Biomedical subjects

M Uchida

Publications and source records attributed to M Uchida.

At least 73 records · Page 4Linked to original sources

PubMed2000

A member of Forkhead family transcription factor, FKHRL1, is one of the downstream molecules of phosphatidylinositol 3-kinase-Akt activation pathway in erythropoietin signal transduction.

The phosphatidylinositol 3-kinase (PI3K) signaling pathway is important for the regulation of a number of cellular responses. Serine/threonine kinase Akt (protein kinase B; PKB) is downstream of PI3K and activated by growth factors. This study found that erythropoietin (EPO) induced tyrosine phosphorylation of Akt in a time- and dose-dependent manner in EPO-dependent human leukemia cell line UT-7/EPO. In vitro kinase assay using histone H2B and glucose synthase kinase as substrates demonstrated that Akt was actually activated by EPO. EPO-induced phosphorylation of Akt was completely blocked by a PI3K-specific inhibitor, LY294002, at 10 micromol/L, indicating that activation of Akt by EPO is dependent on PI3K activity. In addition, overexpression of the constitutively active form of Akt on UT-7/EPO cells partially blocked apoptosis induced by withdrawal of EPO from the culture medium. This finding suggested that the PI3K-Akt activation pathway plays some role in the antiapoptotic effect of EPO. EPO induced phosphorylation of a member of the trancription factor Forkhead family, FKHRL1, at threonine 32 and serine 253 in a dose- and time-dependent manner in UT-7/EPO cells. Moreover, results showed that Akt kinase activated by EPO directly phosphorylated FKHRL1 protein and that FKHRL1 phosphorylation was completely dependent on PI3K activity as is the case for Akt. In conjunction with the evidence that FKHRL1 is expressed in normal human erythroid progenitor cells and erythroblasts, the results suggest that FKHRL1 plays an important role in erythropoiesis as one of the downstream target molecules of PI3K-Akt.

DNA-Binding Proteins↗

PubMed2000

Inhibition by 9alpha-fluoromedoroxyprogesterone acetate (FMPA) against mammary carcinoma induced by dimethylbenz[a]anthracene in rats and angiogenesis in the rabbit cornea - comparison with medroxyprogesterone acetate (MPA).

Medroxyprogesterone acetate (MPA) is currently used therapeutically in the treatment of mammary and endometrial carcinomas. In order to develop a more potent and useful drug, we synthesized the novel compound, 9alpha-fluoromedoroxyprogesterone acetate (FMPA), by fluorinating MPA, and we also previously reported that FMPA displays more potent anti-angiogenic activity in the chorioallantoic membrane assay than MPA. In the present study, we investigated (1) the effects of FMPA on rat mammary carcinomas induced by dimethylbenz[a]anthracene (DMBA) to determine the anti-tumor activity, (2) the effect on angiogenesis in rabbit corneal assays, and (3) compared these results with those for MPA. FMPA inhibited the growth of mammary carcinomas in a dose-dependent manner (7.5, 30 and 120 mg/kg). Almost complete involution of the carcinomas was observed at doses of 30 and 120 mg/kg. MPA also inhibited the growth of carcinomas at doses of 30 and 120 mg/kg, but no involution of carcinomas was observed even at 120 mg/kg. FMPA significantly and MPA to a lesser degree inhibited carcinogenesis at 120 mg/kg within their treatments. In rabbit corneal assays, FMPA significantly inhibited angiogenesis (IC50 value=0.085 microg/pellet). MPA also significantly inhibited angiogenesis (IC50 value=0.60 microg/pellet). From these results, we conclude that FMPA is potentially more effective in the treatment of mammary carcinomas than MPA.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed2000

Toward new materials for organic electroluminescent devices: synthesis, structures, and properties of a series of 2,5-diaryl-3,4-diphenylsiloles

A series of 2,5-diaryl-3,4-diphenylsiloles, with various mono-substituted phenyl groups, extended pi-conjugated groups, and heteroaryl groups as aryl groups at the 2,5-positions, has been prepared by a one-pot synthesis from bis(phenylethynyl)silanes based on the intramolecular reductive cyclization followed by the palladium-catalyzed cross-coupling with aryl halides. Crystal structures and chemical reactivities toward the alkaline desilylation reactions have been studied on the 2,5-bis(p-mono-substituted phenyl)silole derivatives to elucidate the effects of the p-substituents. The UV-visible absorption and fluorescence spectra, and cyclic voltammetry of the 2,5-diarylsiloles have been systematically evaluated. Their photophysical properties as well as their electronic structures significantly depend on the nature of the 2,5-aryl groups.

