Biomedical subjects

T Iida

Publications and source records attributed to T Iida.

At least 109 records · Page 6Linked to original sources

PubMed2000

A filamentous phage associated with recent pandemic Vibrio parahaemolyticus O3:K6 strains.

A specific serotype, O3:K6, of Vibrio parahaemolyticus has recently been causing epidemics of gastroenteritis in Southeast Asia, Japan, and North America. To examine whether the new O3:K6 strains possess characteristics that may exacerbate outbreaks, we compared V. parahaemolyticus O3:K6 strains with non-O3:K6 strains using strains isolated from individuals with traveler's diarrhea at Kansai Airport Quarantine Station, Osaka, Japan. All 24 O3:K6 strains possessed a common plasmid, pO3K6 (DNA size, 8,782 bp, with 10 open reading frames [ORFs]). The gene organization of pO3K6 was similar to that of Vf33, a filamentous phage previously described in V. parahaemolyticus. We isolated a phage (phage f237) from the culture supernatant of V. parahaemolyticus O3:K6 strain KXV237, which formed a turbid plaque on an indicator strain. The genome of f237 was single-stranded DNA, and the double-stranded DNA obtained by treatment of the genome with DNA polymerase was identical to that of pO3K6 when analyzed by agarose gel electrophoresis after HindIII digestion. Furthermore, the N-terminal amino acid sequence of the f237 major coat protein was found in ORF4 of pO3K6. Our results showed that pO3K6 is a replicative form of f237. Among the ORFs found in the f237 genome, the sequence of ORF8 had no significant homology to those of any proteins in databases. ORF8 was located on a region corresponding to the distinctive region of Vf33, and its G+C content was apparently lower than that of the remaining DNA sequence of f237. By colony hybridization, ORF8 was detected only in O3:K6 strains isolated since 1996 and was not found in O3:K6 strains isolated before 1996 and clinical V. parahaemolyticus strains other than those of serotype O3:K6. Thus, this study shows that f237 is exclusively associated with recent V. parahaemolyticus O3:K6 strains. The ORF8 gene can be a useful genetic marker for the identification of the recently widespread O3:K6 strains of V. parahaemolyticus.

Diarrhea↗

PubMed2000

Redox regulation of the nuclear receptor.

Redox regulation is currently considered a mode of signal transduction for coordinated regulation of a variety of cellular processes. Transcriptional regulation of gene expression is also influenced by the cellular redox state, for example, through the oxidoreductive modification of transcription factors. The nuclear receptors act as ligand-dependent transcription factors and are considered to be one of the key regulators of essential biological processes. Using a glucocorticoid receptor as a model, we show that redox regulation of receptor function is one of the dynamic cellular responses to environmental stimuli.

Animals↗

PubMed2000

Cardiac arrhythmias and left ventricular hypertrophy in dipper and nondipper patients with essential hypertension.

To evaluate the behavior of cardiac arrhythmias in dipper and nondipper hypertensive patients, 48-h ambulatory blood pressure monitoring, 24-h Holter electrocardiogram recording and echocardiographic studies were performed in 56 untreated outpatients with essential hypertension. These patients were divided into 2 groups according to the presence (dipper, n=33) or absence (nondipper, n=23) of reduction of both systolic and diastolic blood pressure during nighttime by an average of more than 10% of daytime blood pressure. Mean 48-h systolic and diastolic blood pressures did not differ between the 2 groups. Nondipper patients had a significantly larger left atrial dimension (31.9+/-3.8 vs 35.6+/-3.7 mm; p<0.01), left ventricular mass index (114+/-26 vs 136+/-36 g/m2; p<0.05), as well as a larger number of total supraventricular (16+/-19 vs 89+/-197 beats; p<0.05) and ventricular ectopic beats (7+/-14 vs 47+/-96 beats; p<0.05) during daytime as compared with dippers. In conclusion, nondipper hypertensive patients are likely to experience supraventricular and ventricular arrhythmias more frequently than dippers. A blunted nocturnal blood pressure fall may be involved in the appearance of cardiac arrhythmias in patients with essential hypertension.

Adult↗

PubMed2000

Plasma levels of the chemokine RANTES in macaque monkeys infected with pathogenic and non-pathogenic SIV/HIV-1 chimeric viruses at an early stage of infection.

