Search PubMed⌕ Search

Biomedical subjects

E Ono

Publications and source records attributed to E Ono.

At least 19 recordsLinked to original sources

The first immunoglobulin-like domain of porcine nectin-1 is sufficient to confer resistance to pseudorabies virus infection in transgenic mice.

Nectin-1 is an alphaherpesvirus receptor that binds to virion glycoprotein D (gD). Porcine nectin-1 mediates entry of pseudorabies virus (PRV), herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), and bovine herpesvirus type 1 (BHV-1). The gD-binding domain of nectin-1 is the first or N-terminal immunoglobulin (Ig)-like domain of the entire ectodomain. Here, we generated three transgenic mouse lines expressing a fusion protein consisting of the first Ig-like domain of porcine nectin-1 and the Fc portion of porcine IgG1 to assess the antiviral potential of the first Ig-like domain of nectin-1 in vivo. All of the transgenic mouse lines showed significant resistance to PRV infection via intraperitoneal inoculation (survival rates of 67% to 100%). In the intranasal challenge, a lower but still significant protection was observed; 21% to 55% of the animals from the three transgenic mouse lines survived. The present results demonstrate that a soluble form of the first domain of porcine nectin-1 is able to exert a significant antiviral effect against pseudorabies virus infection.

Animals↗

Cerebellar hypoplasia associated with an avian leukosis virus inducing fowl glioma.

Fowl glioma-inducing virus (FGV), which belongs to subgroup A of avian leukosis virus (ALV), shows tumorigenicity and pathogenicity, mainly in the nervous system, and causes astrocytoma and perineurioma. Apart from these neoplasms, cerebellar anomaly was found in chickens infected with FGV in ovo. The study reported here describes the morphologic characteristics of the affected cerebellum. Specific-pathogen-free chickens (C/O) were inoculated with FGV through the yolk sac on the 7th day of incubation. The cerebellar anomaly included diffuse depletion of granular cells of the internal granular layer (IGL), remnants of the external granular layer (EGL), and disorganization of the Purkinje cell layer. These cerebellar changes were observed in all birds except one. In the infected embryos, the EGL was thicker and had an irregular arrangement with a thin molecular layer (ML) and IGL, compared with the control. The granular cells were immunohistochemically positive for ALV common antigen. Immunohistochemical analysis for vimentin revealed disarrangement and decreased number of Bergmann's fibers. Use of the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method and electron microscopy indicated that apoptotic granular cells were frequently observed in the EGL and ML. These results suggested that the cerebellar anomaly was hypoplasia, principally resulting from the apoptosis of granular cells in the EGL and ML caused by FGV infection and that the cell loss induced obstruction of granular cell migration and disarrangement of Bergmann's fibers in the ML.

Animals↗

Immunogenicity and tolerability of hepatitis A vaccine in HIV-infected children.

The immunogenicity and tolerability of hepatitis A virus vaccine was evaluated in a group of 32 children with human immunodeficiency virus (HIV) infection and 27 children with seroreversion. After 2 doses of vaccine, 100% of children experienced seroconversion with good toleration of the vaccine. There were no differences in variation of virus load between immunized HIV-positive children and a group of 31 nonimmunized HIV-positive children with similar characteristics.

Antiretroviral Therapy, Highly Active↗

Nested polymerase chain reaction for detection of the avian leukosis virus causing so-called fowl glioma.

The complete nucleotide sequence of the avian leukosis virus causing so-called fowl glioma has been previously determined. Primers were designed for detection of the fowl glioma-causal virus (FGV) based on the 3' untranslated region of the viral genome. The provirus and viral RNA of FGV were specifically detected in various organs and tissues, including feather pulp, from experimentally infected birds using nested polymerase chain reaction (PCR) and reverse transcription nested PCR. The prevalence of FGV was evaluated in 131 Japanese fowls of a zoological garden in Japan based on the detection of the FGV genome in feather pulp using PCR and the detection of viral antigen in faeces by enzyme-linked immunosorbent assay. FGV proviral DNA was detected in feather pulp of 52 birds (39.7%) by nested PCR. Later, nine dead birds from among the 52 were histologically diagnosed as having fowl glioma and found to have the proviral DNA in the affected brain. These results demonstrated that the PCR-based detection of FGV in feather pulp is useful for epidemiological studies on fowl glioma.

Animals↗

Varicella zoster antibodies in healthcare workers from two neonatal units in São Paulo, Brazil--assessment of a staff varicella policy.

