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Biomedical subjects

K Shinozaki

Publications and source records attributed to K Shinozaki.

At least 19 recordsLinked to original sources

Detection and sequence-based typing of human adenoviruses using sensitive universal primer sets for the hexon gene.

The universal primer sets for identification of human adenovirus (HAdV) targeting hexon gene were designed and applied to 121 clinical samples suspected of HAdV infection. The primer sets amplified at least 20 HAdV reference strains of six species. Of these clinical samples, 81 (66.9%) samples were positive for HAdV. They were classified into 11 serotypes belonging to 5 HAdV species (B-F). The primer sets described here are sensitive and reactive to the broad spectrum of HAdV and are useful for rapid diagnosis of various HAdV infections.

Adenovirus Infections, Human↗

The detection of human sapoviruses with universal and genogroup-specific primers.

Sapovirus (SV) causes gastroenteritis in humans and comprises genetically divergent viruses. A nested reverse transcription-polymerase chain reaction (RT-PCR) targeting the capsid-protein-coding region was developed using universal and genogroup-specific primer sets. The universal primers were capable of detecting human SV genogroups I, II, IV and V. Genetic analysis of the amplified products enabled us to phylogenetically determine the genotypes of the viruses. In addition, genogroup-specific primers that amplified different lengths of the amplicon depending on the genogroup were developed. These genogroup-specific primers were also used as inner primers for the nested PCR. These two simple RT-PCR methods are powerful tools for both detection and epidemiological studies of human SV.

Caliciviridae Infections↗

In-plane and out-of-plane ferroelectric instabilities in epitaxial SrTiO3 films.

The in-plane and out-of-plane ferroelectric instabilities in compressed (100)-epitaxial SrTiO3 films were examined by infrared reflection spectroscopy. The strongly stiffened in-plane soft mode frequency softened very slowly on cooling. On the other hand, the silent mode appeared at around 150 K, indicating an out-of-plane ferroelectric transition. This behavior points to a split of in-plane and out-of-plane ferroelectric instability temperatures due to the lowered symmetry of the SrTiO3 lattice caused by mechanical misfit strain. Infrared spectroscopy provides a possibility to detect such an effect in the strained epitaxial ferroelectric films.

Journal Article↗

Genetic analysis of noroviruses associated with fatalities in healthcare facilities.

Norovirus outbreaks occurred in 236 healthcare facilities for the elderly in Japan during the winter of 2004-2005. Three norovirus strains associated with three fatal clinical courses were isolated from geographically separate facilities and genetically analyzed along with three strains from non-fatal cases in the same season. All six isolates were classified as the GII-4 genotype. No new variant strains like those observed in Europe in 2002 and 2004 were found in fatal cases, and the three outbreaks were deemed to have been caused by genetically close conventional norovirus GII-4 strains.

Aged, 80 and over↗

Efficient infection of tumor endothelial cells by a capsid-modified adenovirus.

Targeted antiangiogenic gene therapy is an attractive approach to treat metastatic cancer. However, the relative paucity of the receptors of the commonly used adenovirus serotype 5 in endothelial cells as compared with liver cells undermines the use of this vector for targeting the endothelial cells in tumors. To overcome this problem, we analyzed the ability of a hybrid Ad5/35 virus, where the serotype 5 fiber has been replaced with the fiber from serotype 35, to target tumor vasculature. Infection of human umbilical vein endothelial cells (HUVECs) with Ad5/35 at MOI 120 infected 100% of cells. In contrast, infection with Ad5 at the same MOI infected only 10% HUVECs. Ad5/35 was even more effective in transducing human aortic endothelial cells (HAECs), as infection with Ad5/35 at MOI 3.6 was sufficient to transduce 95% of cells. Gene expression analyses demonstrated that infection of HUVECs and HAECs with Ad5/35 resulted in between 1 and 3 orders of magnitude higher gene expression than infection with Ad5. Furthermore, various liver-derived cells were less infectable with Ad5/35 than Ad5, indicating a favorable toxicity profile for this virus. In a rat colon carcinoma tumor model, Ad5 was located mainly in the liver parenchyma after hepatic artery administration. In contrast, Ad5/35 was found only in the angiogenesis-rich border region of the tumor. Double immunostaining revealed that Ad5/35 colocalized with CD31 and Flk-1 positive endothelial cells. These results indicate that Ad5/35 may be useful in anticancer strategies targeting tumor endothelial cells.

Adenoviruses, Human↗

Cystadenoma of the liver with ovarian-like stroma: case report.

Cystadenomas are rare benign tumors with malignant potential. A 36-year-old woman presented with a 20-cm cystic mass in the left lobe of the liver. Surgery confirmed the diagnostic imaging findings. We present a case of cystadenoma of the liver with ovarian-like stroma and its associated radiologic characteristics.