Journal Article↗

PubMed2000

Reduction of Iron(III) Ion by Activated Carbon Fiber.

The mechanisms of adsorption of iron(II) ion, iron(III) ion, and reduced iron(III) ion onto an activated carbon fiber and the ability of carbon fibers to reduce iron(III) ion were investigated on the basis of the amounts of iron ion adsorbed. The amount of iron(II) ion adsorbed onto the activated carbon fiber increased with increasing adsorption temperature. Iron(II) ion was more easily removed by the activated carbon fiber than iron(III) ion. Iron(III) ion was adsorbed onto the activated carbon fiber after being reduced to iron(II) ion. The reduction ability of A-20 was stronger than that of A-10 because the hydrophilic groups of A-20 were larger than those of A-10. It is concluded that the activated carbon fiber has a reduction effect on iron(III) ion and that the reduction effect of the activated carbon fiber depended on the number of hydrophilic groups on the activated carbon fiber. Copyright 2000 Academic Press.

Journal Article↗

PubMed2000

Induction of superovulation and recovery of fertilized oocytes in prepubertal miniature pigs after treatment with PG600.

The purpose of this study was to examine whether superovulation can be induced by hormonal treatment with PG600 (400 IU eCG and 200 IU hCG) at the prepubertal stage in miniature pigs. In Experiment 1, 14 prepubertal miniature pigs received 1, 1/2 or 1/4 vial of PG600, im on Day 0 (the first day of treatment). Presentation of estrus was monitored thereafter. On Days 10 to 13 (i.e., 6 to 8 d after estrus), the number of corpora lutea (CL) and residual follicles was counted by an exploratory laparotomy. Injection of 1/2 vial of PG600 effectively induced estrus and ovulation in the pigs. In Experiment 2, 15 prepubertal miniature pigs that received 1/2 vial of PG600 were artificially inseminated into the uterus by an exploratory laparotomy at 100 to 104 h after PG600 injection. Oocytes were recovered from the oviducts at 121 to 145 h after PG600 administration. The oocyte recovery rate was 66% (15 oocytes/pig, average), and 84% of these were at the 1-cell stage. In Experiment 3, 5 prepubertal miniature pigs that received 1/2 vial of PG600, followed by 100 IU hCG 70 h later, were artificially inseminated into the uterus. Oocytes were recovered synchronously at 120 to 122 h after PG600 treatment. The recovery rate was 80% (17 oocytes/pig, average) and 90% of the oocytes recovered were at the 1-cell stage. These results suggest that superovulation of prepubertal miniature pigs can be induced by 1/2 vial of PG600 injection, and by the combined treatment with PG600 and hCG injection, the fertilized ova can be synchronously recovered at around 120 h after PG600 injection. This procedure may provide a useful system for biomedical research using the miniature pigs, especially for producing transgenic animals for use in human disease models.

Animals↗

PubMed2000

Regulation of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) by bone resorptive factors in osteoblastic cells.

