Biomedical subjects

Y Setoguchi

Publications and source records attributed to Y Setoguchi.

At least 37 records · Page 2Linked to original sources

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↗

PubMed1996

[Transfer of endothelial nitric oxide synthase gene in the purpose of gene therapy for pulmonary arterial hypertension].

Nitric oxide (NO) is a messenger molecule involved in diverse processes in many tissues. For example, NO is responsible for the bactericidal activities of macrophages, and in blood vessels it accounts for endothelium-derived relaxing factor activity. Recently, inhalation of NO gas was reported to improve the acute pulmonary arterial hypertension. Based on this knowledge, recombinant expression of endothelial nitric oxide synthase (eNOS) in lung may have profound effects on pulmonary vasomotor function and pulmonary arterial smooth muscle proliferation and platelet adhesion. On the basis of this concept, we evaluate the feasibility of gene therapy for chronic pulmonary arterial hypertension using hypoxia regulatable adenoviral vector coding eNOS cDNA.

Animals↗

PubMed1996

Lung cancer-reacting human recombinant antibody AE6F4: potential usefulness in the sputum cytodiagnosis.

Human monoclonal antibody (hMAb) AE6F4 has been shown to be potentially useful for immunocytological detection of lung cancer cells in sputum. By recombinant DNA technology, IgM type hMAb AE6F4 was switched to lgG. The IgG mimic recombinant AE6F4 antibody expression plasmid was assembled using the antibody heavy chain gene, which ligated the gene encoding VH and CH1(mu) domains of hMAb AE6F4 heavy chain to the gene encoding CH2(gamma 1) and CH3(gamma 1) domains of human IgG heavy chain, and the antibody light chain gene of hMAb AE6F4. The recombinant antibody expressed by baby hamster kidney (BHK)-21 cells showed molecular size equivalence to IgG, and consisted of human mu-gamma hybrid heavy and kappa light chains. The immunological specificity of the recombinant antibody was the same as that of hMAb AE6F4 by immunoblotting analysis to the 14-3-3 protein, the putative antigen of hMAb AE6F4, and by immunohistochemical and immunocytological analyses using tissue sections and sputa of lung cancer patients. The transfected BHK-21 cells produced the recombinant antibody persistently and the productivity was greater than 20 times that by human-human hybridoma producing hMAb AE6F4.

14-3-3 Proteins↗

PubMed1995

Molecular cloning of the 31 kDa cytosolic phospholipase A2, as an antigen recognized by the lung cancer-specific human monoclonal antibody, AE6F4.

The human monoclonal antibody AE6F4 specifically reacts with human lung cancer tissues but does not with normal tissues. This monoclonal antibody recognizes a cytosolic 31 kDa antigen in the cancer cells. In a previous study, we elucidated that the 31 kDa antigen belonged to a family of proteins collectively designated as 14-3-3 proteins, which were known as protein kinase-dependent activators of tyrosine/trytophan hydroxylases, or protein kinase C inhibitor proteins. Here we report molecular cloning of the 31 kDa antigen from the human lung adenocarcinoma cell line, A549. Sequencing analysis indicates that the cloned cDNA is identical to that of previously reported human placental cytosolic phospholipase A2 (cPLA2), which is also a member of the 14-3-3 protein family. Western analysis demonstrated that a 31 kDa recombinant cPLA2 expressed in monkey COS cells was recognized by the AE6F4 monoclonal antibody. Binding of the monoclonal antibody to the recombinant cPLA2 was abolished when treated with sodium periodate, suggesting that not only are carbohydrate chains associated with the cPLA2, but they also play a crucial role in antigen recognition by the monoclonal antibody.

14-3-3 Proteins↗

PubMed1995

Serum lipoprotein(a) levels before and after subtotal thyroidectomy in subjects with hyperthyroidism.

