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Masato Ikeda

Publications and source records attributed to Masato Ikeda.

At least 37 records · Page 2Linked to original sources
PubMed2005

Expression and characterization of N-terminal domain of Epstein-Barr virus latent membrane protein 2A in Escherichia coli.

Latency of Epstein-Barr virus (EBV) is maintained by the transmembrane protein latent membrane protein (LMP) 2A, which mimics the B-cell receptor (BCR) and perturbs BCR signaling. LMP2A contains a cytoplasmic N-terminal domain composed of 119 amino acids, which provides signals that are responsible for the association with various signal molecules, resulting in negative regulation of B-cell signaling and the EBV lytic cycle. In the present study, to obtain N-terminal domain of LMP2A (LMP2A NTD, 13 kDa) in Escherichia coli for structural analysis, a strategy for obtaining the unfused form of LMP2A NTD without any fusion partners was proposed. Recombinant LMP2A NTD has previously been expressed using the GST fusion system in E. coli [Virology 268 (2000) 178, J. Virol. 71 (1997) 4752, Mol. Cell. Biol. 20 (2000) 8526]. However, we were unable to obtain untagged LMP2A NTD from this construct because of rapid proteolysis by thrombin. To overcome the proteolysis by thrombin, C-terminal His-tagged LMP2A NTD and intein-fused LMP2A NTD were prepared. As a result, LMP2A NTD without a fusion partner could be successfully obtained using non-enzymatic cleavage. The secondary structure of the recombinant LMP2A NTD was analyzed using circular dichroism. In aqueous solution, LMP2A NTD adopts an unordered structure, which was not affected by varying pH and salt concentration. In addition, any secondary structural components of LMP2A NTD were not induced in the membrane-mimicking environments, suggesting that LMP2A NTD may intrinsically have a random coil-like structure. The biological activity of recombinant LMP2A NTD was monitored by chemical shift perturbation in HSQC spectra of LMP2A NTD with or without WW domains, which result supports that the structural change induced by WW domains is restricted within narrow region.

Amino Acid Sequence↗
PubMed2005

Pre-B-cell colony formation assay.

Latent membrane protein 2A (LMP-2A) of Epstein-Barr virus (EBV) mimics a constitutively active B-cell receptor (BCR) and plays a key role in viral latency and EBV pathogenesis. By functioning as a BCR mimic, LMP-2A drives B-cell development, resulting in the bypass of normal B-cell developmental checkpoints. To assess the function of LMP-2A, we have utilized a colony formation assay for the progenitor B cells that uses the B-cell proliferation factor IL-7.

Animals↗
PubMed2005

[Dialysis as bridge therapy for renal transplantation: single center experience, a comparison of hemodialysis and continuous ambulatory peritoneal dialysis].

BACKGROUND: Kidney transplantation is the most ideal treatment in renal replacement therapy for patients with end-stage renal disease. However, the prevalence of transplantation is extremely low and most patients with ESRD should continue dialysis for their whole life. Recently, high transposition rate of renal transplantation from peritoneal dialysis (PD) was reported, however, it was unclear whether a difference in dialytic modality can influence the outcome. Therefore, we evaluated the influence of dialytic modality on the rate of kidney transplantation and outcome in our single center. METHODS: Forty-two kidney transplants were carried among 1,573 dialysis patients from the years 1986 to 2004 in our center. Transposition rates from two modalities (HD and PD) and graft survival were compared. The incidence of acute rejection episode, complications after receiving transplantations, and coexisting diseases were also evaluated between the two modalities prior to transplantation. RESULT: The number of patients who received HD was larger than PD (HD 77.1%, PD 22.9%, respectively). Forty-two patients undergoing dialytic therapy received a living-donor kidney transplantation. Overall graft survival was 92% at 5 years and 75% at 10 years. Among these cases, dialytic modality prior to transplantation was 57.1% in HD, and 42.9% in PD. The transfer rate from PD to transplantation was significantly (p = 0.0036) higher (4.7%) than that of HD (1.9%). The reason for the high transfer rate of PD patients might be cooperation with their family and the provision of relevant information by nephrologists during PD. There were no differences between the two modalities prior to transplantation in the graft survival rate, incidence of acute rejection, and complications before and after transplantation. CONCLUSION: Difference in pretransplant dialysis modality did not affect the outcomes, however, the transfer rate from PD was significantly higher than from HD. Accordingly, PD is useful compared to HD as bridge therapy for kidney transplantation from the high feasibility of living-donor kidney transplantation.

