Biomedical subjects

K Iida

Publications and source records attributed to K Iida.

At least 73 records · Page 4Linked to original sources

PubMed2000

Human herpesvirus 6 infection after living related liver transplantation.

The aim of the study was to investigate human herpesvirus-6 (HHV-6) infection after liver transplantation from living related donors, and to evaluate the reliability of the presence of HHV-6 DNA in plasma by the polymerase chain reaction (PCR) for monitoring active HHV-6 infection. EDTA peripheral blood was collected from 47 donor and recipient (16 males and 31 females, age 1-320 months) pairs at the time of transplantation and biweekly from these recipients after transplantation until 2 months after operation. Isolation of HHV-6 and serological assays were carried out to evaluate active HHV-6 infection in this study. The presence of the viral DNA in plasma was tested by nested PCR. Four clinical events, such as unexplained fever, thrombocytopenia, rejection, and central nervous system (CNS) involvement, were evaluated for clinical features of the virus infection. Risk factors for the virus activity after liver transplantation were also examined. HHV-6 activity was detected in 23 (49%) of the 47 recipients approximately 2-4 weeks after transplantation. All 9 isolates were HHV-6 variant B. The presence of the viral DNA in plasma correlated well with virus isolation and serology (P < 0.01). Only unexplained fever was associated statistically with HHV-6 activity after liver transplantation (P < 0. 01). If the recipient was seronegative to HHV-6 before transplantation, the recipient was more likely to develop the active virus infection after liver transplantation (P = 0.11). HHV-6 activity occurred in one-half of the recipients approximately 2-4 weeks after liver transplantation, and there was a close association between HHV-6 activity and unexplained fever following transplantation. Detection of the viral DNA in plasma by PCR is useful for monitoring active HHV-6 infection in these patients. Seronegative recipients were more likely to have evidence of active HHV-6 infection after liver transplantation.

Adolescent↗

PubMed2000

[Leiomyosarcoma of the scrotum: a case report].

Leiomyosarcoma of the scrotum is a rare tumor. Up to 1999, only 29 cases have been reported in the literature worldwide. A 27-year-old man presented with a mass on the left side of the scrotum which had been painless and had gradually enlarged over the previous 10 years. During the following 3 months, however, it became painful and he was then referred to our hospital. Physical examination revealed a solid mass on the left side of the scrotum, measuring 7 cm in diameter, which was not adhering to the testis or to the vas deferens. The tumor was surgically resected. The histological examination confirmed the diagnosis of well differentiated leiomyosarcoma. Adjuvant therapy was considered unnecessary. The follow-up at 41 months revealed no local recurrence or distant metastasis.

Adult↗

PubMed1999

Growth hormone stimulates tyrosine phosphorylation of focal adhesion kinase (p125(FAK)) and actin stress fiber formation in human osteoblast-like cells, Saos2.

Bone is one of the essential target tissues of growth hormone (GH). In bone remodeling, cell-matrix attachment is important where focal adhesion kinase (FAK) is involved. FAK plays a central role in determining the shape and motility of cells in response to the extracellular matrix stimuli. In the present study, we have demonstrated that GH stimulated tyrosine phosphorylation of FAK in human osteoblast-like cells, Saos2. Moreover, GH rapidly enhanced the formation of actin stress fibers. In Saos2, Jak2 was tyrosine phosphorylated by GH stimulation, and AG490, a Jak2 specific inhibitor, inhibited GH-induced tyrosine phosphorylation of FAK and actin stress fiber reorganization. These results suggest that GH activates FAK via Jak2, and stimulates the formation of actin stress fibers in Saos2. Activation of FAK and actin stress fiber formation induced by GH seem to be important for the physiological role of osteoblast.

Actins↗

PubMed1999

Elevated levels of pro-inflammatory cytokines in coronary artery thrombi.

