Biomedical subjects

Y Setoguchi

Publications and source records attributed to Y Setoguchi.

At least 19 recordsLinked to original sources

PubMed1999

Neuron-targeted gene transfer by adenovirus carrying neural-restrictive silencer element.

Adenovirus transfers genes to a wide range of cell types, but its application to neurons has been hampered by its reduced efficiency of infection as compared with that for glia. To achieve neuron-targeted gene transfer, we have produced an adenovirus carrying the reporter lacZ gene driven by the SCG10 minimum promoter containing the neural-restrictive silencer element (NRSE), which element selectively represses the transcription of genes in non-neuronal cells. When rat hippocampal slice cultures were infected with NRSE-bearing adenovirus, beta-galactosidase-positive cells were mostly pyramidal and granular neurons, whereas infection with virus carrying a mutated NRSE resulted in beta-galactosidase expression in both neurons and glia. The results suggest that the adenovirus carrying NRSE to be a useful tool for neurontargeted gene transfer.

Adenoviridae↗

PubMed1999

Toddlers with limb deficiency: conceptual basis and initial application of a functional status outcome measure.

OBJECTIVE: To describe the conceptual foundation, development, and initial psychometric analyses of a new outcome measure of functional status in toddlers with limb deficiency. DESIGN: Parents of children with limb deficiency completed self-report measures during a routine medical clinic visit. SETTING: Outpatient orthopedic pediatric clinic. PARTICIPANTS: Twenty parents (mothers) of children (ages 1 to 4 years) with acquired or congenital limb deficiency. MAIN OUTCOME MEASURE: The newly developed Child Amputee Prosthetics Project-Functional Status Inventory for Toddlers (CAPP-FSIT). RESULTS: Estimates of internal consistency reliability of the measure are high, suggesting conceptual congruence among the items. Initial validity studies confirm the CAPP-FSIT differentiates between toddlers with upper limb deficiency and lower limb deficiency in terms of functional activity and prosthesis use. The new measure does not appear to be contaminated by gender or socioeconomic status. CONCLUSION: The CAPP-FSIT is a promising measure for assessing functional abilities in toddlers with limb deficiency.

Activities of Daily Living↗

PubMed1999

Gene therapy for renal anemia in mice with polycystic kidney using an adenovirus vector encoding the human erythropoietin gene.

BACKGROUND: Recombinant human erythropoietin (rHuEPO) is primarily used for patients with anemia associated with end-stage renal disease. We evaluated the efficacy of EPO gene therapy using adenovirus vector for chronic renal failure mice expressing severe renal anemia. METHODS: Recombinant HuEPO gene transfer to mesothelial cells was performed in vitro and in vivo. Recombinant replication-deficient adenoviruses containing rHuEPO cDNA (AdCMVEPO), E. coli lacZ gene (AdCMVlacZ), or an nonexogenous gene (AdNull as control vector) driven by the cytomegalovirus promotor/enhancer were constructed. The oligosaccharides associated with the rHuEPO from AdCMVEPO-treated mesothelial cells were analyzed. For in vivo study, the DBA/2FG-pcy mouse, a model for human autosomal recessive polycystic kidney disease resulting in chronic renal failure with progressive anemia, was used. RESULTS: The sialylated oligosaccharides associated with the rHuEPO produced in AdCMVEPO-treated mesothelial cells occupied 78 +/- 0.7% of the total oligosaccharide pool. A single intraperitoneal administration of AdCMVEPO induced rHuEPO synthesis in the peritoneal cells and a marked increase in erythrocyte production. The maximal increase in hematocrit (43 +/- 4%) was observed on day 28, and it remained elevated for 40 days. CONCLUSION: These results indicate that intraperitoneal administration of AdCMVEPO improves renal anemia in mice with chronic renal failure and that the mesothelial cell is an appropriate target cell for gene transfer.

Adenoviridae↗

PubMed1998

Induction of DNA fragmentation and HSP72 immunoreactivity by adenovirus-mediated gene transfer in normal gerbil hippocampus and ventricle.