In addition to their stimulating function on osteoclastic bone resorption, bone resorptive factors may regulate proteinases and related factors in osteoblastic cells to degrade bone matrix proteins. This study investigated the regulation of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) by bone resorptive factors in the cultures of mouse osteoblastic MC3T3-E1 cells, mouse primary osteoblastic (POB) cells, and neonatal mouse calvariae. Expression of either MMP-2, -3, -9, -11, -13, and -14 or TIMP-1, -2, and -3 was detected in MC3T3-E1 cells and POB cells. When the bone resorptive factors parathyroid hormone, 1,25-dihydroxyvitamin D(3), prostaglandin E(2), interleukin-1beta (IL-1beta), and tumor necrosis factor-alpha (TNF-alpha) were added to the cell cultures, MMP-13 mRNA levels were found predominantly to increase by all resorptive factors in the three cultures. mRNA levels of either MMP-3 and -9 or TIMP-1 and -3 were found to increase mainly by the cytokines IL-1beta and TNF-alpha. BB94, a nonselective MMP inhibitor, neutralized the (45)Ca release stimulated by these resorptive factors to an extent similar to that of calcitonin, strongly suggesting that bone resorptive factors function at least partly through MMP formation. We propose that MMP-13 mRNA expression in osteoblastic cells may play an important role in stimulating matrix degradation by both systemic and local resorptive factors, whereas either MMP-3 and -9 or TIMP-1 and -3 might modulate matrix degradation by local cytokines only.

Animals↗

PubMed2000

The effect of alcohol on convulsions and nitric oxide levels in seizure-susceptible el mice.

To evaluate the influences of ethanol intake on convulsive seizures and brain nitric oxide (NO) production, EL mice, a strain highly susceptible to seizures, were given a 10% ethanol solution ad libitum. In mice consuming ethanol for 4, 8, and 12 weeks, seizures were not suppressed by zonisamide (75 mg/kg ip). Brain NO metabolite levels in mice after 12 weeks of consumption were significantly lower than those in control mice and those consuming ethanol for 4 weeks. Numbers of NADPH diaphorase-positive neurons in the hippocampal formation and parietal cortex of mice consuming for 4 and 12 weeks were significantly higher than in controls. These results suggested that increasing of numbers of NADPH diaphorase-positive neurons in the hippocampal formation and parietal cortex were assumed to develop in compensation for reduction in whole-brain NO metabolite levels of EL mice exposed to ethanol.

Journal Article↗

PubMed2000

Usefulness and pitfalls of intraoperative spinal motor evoked potential recording by direct cortical electrical stimulation.

OBJECTIVE: The aim of this study is to investigate the usefulness and problems with spinal motor evoked potential (MEP) recording, especially the reasons for failed recording. We report our personal experience over the last 8 years in patients with lesions adjacent to the primary motor cortex. METHODS: MEP records of 50 consecutive patients were retrospectively reviewed. MEP was recorded by a catheter electrode inserted in the cervical epidural space. Stimulation electrodes were placed on the cortical surface during surgery. SEP recording was also performed in 29 of 50 patients. RESULTS: MEP was obtained in 40 cases, and SEP was recorded in all 29 cases. The central sulcus was identified in 93% of patients in whom both MEP and SEP were performed, whereas in only 86% of patients who underwent only MEP. The main reason for MEP failure were inadequate exposure of the motor cortex, pre-existing hemiparesis and technical errors. Postoperative deterioration of motor function was closely related to intra-operative MEP changes. CONCLUSION: MEP is a useful tool to determine the motor cortex and to predict postoperative motor function. However, precise pre-operative craniotomy planning and combination with intra-operative SEP is essential to reduce the MEP failure.

Adolescent↗

PubMed2000

Spatiotemporal change of rat collagenase (MMP-13) mRNA expression in the development of the rat femoral neck.

The interepiphyseal region between the greater trochanter and the capital femoral epiphysis and the medioproximal portion of the femoral neck exhibit extensive morphological changes during the first 4 weeks after birth in rats. Previous reports show that matrix metalloproteinase-13 (MMP-13, rat collagenase) mRNA is expressed in bone and cartilage during embryonal development and fracture healing. We examined MMP-13 mRNA expression and compared it with the distribution of osteopontin and osteocalcine mRNA in the femoral neck. Moreover, we examined histomorphometric analysis in the femoral neck where the morphology changes rapidly. Histomorphometric analysis of the 4-week-old rat femoral neck showed a high rate of bone formation and resorption in the region where shape changed rapidly. Osteopontin mRNA was expressed diffusely along the endosteum. In contrast, MMP-13 mRNA expression was restricted to the medial endosteal portion near the cartilage-bone interface of the femoral neck in 15- and 28-day-old rats and in the deepest endosteal interepiphyseal region of 15-day-old rats. MMP-13 mRNA-expressing osteoblastic cells were also expressing osteopontin but not osteocalcin mRNA. MMP-13 mRNA-expressing cells differ from tartrate-resistant acid phosphatase (TRAP)-positive cells, and MMP-13 mRNA-positive cells are located adjacent to TRAP-positive cells. The results of the site- and cell-specific expression of MMP-13, taken together with its enzymatic property, suggest that MMP-13 plays an important role in morphological changes in the rat femur, at least during the third and fourth week after birth, and that MMP-13 itself is involved in the interaction between osteoblastic and TRAP-positive cells.

