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Biomedical subjects

Satoshi Inoue

Publications and source records attributed to Satoshi Inoue.

At least 55 records · Page 3Linked to original sources
PubMed2005

Production of rabies neutralizing antibody in hen's eggs using a part of the G protein expressed in Escherichia coli.

In an attempt to produce anti-rabies immunoglobulin affordable for people living in developing countries, we have immunized layer chickens with a part of the G protein of rabies virus expressed in Escherichia coli. Immunoglobulin (IgY) was purified from the yolks of eggs layed by immunized hens. It was revealed in vitro that the antibody specifically bound to virions as well as cells infected with rabies virus. Moreover, the antibody apparently neutralized rabies virus infectivity. Inoculation of the antibody into mice infected with rabies virus reduced the mortality caused by the virus, suggesting that IgY directed to the part of the G protein expressed in E. coli could serve as a possible alternative to currently available anti-rabies human or equine immunoglobulins.

Animals↗
PubMed2005

Nephrotoxicity of a novel antineoplastic platinum complex, nedaplatin: a comparative study with cisplatin in rats.

The present study was designed to characterize the nephrotoxicity induced by the antineoplastic platinum complex nedaplatin (NDP) in rats of different ages in comparison with cisplatin (CDDP). A single dose of 15 mg/kg NDP or 7.5 mg/kg CDDP was administered intravenously to 8-, 11-, or 15-week-old male and female SD rats, which were then sacrificed after ten days. Body weight decreases were observed for both drugs, in direct relation to age. CDDP treatment markedly increased urinary excretion of NAG, gamma-GTP, LDH and protein, with peaks on day 4 and complete or partial recovery on day 7; NDP increased NAG, LDH and protein excretion, but to a lesser extent, and these elevations were generally more marked for females. CDDP increased plasma creatinine and BUN in males and females of all age groups at necropsy. No apparent changes were seen following NDP treatment except in the 15-week-old rats. These results also show that NDP is less nephrotoxic than CDDP. CDDP-treated rats showed remarkable proximal tubular lesions in the renal cortex and corticomedullary region, and the papillary lesions were minor. On the other hand, the NDP-induced nephrotoxicity was morphologically characterized by hyaline droplet changes (electron microscopically, hyperplasia of lysosomes), necrosis or hyperplasia of the collecting duct epithelium in the renal papilla and the epithelium covering the papilla. Cortical lesions, indicated by slight tubular dilatation, were found only in the animals with papillary lesions. In summary, NDP is a promising second-generation platinum complex with reduced nephrotoxicity.

Animals↗
PubMed2005

Characterization of P gene-deficient rabies virus: propagation, pathogenicity and antigenicity.

The RNA polymerase of rabies virus (RV) is a two-protein complex composed of L (a large catalytic component) and P (a non-catalytic phosphoprotein cofactor) proteins. We generated a gene-deficient RV lacking the entire P gene from HEP-Flury (HEP) strain, one of the most attenuated RV strains, by the method of reverse genetics. This P gene-deficient (def-P) virus could replicate and produce progeny viruses with a slightly retarded rate in the cell lines that constitutively express the P protein. The def-P virus could perform the primary RNA transcription by the virion-associated polymerase even in the infected host without de novo P protein synthesis. However, the def-P virus required the newly synthesized P protein for the secondary RNA transcription and genome RNA replication of virus. No progeny virus was produced in the infected host that did not express P protein. The def-P virus was apathogenic in adult and suckling mice even when inoculated intracranially. On the other hand, inoculation of the def-P virus into mice induced a high titer of virus-neutralizing antibody and protected mice from lethal challenge with the CVS strain. These results demonstrated that the def-P virus could induce strong protective immunity against rabies virus without the production of progeny virus and the severe host damage. The def-P virus would be a potential resource of safe live-attenuated rabies vaccine.

Animals↗
PubMed2005

Effect of double-application of all-in-one adhesives on dentin bonding.