Plasma levels of the chemokine RANTES were examined in monkeys infected with either a pathogenic simian and human immunodeficiency chimeric virus (SHIV) or a non-pathogenic SHIV to determine whether RANTES levels were related to the pathogenicity of the virus, the plasma viral load, or the kinetics of CD4+ T-cells. In the results no significant correlation was found between the RANTES kinetics and changes in the CD4+ T-cell numbers nor the plasma viral loads in any of the monkeys, although a transient decrease of the RANTES level was observed in the pathogenic virus-infected monkeys. At least, the plasma RANTES level can not be used as an index of the pathogenicity of the virus at the early stage of infection.

Animals↗

PubMed1999

Phylogenetic relationships of symbiotic spirochetes in the gut of diverse termites.

Phylogenetic relationships of symbiotic spirochetes in the gut of diverse termites were analyzed without cultivation of these microorganisms. A portion of the 16S rDNA (ca. 850 bp) was amplified directly from DNA of the mixed population in the gut by PCR and cloned. A total of 30 spirochetal phylotypes affiliated with the treponemes were identified from four termite species and they were compared with those already reported from other termites. They represented separate lines of descent from any known species of Treponema, and they were divided into two discrete clusters; one was related to Spirochaeta stenostrepta and S. caldaria, and the other was grouped together with members of the Treponema bryantii subgroup. Although some sequences from evolutionarily related termites showed close similarity, most of the sequences of spirochetes were dissimilar among different termite species, and spirochetal sequences from a single termite species occurred in several distinct phylogenetic positions. These findings suggest that termites constitute a rich reservoir of novel spirochetal diversity and that evolution of the symbiosis is not simple.

Animals↗

PubMed1999

Occurrence of D-amino acids in a few archaea and dehydrogenase activities in hyperthermophile Pyrobaculum islandicum.

The contents of D-enantiomers of serine, alanine, proline, glutamate (glutamine) and aspartate (asparagine) were examined in the membrane fractions, soluble proteins and free amino acids from some species of archaea, Pyrobaculum islandicum, Methanosarcina barkeri and Halobacterium salinarium. Around 2% (D/D+L) of D-aspartate was found in the membrane fractions. In the soluble proteins, the D-amino acid content was higher in P. islandicum than that in the other archaeal cells: the concentrations in P. islandicum were 3 and 4% for D-serine and D-aspartate, respectively. High concentrations of free D-amino acids were found in P. islandicum and H. salinarium; the concentrations of D-serine (12-13%), D-aspartate (4-7%) and D-proline (3-4%) were higher than those of D-alanine and D-glutamate. This result showed a resemblance between these archaea and not bacterial, but eukaryotic cells. The presence of D-amino acids was confirmed by their digestion with D-amino acid oxidase and D-aspartate oxidase. The occurrence of D-amino acids was also confirmed by the presence of activities catalyzing catabolism of D-amino acids in the P. islandicum homogenate, as measured by 2-oxo acid formation. The catalytic activities oxidizing D-alanine, D-aspartate and D-serine at 90 degrees C were considerably high. Under anaerobic conditions, dehydrogenase activities of the homogenate were 69, 84 and 30% of the above oxidase activities toward D-alanine, D-aspartate and D-serine, respectively. Comparable or higher dehydrogenase activities were also detected with these D-amino acids as substrate by the reduction of 2, 6-dichlorophenolindophenol. No D-amino acid oxidase activity was detected in the homogenates of M. barkeri and H. salinarium.

Amino Acids↗

PubMed1999

Cytotoxic T lymphocyte antigen 4 (CTLA-4) engagement delivers an inhibitory signal through the membrane-proximal region in the absence of the tyrosine motif in the cytoplasmic tail.