To validate the hypothesis that artery sites occluded with thrombi release pro-inflammatory cytokines, we measured concentrations of interleukin (IL)-6 and IL-8 in infarct-related coronary artery thrombi and atherosclerotic plaque specimens obtained with a transluminal extraction catheter (TEC) from cases of acute myocardial infarction (MI). Fifteen patients (group I) were enrolled in the study and four sets of samples were obtained (taken from the right atrium both before and after angioplasty, from infarct-related coronary artery thrombi and atherosclerotic plaque aspirated with a TEC and from the thoracic aorta aspirated with a TEC). Ten patients undergoing elective TEC served as controls (group II). IL-6 and IL-8 were measured in all patients by means of an enzyme-linked immunosorbent assay. Both IL-6 and IL-8 levels of infarct-related coronary artery samples in group I were significantly higher than in group II (mean +/- SEM, 15.3+/-4.5 vs. 3.8+/-1.2 pg/ml; P<0.01 and 44.0+/-2.4 vs. 15.6+/-0.6 pg/ml; P<0.01, respectively). The results suggest that pro-inflammatory cytokines originate from occluded coronary arteries in acute MI.

Angioplasty, Balloon, Coronary↗

PubMed1999

Characterization of the promoter region of the human RFP gene.

The RFP gene encodes a Ring finger protein that has a tripartite motif consisting of a Ring finger, a B-box finger and a coiled-coil domain. In the present study, we cloned and characterized the promoter region of the human RFP gene. The nucleotide sequence of the promoter was GC-rich and had no typical TATA and CAAT boxes. Instead, it contained one AP2 and two Sp1 binding sites within 100 base pairs upstream of the transcription initiation site. Analysis by the luciferase assay revealed that the activity of this promoter region is very strong in both human and mouse cell lines, although the activity in human cells was approximately 10-15 fold higher than that in mouse cells. In addition, the AP2 and Sp1 binding sites appeared to synergistically function for the promoter activity. Thus, the promoter of the RFP gene could be useful for high levels of expression of various genes in culture cells.

3T3 Cells↗

PubMed1999

AlphaB-crystallin in the rat lens is phosphorylated at an early post-natal age.

We determined the developmental changes in the phosphorylation state of alphaB-crystallin in lenses from rats at various post-natal ages by isoelectric focusing gel electrophoresis or sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a subsequent Western blot analysis of extracts of lenses using antibodies that recognized the carboxy-terminal sequence or each of the three phosphorylated serine residues (Ser-19, Ser-45 and Ser-59) in alphaB-crystallin. Phosphorylated forms of alphaB-crystallin were barely detected at birth but they became detectable at 3 weeks of age and reached plateau levels at 8 weeks of age. The phosphorylation of alphaB-crystallin at Ser-45 was observed preferentially. The active form of p44/42 MAP kinase, which is responsible for the phosphorylation of Ser-45 in alphaB-crystallin, also increased in a development-dependent manner. Thus we found that the developmental increase of the phosphorylation at Ser-45 of alphaB-crystallin in the rat lens was due to the developmental activation of p44/42 MAP kinase.

Animals↗

PubMed1999

Strong expression of glutathione S-transferase placental form in early preneoplastic lesions and decrease with progression in hamster buccal pouch carcinogenesis.

The objective of this work was to investigate the expression of glutathione S-transferase placental form (GST-P) in 9,10-dimethyl-1,2-benzanthracene (DMBA)-induced hamster buccal pouch carcinogenesis. Lesions were classified histopathologically into four categories, simple hyperplasia, papillary and nodular (PN) hyperplasia, papilloma and squamous cell carcinoma (SCC). Respective mean percentage GST-P positive areas were 81.6 +/- 7.3%, 76.1 +/- 7.3%, 25.8 +/- 4.9% and 1.9 +/- 1.2%, with significant (P < 0.001) differences confirmed between each of the lesions. These results indicate that GST-P is a useful positive marker for neoplastic lesions and that a decreased expression occurs with progression so that it may be predictive of future development of malignancy.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1999

Promotional effects of CO2 laser on DMBA-induced hamster buccal pouch carcinogenesis as shown by immunohistochemistry of the placental form of glutathione S-transferase.

