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Biomedical subjects

Y Setoguchi

Publications and source records attributed to Y Setoguchi.

At least 55 records · Page 3Linked to original sources
PubMed1994

Stimulation of erythropoiesis by in vivo gene therapy: physiologic consequences of transfer of the human erythropoietin gene to experimental animals using an adenovirus vector.

Erythropoietin (Epo), a 30.4-kD glycoprotein, is the principal regulator of erythropoiesis. To evaluate the concept that in vivo gene transfer might be used as an alternative to recombinant human Epo (rhEpo) in applications requiring a 1- to 3-week stimulation of erythropoiesis, the replication-deficient recombinant adenovirus AdMLP.Epo was constructed by deleting the majority of E1 from adenovirus type 5, and replacing E1 with an expression cassette containing the adenovirus type 5 major late promoter (MLP) and the human Epo gene, including the 3' cis-acting hypoxia response element. In vitro studies showed that infection of the human hepatocyte cell line Hep3B with AdMLP.Epo resulted in a 15-fold increase in Epo production in 24 hours that was enhanced to 116-fold in the presence of a hypoxic stimulus. One-time in vivo administration of AdMLP.Epo (7 x 10(9) plaque-forming units/kg) to the peritoneum of cotton rats caused a marked increase in red blood cell production, with a 2.6-fold increase in bone marrow erythroid precursors by day 4, and sevenfold increase in reticulocyte count by day 7. The hematocrit increased gradually, with a maximum of 64% +/- 4% at day 14 (compared with an untreated baseline of 46% +/- 2%), and a level of 55% +/- 1% at day 24. Furthermore, one-time subcutaneous administration of AdMLP.Epo caused an increase in hematocrit that peaked at 14 days (57% +/- 2%) and was still elevated at day 42. Hematocrit level in animals receiving subcutaneous administration of AdMLP.Epo sustained a long-term increase compared with animals receiving intraperitoneal administration. In the context of these observations, gene therapy with a single administration of an adenovirus vector containing the human EPO gene may provide a means of significantly augmenting the circulating red blood cell mass over the 1- to 3-week period necessary for many clinical applications.

Adenoviridae↗
PubMed1994

Cloning and expression of cDNA of human delta 4-3-oxosteroid 5 beta-reductase and substrate specificity of the expressed enzyme.

The enzyme delta 4-3-oxosteroid 5 beta-reductase (3-oxo-5 beta-steroid: NADP+ oxidoreductase and 4,5 beta-dihydrocortisone: NADP+ delta 4-oxidoreductase) catalyzes the reduction of the delta 4 double bond of bile acid intermediates and steroid hormones carrying the delta 4-3-one structure in the A/B cis configuration. Human delta 4-3-oxosteroid 5 beta-reductase cDNA was isolated from a liver cDNA library by cross hybridization with a previously cloned rat cDNA, which was used as a probe [Onishi, Y. Noshiro, M., Shimosato, T. & Okuda, K.-I. (1991) FEBS Lett. 283, 215-218]. DNA sequence analysis of a hybridization-positive clone predicted the human delta 4-3-oxosteroid 5 beta-reductase to contain 326 amino acids. The amino acid sequence of the human delta 4-3-oxosteroid 5 beta-reductase had 79% overall identity to the rat enzyme sequence. It also showed 54% and 50% overall identity with rat 3 alpha-hydroxysteroid dehydrogenase and human aldose reductase, respectively. RNA blotting analysis demonstrated the existence of a single delta 4-3-oxosteroid 5 beta-reductase mRNA of approximately 2.7 kb in human liver. Transfection of the cDNA into COS cells resulted in the expression of an active enzyme with a high activity toward the bile acid intermediates 7 alpha,12 alpha-dihydroxy-4-cholesten-3-one and 7 alpha-hydroxy-4-cholesten-3-one. In addition, the expressed enzyme showed a small but significant 5 beta-reduction activity toward 11 beta,17 alpha,21-trihydroxy-delta 4-pregnene-3,20-dione (cortisol) and 17 beta-hydroxy-delta 4-androsten-3-one (testosterone) whereas no activity was observed toward delta 4-pregnene-3,20-dione (progesterone) or delta 4-androstene-3-17-dione (androstenedione). The substrate specificity of the human enzyme is considerably narrower than that of the rat enzyme, and the enzyme seems to be more important for bile acid biosynthesis than for metabolism of steroid hormones.