Although frequently reported in the literature, a staff varicella policy is not standard in many hospitals even in developed countries. In the present study, we assessed varicella zoster immunity in staff from two neonatal units from hospitals in São Paulo, Brazil. Ninety-seven percent of all staff working in both units agreed to participate. A simple and cost-effective varicella policy was subsequently set up, based on costs and data from serology and a history of previous varicella infection. Our results confirm that a varicella vaccination programme can be implemented in a healthcare facility, even in developing countries.

Adult↗

Genome sequence analysis of the avian retrovirus causing so-called fowl glioma and the promoter activity of the long terminal repeat.

So-called fowl glioma is a retroviral infectious disease caused by avian leukosis virus subgroup A (ALV-A). We determined the complete nucleotide sequence of the virus genome. The full-length sequence was consistent with a genetic organization typical of a replication-competent type C retrovirus lacking viral oncogenes. The coding sequences were well conserved with those of replication-competent viruses, but the 3' noncoding regions including LTR were most related to those of replication-defective sarcoma viruses. The U3 region of the LTR had a few deletions and several point mutations compared to that of other ALVs. The promoter activities of the LTRs of glioma-inducing ALV and ALV-A standard strain, RAV-1, were equivalent in chick embryo fibroblasts (CEF), while that of glioma-inducing ALV was significantly lower than that of RAV-1 in human astrocytic cells. These subtle differences of the promoter activity of the LTR may be related to the induction of glial neoplasm.

Alpharetrovirus↗

The pseudorabies virus immediate-early promoter directs neuronal tissue-specific expression in transgenic mice.

Pseudorabies virus (PRV) immediate-early (IE) gene product is required for expression of the viral early and late genes as a transactivator. The IE gene is expressed as the first gene among the viral genes after the infection. To examine the activity of the IE promoter in vivo, we have generated transgenic mice expressing transgenes under the control of the IE promoter. To analyze the tissue specificity of the transgene expression, mRNA of the transgene was monitored in various tissues from the transgenic mice by reverse transcriptase (RT)-polymerase chain reaction (PCR) analysis. A strong transgene expression was observed in the neuronal tissues by the RT-PCR analysis. These neuronal tissues included cerebrum, cerebellum and trigeminal nerve. Although the PCR product was hardly detected in other tissues by the RT-PCR analysis, specific PCR bands were detected in multiple organs (skin, skeletal muscles, heart muscles, lung, liver, spleen, small intestine and kidney) by Southern blot analysis using the RT-PCR products. These results indicate that although the IE promoter acts as a pan-specific promoter in vivo, it is capable of driving a high level of transgene expression in neuronal tissues.

Animals↗

A pan-specific promoter activity of the 213bp segment of the pseudorabies virus early protein 0 gene in transgenic mice.

Pseudorabies virus (PrV) early protein 0 (EP0) is a transactivator that plays important roles in the viral gene expression. To examine a promoter regulatory element for the EP0 gene expression in vivo, we have generated two transgenic mouse lines expressing the EP0 gene under the control of a 213bp 5'-flanking sequence of the EP0 gene. To analyze the tissue specificity of transgene expression, mRNA of the EP0 gene was monitored in various tissues from the transgenic mice by the reverse transcriptase (RT)-polymerase chain reaction (PCR) analysis. The EP0 gene expression was demonstrated in all tissues tested by the RT-PCR analysis. These tissues included skin, muscles (skeletal and heart), lung, liver, spleen, small intestine, kidney, brain and testis. These results indicated that the 213bp sequence of the 5'-flanking region of the EP0 gene is capable of driving expression of the EP0 gene in vivo and the promoter is pan-specific.

Animals↗

Inhibition of pseudorabies virus replication by a dominant-negative mutant of early protein 0 expressed in a tetracycline-regulated system.

Pseudorabies virus (PRV) early protein 0 (EP0) consisting of 410 amino acids is a transactivator of viral genes. A mutant consisting of amino acids 1-113 exhibits dominant-negative properties. In order to assess the antiviral potential of the EP0 mutant, Vero cells were transformed with the EP0 mutant gene expressed in a tetracycline-regulated system. The transformed cell lines showed marked resistance to PRV infection when expression of the EP0 mutant gene was induced. In the transformed cell line infected with PRV, synthesis of the immediate-early protein (IE180) and of EP0 was inhibited, whereas the levels of IE and EP0 messenger RNA (mRNA) were not decreased, as compared with those of the control cell line. The present results suggest that the EP0 mutant may not alter the efficiency of the viral gene transcription but rather translation efficiency of the viral mRNA.

Animals↗

Essential role of the small GTPase Rac in disease resistance of rice.