Adult↗
PubMed2004

Monoclonal antibodies against regions topologically surrounding the homodimeric beta-barrel interface of Epstein-Barr virus nuclear antigen-1.

Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) is essential for maintenance of EBV latency. Four mouse monoclonal antibodies (mAbs) against the part of the EBNA-1 sequence (amino acids 451-641) containing the domain that forms a homodimeric eight-stranded beta-barrel were generated and characterized, examined for immunocytochemical staining, immunoblotting and isoelectric focusing of EBNA-1 proteins, and used to examine interactions between EBNA-1 polypeptides by far-Western blot assays. Far-Western blot analyses using the mAbs suggest that both the beta-strand (aa 593-604) and alpha helix (aa 568-582) are essential for EBNA-1 dimerization, consistent with yeast two-hybrid studies of mutant EBNA-1 polypeptides. These mAbs should be useful for studies on the structure and function of EBNA-1 proteins.

Animals↗
PubMed2004

Latent membrane protein 2A, a viral B cell receptor homologue, induces CD5+ B-1 cell development.

The latent membrane protein 2A (LMP2A) of EBV plays a key role in regulating viral latency and EBV pathogenesis by functionally mimicking a constitutively active B cell Ag receptor. When expressed as a B cell-specific transgene in mice, LMP2A drives B cell development, resulting in the bypass of normal developmental checkpoints. In this study, we have demonstrated that expression of LMP2A in transgenic mice results in B cell development that exclusively favors B-1 cells. This switch to B-1 cell development occurs at the pre-B-cell stage of normal B cell development in the bone marrow, a B cell stage much earlier than appreciated for B-1 commitment. This finding indicates that all pre-B cells have the capacity to assume a B-1 cell phenotype if they encounter the appropriate signal during normal development. Furthermore, these studies offer insight into EBV latency and pathogenesis in the human host.

Animals↗
PubMed2004

Rod-like architecture and helicity of the poly(C)/schizophyllan complex observed by AFM and SEM.

Microscopic studies of the complex between poly(C) and schizophyllan (SPG), employing both AFM and SEM, revealed that the complex takes the same rod-like architecture on the mica surface as those of the renatured SPG and the original triple helix of SPG, indicating that the complex also has a helical structure. The SEM observations showed the helical pattern on the rod surface, only when the sample was metal shadowed. The pitch evaluated from the image is comparable with that obtained from crystallographic data. The ability to visualize the helical structure can be explained from the hypothesis that the platinum grains may assemble on the sample using the molecular surface of the SPG (or complex) as the template.

Carbohydrate Conformation↗
PubMed2004

A 3-year follow-up study of anti-Legionella antibodies in users of Japanese 24-hour hot water baths.