Amino Acid Sequence↗
PubMed1994

Ex vivo and in vivo gene transfer to the skin using replication-deficient recombinant adenovirus vectors.

The skin has the potential for a variety of gene therapy applications. In addition to local delivery, it is the largest organ of the body, and highly vascular, and thus is an ideal site for systemic delivery of gene products. To evaluate the potential for adenovirus-mediated skin gene transfer, the replication-deficient recombinant adenovirus vectors Ad.RSV beta gal (coding for Escherichia coli beta-galactosidase) and Ad alpha 1AT (coding for human alpha 1-antitrypsin) were used in both ex vivo and in vivo approaches. Following in vitro infection with Ad.RSV beta gal, murine keratinocytes expressed beta-galactosidase. Parallel in vitro studies with Ad alpha 1AT documented de novo synthesis and secretion of human alpha 1AT as shown by [35S]methionine labeling and immunoprecipitation. Quantification of human alpha 1AT in the culture supernatants demonstrated 0.1-0.3 microgram human alpha 1AT secreted/ml-24 h. Evaluation of the serum of mice receiving transplants (10(5) cells/mouse) of Ad alpha 1AT-infected syngeneic keratinocytes demonstrated human alpha 1AT for at least 14 d with maximum levels of 41 ng/ml. To demonstrate the feasibility of direct adenovirus-mediated in vivo transfer of genes to the skin, Ad.RSV beta gal or Ad alpha 1AT were administered subcutaneously to mice. Histologic evaluation after 4 d demonstrated expression of beta-galactosidase in various types of skin cells. Quantification of human alpha 1AT in serum of animals infected subcutaneously with Ad alpha 1AT showed levels of 53 ng/ml at day 4, with human alpha 1AT detectable for at least 14 d. These observations support the feasibility of ex vivo and in vivo gene transfer to the skin mediated by replication-deficient adenovirus vectors.

Adenoviridae↗
PubMed1994

Intraperitoneal in vivo gene therapy to deliver alpha 1-antitrypsin to the systemic circulation.

The utility of replication-deficient recombinant adenovirus vector-mediated transfer and expression of the alpha 1-antitrypsin (alpha 1AT) cDNA to peritoneal mesothelial tissues was evaluated as a means of delivering alpha 1AT to the systemic circulation. Preliminary studies with Ad.RSV beta gal, an adenovirus vector expressing the Escherichia coli lacZ gene (beta-galactosidase), showed that intraperitoneal injection of 10(9) plaque-forming units (pfu) to cotton rats resulted in beta-galactosidase activity in mesothelial cells lining the peritoneal cavity. After intraperitoneal administration of 10(9) pfu of Ad alpha 1AT (an adenovirus vector containing the human alpha 1AT cDNA), human alpha 1AT was detectable in serum for up to 24 days, with a maximal level of 3.4 micrograms/ml at 4 days. Expression of the exogenous gene was localized to the peritoneal mesothelium as PCR analyses detected no evidence of expression of the exogenous gene in any other tissues evaluated. Anti-adenovirus vector antibodies were detectable in serum after intraperitoneal administration of the recombinant vectors, including antibodies with neutralizing activity. Repeat administrations of adenovirus vectors to the peritoneal cavity at 1 wk and 1 mo after the initial dose failed to show gene expression, but repeat administration 3 mo after demonstrated measurable gene transfer and expression. Together these observations suggest replication-deficient adenovirus-mediated gene transfer to the peritoneal mesothelium offers a promising means to transfer alpha 1AT to the systemic circulation, although immunity induced against the adenovirus may limit frequent repetitive dosing.