Foreign genes have been successfully transferred and expressed in experimental animal brains using adenoviral vectors. However, it is not fully understood whether adenovirus-mediated gene transfer causes stressful or cytotoxic injury in brain. A replication-defective adenoviral vector containing the Escherichia coli lacZ gene (AdCMVnLacZ) was directly injected into right hippocampus and lateral ventricle of normal gerbil brains. Temporal and spatial profiles of the expression of lacZ gene products, DNA fragmentation detected by terminal deoxynucleotidyl d-UTP nick end labeling (TUNEL) staining, and heat shock protein 72 (HSP72) immunoreactivity were examined until 21 days after the injection. In the ventricle, lacZ gene was immediately and strongly expressed at 8 hr after the injection of AdCMVnLacZ, with a peak at 1-3 days, and disappeared by 21 days. Although a small number of choroid plexus cells were TUNEL positive at 3 and 7 days, no HSP72 immunostaining was observed in the ventricle. Small-to-moderate expression of lacZ gene was found in the needle route from 8 hr to 3 days after the injection, and a small number of TUNEL-positive cells were detected at the needle track at 1-3 days. In the hippocampus, lacZ gene was markedly expressed around the dentate gyrus (DG) at 8 hr to 3 days with a peak at 1 day. Large number of TUNEL or moderate-to-dense HSP70 staining cells were also detected in the same area. CA1 neuronal cells just adjacent to the needle route showed TUNEL positivity at 1 to 3 days. However, the TUNEL staining was not associated with lacZ gene expression. The majority of lacZ-expressing cells were discriminated from the TUNEL-positive cells, whereas some were double-positive with HSP72 staining in DG. Cellular loss was observed in the CA1 layer around the needle route. An apoptotic change was morphologically observed in the marginal region of the DG at 1-3 days and in the ventricle at 3-7 days. In the sham control group, TUNEL-positive or HSP72-staining cells were only detected around the needle track including CA1 cells adjacent to the needle route. These data suggest that adenoviral gene transfer may induce direct traumatic injury in the CA1 sector near the needle route, indirect apoptotic cell loss in the DG and ventricle, and stressful effect on the dentate granule cells in association with adenovirus infection in normal gerbil brain.

Adenoviridae↗

PubMed1998

Expression of adenovirus-mediated E. coli lacZ gene in skeletal muscles and spinal motor neurons of transgenic mice with a mutant superoxide dismutase gene.

A replication-defective recombinant adenoviral vector containing E. coli lacZ gene was injected into the right biceps brachii muscles of transgenic mice carrying mutant human Cu/Zn superoxide dismutase (SOD1) gene and non-transgenic wild-type mice at 27 weeks of age. Although the transgenic mice showed remarkable neurogenic muscular changes and a marked motor neuron loss in the anterior horn of spinal cord, the lacZ gene was widely expressed in all the injected muscles of transgenic mice as well as of wild-type mice at 7 days after the injection. In one transgenic and two wild-type mice, the lacZ gene expression was first detected in a few motor neurons of right lower cervical cord (C5-C6). These results demonstrate that an adenovirus-mediated foreign gene is transferred and expressed in skeletal muscles both of normal and transgenic mice model for familial amyotrophic lateral sclerosis (FALS), and also, in the spinal motor neurons, may be transferred by retrograde transport from innervated muscles.

Adenoviridae↗

PubMed1998

Functional status in limb deficiency: development of an outcome measure for preschool children.

OBJECTIVE: To develop an outcome measure of functional status in preschool children with limb deficiency. DESIGN: Parents of preschool children with limb deficiency completed self-report measures during a routine medical clinic visit. SETTING: Outpatient pediatric clinic. PARTICIPANTS: Fifty-two parents of children (ages 4 to 7) with acquired or congenital limb deficiency. MAIN OUTCOME MEASURE: The newly developed Child Amputee Prosthetics Project-Functional Status Inventory for Preschool children (CAPP-FSIP). RESULTS: Initial psychometric studies indicate high internal consistency reliability of the new instrument. Content validity was established through expert clinician review. Initial construct validity analyses confirm that the CAPP-FSIP discriminates between children with upper and lower deficiency and their prosthesis use for functional activities. CONCLUSION: The CAPP-FSIP is a promising new measure for evaluating preschool age children with limb deficiency.

Activities of Daily Living↗

PubMed1998

DNA fragmentation and HSP72 gene expression by adenovirus-mediated gene transfer in postischemic gerbil hippocampus and ventricle.