Lipoprotein(a) [Lp(a)], a lipoprotein that structurally resembles low-density lipoprotein (LDL), contains apolipoprotein(a) [apo(a)] and apolipoprotein B-100 (apo B). There exists a close inverse correlation between serum concentrations of LDL or apo B and concentrations of thyroid hormone in patients with thyroid disease, probably due to a change in LDL receptor activity. To clarify the relations between thyroid hormone and Lp(a), we measured serum Lp(a) levels in 13 hyperthyroid subjects before treatment (stage H), during the euthyroid stage induced immediately before performing a subtotal thyroidectomy (stage E), and during the hypothyroid stage observed transiently after the operation (stage L). The mean serum concentration of Lp(a) increased significantly (P = .01) from 9.4 mg/dL in stage H to 26.8 in stage L through the level of 15.5 mg/dL in stage E. There was no significant difference between the mean serum concentration of Lp(a) in these patients in stage E and healthy controls (14.2 mg/dL). There was a low but statistically significant negative correlation between the Lp(a) level and the serum free thyroxine (fT4) concentration (r = .31, P < .05). The results suggest that thyroid hormone is a potent modulator of Lp(a) metabolism.

Adult↗

PubMed1995

Pharmacological effects of dai-saiko-to on lipid biosynthesis in cultured human hepatocyte HepG2 cells.

The pharmacological effects of Dai-saiko-to, a Japanese and Chinese traditional medicinal mixture (Kampohozai), on lipid biosynthesis were investigated in cultured human hepatocyte HepG2 cells. The addition of Dai-saiko-to (0.5 mg/ml), which had no significant effect on cell proliferation, caused a marked decrease in the intracellular triglyceride content with no significant changes in the other lipid fraction. At the same time, the incorporation of 14C-acetate or 3H-glycerol into the triglyceride or diglyceride fractions also decreased significantly. These results suggest that Dai-saiko-to decreases hepatic triglyceride biosynthesis, which might contribute to a reduction in plasma VLDL levels.

Cell Division↗

PubMed1995

Localized myxedema, associated with increased serum hyaluronic acid, and response to steroid pulse therapy.

A 66-year-old man presented with Graves' disease and widespread localized myxedema. Extensive lesions were present on the legs, feet, hands, and face. TSH receptor antibody (TBII) was markedly positive and the serum hyaluronic acid level was very high. Intravenous steroid pulse therapy was followed by oral therapy with gradual tapering. This regimen markedly improved the skin lesions and resulted in a decrease of the serum hyaluronic acid level. The findings suggest that steroid pulse therapy is effective for the treatment of extensive localized myxedema. In addition, the serum hyaluronic acid level may be a useful parameter for the follow-up of patients with this condition.

Administration, Oral↗

PubMed1995

Is body powered operation of upper limb prostheses feasible for young limb deficient children?

The investigators measured efficiencies of body powered prehensors and cable control components of prostheses available for young children. Results indicated that the cable control systems and hook type prehensors have moderate to high efficiencies, but children's body powered hands have very low efficiencies. Measures of arm and shoulder strength of 3-5 year-old limb deficient children, both on the limb deficient and sound sides, were less than that reported in the sound sides, were less than that reported in the literature for normal children. The findings were examined in relation to children's strength available compared with prosthesis strength requirements. The comparison demonstrates a way to establish measurable efficiency targets for new prehensor designs. The article includes detailed findings on children's strength, and findings on efficiencies of the prehensors and cable control systems of children's upper limb prosthses. Sample calculations may be useful to future designers of body powered prehensors for young children. A more efficient body powered hand is especially needed. Preliminary calculations indicate that the use of currently available children's voluntary opening (VO), body powered hands is not feasible, given the low strength of young limb deficient children and the low efficiencies of the VO hands. The use of voluntary closing (VC) hands may be feasible but remains to be tested.

Age Factors↗

PubMed1995

Effects of fluvastatin, a new inhibitor of HMG-CoA reductase, and niceritrol on serum lipids, lipoproteins and cholesterol ester transfer activity in primary hypercholesterolemic patients.