Adenoviruses, Human↗
PubMed1994

[Gene transfer to airway epithelial cells: current status and future direction].

Advances in recombinant DNA technology and molecular and cellular biology have made it feasible to introduce genes into living cells. The most sophisticated gene transduction methods have bee applied to gene therapy strategies for the potential treatment of genetic diseases. In regard to lung diseases, alpha 1-antitrypsin deficiency and cystic fibrosis, the most common hereditary lung disorders in Caucasians, have been targeted for gene therapy. To date, gene therapy studies have been confined to ex vivo strategies for treatment of ADA deficiency with retroviral vectors. However, there are two major obstacles to gene transfer to the bronchial epithelium. First, bronchial epithelium, such as that with ciliated cells, is terminally differentiated, and does not divide rapidly. Second, the complex architecture of the lung precludes replacing the existing bronchial epithelium with cells modified by gene transfer. In the context of these properties of bronchial epithelium, adenovirus vectors have been evaluated for direct introduction of therapeutic genes to bronchial epithelium via the airway in vivo. An in vivo experiment revealed that gene transfer with a replication-deficient adenovirus containing the E. coli lacZ (beta-galactosidase) gene driven by cytomegalovirus promoter (AdCMVlacZ) was 10(4) times more efficient than gene transfer with a plasmid containing the same expression cassette (pCMVlacZ). An experiment based on in vitro data was done to evaluate the distribution of the expression of the exogenous genes transferred by adenovirus vectors. Intratracheal administration of AdCMVlacZ into lungs of experimental animals resulted in a high number of beta-gal-positive epithelial cells in bronchiols, rather than in proximal bronchi. Thus, a replication-deficient adenovirus can be used to transfer exogenous genes to airway epithelial cells in vivo. This technique may be useful in gene therapy for cystic fibrosis. Gene transfer can be thought of as the use of genetic information to modity the milieu of the target organ. In addition, gene transfer may allow the introduction of new genes, or the alteration tion of existing genes in intact animals. Gene transfer could them be used to produce animal models of human lung diseases that are particularly difficult to study.

Animals↗
PubMed1994

The 14-3-3 protein as the antigen for lung cancer-associated human monoclonal antibody AE6F4.

Human monoclonal antibody (MAb) AE6F4, which had been shown potentially useful for the immunocytological detection of lung cancer cells in sputum, was characterized for its antigen(s). Of the three MAb-reacting materials found in A549 cells by the immunoblotting analysis, the cytoplasmic 31-kDa protein extractable with phosphate-buffered saline was evidenced as the most plausible antigen by its highest content and outstanding affinity to the MAb AE6F4-derivatized Sepharose 4B column. This 31-kDa protein was identified by the amino acid sequence analysis of the CNBr-cleaved fragment as the 14-3-3 family of proteins, the members of which are known to play important physiological roles such as in the regulation of neurotransmitter levels and intracellular signal transduction. The purified 14-3-3 protein from bovine brain showed a comparable MAb-reacting activity to that of the 31-kDa protein from A549 cells in the enzyme-linked immunosorbent assay (ELISA). The significant reactivity of bovine 14-3-3 protein by MAb AE6F4, shown by the cross inhibition of antibody binding to the coated 31-kDa antigen in ELISA as well as by the inhibition of immunostaining with lung cancer tissues, consistently demonstrated that the antigen(s) recognized by the MAb was involved in the 14-3-3 protein family. It was found that the expression of the 14-3-3 protein was significantly enhanced in lung cancer tissues compared with the neighboring normal part of the lung as examined by the immunoblotting method. These results implicated that some member(s) of the 14-3-3 protein family can be the tumor marker(s), providing a rational basis for the immunocytological diagnosis of lung cancer with this human MAb.

14-3-3 Proteins↗
PubMed1993

Intrathecal injection of high-dose meglumine amidotrizoate with complete recovery.