Production of reactive oxygen intermediates (ROI) and a form of programmed cell death called hypersensitive response (HR) are often associated with disease resistance of plants. We have previously shown that the Rac homolog of rice, OsRac1, is a regulator of ROI production and cell death in rice. Here we show that the constitutively active OsRac1 (i) causes HR-like responses and greatly reduces disease lesions against a virulent race of the rice blast fungus; (ii) causes resistance against a virulent race of bacterial blight; and (iii) causes enhanced production of a phytoalexin and alters expression of defense-related genes. The dominant-negative OsRac1 suppresses elicitor-induced ROI production in transgenic cell cultures, and in plants suppresses the HR induced by the avirulent race of the fungus. Taken together, our findings strongly suggest that OsRac1 has a general role in disease resistance of rice.

Agrobacterium tumefaciens↗

Translational regulation of platelet-derived growth factor B chain (c-sis) mRNA by short open reading frames in the 5'-untranslated region.

Platelet-derived growth factor B chain (PDGF-B/c-SIS), the product of c-sis proto-oncogene, is a potent mitogen and chemoattractant for cells of mesenchymal origin. Expression of PDGF-B/c-SIS is regulated at the translational level, in addition to at the transcriptional level. The 5'-untranslated region (5'-UTR) of PDGF-B/c-sis mRNA is known to inhibit translation of the downstream coding sequences. The 5'-UTR contains putative influential elements, such as GC-rich elements, stem-loop structures and short open reading frames (SORFs). To clarify the inhibition mechanism of PDGF-B/c-sis mRNA translation, effects of three SORFs in the 5'-UTR on the translational regulation were investigated in transient expression assays. Introducing point mutation(s) in the initiation codons of SORFs affected the reporter gene expression in several cell lines (COS-1, U-2, JEG-3). Abrogation of three SORFs resulted in an increase of the reporter gene expression both in beta-galactosidase assay and Western blot analysis. These results suggest that SORFs in the 5'-UTR sequences have inhibitory effects on the translation of the downstream coding sequences.

5' Untranslated Regions↗

Resistance to pseudorabies virus infection in transgenic mice expressing the chimeric transgene that represses the immediate-early gene transcription.

A chimeric gene encoding a fusion protein consisting of the DNA-binding domain of the immediate-early (IE) protein of pseudorabies virus (PRV) and a tail-truncated VP16 of herpes simplex virus 1, lacking the transcription activation domain, has been shown to repress transcription of the PRV IE gene, resulting in the inhibition of PRV growth in vitro. To assess the antiviral potential of the fusion protein in vivo, transgenic mice containing the chimeric gene under the control of the virus- and interferon-inducible Mx 1 promoter were generated. A transgenic mouse line showed marked resistance to PRV infection when the mice were challenged intranasally with PRV. Inhibition of PRV replication was also observed in monolayers of embryonic cells prepared from the transgenic mice. In the cells infected with PRV, transcription of the PRV IE gene was repressed. The present results indicate that the chimeric gene is able to exert a significant antiviral effect against PRV infection in vivo.

Animals↗

The small GTP-binding protein rac is a regulator of cell death in plants.

Cell death plays important roles in the development and defense of plants as in other multicellular organisms. Rapid production of reactive oxygen species often is associated with plant defense against pathogens, but their molecular mechanisms are not known. We introduced the constitutively active and the dominant negative forms of the small GTP-binding protein OsRac1, a rice homolog of human Rac, into the wild type and a lesion mimic mutant of rice and analyzed H(2)O(2) production and cell death in transformed cell cultures and plants. The results indicate that Rac is a regulator of reactive oxygen species production as well as cell death in rice.

Amino Acid Sequence↗

Cepharanthin enhances thermosensitivity without a resultant reduction in the thermotolerance of a murine mammary carcinoma.

Cepharanthin (Ce) is a biscoclaurine alkaloid extracted from Stephania cepharantha Hayata. The results of our previous in vitro study indicated that Ce reduces thermotolerance by enhancing thermosensitivity. In the present study, we investigated the in vitro and in vivo effects of Ce on thermosensitivity and thermotolerance using a murine mammary carcinoma, MCa, and C3H/HeN mice. Ce enhanced the thermosensitivity of MCa cells for heating at 44 degrees C not only in vitro but also in vivo. The in vivo enhancement ratio +/- SD of Ce at 100 mg/kg for heating at 44 degrees C was 1.3+/-0.3. The fractionated heat treatments at 44 degrees C for 30 and 60 min with an interval time of 0-6 days resulted in the development of remarkable thermotolerance and the expression of heat shock protein 70 in MCa tumors after the first heating. Ce at 100 mg/kg given immediately after the first heating increased the expression of heat shock protein 70 in MCa tumors, and did not reduce the development of thermotolerance. Ce given immediately before the first or second heating also did not inhibit the thermotolerance. The results of this study suggest that Ce enhances the thermosensitivity of MCa tumors as a thermosensitizer, but that this mild thermosensitizing property of Ce might be insufficient to conquer the remarkable thermotolerance in MCa tumors that develops after the first heating.