Cytotoxic T lymphocyte antigen 4 (CTLA-4) is a T cell costimulation receptor that delivers inhibitory signals upon activation. Although the tyrosine-based motif ((165)YVKM) within its cytoplasmic tail has been shown to associate in vitro with Src homology 2 domain-containing tyrosine phosphatase (SHP-2) and phosphatidylinositol 3 kinase upon phosphorylation, the mechanism of negative signaling remains unclear. Here, we report a new mechanism of negative signaling based on the analysis of murine T cell clones transfected with various mutants of CTLA-4. Upon T cell activation by cross-linking with anti-CD3 and anti-CD28 antibodies, CTLA-4 engagement inhibited both proliferation and interleukin 2 production in tyrosine mutants as well as in wild-type CTLA-4 transfectants. Furthermore, the mutant CTLA-4 lacking most of the cytoplasmic region strongly suppressed interleukin 2 production as well. These data suggest that negative signals by CTLA-4 could be mediated through the membrane-proximal region of CTLA-4 but not through the YVKM motif and that the association of CTLA-4 with SHP-2 is not required for CTLA-4-mediated suppression of T cell activation.

Abatacept↗

PubMed1999

Co-expression of gamma-glutamylcysteine synthetase sub-units in response to cisplatin and doxorubicin in human cancer cells.

Glutathione (gamma-glutamylcysteinyl glycine, GSH) is believed to be important in the acquisition of resistance to anti-cancer drugs such as cisplatin (CDDP) and doxorubicin (DOX). gamma-Glutamylcysteine synthetase (gamma-GCS) is a key enzyme for maintaining intracellular GSH levels; it is composed of a catalytic heavy (gamma-GCSh) and a regulatory light (gamma-GCSl) sub-unit. In the present study, the expression of gamma-GCS sub-units was examined in human cancer cell lines. The levels of GSH, the expression of gamma-GCSh and gamma-GCSl mRNAs and proteins in human cancer cells were higher in CDDP-resistant cells and DOX-resistant cells than in the drug-sensitive cells. Treatment of CDDP/DOX-resistant human colonic-cancer cells (HCT8DDP) with CDDP and DOX caused simultaneous induction of the expression of gamma-GCSh and gamma-GCSl mRNAs. The transcriptional regulation of gamma-GCS was studied and it was observed that the DNA-binding activity of activator protein 1 (AP-1) is induced by CDDP and DOX using an electrophoretic mobility shift assay. We constructed chimeric genes containing various regions of the gamma-GCSh-promoter gene and the coding region for luciferase. The HCT8DDP cells transiently transfected with a plasmid containing an AP-1 site of the gamma-GCSh-promoter-luciferase construct showed increased luciferase activity when treated with CDDP and DOX. These results suggest that stimulation of the expression of gamma-GCSh by CDDP and DOX is mediated by AP-1, which relates to the resistance of cancer cells to the drug.

Antineoplastic Combined Chemotherapy Protocols↗

PubMed1999

Sequential analysis of apoptosis induction in peripheral blood mononuclear cells and lymph nodes in the early phase of pathogenic and nonpathogenic SIVmac infection.

To investigate the role of apoptosis in the early phase of HIV infection, we used macaques infected with simian immunodeficiency virus strain mac (SIVmac) as a primate model and examined sequentially the characteristics of apoptosis of lymphocytes in peripheral blood mononuclear cells (PBMCs) and lymph nodes in the early phase of SIVmac infection. Five macaques infected with a pathogenic strain of SIV, SIVmac239, were analyzed during the first 4 weeks after infection. Peripheral CD4+ and CD8+ cells transiently decreased at 1 week postinfection. The percentage of apoptotic cells in cultured PBMCs increased from about 2 weeks postinfection. The number of apoptotic cells in lymph node sections was higher on days 13 and 28 postinfection than before infection and on day 5 postinfection. Fas antigen expression on peripheral lymphocytes was upregulated from day 8 postinfection. These results indicate that apoptosis is induced about 2 weeks after SIVmac239 infection, following the upregulation of Fas antigen expression on lymphocytes. Since apoptosis was induced about 1 week after the decrease in peripheral CD4+ and CD8+ cell counts, it appears that the apoptosis induction does not play an important role in the transient lymphopenia in the early phase of SIVmac infection. In macaques infected with a nonpathogenic derivative of SIVmac239, SIVmac delta nef, apoptosis of lymphocytes was induced as it was in SIVmac239-infected macaques, but to a lesser degree, suggesting a correlation between the extent of apoptosis induction in lymphocytes in the early phase of SIVmac infection and the pathogenicity of SIVmac.

Animals↗

PubMed1999

Application of peptide nucleic acid to the direct detection of deoxyribonucleic acid amplified by polymerase chain reaction.