Rarely we are faced with accidental spinal injection of potentially toxic substances. We present 2 cases in which amidetrizoate, water-soluble ionic contrast medium, was accidentally injected intrathecally. Our treatment consisted of vigorous hydration and barbiturate coma. This report suggests that for water-soluble ionic contrast media increasing cerebrospinal fluid circulation by vigorous hydration may be as effective as spinal lavage in diminishing toxicity.

Adult↗
PubMed1993

Dynamics of below-knee child amputee gait: SACH foot versus Flex foot.

Gait kinematics and dynamics during stance of unilateral, below-knee child amputees were analyzed for self-selected, comfortable (0.9 m s-1) and fast (1.3 m s-1) speeds with the SACH foot and the energy-storing Flex-foot prostheses. The three-dimensional movements of the lower limbs were recorded and synchronized with ground reaction forces for 12 subjects (7 girls and 5 boys, ages 6-16 yr). Each lower limb was modeled as a set of interconnected rigid links (thigh, leg, and foot) with frictionless joints (hip, knee, and ankle) to calculate moment and joint power profiles. Marked asymmetries were noted in ground reaction force, joint moment, and power profiles for the prosthetic versus the natural limb, but with the Flex foot the asymmetries were less pronounced than with the SACH foot. For the amputee wearing the Flex foot, greater moments and power were generated by the natural limb at the comfortable pace as compared to the SACH foot, but during fast walking, the SACH foot required greater output from the natural limb. With both prostheses, for the prosthetic limb the amputees used similar force, moment, and power patterns, but with significantly different amplitudes. At both speeds of walking, the Flex foot returned significantly more energy (66% at comfortable and 70% at fast walking) than the SACH foot (21% at comfortable and 19% at fast walking). Thus, the Flex foot had a greater potential for reducing the energy cost of walking at comfortable and fast speeds for the below-knee child amputee.

Adolescent↗
PubMed1993

Cancer-specific binding of a mouse MAb vs. Candida krusei cytochrome c: an antigen recognized by a cancer-associated human MAb HB4C5.

More than 60 mouse monoclonal antibodies directed to cytochrome c from Candida krusei with different specificities were raised. Most of these monoclonal antibodies, except for three of them, did not cross-react with bovine cytochrome c. By the immunoblotting method, the monoclonal antibodies of clones HCC 5-13, 9-2, and 10-5 reacted with the Candida cytochrome c, which had been transferred onto nitrocellulose membrane, but those of clones HCC 1-22, 6-3, and 17-3 did not, although all these monoclonal antibodies strongly reacted with coated Candida cytochrome c on plastic immunoplates when examined by ELISA. On the contrary, monoclonal antibody activities of clones HCC 1-22, 6-3, and 17-3 in binding to the coated cytochrome c in ELISA were inhibited competitively by the addition of extra Candida cytochrome c, whereas those of clones HCC 5-13, 9-2, and 10-5 were not inhibited. Among these monoclonal antibodies, the antibody of clone HCC 6-3, which showed a good reactivity to added cytochrome c in inhibiting ELISA reaction but was not reactive with the transblotted cytochrome c on nitrocellulose, was found to be reactive with human lung cancer tissues specifically with no reactivity to normal tissues. The immunostaining of lung cancer tissue showed that this mouse monoclonal antibody to Candida cytochrome c reacted to the cytoplasmic fraction of the cancer cells specifically.

Animals↗
PubMed1993

[Clinical investigation of HCV antibodies positive cases in the district with high mortality rate of liver diseases in Saga Prefecture].

The mortality and morbidity rates of chronic liver disease (CLD) have been extremely high for several years at Saga prefecture in Japan. Epidemiological studied revealed districts with high mortality rate of CLD (H-district). We measured C 100-3Ab and HCVCoreAb (CoreAb) in the H-district and observed high discrepancy between C100-3 and CoreAb. In order to examine pathogenesis of this discrepancy, we determined serum HCVRNA in these cases. The positive cases of HCV-Ab among 459 subjects were 33.6% (159). 59 subject were positive for both, 88 were only positive for CoreAb, 7 were only positive for C100-3Ab. Serum HCVRNA was detected in 92% (22/24) of both positive cases, in 67% (6/9) of only CoreAb positive cases, in 43% (3/7) of only C100-3Ab positive cases. These results suggested the cases positive for C-100-3 and CoreAb have chronic liver injury due to HCV infection, while the cases positive for only CoreAb involves healthy carrier or the memory state of HCV infection without liver injury.