Alkaloids↗

A patient with a large hepatocellular carcinoma alive 13 years after repeated transcatheter arterial chemoembolization and hepatectomy.

We describe a rare case of a large hepatocellular carcinoma (HCC) in a patient who survived 13 years after repeated transcatheter arterial chemoembolization (TAE) and hepatectomy. The patient was a 41-year-old woman found in March 1985 to have a large HCC, measuring 14 x 12 cm in diameter and associated with obstruction of the main portal trunk. She underwent TAE 8 times over a period of 1 year. The serum alpha-fetoprotein level decreased from 18342 to 1871 ng/ml. The patient subsequently underwent left hepatectomy on October 9, 1986. Thirteen years after hepatectomy, the patient is being followed up on an outpatient basis and has had no evidence of recurrence.

Adult↗

A case of biliary cystadenocarcinoma of the liver.

We describe a case of biliary cystadenocarcinoma of the liver, a very rare malignant tumor, in a 40-year-old man referred to our hospital because of sporadic abdominal pain. A left lobectomy of the liver was performed, and macroscopic examination of the resected specimen revealed prominent papillary projections in a multiloculated cystic tumor containing mucinous material. Histologic examination of the specimen showed a biliary cystadenocarcinoma without mesenchymal stroma, associated with a cystadenoma. Cystadenoma with mesenchymal stroma (CMS), which occurs exclusively in women, is generally regarded as a precancerous lesion, and patients with biliary cystadenocarcinoma with mesenchymal stroma have a good prognosis. Cystadenoma without mesenchymal stroma arises in both men and women. However, the origin and precancerous lesions of cystadenocarcinoma are unknown. Moreover, the prognosis of patients with biliary cystadenocarcinoma without mesenchymal stroma is poor, especially in men. The cystadenoma without mesenchymal stroma in our patient was considered a precancerous lesion. The present patient has shown no evidence of recurrence in the 8 years after hepatic resection. Previously reported cases of cystadenocarcinoma without mesenchymal stroma, especially those in men, have had a poor outcome. A good outcome in men is very rare.

Adult↗

[Estimation of antibacterial activity of various antibiotics against Pseudomonas aeruginosa by score method].

Antibacterial activity of various antibiotics against Pseudomonas aeruginosa isolated from each hospitals depends on the variety or amount of antibiotics used in each hospital. The antibiotic, which is effective to P. aeruginosa in a certain hospital is not always effective to that in other hospital. The excellent antibiotics in antibacterial activity have low MIC and hard to progress in resistance, and such antibiotics may be effective against P. aeruginosa isolated from any hospitals. Therefore we thought that the antibiotic, which was progress to resistance, would show a great difference in MIC among hospitals, and we investigated MIC and difference of MIC of various antibiotics against P. aeruginosa isolated from six hospitals. Furthermore, we converted the data of MICs and difference of MIC among six hospitals into the score, and tried to estimate antibacterial activity of various antibiotics by using those scores. From the results of analysis in this report, we think the antibiotics actually surpass in antibacterial activity may be imipenem, cefozopran, cefsulodin and amikacin. New analytical method proposed in this report will become one of potential methods to estimate antibacterial activity of antibiotics against bacteria isolated from inpatient with bacterial infections.

Anti-Bacterial Agents↗

Promoter activity of sequence located upstream of the pseudorabies virus early protein 0 gene.

Promoter activity of the 5'-flanking region of the pseudorabies virus (PRV) early protein 0 (EP0) gene was analysed by transient transfection assays employing chloramphenicol acetyl transferase (CAT) reporter constructs. We identified a 213 bp segment of the viral genome that was capable of efficiently driving expression of the EPO gene and a linked reporter gene upon transient transfection into Vero cells. This segment lacked the typical TATA element, and possessed an initiator element and the putative binding sites for the transcription factor Sp1 and immediate-early protein IE180, a strong transactivator of PRV. By analysing 5'-deletion mutants of the segment, a 48 bp segment (from nucleotide positions -65 to -17), which possessed three Sp1 binding sites, was identified to be critical for the promoter activity. Cotransfection of Vero cells with the mutant constructs and an IE180 expression plasmid resulted in transactivation of only those constructs in which the Sp1 sites were present. These results indicate that the EP0 gene may be transcribed from the TATA-less promoter that responds to Sp1.

Animals↗