BACKGROUND AND OBJECTIVE: The purpose of the present study was to investigate the kinetics of the expression of the placental form of glutathione S-transferase (GST-P), a useful marker of premalignant lesions, and cell proliferation after CO2 laser surgery on the carcinogen-initiated epithelium. STUDY DESIGN/MATERIALS AND METHODS: CO2 laser incisions were made on buccal pouch epithelium of 36 hamsters after initiation by 9,10-dimethyl-1,2-benzanthracene (DMBA) (group 1, G1), and scalpel incisions were similarly made on 33 animals (group 2, G2). Twenty animals not treated further after initiation were used as DMBA-treated controls. Incidence of malignant transformation, expression of GST-P, and cell proliferation were examined. RESULTS: The incidence of malignant transformation in G1 and G2 increased significantly (G1: P < 0.001; G2: P < 0.05) compared with that in DMBA-treated controls. GST-P expression of hyperplasia in G1 and G2 decreased significantly (P < 0.001) compared with that in DMBA-treated controls. In hyperplasia, cell proliferation of the GST-P-negative area was significantly (P < 0.001) higher than that of the GST-P-positive area. CONCLUSION: The incisions, particularly by the CO2 laser, on the initiated areas made expression of GST-P decrease and cell proliferation increase in the GST-P-negative areas. These incisions may serve to promote malignant transformation.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1999

Degranulation of eosinophils mediated by intercellular adhesion molecule-1 and its ligands is involved in adhesion molecule expression on endothelial cells-selective induction of VCAM-1.

BACKGROUND: Adhesion molecules and eosinophils may play an important role in the pathogenesis of allergic inflammatory reactions. OBJECTIVE: We attempted to clarify eosinophil activation, such as degranulation, by signaling through adhesion molecule and to determine whether degranulation is involved in adhesion molecule expression on endothelial cells. METHODS: Eosinophils were cultured with or without recombinant soluble intercellular adhesion molecule-1 (ICAM-1), and the levels of eosinophil cationic protein and eosinophil-derived neurotoxin were determined. The influence of these eosinophil granule proteins or supernatant from eosinophil cultured with ICAM-1 on the expression of ICAM-1 or vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells was also examined by flow-cytometric analysis. RESULTS: Supernatant levels of eosinophil granule protein were significantly increased by culture for 4 hourss or 16 hours with recombinant soluble ICAM-1, suggesting degranulation by adherence to ICAM-1. Both granule proteins and the supernatants of eosinophils cultured with recombinant soluble ICAM-1 induced expression of ICAM-1 and VCAM-1 on endothelial cells, with the latter showing a more prominant increase. CONCLUSION: Degranulation mediated through adherence to endothelial cells by ICAM-1 and its ligands may be involved in the expression of adhesion molecules, such as ICAM-1 or VCAM-1, on these cells. Our finding of the selective induction of VCAM-1 expression suggests that eosinophil adherence to endothelial cells, even if it is because of ICAM-1, may be involved in selective eosinophil recruitment and accumulation at sites of allergic inflammation.

Cell Adhesion Molecules↗

PubMed1999

A method for modeling life based on physical attributes.

An incremental method for modeling the essential system of life, in which physical attributes of life are brought together to improve the model of life, is explained. We present a tiny example that uses only four attributes, i.e. locality, metabolism, entropy reduction, and chemotaxis. These four attributes are translated into physical formulae and evaluated in a two dimensional simulation of an autocatalytic reaction. Autocatalysts injected into a vessel begin to flow toward the source of their substrate similarly to chemotaxis. At the front line of this flow, a subsystem associated with the four properties is detected. This kind of subsystem could be increasingly life-like if more critical attributes are used for detection. A database of the attributes is being made.

Catalysis↗

PubMed1999

Elevated levels of beta-chemokines in bronchoalveolar lavage fluid (BALF) of individuals infected with human T lymphotropic virus type-1 (HTLV-1).