Aged↗
PubMed1993

Effects of parental adjustment on the adaptation of children with congenital or acquired limb deficiencies.

Maternal and paternal depression, anxiety, and marital discord were investigated as predictors of depression, anxiety, and self-esteem in 54 children with congenital or acquired limb deficiencies. Higher paternal depression predicted higher child depression and higher anxiety. Higher paternal anxiety predicted higher child depression and anxiety and lower self-esteem. Higher marital discord predicted higher child depression and anxiety and lower self-esteem. Maternal depression and anxiety did not predict child psychological adaptation. Family support had a positive effect on child adaptation, as did parent, classmate, teacher, and friend social support. The findings are discussed in terms of the risk and protective effects of parental distress, marital discord, and social support on the psychological adaptation of children with visible chronic physical disorders. J Dev Behav Pediatr 14:13-20, 1993. Index terms: limb deficiencies, children, adjustment, social support, parental distress, family functioning, marital discord.

Adaptation, Psychological↗
PubMed1992

Evidence for possible structural changes of particular H3 and H2B histone genes in different chicken tissues (cells).

The two chicken histone gene families H3 and H2B contain nine and eight members, respectively. To clarify whether the structures of these genes differ in different tissues (and cells), we analyzed DNAs from chicken lung, kidney, oviduct, and sperm. An H3-specific probe (probe 1.3SS) hybridized with a 10 kb EcoRI fragment carrying two H3 genes (H3-II and H3-III) from the lung, kidney, and oviduct with intensities of about one quarter of that of the fragment from the sperm. On the other hand, the intensities of hybridization of the H2B-specific probes (probes H2B-Ia, H2B-IIb, and H2B-III) with a 12 kb EcoRI fragment carrying two H2B genes (H2B-IV and H2B-V) from the oviduct, lung, and sperm were about a quarter of the intensity of hybridization with this fragment from the kidney. These findings, together with those reported previously, suggest that these particular histone genes H3 and H2B possess inherent abilities to form either a tight or loose structure, and that they exist in a loose form in the sperm and kidney but in a tight form in the other tissues tested.

Animals↗
PubMed1992

Structural change of a particular H2B histone gene possibly results in differences in its transcriptional regulation in different chicken tissues.

The chicken H2B histone gene family consists of eight highly homologous members (H2B-I to H2B-VIII) belonging to two major histone gene clusters. Seven of these genes have been sequenced and shown to encode three different H2B protein variants. Northern analysis with a probe, which mainly consists of the 5'-flanking region containing the sequence for the mRNA leader of H2B-V encoding a particular H2B protein variant, revealed that the mRNA level transcribed from this particular gene was higher in the kidney than in the oviduct and lung. To elucidate whether the structure of the H2B gene differs in the three different tissues, we analyzed DNAs from the oviduct, lung, and kidney. On Southern analysis, various H2B gene-specific probes hybridized with two particular H2B genes (H2B-IV and H2B-V), which are located in close proximity within a 12 kb EcoRI fragment, from the oviduct and lung with an intensity of about one quarter of that from the kidney. These findings suggest that some difference of DNA structure of the H2B-V gene may result in its relatively higher expression in the kidney.

Animals↗
PubMed1992

Screening for behavioral and emotional problems in children and adolescents with congenital or acquired limb deficiencies.