OBJECTIVES: To evaluate the effect of double-application of all-in-one adhesives using the micro-shear bond test. METHODS: The occlusal surfaces of extracted human third molars were ground perpendicular to the long axis of each tooth to expose a flat dentin surface. Three commercially available and one experimental bonding system were used in this study: Adper Prompt L-Pop (APL, 3M ESPE), REACTMER BOND (RB, Shofu), XENO III (Xeno, Dentsply-Sankin) and newly developed OBF-2 (OB2, Tokuyama Dental). These adhesives were applied on the dentin surfaces by either the manufacturers' instructions or by an experimental method (single-application or double-application). Resin composite was then placed and light-cured for 40s. After 24h immersion in water, a micro-shear bond test was carried out and the fractured dentin surfaces were observed microscopically. The data were analyzed by one-way ANOVA and Tukey's HSD tests (p<0.05). RESULTS: The mean bond strengths of APL, RB, Xeno and OB2 with single-application were 22.7, 28.3, 30.3 and 34.6MPa, respectively. The mean bond strengths of APL, RB, Xeno and OB2 with double-application were 29.5, 27.2, 29.6 and 32.5MPa, respectively. There were no statistically significant differences in micro-shear bond strengths between the single- and double-application methods for each adhesive system (p>0.05). The morphological observation of the fractured dentin surfaces revealed differences between the single- and double-application groups especially for APL and OB2. CONCLUSIONS: Micro-shear bond strengths of all-in-one adhesives in this study showed no statistically significant differences between the single-application and the double-application method.

Analysis of Variance↗
PubMed2005

Organization of multisynaptic inputs from prefrontal cortex to primary motor cortex as revealed by retrograde transneuronal transport of rabies virus.

The organization of multisynaptic projections from the prefrontal cortex to the primary motor cortex (MI) was examined in macaque monkeys by retrograde transneuronal transport of rabies virus. In the first series of experiments, the virus was injected into the MI forelimb region, and the time-dependent distribution patterns of transsynaptic labeling were analyzed in the frontal lobe with various survivals (2-4 d). Two days after the viral injection, neuronal labeling emerged in the caudal aspects of the nonprimary motor-related areas that are known to project to the MI directly. At the same time, the motor thalamus contained labeled neurons. On the third day, cortical labeling extended into the rostral motor-related areas and, also, prearcuate area 8. Moreover, a number of labeled neurons were located in the internal pallidum and the cerebellar nuclei. At the 4 d postinjection period, neuronal labeling occurred widely in prefrontal areas as well as in the putamen and the cerebellar cortex. In the second series of experiments, the viral injection was made into the MI hindlimb region, and the distribution pattern of prefrontal labeling on the fourth day was compared with that in the forelimb-injection case. The labeled neurons in each prefrontal area were much fewer in the hindlimb-injection case than in the forelimb-injection case. Whereas ventral area 46 was most densely labeled from the forelimb region, only sparse labeling from the hindlimb region was observed in this prefrontal area. The present results suggest the importance of ventral area 46 in the cognitive control of forelimb movements.

Animals↗
PubMed2005

Programming of CTL with heat-killed Brucella abortus and antigen allows soluble antigen alone to generate effective secondary CTL.

Optimal generation of cytotoxic T cell (CTL) responses continues to be a challenge in the production of vaccines against pathogens such as HIV-1, in part because it is difficult to introduce soluble protein antigens (Ag) into the MHC class I pathway. Using heat-killed Brucella abortus (HKBA) as an adjuvant and ovalbumin (ova) protein as an Ag, we demonstrated that a high dose of Ag was required for systemic and effective CTL. In an adoptive transfer model, primary and secondary ova-specific OT-1 CD8 cell expansion by HKBA plus high dose of ova were partially CD4 T cell-dependent. Interestingly, primary stimulation with HKBA plus ova allowed effective secondary stimulation with ova alone that was equivalent to HKBA plus ova in terms of IFN-gamma production from Ag-specific CD8 cells. Thus a combination of adequate Ag dose, and selection of appropriate adjuvants can meet the threshold not only for primary effective CTL responses to soluble protein Ags but for secondary CTL responses following stimulation with protein Ag alone.