A replication defective adenoviral vector containing the E. coli lacZ gene (AdCMVnLacZ) was directly injected into right hippocampus and lateral ventricle immediately after 5 min of transient global ischemia in gerbils. The relations between the lacZ gene expression and DNA fragmentation or heat shock protein 72 (HSP72) immunoreactivity were examined up to 21 days post ischemia. The lacZ gene was transiently expressed at 1 day in the hippocampus except around the CA1 region, while a large number of the periventricular cells strongly expressed the lacZ gene from 8 h to 7 days. In CA1 layer, terminal deoxynucleotidyl dUTP nick end labeling (TUNEL) positive cells, which were present only adjacent to the needle track at 8 h to 1 day, became more extensive in the whole CA1 layer at 3 to 7 days. TUNEL-positive cells were also detected around the DG at 1 day, around the needle track at 8 h to 3 days, and in the choroid plexus cells at 7 days. HSP72 staining was detected in the subiculum at 1 to 3 days, the dentate granule cells at 8 h to 1 day, and in the CA3 or CA4 pyramidal cells at 1 to 3 days. Some lacZ expressing cells were double-positive with HSP72 in DG, while the majority of those were distinguished from the TUNEL-positive cells. Pyramidal neurons were almost completely lost in the CA1 sector at 7 days after the ischemia. The present study demonstrates the successful LacZ gene transfer into the hippocampus and ventricle of postischemic gerbil brain except in the vulnerable CA1 layer by adenoviral vector injection. However, adenovirus-mediated gene transfer may induce indirect apoptotic cell death in the DG and ventricle, in addition to direct traumatic injury around the needle track.

Adenoviridae↗

PubMed1998

Temporal profile of adenovirus-mediated E. coli lacZ gene expression in normal and post-ischemic gerbil hippocampus and ventricle.

A replication-defective adenoviral vector containing the E. coli lacZ gene was directly injected into normal and post-ischemic gerbil right hippocampus and lateral ventricle, and temporal profiles of the exogenous gene expression were compared. In case of ischemia, common carotid arteries (CCA) were transiently occluded for 5 min, and the adenoviral vector was administered just after the reperfusion. The animals were recovered for 8 h, 1, 3, 7 or 21 days. A small to moderate number of neural cells in the normal hippocampus expressed the gene from 1-3 days except for the cells around dentate gyrus (DG) and the needle route that began to express from 8 h of injection. Some normal hippocampal cells persisted the expression until 7 days. A moderate to large number of ventricular cells expressed the lacZ gene from 8 h to 7 days in the normal brain. On the other hand, no expression of the lacZ gene was observed in the post-ischemic hippocampus at 8 h including cells at DG and the needle route. Hippocampal CA1 neurons, that were selectively lost at 7 days of reperfusion, never expressed the gene throughout the post-ischemic course. The other hippocampal cells such as CA3 and dentate granule cells that survived ischemia expressed the gene only transiently at 1 day. A robust expression of the gene persisted in the ventricular cells from 8 h to 7 days. The majority of brain cells in the hippocampus that expressed the lacZ gene was not the pyramidal neurons, but small neurons at around the pyramidal layers of DG. Some astroglial, but no microglial, cells expressed the lacZ gene in the hippocampus. The present study shows that an expression of the lacZ gene was limited in the post-ischemic gerbil hippocampus especially at the vulnerable CA1 layer in contrast to the strong and persistent expression in the ventricular cells, and that the majority of beta-gal positive cells were not the pyramidal neurons but small neurons at around the cell layer both in the control and post-ischemic gerbil hippocampus.

Adenoviridae↗

PubMed1998

Differential expression of laminin receptors in human hepatocellular carcinoma.