Adult↗

Portal vein occlusion or stenosis in patients with hepatolithiasis: observation by multiphasic contrast-enhanced CT.

AIMS: To analyze the dynamic findings of multiphasic contrast-enhanced CT in hepatolithiasis and to elucidate occlusive changes in portal veins and other associated abnormalities. METHODS: This was a retrospective study of 25 selected patients with hepatolithiasis who underwent various imaging examinations, including multiphasic contrast-enhanced CT. The following CT findings were evaluated in each of 71 hepatic segments: visualization of a calculus; biliary dilation or focal hepatic atrophy of the affected segment; areas that were abnormally enhanced in the hepatic arterial phase; degrees (normal, stenosis, occlusion) of portal vein calibre; and linear delayed enhancement along the bile-duct walls, suggesting cholangitis. RESULTS: On CT, calculi were depicted as a hyperdense structures in 61 of 71 segments (86%). Focal hepatic atrophy, which frequently accompanied CT findings suggesting compensatory hypertrophy of other segments, was seen in 50 of 71 segments (70%). Areas that were abnormally enhanced were recognized in 36 of 71 segments (51%). Stenosis or occlusion of portal venous branches was observed in 59 of 71 segments (83%), including 13 segments with occlusion. Findings indicating cholangitis were noted in 50 of 71 segments (70%). The degrees of portal vein calibre were significantly correlated with the presence of hepatic atrophy or cholangitis. CONCLUSION: Hepatolithiasis is associated with significant rates of stenosis or occlusion of adjacent portal veins as well as hepatic parenchymal changes in the affected area. Chronic deterioration of portal flow may cause these morphological changes.

Adult↗

Polarization of Th1/Th2 in human CD4 T cells separated by CD62L: analysis by transcription factors.

BACKGROUND: T-cell surface antigens that differentiate clearly between Th1 and Th2 have not been identified. Discrimination of Th1/Th2 subpopulations by CD62L expression has been reported. We investigated the expression of transcription factors that regulate Th1/Th2 cytokine synthesis in human CD4+ T-cell subpopulations separated by CD45RO and CD62L, and compared the ratio of CD62L+ to CD62L- cells between healthy individuals and patients with allergic diseases. METHODS: Human peripheral blood samples were obtained from healthy volunteers and patients. CD4+ T cells were isolated by negative selection. Three CD4+ T-cell subpopulations separated by CD45RO and CD62L were isolated using three-color fluorescence. Sorted cells were stimulated with anti-CD3 monoclonal antibody, and the cytokine levels were measured using a Cytometric Bead Array Kit. Transcription factor expression was examined by reverse transcriptase polymerase chain reaction (RT-PCR) and real-time RT-PCR. RESULTS: Interleukin (IL)-4 and IL-5 production levels by CD45RO+CD62L+CD4+ T cells were higher than those of CD45RO+CD62L-CD4+ T cells (P < 0.05), whereas interferon-gamma and tumor necrosis factor-alpha production were lower levels (P < 0.05). T-cell immunoglobulin mucin-3 and T-bet expression were detected in CD45RO-CD62L+ and CD45RO+CD62L- cells following stimulation, but not in CD45RO+CD62L+ cells. However, the ratio of CD62L+ to CD62L- cells was the same in both healthy individuals and patients (P = 0.54). There was no difference in Th1/Th2 cytokine synthesis by CD4+ T cells. CONCLUSION: Analyses of cytokine syntheses and transcription factor expression demonstrated that CD62-negative and -positive subpopulations of human CD45RO+CD4+ T cells represent characteristics of Th1 and Th2, respectively.

CD4-Positive T-Lymphocytes↗

Liver lesions of visceral larva migrans due to Ascaris suum infection: CT findings.

The purpose of this study was to analyze computed tomographic (CT) findings of hepatic lesions due to Ascaris suum infection. CT of the liver in three patients, all of whom had immunoserologically confirmed A. suum infection, were retrospectively reviewed. Twenty-five lesions were identified in total. Two radiologists analyzed CT findings in a consensus fashion, with particular interest in the margin, shape, and location of the lesions. Hepatic lesions were ill-defined (22 of 25), small (3-35 mm; average, 11 mm), and nodular (18 of 25) or wedge (three of 25) in shape. Most were located in periportal (16 of 25) or subcapsular (six of 25) regions. Hepatic nodules due to visceral larva migrans of A. suum were located mainly in periportal or subcapsular regions, which may represent periportal eosinophilic granuloma, its pathologic feature. The results were considered to represent the pathophysiology of this entity.