Adoptive Transfer↗
PubMed2005

Survival versus apoptotic 17beta-estradiol effect: role of ER alpha and ER beta activated non-genomic signaling.

The capability of 17beta-estradiol (E2) to induce the non-genomic activities of its receptors (ER alpha and ER beta) and to evoke different signaling pathways committed to the regulation of cell proliferation has been analyzed in different cell cancer lines containing transfected (HeLa) or endogenous (HepG2, DLD1) ER alpha or ER beta. In these cell lines, E2 induced different effects on cell growth/apoptosis in dependence of ER isoforms present. The E2-ER alpha complex rapidly activated multiple signal transduction pathways (i.e., ERK/MAPK, PI3K/AKT) committed to both cell cycle progression and apoptotic cascade prevention. On the other hand, the E2-ER beta complex induced the rapid and persistent phosphorylation of p38/MAPK which, in turn, was involved in caspase-3 activation and cleavage of poly(ADP-ribose)polymerase, driving cells into the apoptotic cycle. In addition, the E2-ER beta complex did not activate any of the E2-ER alpha-activated signal molecules involved in cell growth. Taken together, these results demonstrate the ability of ER beta isoform to activate specific signal transduction pathways starting from plasma membrane that may justify the effect of E2 in inducing cell proliferation or apoptosis in cancer cells. In particular this hormone promotes cell survival through ER alpha non-genomic signaling and cell death through ER beta non-genomic signaling.

Apoptosis↗
PubMed2005

Association of a single nucleotide polymorphism in the lipoxygenase ALOX15 5'-flanking region (-5229G/A) with bone mineral density.

The 12/15-lipoxygenase gene Alox15 has been identified as a susceptibility gene for bone mineral density (BMD) in mice through combined genetic and genomic analyses. Here we studied the association between bone mineral density and an ALOX15 gene single nucleotide polymorphism to assess the potential involvement of the human ALOX15 gene in postmenopausal osteoporosis. Specifically, we examined the association between a single nucleotide polymorphism at -5299G/A in the ALOX15 5'-flanking region with BMD in 319 postmenopausal Japanese women (66.7 +/- 8.9 years, mean +/- SD). We found that subjects bearing at least one variant A allele (GA + AA; n = 273) had significantly lower Z scores for lumbar spine and total body bone mineral density than did subjects with no A allele (GG; n = 46) (lumbar spine, -0.25 +/- 1.34 versus 0.48 +/- 1.70; P = 0.0014; total body, 0.25 +/- 1.01 vs 0.62 +/- 1.11; P = 0.048). These findings suggest that the ALOX15 gene is one of the genetic determinants of BMD in postmenopausal women. Accordingly, this polymorphism could be useful as a genetic marker for predicting the risk of osteoporosis.

Aged↗
PubMed2005

Effect of cavity configuration and aging on the bonding effectiveness of six adhesives to dentin.

OBJECTIVES: The purpose of this study was to determine the effect polymerization contraction stress may have on bond durability. METHODS: Bonding effectiveness was assessed by micro-tensile bond strength testing (muTBS) and electron microscopy. The muTBS to flat dentin surfaces and in standardized cavities was determined (this after 1 day as well as 1 year water storage). Six adhesives representing all current classes were applied: two etch-and-rinse (OptiBond FL, Kerr; Scotchbond 1, 3M ESPE), two self-etch (Clearfil SE Bond, Kuraray; Adper Prompt, 3M ESPE) and two glass-ionomer (Fuji Bond LC, GC; Reactmer, Shofu) adhesives. RESULTS: The conventional 3-step etch-and-rinse adhesive OptiBond FL bonded most effectively to dentin, and appeared insensitive to polymerization shrinkage stress and water degradation. The 2-step self-etch adhesive Clearfil SE Bond most closely approached this superior bonding effectiveness and only slightly lost bond strength after 1-year water exposure. The 2-step etch-and-rinse adhesive Scotchbond 1 and the 'strong' 1-step self-etch adhesive Adper Prompt appeared very sensitive to cavity configuration and water-aging effects. The 2-step resin-modified glass-ionomer adhesive Fuji Bond LC only suffered from shrinkage stress, but not from 1-year water-exposure. Remarkable also is the apparent repairability of the 'mild' 1-step glass-ionomer adhesive Reactmer when stored for 1 year in water, in spite of the very low 1-day muTBS. SIGNIFICANCE: Simplified bonding procedures do not necessarily imply improved bonding performance, especially in the long term.