Acid Phosphatase↗

PubMed2000

Antibacterial effect of silver-zeolite on oral bacteria under anaerobic conditions.

OBJECTIVES: The purpose of this study was to evaluate the antibacterial effect of silver-zeolite (SZ) against oral bacteria under anaerobic conditions. METHODS: The antibacterial activity of SZ was evaluated by determining the minimum inhibitory concentrations (MICs) using two-fold serial dilutions of SZ in Brain Heart Infusion broth. Release of Ag+ into the broth was measured by an atomic absorption technique. RESULTS: SZ inhibited the growth of the bacteria tested under anaerobic conditions. The MIC of SZ ranged between 256 and 2048 micrograms/ml, which corresponded to a range of 4.8-38.4 micrograms/ml of Ag+. All strains grew in broth containing 16,384 micrograms/ml of type-A zeolite. SIGNIFICANCE: These results suggested that SZ may be a useful vehicle to provide antibacterial activity to dental materials used even under anaerobic conditions such as deep in the periodontal pocket.

Anaerobiosis↗

PubMed2000

Age-related alterations of nitric oxide production in the brains of seizure-susceptible EL mice.

We evaluated age-related changes in nitric oxide (NO) production in the brains of EL mice, a strain highly susceptible to seizures. A group of EL(s) mice were tossed up weekly to induce convulsive seizures, while in a nonstimulated EL(ns) group induction of convulsive seizures was avoided. Brain levels of nitrite plus nitrate (NOx) in EL(ns) mice were significantly higher than in nonstimulated mice at 10 days, and also higher than levels at 15 and 50 weeks in either EL(s) or EL(ns) mice. A significantly higher number of NO-producing cells were demonstrated in the hippocampus and parietal cortex by staining for nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase in EL(s) mice at the ages of 15 and 50 weeks than in EL(ns) mice at the age of 6 weeks. In EL(ns) mice, significantly fewer neurons showed NADPH-diaphorase staining in the hippocampus, striatum and parietal cortex at the age of 50 weeks than at 6 weeks. The present results suggest that whole-brain NOx levels in EL(ns) and EL(s) mice and numbers of NADPH-diaphorase-positive neurons in EL(ns) mice decreased with aging, while increasing of numbers of such neurons in EL(s) mice were assumed to develop in compensation for reduction in whole-brain NOx levels.

Aging↗

PubMed2000

Twelve-fold quasicrystal and its approximant of Ta62Te38 interpreted as modulated crystals

A transmission electron microscopy study reveals that the twelve-fold quasicrystal and its approximant in Ta62Te38 are crystals subjected to the structure modulation. It is composed of two modulated layers rotated by 30 degrees (or 90 degrees) to each other about their normal. Structures of the twelve-fold quasicrystal and its approximant can be related by modulation waves with the same directions but with slightly different wavelengths. The modulation is considered to be due to the rearrangement of atomic vacancies as a response to the occurrence of charge density waves.

Journal Article↗

PubMed2000

Liver-infiltrating CD56 positive T lymphocytes in hepatitis C virus infection.