Effects of a combination therapy of fluvastatin, a new inhibitor of HMG-CoA reductase, and niceritrol on lipid metabolism were investigated measuring a wide range of parameters in 42 patients with primary hypercholesterolemia. After a wash-out period patients were randomly allocated to 1 of the 2 groups, the fluvastatin-preceding group (G-1) and the niceritrol-preceding group (G-2). In G-1 fluvastatin monotherapy (30 mg/day) significantly decreased total cholesterol (TC) and LDL-cholesterol (LDL-C). There was no significant change in HDL-cholesterol (HDL-C), triglyceride (TG) and lipoprotein (a) (Lp(a)). Further effect in HDL-C and TG was observed after the addition of niceritrol (750 mg/day). On the other hand, in G-2, while niceritrol alone (750 mg/day) produced no significant change in TC, LDL-C, HDL-C, TG and Lp(a), the addition of fluvastatin (30 mg/day) reduced TC and LDL-C levels significantly. Cholesterol ester transfer (CET) activity was significantly reduced by niceritrol monotherapy. After the concomitant use of the 2 drugs CET activity was significantly reduced only in G-2. No significant change in lipoprotein lipase and hepatic triglyceride lipase activities were observed in the 2 groups at either point in time. No serious adverse effect was observed in this study. It is concluded that fluvastatin is an effective drug for lowering LDL-cholesterol and causes no adverse alteration in lipid metabolism. Combination with niceritrol at a dose of 750 mg/day dose not appear to augment or attenuate beneficial effects of fluvastatin.

Anticholesteremic Agents↗

PubMed1995

Synchronous circadian rhythms of mRNA levels and activities of cholesterol 7 alpha-hydroxylase in the rabbit and rat.

Cholesterol 7 alpha-hydroxylase, the key enzyme in a series of metabolic changes for the production of bile acids from cholesterol, shows circadian rhythms. The mechanism of the circadian rhythms is the subject of controversy; moreover, few pieces of information are presently available. Because the rabbit is a good animal model for the study of human cholesterol-bile acid metabolism, in the present study we isolated a complete cDNA encoding rabbit cholesterol 7 alpha-hydroxylase (3022 base pairs (bp) long and 1503 bp open-reading frame encoding 501 amino acid residues). Using this isolate rabbit cDNA and the rat cDNA as probes, the circadian rhythms of this enzyme were studied in normal male rabbits and rats. The animals were maintained on a 12-h light-dark cycle. Three subjects each were then killed every 3 h for 1 day. The livers were harvested, and the mRNA levels and the activities of the enzyme were measured. Both mRNA levels and activities of the enzyme showed remarkable circadian rhythms, with higher values during the dark phase and lower values reaching minimum during the light phase. The enzyme activities correlated well with the mRNA levels. There was no shift in the cycles of the enzyme activities and the mRNA levels. These findings suggest that pretranslational regulation is the most likely mechanism for the circadian rhythms of cholesterol 7 alpha-hydroxylase. Because the rhythms of mRNA levels and activities of this enzyme are synchronous, the turnover of the enzyme should be as fast as that of the mRNA, and we can hereafter measure the mRNA levels instead of the enzyme.

Amino Acid Sequence↗

PubMed1995

Immunocytochemical detection of lung cancer cells with monoclonal antibodies to 14-3-3 proteins.

Murine monoclonal antibodies were raised against 14-3-3 proteins, the antigen of human monoclonal antibody AE6F4 which had been shown potentially useful for the immunochemical diagnosis of lung cancer via sputum cytology. Enzyme-linked immunosorbent assays of the murine anti-14-3-3 monoclonal antibodies with isolated bovine brain 14-3-3 isoforms showed that the antibodies were classified into four different profiles of isoform reactivity. The comparison of 14-3-3 isoform and lung cancer tissue on the reactivity with murine monoclonal antibodies indicated that beta isoform can be responsible for cancer recognition, whereas human monoclonal antibody AE6F4 showed preferential binding to zeta isoform. No murine monoclonal antibody of the same isoform specificity as human monoclonal antibody AE6F4 was obtained. Since murine monoclonal antibodies with different isoform specificities could immunostain lung cancer cells in sputum successfully, the combination use of murine monoclonal anti-14-3-3 antibodies with human monoclonal antibody AE6F4 is potentially useful for facilitating the sputum cytodiagnosis of lung cancer.

14-3-3 Proteins↗

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↗