Acid Etching, Dental↗
PubMed2005

Effect of fracture strength of primer-adhesive mixture on bonding effectiveness.

OBJECTIVE: The purpose of this study was to measure the micro-tensile bond strength (microTBS) of two adhesives to dentin and to correlate this microTBS with the micro-tensile fracture strength (microTFS) of primer-adhesive mixtures of the two adhesives. METHODS: The three-step etch&rinse adhesive OptiBond FL (Kerr) and the two-step self-etch adhesive Clearfil SE Bond (Kuraray) were used to bond Filtek Z100 (3M ESPE) to twenty mid-coronal dentin surfaces. The microTBS was determined at 1 and 24 h after light-curing. In addition, the microTFS of primer-adhesive mixtures (primer:adhesive=1:3 by weight) and of the pure adhesives (controls) were measured after the same time intervals. RESULTS: The microTBS to dentin of OptiBond FL at 24 h was significantly higher than at 1 h and than the microTBS of Clearfil SE Bond at 24 h. The mean microTFS of the primer-adhesive mixtures was always lower than the microTFS of the respective adhesives. The highest reduction in microTFS due to mixing primer with the adhesive resin was measured for Clearfil SE Bond. Both the microTFS of the primer-adhesive mixtures and the pure adhesives of OptiBond FL and Clearfil SE Bond increased with time. SIGNIFICANCE: The early 1 h microTBS to dentin was lower than the 24 h microTBS for OptiBond FL, but not for Clearfil SE Bond. Differences in polymerization efficiency as well as in the resulting interfacial strength may explain such varying early bond strengths to dentin.

Composite Resins↗
PubMed2005

A fibroin secretion-deficient silkworm mutant, Nd-sD, provides an efficient system for producing recombinant proteins.

The silkworm Nd-s(D) mutant is silk fibroin-secretion deficient. In the mutant, a disulfide linkage between the heavy (H) and light (L) chains, which is essential for the intracellular transport and secretion of fibroin, is not formed because of a partial deletion of the L-chain gene. To utilize the inactivity of the mutant L-chain, we investigated the possibility of using the Nd-s(D) mutant for the efficient production of recombinant proteins in the silkworm. A germ line transformation of the mutant with a normal L-chain-GFP fusion gene was performed. In the transgenic mutant, normal development of the posterior silk gland (PSG) was restored and it formed a normal cocoon. The biochemical analysis showed that the transgenic silkworms expressed the introduced gene in PSG cells, produced a large amount of the recombinant protein, secreted it into the PSG lumen, and used it to construct the cocoon. The molar ratio of silk proteins, H-chain:L-chain-GFP:fibrohexamerin, in the lumen and cocoon in the transgenic silkworm was 6:6:1, and the final product of the fusion gene formed about 10% of the cocoon silk. This indicates that the transgenic mutant silkworm possesses the capacity to produce and secrete the recombinant proteins in a molar ratio equal to that of the fibroin H-chain, contributing around half molecules of the total PSG silk proteins.

Animals↗
PubMed2005

Geographical distribution of judo therapists and orthopedists in Japan.