BACKGROUND: Laminin receptors are involved in cell-extracellular matrix interactions in malignant cells that show invasion and metastasis. Hepatocellular carcinoma frequently shows early invasion into blood vessels, and intrahepatic and extrahepatic metastases. However, the role of laminin receptors in hepatocellular carcinoma is unknown. AIMS: To examine the expression of mRNA for laminin receptors and their isoforms in hepatocellular carcinoma. METHODS: The expression of several laminin receptors, including alpha1 integrin, alpha6 integrin and its isoforms alpha6A and alpha6B, beta1 integrin and its isoforms beta1A and beta1B, and 32kD/67kDa laminin binding protein was examined in human hepatocellular carcinomas and non-cancerous liver tissues using the reverse transcription polymerase chain reaction. RESULTS: Alpha6 Integrin, beta1 integrin, and laminin binding protein showed notably increased expression in hepatocellular carcinoma, compared with non-cancerous liver tissue, although the alpha1 integrin did not show a significant change. Furthermore, beta1B integrin, a splicing variant of beta1 integrin, was overexpressed in hepatocellular carcinoma while the beta1A integrin isoform did not show significant changes between hepatocellular carcinoma and surrounding non-cancerous liver tissue. CONCLUSIONS: The differential upregulation of laminin receptors and their splicing isoforms was shown in hepatocellular carcinoma, suggesting that certain laminin receptors and their isoforms may be involved in the development and progression of hepatocellular carcinoma.

Aged↗

PubMed1998

Efficient generation of autologous peripheral blood-derived cytotoxic T lymphocytes against poorly immunogenic human tumors using recombinant CD80-adenovirus together with interleukin 12 and interleukin 2.

To generate CTLs against poorly immunogenic human tumor cells, we transfected the human CD80 gene into the tumor cells using a replication-deficient adenovirus (Ad) vector. The successful surface expression of CD80 was obtained in both cultured tumor cell lines and primary cultured tumor cells. Transduction of CD80 alone was not sufficient to induce cytotoxicity of peripheral blood lymphocytes against allogeneic tumor cell lines except for melanoma cells. We, therefore, investigated a combined effect of CD80-Ad-infected tumor cells and interleukin 12 (IL-12). Although 7-day cultivation of autologous or allogeneic lymphocytes with CD80-Ad-infected tumor cells and IL-12 slightly enhanced cytotoxicity against some allogeneic tumor cells, no substantial cytotoxicity was observed against autologous tumor cells. When we extended the culture period to 14 days in the presence of IL-2, a prominent enhancement of cytotoxicity was observed against both allogeneic and autologous tumor cells. Cytotoxicity against autologous tumor cells, but not against allogeneic tumor cells, was efficiently inhibited by anti-CD3 monoclonal antibody. Furthermore, the selective cytotoxicity against a panel of targets indicated that the induced CTLs recognize specific antigens on autologous tumor cells. These results suggest that stimulation with a combination of IL-12- and CD80-modified tumor cells and subsequent expansion with IL-2 may efficiently generate tumor-specific CTLs from autologous peripheral blood lymphocytes. Our data imply that the combination of CD80 transduction and suitable cytokines is useful for enhancing antitumor immunity to poorly immunogenic human tumors.

Adenocarcinoma↗

PubMed1997

In vivo adenovirus-mediated gene transfer and the expression in ischemic and reperfused rat brain.

In an attempt to study whether ischemic brain could express a foreign gene in vivo, a replication-defective adenoviral vector containing the Escherichia coli lacZ gene was directly injected into the ischemic or reperfused cerebral cortex of rat, and temporal and spatial profiles of the exogenous gene expression were compared with that of the control brain. Right middle cerebral artery (MCA) of rat was continuously occluded by an insertion of nylon thread for 2 days, or only transiently occluded for 90 min and then the blood flow was restored for 21 days. The adenoviral vector was administered just after the MCA occlusion or reperfusion in the case of continuous ischemia and reperfusion, respectively. Adenoviral vector was transferred into the continuous ischemic brain, and the lacZ gene was expressed until 2 days of the occlusion in the cerebral cortex of the occluded MCA territory with the number of expressing cells smaller and the staining just weaker than that of the control brain. In contrast, expression of the lacZ gene was not or only minimally observed in the reperfused brain until 2 days. However, the expression dramatically exploded at 7 days of reperfusion at a level similar to that of the control, and the expression diminished by 21 days. A few neurons in the ipsilateral thalamus, hypothalamus, and basal ganglia, and in the contralateral cerebral cortex expressed the lacZ gene at 7 days after reperfusion, a phenomenon similar to the case of the control. The majority of brain cells that expressed the lacZ gene were neurons, and a part (5-10%) were astroglial cells. Traumatic injury and immunological response in the brain were minimal both in the cases of control and ischemia/reperfusion. The present study shows an effective gene transfer and the expression in neural cells of ischemic and reperfused brains in vivo, and suggests a great potential of the gene therapy for ischemic stroke patients in the future.