Pulmonary complications are known to develop in HTLV-1 carriers, including T lymphocytic alveolitis, and increased IL-2 receptor alpha (CD25)-bearing T cells have been found in BALF. Several chemokines may contribute to accumulation of T lymphocytes in the lungs of HTLV-1 carriers. Here, we compared the distribution of T lymphocyte subsets and beta-chemokines, such as macrophage inflammatory peptide-1alpha (MIP-1alpha), regulated on activation normal T expressed and secreted (RANTES), and macrophage chemoattractant protein-1 (MCP-1), in BALF and peripheral blood between HTLV-1 carriers and non-infected healthy normal subjects. Flow cytometric analysis with MoAbs to cell surface antigens was used to identify T lymphocyte subsets in BALF samples from HTLV-1 carriers (n = 13) and non-infected healthy controls (n = 10). The levels of different beta-chemokines were estimated by ELISA. High percentages of CD3+ cells, CD3 expressing HLA-DR antigen and CD3+CD25+ cells were detected in BALF of HTLV-1 carriers compared with non-infected controls. The concentration of MIP-1alpha in BALF of patients was significantly higher than in non-infected healthy controls and correlated well with the percentage of CD3+CD25+ cells. The level of RANTES in BALF was also significantly high in HTLV-1 carriers, but did not correlate with the percentage of CD3+CD25+ cells. On the other hand, the level of MCP-1 in BALF of HTLV-1 carriers was not different from that of controls. Our results suggest a possible interaction between activated T cells bearing CD25 and beta-chemokines, especially MIP-1alpha, which may contribute to the pulmonary involvement in HTLV-1 carriers.

Adult↗

PubMed1999

Cooperation of two actin-binding proteins, cofilin and Aip1, in Saccharomyces cerevisiae.

BACKGROUND: Cofilin is a low-molecular weight actin-modulating protein, and is structurally and functionally conserved among eukaryotes. Cofilin is encoded by COF1 in Saccharomyces cerevisiae, and is essential for cell viability. Cofilin binds to and severs actin filaments in vitro, and also enhances their depolymerization. A partner protein that cooperates with cofilin in vivo has not been identified. RESULTS: When COF1 was over-expressed in yeast cells under the GAL1 promoter in a medium containing galactose as a sole carbon source, the cells did not survive. These results indicate that cells can grow only when the expression of cofilin is appropriately regulated. Several temperature sensitive (ts-) mutants were independently created by the random mutagenesis of COF1 with hydroxylamine. Mutated amino acids in ts-mutants were mapped in the sequences that were presumed to be involved in actin binding. A gene on a multicopy plasmid which suppresses the ts-phenotype of cof1-101, a typical ts-cofilin mutant, was isolated. The suppressor gene, SCF1, was found to be identical to AIP1, a gene encoding an actin-interacting protein. Although SCF1/AIP1 is not essential for cell viability, a combination of cof1-101 and Deltascf1/aip1 is synthetic lethal. Immunofluorescence staining of a wild-type strain using anti-Aip1 antibodies revealed that Aip1 was distributed in cortical actin patches where cofilin was also co-localized. Thick and long fibres stained with anti-cofilin antibody were detected in Deltascf1/aip1 cells, but not in SCF1/AIP1 cells. CONCLUSIONS: These results suggest the cooperative modulation of the actin cytoskeleton by cofilin and Aip1.

Actin Depolymerizing Factors↗

PubMed1999

A case of pelvic arteriovenous malformation in a male.

BACKGROUND: Congenital pelvic arteriovenous malformation (AVM) is rare especially in males. We present a case of pelvic AVM in a 38-year-old male. He was admitted to our hospital with left flank abdominal pain and painless gross hematuria. METHODS/DISCUSSION: Pelvic computed tomography demonstrated a homogeneous contrast-enhanced mass, 6 cm in diameter. For diagnosis, non-invasive magnetic resonance imaging (MRI) and color Doppler ultrasonography were very useful in our case. Though the treatment is controversial, we decided to monitor his clinical course.

Abdominal Pain↗

PubMed1999

Development of a simple and highly sensitive enzyme immunoassay for hepatitis C virus core antigen.