BACKGROUND: Judo therapy is a unique and long-established Japanese comedical profession, specialized in manually treating fractures and sprains in practitioners' offices. Having the overlapped scopes of services between judo therapists and orthopedists, disputes between the 2 have gradually emerged recently. OBJECTIVE: To examine the relationship between judo therapists and orthopedists by comparing their geographic distributions at the whole country level and at prefecture level. STUDY DESIGN: Cross-sectional study. METHODS: The numbers of judo therapist facilities and orthopedists per 100,000 population in each municipality were obtained through web yellow pages and from the Japanese census data. Lorenz curves and Gini indices were calculated to show the distributions and to compare the numbers of judo therapist facilities and orthopedists per 100,000 population. RESULTS: The median number of judo therapist facilities per 100,000 population in all municipalities was 12.0 (range 0-135) and that for orthopedists was 4.3 (0-186). The Gini indices of the nationwide judo therapist facilities and those for orthopedists were 0.29 and 0.36, respectively. CONCLUSION: These data show the relatively homogeneous distribution of judo therapist facilities compared with orthopedists. In smaller municipalities, judo therapist facilities may fill in the absence of orthopedists. As for lager municipalities, further study is needed to examine whether the relationships between them are competitive or complementary.

Complementary Therapies↗
PubMed2005

Changes in medical and nursing care in cancer patients transferred from a palliative care team to a palliative care unit.

The primary aim of this study was to explore the changes in medical/nursing care in patients who transferred from a palliative care team (PCT) to a palliative care unit (PCU) in the same hospital, and to explore the reasons why new or modified interventions were required. This was a retrospective study of 50 consecutive patients who were transferred from PCT to PCU in a 750-bed general hospital. A trained nurse performed a chart review and recorded the changes in 1) medical/nursing care, 2) help with decision-making within 48 hours after PCU admission, and 3) documentation of family and psycho-existential care. Group discussions among PCT and PCU staff members explored the potential reasons for the changes. Five patients were excluded due to admission periods of shorter than 48 hours. The number of new/modified medical and nursing care interventions after PCU admission averaged 1.9 +/- 1.5 and 1.5 +/- 1.3 per patient, respectively. The most common medical treatments were: reduction in hydration volume, dose titration of opioids, change in opioid administration device, opioid rotation, and addition of NSAIDs and steroids. The most common nursing interventions were: allowing patient to take a bath, changing mattress, use of massage, and discontinuation of bronchial suctioning. In addition, PCU staff newly coordinated opportunities to discuss preferred end-of-life care with primary caregivers in 38% of the cases, family members other than primary caregivers in 16%, and patients in 6.7%. The chart documentation of family care and psycho-existential care increased considerably after PCU admission. The chief reasons for these changes were: under-recognition of the problems and unavailability of treatments (pharmacological treatments), no intention to intervene and recommendations not followed by primary physicians (rehydration therapy), no intention to intervene (nursing care), and no intention to intervene and under-recognition of the problems (help with decision-making). These data demonstrate that many patients under PCT consultation receive new or modified interventions after PCU admission. Potentially useful strategies to strengthen the PCT interventions are: modification of intervention structure to minimize under-recognition of symptoms and decision making problems (e.g., use of standardized assessment tools, regular conferences), changes in the health care system to allow unlicensed drugs, clinical studies to clarify the benefits of artificial hydration therapy, and greater efforts to intervene in the areas of nursing care, help with decision making, family care, and psycho-existential care.

Aged↗
PubMed2005

Inhibition of Src kinase activity by Ad-mda7 suppresses vascular endothelial growth factor expression in prostate carcinoma cells.