Double-stranded DNA amplified by polymerase chain reaction (PCR) was detected by peptide nucleic acid (PNA) using a BIAcore 2000 biosensor based on surface plasmon resonance (SPR). PNA is an artificial oligo amide that is capable of forming highly stable complexes with complementary oligonucleotides. We succeeded in the direct detection of double-stranded DNA, amplified by PCR with high-sequence specificity. It was shown that the target DNA was available for detection over the range of 40-160 nM. Therefore, the detection limit was 7.5 pmol of the target DNA (143 bases, applied volume 30 microliters). Our DNA detection system, the combination of BIAcore and the probe PNA, could detect the target DNA with good reproducibility. In this report, we show that our system is a powerful tool for the diagnosis of pathologically significant DNA.

Bacterial Toxins↗

PubMed1999

Detection of PCR products in solution using surface plasmon resonance.

Polymerase chain reaction (PCR) products were detected using a flow injection-type sensor based on surface plasmon resonance. Asymmetric PCR was used to amplify the target DNA sequence, and two products with different length were produced. The novelty of our DNA detection system was that our target DNA was double stranded but the probe binding site, located in the 3'-terminus, was single stranded. This avoids the formation of intra- and intermolecular complexes. This novel design permitted us not only to detect PCR product but also to develop a rapid detection system for the detection of the verotoxin 2 gene of Escherichia coli O157:H7.

Bacterial Toxins↗

PubMed1999

Clonal expansion of T cells that are specific for autologous ovarian tumor among tumor-infiltrating T cells in humans.

OBJECTIVE: The purpose of this study was to determine whether oligoclonally expanding tumor-infiltrating lymphocytes (TIL) were tumor-specific. STUDY DESIGN: Peripheral blood lymphocytes (PBL) from an ovarian tumor-bearing patient were stimulated in vitro with an autologous cancer cell line (SMOV-2). Then genes coding for the third complementarity-determining region of the T cell receptor (TCR) beta chain were amplified by reverse transcription-polymerase chain reaction and separated by single-strand conformation polymorphism. Accumulated TCR clonotypes in vitro and in vivo in TIL were compared. RESULTS: Clonal expansion of T cells was generated from PBL by stimulation with SMOV-2. A portion of the proliferated clonotypes was found to be identical to those accumulated in TIL in vivo. CONCLUSION: This is the first demonstration that accumulating T cell clones in TIL recognize antigen(s) on an autologous tumor. Further characterization of such T cell clonotypes may lead to tumor antigen-specific immunotherapy.

Base Sequence↗

PubMed1999

Plasminogen activator inhibitor 1 in central serous chorioretinopathy.

PURPOSE: To describe the plasma level of plasminogen activator inhibitor 1, the major antifibrinolytic agent, in eyes with central serous chorioretinopathy, in which choroidal thrombosis is suspected as the underlying condition based on the findings of choroidal hyperpermeability in indocyanine green angiograms. METHODS: Plasminogen activator inhibitor 1 concentrations in the plasma of 17 patients with central serous chorioretinopathy were measured by enzyme-linked immunosorbent assay and compared with those in 12 age-matched normal volunteers. RESULTS: The plasma levels of plasminogen activator inhibitor 1 in patients with central serous chorioretinopathy (range, 25 to 439 ng/ml; median, 87 ng/ml) were significantly increased compared with those in normal volunteers (range, 7 to 84 ng/ml; median, 36 ng/ml) (P = .0013, Mann-Whitney U test). CONCLUSION: Increased plasminogen activator inhibitor 1 concentrations in patients with central serous chorioretinopathy support the hypothesis that the choroidal hyperpermeability disclosed by indocyanine green angiography is caused from impaired fibrinolysis and the resulting thrombotic occlusion in the choroidal veins.

Adult↗

PubMed1999

Recent trend of polychlorinated dibenzo-p-dioxins and their related compounds in the blood and sebum of Yusho and Yu Cheng patients.

We investigated the levels of polychlorinated dibenzo-p-dioxins (PCDDs) and their related compounds in blood and sebum from Yusho and Yu-Cheng patients and findings compared with those of normal subjects. Concentrations of these compounds in blood and sebum still were obviously reflected the chemical concentration in the causal rice oil ingested 17 or 26 years since the outbreak. In addition, sebum form the body surface was the most useful sample for monitoring the amount of chemicals contaminating the human body.