A highly sensitive enzyme immunoassay (EIA) for the hepatitis C virus (HCV) core antigen (HCVcAg) was developed, and its performance was compared with that of the AMPLICOR HCV test (Roche Molecular Systems). The developed one-step pretreatment method, 30-min incubation of the specimen with a solution containing three different types of detergents (Triton X-100, 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate [CHAPS], and sodium dodecyl sulfate), does not require any special device. Because the interfering anti-core antibody in the sample was sufficiently inactivated by the pretreatment, HCVcAg in the sample could be detected. The immunoreactivity on gel filtration was shifted from void fractions to those corresponding to the molecular mass range from 20 to 25 kDa, which is equal to the estimated molecular mass of HCVcAg, after the pretreatment. By the recovery test with HCVcAg-positive serum, the recovery rate was 93.5 to 106. 5%. There was no interference with the EIA by anticoagulants or blood components in the serum. When the cutoff value was tentatively set at 0.5 mU/ml based on the distribution of healthy subjects' sera, the sera of all healthy subjects (n = 125) and patients with hepatitis B (n = 50) were negative. HCVcAg was detected in sera from 57 of 73 individuals (78.1%) with anti-HCV antibody. Similarly, HCV RNA was detected in sera from 59 individuals (80.8%) with the AMPLICOR HCV as the qualitative test (AMPLICOR HCV test) and in sera from 54 individuals (74.0%) by the AMPLICOR HCV Monitor as the quantitative test (AMPLICOR Monitor test). Concentrations of HCVcAg and HCV RNA (measured by the AMPLICOR Monitor test) correlated significantly (r = 0.8, P < 0.001). On seroconversion panels, HCVcAg was detected during the early stage of infection, when anti-HCV antibodies had not been produced. This assay for HCVcAg is simpler than assays for HCV RNA based on gene technology and shows specificity and sensitivity equivalent to those of the AMPLICOR HCV test.

Antibodies↗

PubMed1999

Thymidine phosphorylase expression in progression of cervical cancer: correlation with microvessel count, proliferating cell nuclear antigen, and apoptosis.

AIMS: To determine how epithelial and stromal thymidine phosphorylase expression affects angiogenesis, rapid tumour growth, and decreased apoptotic activity in cervical cancer at varying stages of progression. METHODS: Epithelial and stromal thymidine phosphorylase expression, the microvessel count (reflected by factor VIII related antigen), and proliferating cell nuclear antigen (PCNA) were assessed immunohistochemically in 25 specimens of normal cervical epithelium, 35 of carcinoma in situ (CIS), 34 of microinvasive carcinoma, and 34 of invasive cervical squamous cell carcinoma. Apoptosis was evaluated by the terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labelling (TUNEL) method. The relation of epithelial and stromal thymidine phosphorylase expression to microvessel count, PCNA index, and apoptotic index was examined. RESULTS: Epithelial and stromal thymidine phosphorylase expression progressively increased along a continuum from normal epithelium to invasive squamous cell carcinoma. Epithelial and stromal thymidine phosphorylase expression showed a significant positive correlation with microvessel counts. Within each histological stage, CIS cases with high stromal thymidine phosphorylase expression, invasive squamous cell carcinoma cases with high epithelial thymidine phosphorylase expression, and microinvasive carcinoma cases with high thymidine phosphorylase expression in both epithelium and stroma had a significantly higher microvessel count. High epithelial thymidine phosphorylase expression was associated with a significantly higher PCNA index in CIS and microinvasive carcinoma, but not in invasive squamous cell carcinoma. No significant correlation was seen between apoptotic index and either epithelial or stromal thymidine phosphorylase expression or microvessel count. CONCLUSIONS: Epithelial and stromal thymidine phosphorylase expression may combine to promote angiogenesis during progression of cervical cancer, and epithelial thymidine phosphorylase expression may stimulate tumour cell proliferation in the early stages.

