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Hiroyuki Fujimoto

Publications and source records attributed to Hiroyuki Fujimoto.

29 records · Page 2Linked to original sources

Hypermethylation of an E-cadherin (CDH1) promoter region in high grade transitional cell carcinoma of the bladder comprising carcinoma in situ.

PURPOSE: We elucidated the role of methylation in the promoter region of the 1 gene in bladder carcinogenesis, particularly in those comprising carcinoma in situ. MATERIALS AND METHODS: A total of 49 cases of transitional cell carcinoma of the bladder obtained from transurethral resection were examined. Methylation status of the 1 promoter region was analyzed by methylation specific polymerase chain reaction from chemically modified DNA after Na-bisulfite treatment. Loss of heterozygosity on 16q was examined by blunt end single strand DNA conformation polymorphism using 4 tetranucleotide repeat microsatellite markers assigned on 16q13 to 22.1. E-cadherin expression was evaluated by immunostaining on formalin fixed, paraffin embedded tissue sections using anti E-cadherin murine monoclonal antibody, HECD1 and standard avidin-biotin immunoperoxidase complex technique. RESULTS: Analysis of the 49 bladder transitional cell carcinoma samples showed 1 promoter methylation in 23 (47%). Methylation of the 1 gene did not correlate with tumor stage (p = 0.2097) but with high grade transitional cell carcinoma (p = 0.0416). 1 promoter methylation was observed at a significantly higher frequency in the carcinoma in situ positive group than in the carcinoma in situ negative group (16 of 18 cases or 89% versus 7 of 31 or 23%, p <0.0001) and it strongly correlated with abnormal E-cadherin expression (p <0.0001). We found 16q loss of heterozygosity in 16 of 47 cases (34%), which correlated with higher histological grade (p = 0.0069) but not with the presence of the carcinoma in situ component (p = 0.1235). CONCLUSIONS: This study showed that 1 gene promoter methylation is strongly associated with bladder transitional cell carcinoma comprising carcinoma in situ.

Alleles↗

Automated analysis of differentiation-induced leukemic cells during all-trans retinoic Acid therapy of acute promyelocytic leukemia.

During differentiation-induction therapy of acute promyelocytic leukemia (APL) patients with all-trans retinoic acid (ATRA), a variety of APL-derived bizarre granulocytic cells appear in the peripheral blood. To evaluate the differentiation induction of leukemic cells, we have developed a new scattergram analyzing program with an automated hematology analyzer and compared the data with the flow cytometry measuring the expression of differentiation-associated cell surface antigens, CD11b and CD16. We used the fluorescence intensity and side scatter as parameters of granulocytic maturation in the analysis with the automated hematology analyzer. The analysis of 2 ATRA-treated APL patients and in vitro study using HL-60 cells demonstrated that the levels of fluorescence intensity and side scatter decreased as accompanied with granulocytic maturation, and these changes were parallel with the results of flow cytometry. Our automated scattergram analysis of cell differentiation will contribute to general, objective, and real-time evaluation of differentiation-induction therapy of APL with ATRA.

Adult↗

Evaluation of immature granulocyte counts by the XE-IG master: upgraded software for the XE-2100 automated hematology analyzer.

We evaluated an automated immature granulocyte (IG) count in the peripheral blood with the XE-IG Master (Sysmex Corporation). The XE-IG Master is the upgraded software package for the XE-2100 automated hematology analyzer. Reproducibility tests demonstrated a mean coefficient of variation of 7.02% for the IG percentage (IG%) and 6.93% for the absolute IG count. Correlations of the IG counts were assessed in two ways. A flow cytometric IG count using CD11b, CD16, and CD45 monoclonal antibodies and a manual differential count were used as reference methods. The regression equation and the correlation coefficient of the IG% for the flow cytometric reference count versus results with the XE-IG Master were: y = 0.91x + 0.10; r = 0.96. For the comparison with the manual differential count of promyelocytes, myelocytes, and metamyelocytes, the regression equation and correlation coefficient were: y = 0.81x + 1.27; r = 0.78. Samples were found to be stable up to 48 hours both at room temperature and when refrigerated. We investigated the clinical significance of the IG count as a new marker of acute inflammation. In this preliminary study, most samples with a high IG count had positive values for C-reactive protein and the erythrocyte sedimentation rate (positive sample rates were 84.0% and 95.0%, respectively) despite neutrophil counts within the normal range. Elevated IG counts correlated most closely with CD64 expression on polymorphonuclear cells and less so with the concentration of interleukin 6. Compared with other available inflammation markers, the IG count is rapidly generated with each full blood count at no extra cost and with no delay in sample analysis.

Acute Disease↗

Prognostic impact of extensive parenchymal invasion pattern in pT3 renal pelvic transitional cell carcinoma.

BACKGROUND: Pathologic T3 renal pelvic transitional cell carcinoma exhibits various patterns of invasion. The authors investigated the prognostic impact of three patterns of invasion of pT3 renal pelvic transitional cell carcinoma. METHODS: Of 212 patients who underwent surgery for renal pelvic transitional cell carcinoma, 70 with pT3 disease were eligible for the main analyses. The candidate predictors of prognosis included patient age, gender, lesion laterality, tumor grade, perioperative cisplatin-based systemic chemotherapy, lymph node involvement, vascular involvement, and patterns of invasion. Invasion patterns were classified as fat invasion, ductal involvement, or parenchymal invasion. RESULTS: Mean postoperative followup was 33.5 months (range, 1-136 months). On univariate analysis, gender, lymph node involvement, vascular involvement, and extensive parenchymal invasion each had a significant impact on the cause specific survival rate. A multivariate analysis using Cox stepwise regression revealed that extensive parenchymal involvement was the strongest prognostic predictor (P = 0.0004, hazard ratio = 5.59). Lymph node involvement (P = 0.0175, hazard ratio = 3.14) and gender (P = 0.0361, hazard ratio = 2.42) were other weaker predictors. Statistically, pT3 disease without extensive parenchymal invasion had a prognosis similar to that of lower stage disease, and pT3 disease with extensive parenchymal invasion had a prognosis similar to that of pT4 disease. CONCLUSIONS: Extensive parenchymal invasion has a strong prognostic impact in renal pelvic transitional cell carcinoma. pT3 disease should be subclassified into two separate entities, that with and that without extensive parenchymal invasion, in view of prognosis.