The ability of an adenoviral vector expressing the melanoma differentiation-associated gene-7 (Ad-mda7) to mediate inhibition of vascular endothelial growth factor (VEGF) has recently been reported. However, the molecular mechanism by which Ad-mda7 inhibits VEGF is unknown. In an attempt to elucidate this mechanism, we studied the effects of Ad-mda7 on VEGF expression using human prostate cancer cells as a model. We found that Ad-mda7 treatment of prostate cancer cells (LNCaP and DU145) in vitro resulted in a significant (P < 0.05) inhibition of VEGF expression. Analysis of the VEGF signaling pathway showed that Ad-mda7 inhibited c-Src kinase activity and abrogated STAT-3 binding to the VEGF promoter. Correlating with these observations were reductions in VEGF mRNA and protein levels in Ad-mda7-treated cells. Furthermore, Ad-mda7 inhibited VEGF in Src(+/+) but not in Src(-/-) mouse embryo fibroblasts. These results showed that Ad-mda7 inhibited VEGF by inhibiting the Src signaling pathway. Finally, conditioned medium from Ad-mda7-treated tumor cells containing reduced VEGF inhibited VEGF receptor signaling, resulting in reduced endothelial cell proliferation and apoptosis. Our results provide evidence for the mechanism by which Ad-mda7 inhibits VEGF in tumor cells and of the effects of this VEGF inhibition on endothelial cell proliferation, a requirement for angiogenesis. Our findings demonstrate that MDA-7 protein, in addition to inhibiting tumor angiogenesis directly, inhibits angiogenesis indirectly by inhibiting VEGF production by tumor cells.

Cell Line, Tumor↗
PubMed2005

Selective induction of cell cycle arrest and apoptosis in human prostate cancer cells through adenoviral transfer of the melanoma differentiation-associated -7 (mda-7)/interleukin-24 (IL-24) gene.

We have previously reported that overexpression of the melanoma differentiation-associated gene -7 (mda-7) using a replication-defective adenovirus (Ad-mda7), results in tumor-specific growth suppression and induction of apoptosis in wide variety of cancer cells. In the present study, we investigated the antitumor activity of Ad-mda7 and the underlying mechanism in human prostate cancer cells and normal prostate epithelial cells. Overexpression of MDA-7 induced significant (P=.001) suppression of cell growth and apoptosis in prostate cancer cells (DU 145, LNCaP, and PC-3). In normal prostate epithelial cells (PrEC) some degree of growth inhibition but not apoptosis was observed. However, the inhibitory effects in normal cells were less compared to tumor cells. Growth inhibitory effects were mediated by the intracellular and not by extracellular MDA-7 protein. Molecular effectors that are involved in Ad-mda7-mediated tumor killing included activation of the caspase cascade, and the induction of G2 phase cell cycle arrest through the inhibition of Cdc25C pathway. These results demonstrate the mechanisms by which Ad-mda7 exerts its antitumor activity in human prostate cancer cells. The antitumor activity combined with previously reported antiangiogenic and proimmune properties of Ad-mda7 can serve as a potential therapeutic agent for treatment of primary and disseminated prostate cancer.

Adenoviridae↗
PubMed2005

In vivo degradation of resin-dentin bonds produced by a self-etch and an etch-and-rinse adhesive.

Resin-dentin bonds are known to degrade in the relatively aggressive oral environment. In order to obtain greater insight into the interfacial degradation process, we examined, by using transmission electron microscopy (TEM), the interfacial ultrastructure of two adhesives bonded to dentin after 1 yr in vivo. Class V cavities were prepared on the buccal surfaces of 14 intact teeth of two monkeys and then restored by using either the two-step self-etch adhesive, Unifil Bond, or the two-step etch-and-rinse adhesive, Single Bond, in combination with the restorative microhybrid composite, Z250. After 1 yr, 10 other teeth were restored by using the same materials (controls). One day later, the monkeys were killed, following which the microtensile bond strength to dentin was determined and the interfacial ultrastructure was examined by TEM. Whereas no noticeable changes in the morphology of the resin-dentin interface were observed between the 1-d and 1-yr specimens for Unifil Bond, Single Bond exhibited signs of interfacial degradation, in particular at the bottom of the 3 microm-deep hybrid layer. In conclusion, the adhesive interface produced by the etch-and-rinse adhesive was less resistant to degradation than that produced by the self-etch system.

Acid Etching, Dental↗