AIM: Hepatitis C virus (HCV) is a major cause of post-transfusional and sporadic hepatitis, and leads to chronic liver disease. It has been suggested that virus-specific cytotoxic T lymphocytes are responsible for liver injuries that occur in HCV-infected patients. However, the detailed characteristics of these lymphocytes have not yet been defined. We have previously reported that CD56+ T lymphocytes, as intermediates between natural killer cell and T lymphocytes, predominantly infiltrated the liver and were increased in patients with chronic hepatitis related to HCV (CH-C). MATERIAL AND METHODS: We obtained peripheral blood and liver tissues from 32 patients diagnosed as having CH-C, and 10 other liver disease patients (5 chronic hepatitis related to HBV, 5 alcoholics), and analyzed peripheral blood and liver-infiltrating lymphocytes using flow cytometric and immunohistochemical techniques. RESULTS: The CD56+ T lymphocyte ratio in the liver of patients with a high histology activity index (HAI) score for chronic hepatitis was higher than that of patients with a low HAI score and patients with other liver diseases. In addition, T lymphocytes from patients with chronic hepatitis with a high HAI score carried mostly gamma delta-TCR. There was a correlation between the ratio of CH-C and serum alanine aminotransferase, category I (periportal inflammation and necrosis), and IV (fibrosis) of the HAI scoring system. The ratio was highest in zone 1 of the hepatic lobules. CONCLUSION: The correlation between CD56+ T lymphocyte ratios and hepatocellular damage was examined. These findings suggest strongly that liver-infiltrating CD56+ T lymphocytes play an important pathologic role in hepatocellular injury in CH-C.

Adult↗

PubMed2000

Liposome-encapsulated OK-432 specifically and sustainedly induces hepatic natural killer cells and intermediate T cell receptor cells.

BACKGROUND: OK-432 is a biological response modifier used in Japan to augment host immunity and is known to increase the host antitumour response. By using liposomes, which are vesicles made from phospholipids that have a structure resembling the cell membrane, we encapsulated OK-432. METHODS AND RESULTS: Encapsulated OK-432 was injected into the tail veins of mice, and its effect was compared with that of unencapsulated OK-432 given intravenously. In mice that received either form of OK-432, both the number of natural killer (NK) and intermediate T cell receptor (intTCR) cells (intrahepatic T cells generated by extrathymic differentiation) increased markedly in the liver, with the peak level occurring 3 days after administration. Both forms of OK-432 also increased cytotoxic activity against Yac-1 cells. The increase in numbers of cells and in cytotoxic activity in the liver persisted for longer in mice that received encapsulated OK-432 than in animals that received unencapsulated OK-432. CONCLUSIONS: Because it has been shown that both NK and intTCR cells play an important role in tumour immunity, an increase in the number of such cells can be considered likely to have an increased antitumour effect. Encapsulated OK-432 elicited liver-specific augmentation of cytotoxic activity and the effect was more persistent than that produced by OK-432 given in the conventional form; therefore, it may be useful for the treatment of tumours, particularly those arising in the liver.

Animals↗

PubMed2000

Detection of intracellular interleukin-2 production in peripheral T lymphocytes by flow cytometry in patients with pancreatobiliary malignancies.

BACKGROUND AND AIMS: To date, it has been reported that cellular immunity is decreased in patients with cancer and investigations into cytokine production has been insufficient. Therefore, we examined intracellular cytokine production by using flow cytometry in patients with cancer and discussed the reasons for the impairment of their immune system. METHODS: Eleven patients with hepatobiliary malignancies (68.5+/-11.8 years of age), eight age-matched controls (70.0+/-12.0 years of age) and 10 young volunteers (31.9+/-3.1 years of age) were used in the present study. Stimulated peripheral blood mononuclear cells from these patients were stained with fluorescence-labeled anticytokine monoclonal antibodies and analyzed with a Fluorescence activated cell sorter (FAC)Scan. RESULTS: The percentage of positively stained T cells was calculated and compared with controls. Repeated measured ANOVA was used for statistical analysis. Interleukin (IL)-2 production was significantly decreased in patients with cancer compared to controls (P=0.0122), and it may suggest decreased cellular immune activity of the patients. Simultaneously, spontaneous intracellular IL-4 production was observed in patients and age-matched controls, but levels were significantly increased when compared with the young volunteers (P=0.0052, P=0.031, respectively). CONCLUSIONS: It was of interest that spontaneous intracellular IL-4 production was detected in elderly subjects.