Adult↗

Telomerase-dependent oncolytic adenovirus for cancer treatment.

Conditionally replicative adenovirus (CRAD) is an attractive anticancer agent as it can selectively replicate in tumor cells. Expression of telomerase reverse transcriptase (TERT) is a unique tumor cell characteristic, being absent in normal postmitotic cells. Thus, we constructed a TERT promoter regulated CRAD for tumor-specific oncolysis by replacing the endogenous adenovirus E1A promoter with that of human TERT (Adv-TERTp-E1A). We showed that its replication was severely attenuated in TERT-negative cells, but that it replicated almost as efficiently as wild-type adenovirus in TERT-positive cells. Accordingly, Adv-TERTp-E1A conferred cytopathicity to TERT-positive, but not TERT-negative, cells. In vivo replication of Adv-TERTp-E1A after local administration into a xenograft model of human hepatocellular carcinoma in nude mice was demonstrated by an increase in adenovirus titers in tumor extracts by several orders of magnitude between 6 h and 3 days postvector injection. Furthermore, significant inhibition of tumor growth with substantial necrotic tumor areas staining positively for adenovirus was observed with Adv-TERTp-E1A, but not with a control replication-deficient adenovirus. There was also the absence of hepatotoxicity in tumor-bearing animals after intratumoral delivery of the CRAD. The results indicate that the TERT promoter-driven CRAD is capable of tumor-selective replication and oncolysis in vitro and in vivo, and can be utilized as an adjuvant treatment agent for cancer.

Adenoviridae↗

Safety and growth suppressive effect of intra-hepatic arterial injection of AdCMV-p53 combined with CDDP to rat liver metastatic tumors.

Surgical resection is thought to be the best treatment for liver carcinoma, including hepatocellular carcinoma and metastatic liver carcinoma if there are a small number of tumors. Liver carcinoma is one of the main causes of death from cancer worldwide. The prognosis of liver carcinoma is still poor. Mutation of p53, which is well known as a tumor suppressor gene, is observed in many cases of advanced liver carcinoma. Cancer gene therapy using p53, which transduces the wild-type p53 gene in the tumor, is a promising new strategy for treating liver carcinoma. Selective and less invasive gene delivery to the liver tumor is necessary for clinical liver tumor gene therapy. The first purpose of the current study was to determine the best way to deliver the gene of interest to the liver tumor selectively. The second purpose was to study the tumor suppressive effect of intrahepatic arterial injection of an adenovirus vector with the p53 gene (AdCMV-p53), followed by administration of CDDP and noting its side effects. We injected AdCMV-LacZ via hepatic arteries of rats bearing RCN-9 colon cancer metastasis in the liver. Injection via the hepatic artery resulted in more successful gene transduction to the liver tumor in a tumor-selective manner than did injection via the portal vein. At 48 hrs after arterial injection of AdCMV-p53, CDDP (3 mg/kg) was administered in the peritoneal cavity of each rat. The use of CDDP with arterial injection of AdCMV-p53 resulted in more extensive apoptosis in the rat liver tumors without any deterioration in liver function. In conclusion, hepatic arterial injection of an adenovirus vector is better than portal vein injection for gene transduction efficiency, and causes no liver function disorder even when the injection is combined with CDDP.

Adenoviridae↗

Molecular epidemiology and phylogenetic analysis of Sapporo-like viruses.

Sapporo-like viruses (SLV) are a causative agent of gastroenteritis in humans. SLV-specific primers were newly designed in capsid protein-coding region and 529 fecal samples collected from gastroenteritis patients were tested. Thirty-five samples (6.6%) were found to be positive by reverse transcriptase-polymerase chain reaction (RT-PCR). Out of 35 positive samples, 25 were classified into a genogroup SG-I, typified by the Sapporo virus, and 9 were classified as a genogroup SG-II, typified by the London virus. Interestingly, one sample could not be classified into any genogroup, which suggests that it may be part of a new SLV genogroup. The RT-PCR used in this study appeared to be capable of widely detecting SLV genogroups, and it was seen to be powerful enough for molecular epidemiological as well as phylogenetic studies on SLV.

Adolescent↗

Involvement of CD27/CD70 interactions in antigen-specific cytotoxic T-lymphocyte (CTL) activity by perforin-mediated cytotoxicity.