Adult↗

PubMed1999

Polychlorinated dibenzo-p-dioxins and related compounds in breast milk of Japanese primiparas and multiparas.

We analyzed polychlorinated dibenzo-p-dioxins and related compounds in the breast milk of primiparas and multiparas, and estimated the levels transferred to newborns by breast milk in Western Japan. 2,3,7,8-TeCDD equivalents (TEQ) of the chemicals in primiparas decreased slightly from 1994 to 1996. In particular, decreases of the TEQs of total PCDDs and total coplanar PCBs were higher than that of total PCDFs. In 2,3,7,8-TeCDD, 1,2,3,7,8-PeCDD, 1,2,3,6,7,8-HxCDD, 1,2,3,7,8,9-HxCDD, 2,3,4,7,8-PeCDF, 1,2,3,6,7,8-HxCDF, 3,3',4,4',5-PeCB and 3,3',4,4',5,5'-HxCB concentrations, those in the breast milk of multiparas were significantly lower than those in the breast milk of primiparas (p < 0.05, lipid basis). Based on the assumption that newborns ingest 120 g of breast milk per kg body weight per day, the amounts converted to TEQ values were 121 pg/kg/day (primiparas) and 97.2 pg/kg/day (multiparas).

Adult↗

PubMed1999

Polychlorinated dibenzo-p-dioxins and related compounds: correlations of levels in human tissues and in blood.

We investigated PCDDs and related compounds levels in human tissue and blood obtained from the general population, and the correlation factor was calculated from the findings. None of the congeners in brain, muscle and lung were correlated except for 2,3,7,8-tetrachlorodibenzo-p-dioxin and octachlorodibenzo-p-dioxin in the brain (p < 0.05). In other tissues, all congeners had relatively good correlations to those in blood (r > 0.707). These congeners detected in blood were at high concentrations in the environment and human body. Therefore, we concluded that these congener levels in the blood might be useful for estimating these congener levels in human tissue.

Adult↗

PubMed1999

Polychlorinated dibenzo-p-dioxins and related compounds: the blood levels of young Japanese women.

We investigated PCDDs and related compounds in the blood of young Japanese women, approximately 20 years of age, who had not yet had children, and discussed how the TEQ level of PCDDs and related compounds in their blood may affect the next generation. Means of total TEQ levels were 0.063 pg/g for whole blood basis and 21 pg/g for lipid basis. TEQ of PCDDs, PCDFs and coplanar PCBs accounted for about 43, 34 and 23% of the total TEQ in the whole blood basis, respectively. In the lipid basis, their values were about 44, 34 and 22%, respectively. Previously, we investigated PCDDs and related compounds levels in mother's breast milk, lymphocyte subpopulation and thyroid function of their children, and found negative correlations between the TEQ level of PCDDs and related compounds and CD4+/CD8+, and/or the TEQ level of PCDDs and related compounds and the T4 level in 36 mothers and children. Of these cases, the average age was approximately 28 years. PCDDs and related compounds may be related to immunopathy, such as atopic dermatitis. The effects of PCDDs and related compounds on babies of young Japanese women are important and must be further evaluated.

Adult↗

PubMed1999

Analysis of localization and function of the COOH-terminal corresponding site of cytochrome b558 in fish neutrophils.

Using an antibody against the synthetic peptide corresponding to the COOH-terminal region of human cytochrome b558 large subunit, a broad band was specifically detected in neutrophil lysates from 6 marine fish and 2 freshwater fish by western blotting. Immunofluorescence assay showed that the antibody recognized the epitopes in eel and tilapia neutrophils permeabilized with detergent. These results suggest that the cytochrome b large subunit universally exists in fish neutrophils and that the epitopes are exposed to the cytoplasmic side of fish neutrophils as well as human neutrophils. Furthermore, a synthetic peptide corresponding to the COOH-terminus of the large subunit apparently blocked superoxide production in a specific and dose-dependent fashion in eel and tilapia neutrophils, indicating that the region equivalent to the COOH-terminus of cytochrome b large subunit is responsible for superoxide generation in fish neutrophils.

Amino Acid Sequence↗