Children with chronic physical handicaps have been found to be at risk for psychological and social adjustment problems. Accurately identifying in a timely manner those physically handicapped children who are functioning at clinically significant levels of maladjustment may aid in preventing further psychosocial morbidity. The parents of 111 children and adolescents with congenital/acquired limb deficiencies completed the Child Behavior Checklist as a screening instrument to facilitate the identification of behavioral and emotional problems and social incompetence. Based on normalized T scores, the children manifested significantly greater behavioral and emotional problems and lower social competence than the normative community sample. Twenty-three percent of the children were reported to function in the clinically significant maladjustment range for behavioral and emotional problems; 14% were reported in the social maladjustment range. Correlations between parent report and child, adolescent, and teacher reports of adjustment ranged from r values of .23 to .41. The findings are discussed in terms of the "new hidden morbidity" in pediatric practice.

Adolescent↗
PubMed1992

Psychological adjustment and perceived social support in children with congenital/acquired limb deficiencies.

The negative impact on psychological adjustment from the chronic strain of living with limb deficiencies appears to be mediated by perceived social support. A multifactorial investigation was conducted to identify empirically psychological adjustment correlates of perceived social support in 49 children with congenital/acquired limb deficiencies. A multiplicity of adjustment factors (depression, trait anxiety, self-esteem) was variously related to perceived parent, teacher, classmate, and friend social support. Hierarchical multiple regression analyses provide initial evidence of the potentially powerful effects of the social environment of the school setting, with perceived classmate social support the only significant predictor variable across depressive symptomatology, trait anxiety, and general self-esteem. The results are discussed as the first step in identifying the potential correlates of multiple perceived social support domains, which may provide empirical guidance for future intervention studies designed to modify adjustment factors in chronically ill and handicapped children.

Adaptation, Psychological↗
PubMed1992

Overlap and discrepancy between tests for anti-C100, anti-GOR and anti-CP9 in patients with chronic liver disease and inhabitants in Saga, Japan.

The authors evaluated the clinical significance of anti-C100, anti-GOR and anti-CP9 in hepatitis C virus (HCV)-related liver disease in two populations: 459 healthy subjects and 385 patients with chronic liver disease (CLD). Previously we reported high rates of mortality and morbidity (5.3%) of CLD in subjects in Saga, Japan. This was ascribed to the high prevalence (10.8%) of anti-HCV among randomized populations, as detected by the C100 ELISA test system, as compared with a finding of 2-3% in Japanese blood donors in the same decade. The incidence of anti-C100, anti-GOR and anti-CP9 detected by ELISA test system in the healthy population currently surveyed was 17.0%, 19.2% and 32.0% respectively, as compared with 75.3%, 60.3% and 73.0% respectively, in those with CLD. The incidence of positivity for at least one of the three antibodies was high (36.4%) among healthy subjects, and even higher (86.5%) among the patients with CLD. In the healthy subjects, incidence of positivity increased with age. The healthy and CLD populations differed in the proportion of cases positive for all three antibodies vs. those positive for at least one antibody: healthy subjects, 52/167, 31.1%, vs. CLD patients, 197/333, 59.2%; P less than 0.01. Among the anti-C100-positive healthy cases, these was a significantly high level of AST, ALT, ZTT and gamma GTP compared with negative cases, with or without anti-GOR and anti-CP9 (P less than 0.01-0.05). These observations suggest that the presence of anti-C100 may be related to the active state of HCV-related liver disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1992

Adenovirus-mediated in vivo gene transfer and expression in normal rat liver.

Replication deficient, recombinant adenovirus (Ad) vectors do not require target cell replication for transfer and expression of exogenous genes and thus may be useful for in vivo gene therapy in hepatocytes. In vitro, primary cultures of rat hepatocytes infected with a recombinant Ad containing a human alpha 1-antitrypsin cDNA (Ad-alpha 1AT) synthesized and secreted human alpha 1AT for 4 weeks. In rats, in vivo intraportal administration of a recombinant Ad containing the E. coli lacZ gene, was followed by expression of beta-galactosidase in hepatocytes 3 days after infection. Intraportal infusion of Ad-alpha 1AT produced detectable serum levels of human alpha 1AT for 4 weeks. Thus, targeted gene expression has been achieved in the liver, albeit at low levels, suggesting that adenovirus vectors may be a useful means for in vivo gene therapy in liver disorders.

Adenoviruses, Human↗