CD27 molecules are shown to be essential in the regulation of the death, activation and differentiation of T and B cells. However, the influence of CD27 on cytotoxic T-cell function remains obscure. Autologous EBV transformed B-cell lines (LCL), which highly express CD27 ligand CD70, here stimulated T cells and induced the cytotoxic T-lymphocyte (CTL) activity via T-cell antigen receptors (TCR). The cytotoxicity against LCL was diminished when anti-CD70 blocking MoAb was added initially in the culture. Resting T cells killed more CD70-transfected P815 cells than wild type P815 cells in the presence of anti-CD3 MoAb as measured by a 4-h 51Cr release assay, and the cytotoxicity of both of the cell populations completely disappeared in the presence of concanamycin A (CMA). The expression of the perforin by the LCL-induced CTL in the presence of anti-CD70 blocking MoAb was diminished as compared with that without the blockage of CD27/CD70 interactions. The CTL induced by LCL did not kill Fas-transfected WR cells. CD27 signalling in the T cells did not affect Fas ligand (FasL) mRNA expression, LAK activity and IFN-gamma synthesis in humans. Our data demonstrate that CD27/CD70 interactions enhance the cytotoxicity of CTL in the induction phase through enhancement of killing activity induced via the perforin-dependent mechanism, but not via the Fas/FasL system.

Antigens, CD↗

Plant histidine kinases: an emerging picture of two-component signal transduction in hormone and environmental responses.

In the Arabidopsis thaliana genome, 11 genes encode bacterial-type two-component histidine kinases. Genetic and biochemical analyses indicate that five two-component histidine kinase-like proteins (ETR1, ETR2, EIN4, ERS1, and ERS2) function as ethylene receptors. A hybrid histidine kinase, CRE1 (also known as AHK4), acts as a cytokinin receptor, and a set of response regulators may be involved in cytokinin signal transduction. In addition to CRE1, histidine kinases CKI1 and CKI2 are likely to play important roles in cytokinin signaling. A database search of the entire Arabidopsis genome sequence has identified two additional homologs of CRE1. Arabidopsis seems to employ a hybrid histidine kinase, ATHK1, as an osmosensor. Plants widely use two-component systems in the detection of, and signal transduction by, the growth regulators ethylene and cytokinin, as well as in their responses to environmental stimuli.

Arabidopsis↗

An essential role of a TatC homologue of a Delta pH- dependent protein transporter in thylakoid membrane formation during chloroplast development in Arabidopsis thaliana.

At least three transport systems function in targeting nuclear-encoded chloroplast proteins to the chloroplast thylakoid membrane. One of these systems requires a thylakoid pH gradient and is named the DeltapH-dependent protein transport system. A similar DeltapH export system of Escherichia coli contains four components, twin arginine translocation A (TatA), TatB, TatC, and TatE. TatC is a major component of the DeltapH-dependent protein transporter in E. coli and functions in the translocation of tightly folded proteins across membranes. We have isolated four transposon-inserted albino mutants named albino and pale green 2 (apg2) from Arabidopsis thaliana and showed that the transposons were inserted into different sites of a single gene. The APG2 gene product (named cpTatC) has sequence similarity with bacterial TatC and contains six putative transmembrane domains, including bacterial TatC proteins and a transit peptide in its N terminus. apg2 mutants showed albino phenotypes and could not grow in soil. The apg2 plastids were highly vacuolated, lacked internal membrane structures and lamellae of the thylakoid membrane, and contained many densely stained globule structures, like undifferentiated proplastids. Immunoblot analysis detected no thylakoid membrane proteins such as D1, light-harvesting complex, and OE23 in apg2 plastids, whereas soluble proteins such as rubisco large and small subunits were not decreased. These results indicate an essential role of cpTatC in chloroplast development, especially in thylakoid membrane formation.

Amino Acid Sequence↗

SH2D1A mutations in Japanese males with severe Epstein-Barr virus--associated illnesses.

X-linked lymphoproliferative disease (XLP), a genetic disorder characterized by immunodeficiency to Epstein-Barr virus (EBV) infection, has been linked to mutations in the SH2D1A gene. To search for the occurrence of SH2D1A mutations in Japan, we performed genetic analysis of the SH2D1A gene in 40 males presenting with severe EBV-associated illnesses, including fulminant infectious mononucleosis, EBV-positive lymphoma, and severe chronic active EBV infection. SH2D1A mutations were detected in 10 of these 40 patients. Five of these 10 cases were sporadic. Patients with SH2D1A mutations displayed severe acute infectious mononucleosis with hyperimmunoglobulin M, hypogammaglobulinemia, and B-cell malignant lymphoma. By contrast, chronic active EBV infection was not associated with SH2D1A mutations. XLP survivors exhibited normal levels of circulating EBV-DNA during convalescence, suggesting that SH2D1A protein is not directly responsible for control of EBV replication. Thus, genetic analysis of the SH2D1A gene is particularly useful in the diagnosis of sporadic cases and carriers of XLP. (Blood. 2001;98:1268-1270)

Adolescent↗