and hepatic venous glutamic-oxaloacetic transaminase (GOT) in Group 2 were higher after revascularization. However, there were no differences between the 2 groups in arterial and hepatic venous HA levels. HA clearance by the graft also showed no differences between the groups. Although GOT reflected the degree of warm ischemia, HA and its hepatic clearance were not influenced by warm ischemic damage. In conclusion, HA was not thought to serve as a marker of liver injury when the graft suffered from warm ischemia.

Animals↗

Ouabain increases aldosterone release from bovine adrenal glomerulosa cells: role of renin-angiotensin system.

To evaluate the potential physiological significance of ouabain or a ouabainlike substance, we investigated the effect of nanomolar concentrations of ouabain on aldosterone release by cultured bovine adrenal glomerulosa cells. Ouabain (10 nM) increased aldosterone release from 0.35 to 0.89 ng.mg-1.4 h-1 in the serum-containing medium. Losartan prevented this increase. When angiotensinogen was added to the nonserum medium, 10 nM ouabain enhanced the aldosterone release. Losartan again blocked the increase. These findings together with a stimulation of renin release by ouabain indicate that angiotensin II generated by the adrenal cell renin-angiotensin system in the presence of exogenous serum or exogenous angiotensinogen is necessary for the ouabain-induced stimulation of aldosterone release. Ouabain (10 nM) enhanced the intracellular calcium concentration increase elicited by 0.1 nM angiotensin II severalfold. Addition of 1 nM ouabain enhanced the aldosterone secretion resulting from the addition of 1 nM angiotensin II. Nanomolar levels of ouabain, therefore, interact with both locally formed and exogenous angiotensin II to stimulate aldosterone production. A suggested mechanism is that ouabain increases calcium stores in the endoplasmic reticulum, thereby increasing the agonist response.

Aldosterone↗

Effects of docosahexaenoic acid and sardine oil diets on the ultrastructure of jejunal absorptive cells in adult mice.

The influence of docosahexaenoic acid (DHA) and sardine oil diets on the ultrastructure of jejunal absorptive cells was studied. Adult male Crj:CD-1 (ICR) mice were fed a fat-free semisynthetic diet supplemented with 5% (by weight) purified DHA ethyl ester, refined sardine oil, or palm oil. The mice received the DHA or palm oil diets for 7 days (groups 1 and 2) and the refined sardine oil or palm oil diets for 30 days (groups 3 and 4). There were significant ultrastructural changes in the jejunal absorptive cells between the mice fed on the palm oil diet and those receiving the DHA and sardine oil diets. The endoplasmic reticulum and Golgi apparatus of some jejunal absorptive cells in the mice fed on the palm oil diet for 7 and 30 days developed vacuolation on the upper site of the nucleus. In contrast, many granules, which appeared to be lipid droplets, were observed in the endoplasmic reticulum and Golgi apparatus of the jejunal absorptive cells in the DHA and sardine oil diet groups. These results suggest that ultrastructural differences in the jejunal absorptive cells between mice in the omega-3 fatty acid and palm oil diet groups may be associated with the changes in lipid metabolism.

Animals↗

Osteoclast function is activated by osteoblastic cells through a mechanism involving cell-to-cell contact.

We have established a method for obtaining an enriched preparation of functionally active osteoclast-like multinucleated cells (enriched OCLs) from co-cultures of mouse primary osteoblasts and bone marrow cells. Using these enriched OCLs, the effect of osteoblastic cells on osteoclast function was examined in two assays: a pit formation assay and an assay for actin ring formation. The enriched OCLs cultured for 24 h on dentine slices formed only a few resorption pits. When various numbers of primary osteoblasts were added to the enriched OCLs, the areas of the resorption pits increased proportionally to the number of osteoblasts added. Like primary osteoblasts, the established cell lines of osteoblastic cells (MC3T3-E1 and KS-4) and bone marrow-derived stromal cells (MC3T3-G2/PA6 and ST2) potentiated the pit formation caused by enriched OCLs. In contrast, the fibroblastic cell lines NIH3T3 and C3H10T1/2) and the myoblastic cell line (C2C12) failed to activate OCL function. When cell-to-cell contact between MC3T3-E1 cells and enriched OCLs was prevented, only a few resorption pits were formed. Pit formation by enriched rat osteoclasts placed on dentine slices was also stimulated by adding MC3T3-E1 cells. Actin ring formation and pit forming activity were well correlated in either culture of enriched mouse OCLs or authentic rat osteoclasts on dentine slices. These results indicate that osteoclast function is activated by osteoblastic cells through a mechanism involving cell-to-cell and/or cell-to matrix contact.

3T3 Cells↗

Osteoclast function is activated by osteoblastic cells through a mechanism involving cell-to-cell contact.

We have established a method for obtaining an enriched preparation of functionally active osteoclast-like multinucleated cells (enriched OCLs) from co-cultures of mouse primary osteoblasts and bone marrow cells. Using these enriched OCLs, the effect of osteoblastic cells on osteoclast function was examined in two assays: a pit formation assay and an assay for actin ring formation. The enriched OCLs cultured for 24 h on dentine slices formed only a few resorption pits. When various numbers of primary osteoblasts were added to the enriched OCLs, the areas of the resorption pits increased proportionally to the number of osteoblasts added. Like primary osteoblasts, the established cell lines of osteoblastic cell (MC3T3-E1 and KS-4) and bone marrow-derived stromal cells (MC3T3-G2/PA6 and ST2) potentiated the pit formation caused by enriched OCLs. In contrast, the fibroblastic cell lines (NIH3T3 and C3H10T1/2) and the myoblastic cell line (C2C12) failed to activate OCL function. When cell-to-cell contact between MC3T3-E1 cells and enriched OCLs was prevented, only a few resorption pits were formed. Pit formation by enriched rat osteoclasts placed on dentine slices was also stimulated by adding MC3T3-E1 cells. Actin ring formation and pit forming activity were well correlated in either culture of enriched mouse OCLs or authentic rat osteoclasts on dentine slices. These results indicate that osteoclast function is activated by osteoblastic cells-through a mechanism involving cell-to-cell and/or cell-to-matrix contact.

Animals↗