Apoptosis↗

PubMed1999

The role of amino acids surrounding tyrosine 1062 in ret in specific binding of the shc phosphotyrosine-binding domain.

We investigated the role of the I-E-N-K-L (amino acids 1057-1061) sequence amino-terminal to Tyr1062 in Ret for binding of the Shc phosphotyrosine-binding (PTB) domain. Substitution of Ser for Ile1057 (I1057S), Ala for Asn1059 (N1059A), or Pro for Leu1061 (L1061P) in this sequence significantly decreased the transforming activity of Ret with the multiple endocrine neoplasm type 2A (MEN2A) mutation as well as the binding affinity of Shc to it in vivo and in vitro, indicating that these three amino acids play a role in Shc binding. In addition, as the RET protooncogene is translated as three isoforms of 1114 amino acids (Ret 51), 1106 amino acids (Ret 43), and 1072 amino acids (Ret 9) that differ from one another in the sequence carboxyl-terminal to Tyr1062, we examined whether these sequence differences influence the binding affinity of Shc to Ret. As a result, we found that the transforming activity of Ret 43 isoform with the MEN2A mutation and the binding affinity of Shc to it were very low, although the consensus sequence for the binding of the Shc PTB domain is conserved in the Ret 43 isoform. This finding suggested that the sequence carboxyl-terminal to Tyr1062 in Ret could also influence the binding affinity to Shc.

3T3 Cells↗

PubMed1999

The C422F mutation of the growth hormone receptor gene is not responsible for short stature.

A missense mutation, C422F, was identified in the intracellular domain of GH receptor (GHR) in a Japanese short boy. Although this mutation was previously reported in a patient with GH insensitivity syndrome (GHIS), it has not been clear whether this mutation causes GH insensitivity. To clarify the effect of this mutation on GH signal transduction, mutant GHR was expressed in CHO cells, and its functional properties were investigated. There were no significant differences in GH-induced tyrosine phosphorylation of STAT5b (signal transducer and activator of transcription) between wild-type GHR (GHR-wt)- and mutant GHR (GHR-C422F)-expressing cells. Moreover, STAT5-mediated transcriptional activation of GHR-C422F-expressing cells was comparable to that of GHR-wt-expressing cells. These findings indicated that the C422F mutation of GHR affected neither GH-induced tyrosine phosphorylation nor the transcriptional activation of STAT5. In addition, the analysis of genotypes and phenotypes of his family revealed that body heights of family members with heterozygous or homozygous C422F mutations were all within normal ranges, with the single exception of the proband. These in vitro and in vivo results indicate that the C422F missense mutation of GHR is a polymorphism that does not result in GHIS.

Animals↗

PubMed1999

Functional characterization of truncated growth hormone (GH) receptor-(1-277) causing partial GH insensitivity syndrome with high GH-binding protein.

We have previously reported a novel heterozygous donor splice site mutation in intron 9 of the GH receptor (GHR) gene in Japanese siblings who showed partial GH insensitivity and high serum GH-binding protein (GHBP) levels. This mutation caused the splicing abnormality and produced the truncated GHR consisting of 277 amino acids (GHR-277), which lacked most of the intracellular domain of GHR, including both boxes 1 and 2. In this study, we have characterized the function of GHR-277 expression in COS-7 and CHO cells in vitro. Scatchard analysis revealed that GHR-277 possessed approximately 1.5 times higher affinity to GH and twice the number of binding sites compared to wild-type full-length GHR (GHR-fl). The GHBP level in culture medium of GHR-277-expressing cells was approximately 3 times higher than that in GHR-fl-expressing cells. Interestingly, the ligand-induced internalization of GHR-277 was significantly reduced compared with that of GHR-fl. Moreover, in GH-induced tyrosine phosphorylation of signal transducer and activator of transcription-5 (STAT5), GHR-277 exerted a dominant negative effect when GHR-277 and GHR-fl were cotransfected. These in vitro data would well explain the clinical characteristics in our patients showing high serum GHBP levels and development of short stature despite a heterozygous mutation of the GHR gene.

Adolescent↗