Although it has been found that legionellae can exist in a 24-h hot water bath (24HHWB), which has been used recently in Japan, whether longer use of the 24HHWB causes legionellosis is unclear. The present longitudinal study was conducted in 2000 to investigate the 3-yr change in antibody titers in association with the continuous use, non-use, or canceling the use of the 24HHWB, and possible factors relating to the antibody changes. Ninety-two subjects (85 males and 7 females), who had had their anti-Legionella pneumophila (Lp) serum antibody titers measured in our initial study in 1997 and consented to blood sampling 3 yr later, were selected as subjects. There were no clinical cases who had experienced Legionnaires' disease or Pontiac fever during the 3 yr. The continuous users showed no significant changes in antibody titers within 3 yr, whereas the continuous non-users had a significant increase in antibody titers against the Lp serogroup (SG) 5 and 6. Eleven ex-users of the 24HHWB showed a significant decrease in antibody titers against Lp SG 6. The changes in the 24HHWB use, job sector, stress coping strategies, and alcohol-drinking habit were associated with the changes in antibody titers against Lp SG 1, 5 or 6. The anti-Lp antibodies were considered to be IgM dominant. In conclusion, this study indicates that 24HHWB use by healthy subjects does not tend to result in a higher onset risk of legionellosis, even if it is continuously used for 3 yr, although 24HHWB use is likely to induce production of antibodies against legionellae.

Adult↗
PubMed2004

Anthropometric, lifestyle and biomarker assessment of Japanese non-professional ultra-marathon runners.

BACKGROUND: Anthropometric characteristics, lifestyle, and baseline biological markers of Japanese non-professional ultra-marathon runners have not been fully assessed. METHODS: We evaluated anthropometric characteristics, lifestyle, and baseline biological markers of 180 Japanese amateur ultra-marathon runners (144 males [mean age: 50.5 +/- 9.4 (standard deviation) years] and 36 females [48.9 +/- 6.9]), and compared them with those of participants in a community heath check-up program and with the figures in the literature. We furthermore evaluated baseline blood indices according to monthly running distance with analysis of variance adjusted for age, body mass index, smoking and alcohol drinking habits. RESULTS: The ultra-marathon runners demonstrated more favorable values for body mass index and bone density, and the proportion of smoking, and undertaking physical activity (for both sexes), eating breakfast (for males), and having daily bowel movements (for females), while greater proportion of alcohol drinking habit (for both sexes), than the comparison group. Average monthly running distances and standard deviations (km) were 257.2 +/- 128.9 for males and 209.0 +/- 86.2 for females. Male runners possessed beneficial markers, including lowered triglyceride and elevated high-density lipoprotein cholesterol, and their values showed hockey-stick (or inverse hockey-stick) patterns depending on their monthly running distance. Some subjects running more than 300 km/month exhibited signs of an over-reaching/training syndrome, including somewhat lowered hemoglobin, ferritin and white blood cell count, and elevated creatine kinase and lactate dehydrogenase. CONCLUSIONS: Together with a desirable lifestyle, Japanese non-professional ultra-marathon runners with vigorous exercise habit demonstrated a preferable health status according to biological indices.

Adult↗
PubMed2003

Depressive state relates to female oxidative DNA damage via neutrophil activation.

This study investigated cancer risk due to psychological stress, particularly depression, and its underlying mechanism, using a biomarker of cancer-related oxidative DNA damage, 8-hydroxydeoxyguanosine (8-OH-dG), in human leukocytes. We performed a cross-sectional study of 156 workers in which we examined the relationships of the 8-OH-dG levels to the scores of the General Health Questionnaire (GHQ) and the Center for Epidemiologic Studies Depression scale (CES-D). We also examined the possibility that 8-OH-dG synthesis might be associated with a stress-related increase in neutrophils, which have been reported to produce superoxide in response to psychological stress. The Severe depression scores of the GHQ in females were strongly and positively associated with the 8-OH-dG levels, whereas the CES-D scores in males were negatively associated. There was a positive correlation between the percentage of neutrophils and the 8-OH-dG levels in females. Psychological depression was related to cancer risk due to oxidative DNA damage in females, possibly via neutrophil activation.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed2003

Expressions of two adenomatous polyposis coli and E-cadherin proteins on human colorectal cancers.