Adenoviridae↗

PubMed1997

Dissociative expression of adenoviral-mediated E. coli LacZ gene between ischemic and reperfused rat brains.

A replication-defective adenoviral vector containing the E. coli lacZ gene was directly injected into the ischemic or reperfused cerebral cortex of rats. An administration of adenoviral vector showed a slight to moderate expression of the lacZ gene in the cerebral cortex of the middle cerebral artery (MCA) region until 2 days after the MCA occlusion. In contrast, expression of the lacZ gene was not observed, or only minimally so, in the reperfused brain until 2 days after a 90 min of transient MCA occlusion. However, the lacZ expression dramatically increased at 7 days after the reperfusion, then diminished by 21 days. The majority of brain cells that expressed the lacZ gene were neurons and a fraction (5-10%) were astroglial cells. The present study showed that an exogenous gene was transferred and expressed in neural cells of ischemic and reperfused brains in vivo, but the temporal profile of the expression is dissociative.

Animals↗

PubMed1997

Prosthesis satisfaction outcome measurement in pediatric limb deficiency.

OBJECTIVE: To describe the development and initial psychometric properties of a new outcome measure to assess satisfaction with prosthesis in children with limb deficiencies. DESIGN: Parents of children with limb deficiency were surveyed during routine outpatient clinic visits. SETTING: Two outpatient pediatric clinics. PARTICIPANTS: Ninety-seven parents of children with limb deficiency aged 1 to 17 years. MAIN OUTCOME MEASURE: The newly developed Child Amputee Prosthetics Project-Prosthesis Satisfaction Inventory (CAPP-PSI). RESULTS: Internal consistency reliability is high. Zero-order correlations with prosthesis wear/use patterns and with parent ratings of prosthesis appearance provide support for the construct validity of the CAPP-PSI. CONCLUSION: The CAPP-PSI is a promising, brief, parent-administered inventory for the assessment of prosthesis satisfaction in children with limb deficiency. It may be useful in future research for predicting prosthesis wear and use patterns in this population.

Activities of Daily Living↗

PubMed1997

cDNA cloning and differential gene expression of three catalases in pumpkin.

Three cDNA clones (cat1, cat2, cat3) for catalase (EC 1.11.1.6) were isolated from a cDNA library of pumpkin (Cucurbita sp.) cotyledons. In northern blotting using the cDNA-specific probe, the cat1 mRNA levels were high in seeds and early seedlings of pumpkin. The expression pattern of cat1 was similar to that of malate synthase, a characteristic enzyme of glyoxysomes. These data suggest that cat1 might encode a catalase associated with glyoxysomal functions. Furthermore, immunocytochemical analysis using cat1-specific anti-peptide antibody directly showed that cat1 encoding catalase is located in glyoxysomes. The cat2 mRNA was present at high levels in green cotyledons, mature leaf, stem and green hypocotyl of light-grown pumpkin plant, and correlated with chlorophyll content in the tissues. The tissue-specific expression of cat2 had a strong resemblance to that of glycolate oxidase, a characteristic enzyme of leaf peroxisomes. During germination of pumpkin seeds, cat2 mRNA levels increased in response to light, although the increase in cat2 mRNA by light was less than that of glycolate oxidase. cat3 mRNA was abundant in green cotyledons, etiolated cotyledons, green hypocotyl and root, but not in young leaf. cat3 mRNA expression was not dependent on light, but was constitutive in mature tissues. Interestingly, cat1 mRNA levels increased during senescence of pumpkin cotyledons, whereas cat2 and cat3 mRNAs disappeared during senescence, suggesting that cat1 encoding catalase may be involved in the senescence process. Thus, in pumpkin, three catalase genes are differentially regulated and may exhibit different functions.

Amino Acid Sequence↗

PubMed1997

Elevated expression levels of the 14-3-3 family of proteins in lung cancer tissues.