Adult↗

Medically and economically appropriate follow-up schedule for prostate cancer patients after radical prostatectomy.

BACKGROUND: Our goal was to determine the optimal frequency and method of follow-up after radical prostatectomy to minimize medical cost without adversely affecting patients. METHODS: Two hundred and twenty-one patients who underwent a radical prostatectomy with or without adjuvant androgen deprivation from 1989 to 1999 were selected for the study. Eighty percent of the patients received postoperative androgen deprivation. Tumor recurrence was strictly defined as detectable serum prostate specific antigen (PSA) and/or clinical findings such as local tumor detection or bone metastasis. Thirty of 221 patients experienced tumor recurrence. Risk of tumor recurrence, procedures for detection of recurrence, and PSA doubling time after biochemical failure were analyzed. RESULTS: None of the 30 patients who were examined showed definitive local recurrence or metastatic sites on the imaging study at the time of initial PSA detection, and there were no observed recurrences in the absence of detectable serum PSA. In patients who showed elevated PSA within 12 months after radical prostatectomy, PSA levels rapidly increased with doubling times ranging from 1.2 to 13.7 months. Excluding those patients, the doubling time of PSA levels ranged from 2.8 to 31.5 months. CONCLUSIONS: Prostate specific antigen screening is sufficient to detect treatment failure after radical prostatectomy, irrespective of adjuvant hormone therapy. Based on the calculated doubling time, the longest advisable interval between checks of PSA levels is estimated to be four months within the first year after radical prostatectomy, and biannually or annually thereafter. Continuously elevated PSA levels or clinical symptoms indicate surveys for local recurrences and distant metastases.

Aged↗

Immature granulocyte count after liver transplantation.

We evaluated the significance of immature granulocyte (IG) count during the clinical course after liver transplantation. We counted IG using the flow cytometric method with CD16, CD11b, and CD45 antibodies. Samples were obtained from 31 patients in the Department of Transplantation and Immunology, and we determined (i) the distribution of IG peak value, (ii) the distribution of IG peak time-points, (iii) the clinical background of patients with high IG, and (iv) the clinical course of high IG cases. We observed the appearance of IG (100/microl or higher) in the majority of the patients (23 out of 31 patients; 74.2%). The IG peak was detected on the 19th day after transplantation. We observed serious complications, such as melena, rejection, or severe infection, in high IG (500/microl or higher) cases. We observed instances of inflammation with low C-reactive protein (CRP) value in the presence of IG. We believe that IG is a useful marker to monitor inflammation.

Biomarkers↗

Total Syntheses of Carazostatin, Hyellazole, and Carbazoquinocins B-F.

Total syntheses of carazostatin (1), hyellazole (2a), and carbazoquinocins B-F (3b-f) have been completed. The cross-coupling reaction between 3-iodoindole 8 and vinylstannane 11b gave the 3-alkenylindole 7. Treatment of 7 with ethynylmagnesium bromide, followed by etherification of the resulting alcohol 12 with MOMCl, yielded the 3-alkenyl-2-propargylindole 6. The compound 6 was treated with t-BuOK in t-BuOH at 90 degrees C to obtain the desired carbazoles 4 together with the N-deprotected carbazole 13 through an allene-mediated electrocyclic reaction. The carbazole 13a, derived from 4a or 4c, was converted into the triflate 24 in two steps. The triflate 24 was subjected to the Suzuki cross-coupling reaction with either 9-heptyl-9-BBN or phenylboronic acid in the presence of a palladium catalyst to produce the 1-heptylcarbazole 25a and the 1-phenylcarbazole 25b. Cleavage of the ether bond of 25a yielded carazostatin (1). Cleavage of the ether bond of 25b followed by O-methylation gave hyellazole (2a). Oxidation of carazostatin (1) with benzene seleninic anhydride afforded carbazoquinocin C (3c). In a similar way, carbazoquinocins B and D-F (3b,d-f) were synthesized, respectively.

Journal Article↗

Fabrication of cell-based arrays using micropatterned alkanethiol monolayers for the parallel silencing of specific genes by small interfering RNA.

The emergence of small interfering RNA (siRNA) opened a new opportunity to study gene functions in a genome. However, large-scale loss-of-function analyses further require cell-based high-throughput methods that allow simultaneous silencing of the huge number of genes by siRNA. In this study, we aim at fabricating the cell-based siRNA arrays that facilitate parallel introduction of multiple siRNAs into cultured mammalian cells. The siRNA arrays were prepared using surface chemical processes including the micropatterning of a self-assembled monolayer and the layer-by-layer assembly of siRNA and cationic lipid. We examined the feasibility of the siRNA array for the sequence-specific gene silencing in an array format. Furthermore, the effects of siRNA loading and culture period after transfection were studied to optimize cell-based assays on the siRNA arrays. The results obtained in this study demonstrated that our method provides the siRNA arrays with spatial specificity in gene silencing, which will serve to obtain a quantitative data set from the cell-based screens on siRNA arrays.

Base Sequence↗