Mutations in the adenomatous polyposis coli (APC) gene contribute to the progression of colorectal tumorigenesis. Despite the importance, few studies regarding the localization of this protein on surgically resected human colorectal cancer specimens using immunohistochemistry have been reported so far because of the unavailability of the antibodies for this use. The goal of this study has been to provide the APC protein expression and to validate the APC molecular studies. We took advantage of an immunohistochemistry procedure of applying the unique detergent-mediated antigen retrieval technique to frozen sections and examined the expressions of one amino (N)-terminal (AC4) and one carboxy (C)-terminal APC antibody (HG2). Further, we compared the stainings of APC antibodies with those of the E-cadherin antibody using a quantitative image analysis. E-cadherin is a critical morphogenetic regulator during embryogenesis and recent evidence strongly suggests that downregulation of E-cadherin expression in cancers is associated with a high rate of invasion and metastasis. The analysis indicated statistically that normal epithelia showed stronger staining than cancer cells ( P<0.05). Further, in normal epithelia, the amino (N)-terminal APC antibody (AC4) showed a positive correlation with another carboxy (C)-terminal APC antibody (HG2). E-cadherin showed no positive correlation with other APCs in either the normal epithelia or cancer cells. This study verified reduced expressions of APCs and E-cadherin proteins in colorectal cancer cells. This suggests that the normal APC and E-cadherin protein expressions in benign epithelium are progressively and independently lost in the sporadic colorectal cancers.

Adenomatous Polyposis Coli Protein↗
PubMed2003

Amino acid production processes.

With the exploitation of new uses and the growing markets of amino acids, amino acid production technology has made large progress during the latter half of the 20th century. Fermentation technology has played crucial roles in this progress, and currently the fermented amino acids represent chief products of biotechnology in both volume and value. This area is highly competitive in the world market and process economics are of primary importance. For cost-effective production, many technologies have been developed to establish high-productive fermentation and recovery processes. The producer organisms used in large-scale, well-established processes have been developed to a high level of production efficiency. The tools of genetic engineering of amino acid-producing organisms have been well developed and are now being applied for enlargement of biosynthetic and transport capacity, which is beginning to have a great impact on the amino acid industry. Furthermore, the rapid strides in genome analysis are bound to revolutionize the strain improvement methodology.

Amino Acids↗
PubMed2003

Seasonal variation in consumption and plasma concentrations of fatty acids in Japanese female dietitians.

OBJECTIVE: To study seasonal variation in intake and plasma concentrations of fatty acids (FAs) in Japanese female dietitians. SUBJECTS AND METHODS: We assessed consumption of FAs based on four season 7 consecutive day weighed diet records from 71 Japanese female dietitians in 1996-1997. Using overnight fasting venous blood, plasma concentrations of FAs were analyzed by gas chromatography. Seasonal variation in consumption and plasma concentrations was examined by ANOVA for repeated values, followed by Tukey's multiple t-test. We calculated Spearman's partial rank correlation coefficients (CCs) between intake and plasma concentrations of FAs. Furthermore, we computed inter-seasonal Spearman's partial rank CCs for consumption and plasma concentrations of FAs. RESULTS: Statistically significant seasonal differences were observed in consumption for most FAs, except for myristic acid, monounsaturated FAs, oleic acid, n-6 polyunsaturated FAs (PUFAs), linoleic acid, gamma-linolenic acid, alpha-linolenic acid, PUFAs/saturated FAs, and n-6 PUFAs/n-3 PUFAs, and for most plasma concentrations, except for stearic acid, gamma-linolenic acid, n-3 PUFAs, alpha-linolenic acid, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and n-3 highly unsaturated FAs (HUFAs). However, statistically significant Spearman's partial rank CCs between intake and plasma concentrations were observed for EPA, DHA, n-3 HUFAs, n-6 PUFAs/n-3 PUFAs and n-6 PUFAs/n-3 HUFAs for almost all seasons. CONCLUSIONS: Seasonal variation exists in consumption and plasma concentrations of FAs, so that this should be taken into account in epidemiological analyses, including case-control and cohort studies.

Adult↗