Immunochemical staining of lung cancer sections with a murine monoclonal anti-14-3-3 antibody showed a sharp discrimination of the cancer tissue from neighboring normal counterparts in 88 of 121 primary lung cancer tissue specimens of all four major lung cancer histologies; specifically, 32 of 48 adenocarcinomas, 36 of 44 squamous cell carcinomas, 10 of 13 large cell carcinomas, and 10 of 16 small cell carcinomas, respectively, were stained positively. Sets of the 10,000 x g supernatants of normal and cancerous lung tissue homogenates, each set prepared from surgically dissected tissues of the cancer and its surrounding normal part, were assayed for 14-3-3 proteins by the sandwich enzyme-linked immunosorbent assay using two different monoclonal antibodies to 14-3-3 proteins. The results of the assay demonstrated 7.2 times higher 14-3-3 protein content in the lung cancer tissue (378 +/- 200 ng ml-1) as compared with the normal lung (54 +/- 35 ng ml-1). These results indicate that the 14-3-3 family of proteins can be an effective marker for lung cancer diagnosis such as sputum cytodiagnosis and that 14-3-3 proteins might be involved in the development of lung cancers.

14-3-3 Proteins↗

PubMed1996

Functional status in children with limb deficiency: development and initial validation of an outcome measure.

OBJECTIVE: To develop a new outcome measure in response to the increasing demands for cost effectiveness analyses and empirically derived outcome instruments in the treatment of pediatric limb deficiency. This article describes the development, refinement, and initial psychometric properties of the Child Amputee Prosthetics Project-Functional Status Inventory (CAPP-FSI). DESIGN: Parents of children with limb deficiency were surveyed during routine clinic visits. SETTING: Two outpatient pediatric clinics. PARTICIPANTS: Seventy-five parents and their children with limb deficiency (ages 8 to 17 years) participated in the study as part of annual physical evaluations or routine follow-up care. MAIN OUTCOME MEASURE: The newly developed CAPP-FSI. RESULTS: Internal consistency reliability (Cronbach's alpha) = .96 for the CAPP-FSI. Content validity is described and initial construct validity is empirically confirmed. CONCLUSION: The CAPP-FSI is a promising assessment instrument for measuring important health outcomes in children with upper or lower limb deficiency.

Activities of Daily Living↗

PubMed1996

Phylogenetic tree-based epidemiological analysis of hepatitis C virus transmission in a region of Japan with a high prevalence of infection.

Local clustering of hepatitis C virus (HCV) infection has been demonstrated in various regions in Japan. HCV genotypes have now been compared between infected individuals from districts of Saga prefecture with either a high (H district) or low (L district) prevalence of HCV-seropositivity. The prevalence of HCV genotype 1b was significantly higher (P < 0.001) in the H district (45/50; 90%) than in the L district (19/36; 52.8%). A phylogenetic tree was constructed based on the genomic sequences of viral isolates from 20 patients infected with genotype 1b in the H district. Almost all these HCV strains clustered in the same regions of the tree. With regard to risk factors for HCV transmission, the percentage of patients with a history of surgery was significantly higher in the H district than in the L district (58 versus 33.3%; P < 0.05). Of 20 patients infected with similar strains of HCV in the H district, 16 (80%) had at least one parenteral risk factor associated with medical care. These results indicate an increased transmission of similar strains of HCV in the H district as a result of nosocomial infection.

Base Sequence↗

PubMed1996

Energy expenditure during walking by children who have proximal femoral focal deficiency.

The energy expended during walking by sixteen children who had proximal femoral focal deficiency was measured to compare the outcomes after Syme amputation (seven subjects) with those after Van Nes rotational osteotomy (nine subjects). Multivariate regression analysis revealed that the subjects who had had a Van Nes procedure had a mean oxygen cost (energy per unit of body mass expended per distance walked) that was 0.12 milliliter per kilogram of body mass per meter lower than that of the subjects who had had a Syme amputation (p = 0.001). The subjects who had had a Van Nes procedure tended to walk faster (p = 0.07). A significant decrease in the oxygen cost as a function of increasing age was observed for both groups (p < 0.0001, r2= 0.79). We believe that the reduced energy expenditure associated with the Van Nes rotational osteotomy is one of several factors to consider when deciding which operation should be done for children who have proximal femoral focal deficiency.

Adolescent↗