Adult↗

PubMed2000

Bacterial interference among nasal inhabitants: eradication of Staphylococcus aureus from nasal cavities by artificial implantation of Corynebacterium sp.

To evaluate the role of normal flora in the nares in preventing Staphylococcus aureus colonization, we conducted a replacement study in vivo. Staphylococcus epidermidis (rate of colonization: 100%), various species of corynebacteria (52.5%) and S. aureus (25.%) were the major bacterial inhabitants in the nares of 156 healthy volunteers. The low incidence of S. aureus colonization in the carriers with corynebacteria (8.5%), compared to non-carriers (44. 5%) indicated the possibility of competition for survival between S. aureus and corynebacteria. To confirm this hypothesis, we artificially implanted a strain of Corynebacterium sp (API Coryne bioprofile; 5100304), denoted as Co304 into the nares of 17 S. aureus carriers. S. aureus was completely eradicated in 71% of carriers by up to 15 inoculations of Co304. However, similar doses of 0.9% NaCl or S. epidermidis into the nares of 10 volunteers did not eradicate S. aureus. No bacteriocin-like activity against S. aureus was detectable, even after mitomycin C stimulation of Co304. Thus Co304 interfered with S. aureus by a different mechanism to a bacteriocin-like activity.

Adult↗

PubMed2000

Protective effect of ginkgo extract on rat brain with transient middle cerebral artery occlusion.

It has been empirically known that Ginkgo extract is useful for reducing many symptoms associated with cerebral blood flow (CBF) insufficiency, but its mechanisms have been uncertain. In the present study, therefore, we gave Ginkgo extract to rats with per os digestion, and investigated its effect on CBF and ischemic brain damage with middle cerebral artery occlusion (MCAO). The treatment with Ginkgo extract (10 mg 100 g-1 rat) increased CBF in the normal condition, but the degree of increase in CBF was lesser during and after MCAO. TTC staining showed that infarct volume was reduced with Ginkgo treatment. TUNEL and HSP72 immunostaining confirmed the protective effect of Ginkgo treatment reducing numbers of TUNEL and HSP72 positive cells. Immunohistochemical analysis showed that caspase-3 expression was less abundant in Ginkgo treated rats. The present results suggest that Ginkgo extract contains a substance which increases normal CBF and reduces ischemic brain damage.

Animals↗

PubMed2000

Vitamin E inhibits cell proliferation and the activation of extracellular signal-regulated kinase during the promotion phase of lung tumorigenesis irrespective of antioxidative effect.

We have already reported that the activation of extracellular signal-regulated kinase (Erk) is critical in the stimulation of cell proliferation during the promotion stage of urethane-induced lung tumorigenesis in mice. Also, we have found that vitamin E suppresses lung tumorigenesis by inhibiting cell proliferation at the promotion stage. However, it is still unclear whether this inhibitory effect at the promotion stage is based on the antioxidative effect of vitamin E or not. In order to address this question, we examined the inhibitory effect of alpha-tocopheryloxybutyric acid (TSE), an ether derivative of vitamin E that cannot act as an antioxidant in vivo, on cell proliferation and the activation of Erk during promotion of lung tumorigenesis. On day 30 after urethane injection (750 mg/kg, i. p.) in A/J mice, TSE or vitamin E at 100 micromol/kg, p.o. was administered. Twenty-four hours after the final administration, the mice were killed to analyze cell proliferation and related parameters. The labeling index of proliferating cell nuclear antigen (a marker of cell proliferation) and ornithine decarboxylase activity (a marker of the promotion stage in lungs) were attenuated by treatment with TSE or vitamin E. TSE or vitamin E treatment also inhibited urethane-induced activation of Erk and suppressed the activation of other essential members of the Erk cascade (Ras, Raf and Mek). These results suggest that vitamin E inhibits cell proliferation and activation of the Erk cascade during promotion of urethane-induced lung tumorigenesis in mice, independent of its antioxidative effect.

Animals↗