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The current paper empirically addresses risk aversion of households and firms toward earthquake risks using a hazard map compiled for the entire region by the Tokyo metropolitan government in 1998. It finds strong evidence for the impact of earthquake risks on land pricing; land prices have been substantially lower in risky areas than in safe areas. That impact became more evident in the 1990s than in the 1980s, indicating that households and firms were becoming more sensitive to earthquake risks. In addition, this paper carefully examines the consistency of the estimated magnitude of earthquake risk premiums within a framework of the expected utility hypothesis.

Objectives: Diagnosing the bladder lesions associated with interstitial cystitis/painful bladder syndrome (IC/PBS) is sometimes difficult for general urologists. We therefore aimed to develop an IC/PBS diagnosis method using a cystoscope with a narrow-band imaging (NBI) system that can detect mucosal angiogenic lesions. Methods: Fifty-two subjects suspected of having IC between October 2006 and June 2007 were included in this study. There were 49 women and three men, ranging in age from 19 through 85 with an average age of 59. First, conventional cystoscopy under spinal anesthesia was performed to examine the ulcerative lesions by a urological specialist. Then, other health care professionals made a separate observation of capillary-rich areas of the superficial layer of the bladder mucosa by cystoscopy with the NBI system. Results: Among the 52 patients, 37 cases were found to have ulcers by conventional cystoscopy, which were also recognized as capillary-rich brownish areas using the NBI system (100% accuracy); 13 cases were found to have NBI-positive areas without ulcer, which were coincided with those with petechial hemorrhages and glomerulations following subsequent hydrodistention; and two cases of normal mucosa were detected. Furthermore, six cases of bladder cancer (carcinoma in situ) were detected by biopsies that were obtained from the ulcerative lesions positively identified by NBI cystoscopy. Conclusions: Examining the urinary bladder mucosa with a flexible cystoscope with the NBI system makes it possible to easily detect ulcers of bladder mucosa and areas with angiogenesis. Therefore, it is considered that the use of a flexible cystoscope with the NBI system is highly practical for the IC/PBS diagnosis.

5-FU is commonly used for treatment of various solid tumors including colon carcinoma. We have previously demonstrated that Egr-1 induced by 5-FU enhanced TSP-1 expression in human colon cancer KM12C cells. In this study, a Genechip analysis of KM12C cells treated with 5-FU revealed down-regulation of 924 genes and up-regulation of 460 genes. The decreased expression of c-Myc mRNA and phosphorylated c-Myc were detected and confirmed by RT-PCR and immunoblotting. Since 5-FU induced the expression of TSP-1, we examined the effect of c-Myc on the TSP-1 promoter. Deletion of the TSP-1 promoter region in which binding sites for c-Myc reside had no effect on the TSP-1 promoter activity induced by 5-FU. Meanwhile, 5-FU dose-dependently decreased the expression of miR-17-92 cluster. These findings suggest that 5-FU decreased the expression of c-Myc and consequently miR-17-92 cluster and increased the expression of TSP-1 mRNA.

5-Fluorouracil (5-FU) is one of the most commonly used anticancer drugs in chemotherapy against various solid tumors. 5-FU dose-dependently increased the expression levels of intrinsic antiangiogenic factor thrombospondin-1 (TSP-1) in human colon carcinoma KM12C cells and human breast cancer MCF7 cells. We investigated the molecular basis for the induction of TSP-1 by 5-FU in KM12C cells. Promoter assays showed that the region with the Egr-1 binding site is critical for the induction of TSP-1 promoter activity by 5-FU. The binding of Egr-1 to the TSP-1 promoter was increased in KM12C cells treated with 5-FU. Immunofluorescence staining revealed that 5-FU significantly increased the level of Egr-1 in the nuclei of KM12C cells. The suppression of Egr-1 expression by small interfering RNA decreased the expression level of TSP-1. Furthermore, 5-FU induced the phosphorylation of p38 mitogen-activated protein kinase (MAPK) and heat shock protein 27 (HSP27). Blockade of the p38 MAPK pathway by SB203580 remarkably inhibited the phosphorylation of HSP27 induced by 5-FU and decreased the induction of Egr-1 and TSP-1 by 5-FU in KM12C cells. These findings suggest that the p38 MAPK pathway plays a crucial role in the induction of Egr-1 by 5-FU and that induced Egr-1 augments TSP-1 promoter activity, with the subsequent production of TSP-1 mRNA and protein. [Cancer Res 2008;68(17):7035–41]

Bladder cancer (BC) is the second most common malignancy of the genitourinary tract and the second leading cause of cancer death in patients with urinary tract malignancies. DNA methylation and histone modifications are important epigenetic mechanisms of gene regulation and play essential roles both independently and cooperatively in tumor initiation and progression. Aberrant epigenetic events such as DNA hypermethylation and altered histone acetylation have both been observed in bladder cancer, in which they affect a large number of genes. Although the list of aberrantly epigenetically regulated genes continues to grow, combination analysis including several candidate genes has given promising results of potential tumor biomarkers for the early diagnosis and risk assessment of bladder cancer. Thus, large-scale screening of aberrant epigenetic events such as DNA hypermethylation is needed to identify bladder cancer-specific epigenetic fingerprints. The reversibility of epigenetic aberrations has made them attractive targets for cancer treatment with modulators that demethylate DNA and inhibit histone deacetylases, leading to the reactivation of silenced genes. In this review, we examine the current literature on epigenetic changes in bladder cancer and discuss the clinical potential of cancer epigenetics for the diagnosis and treatment of this disease.

Secreted frizzled-related protein 2 (sFRP2) is a negative modulator of the Wingless-type (Wnt) signaling pathway, and shown to be inactivated in renal cell carcinoma (RCC). However, the molecular mechanism of silencing of sFRP2 is not fully understood. Our study was designed to elucidate the silencing mechanism of sFRP2 in RCC. Expression of sFRP2 was examined in 20 pairs of primary cancers by immunohistochemistry. Kidney cell lines (HK-2, Caki-1, Caki-2, A-498 and ACHN) were analyzed for sFRP2 expression using real-time RT-PCR and Western blotting. The methylation status at 46 CpG sites of the 2 CpG islands in the sFRP2 promoter was characterized by bisulfite DNA sequencing. Histone modifications were assessed by chromatin immunoprecipitation (ChIP) assay using antibodies against AcH3, AcH4, H3K4 and H3K9. sFRP2 was frequently repressed in primary cancers and in RCC cells. The majority of sFRP2 negative cells had a methylated promoter. Meanwhile, sFRP2 expression was repressed by a hypomethylated promoter in Caki-1 cells, and these cells had a repressive histone modification at the promoter. In Caki-1 cells, sFRP2 was reactivated by trichostatin A (TSA). Repressive histone modifications were also observed in RCC cells with hypermethylated promoters, but sFRP2 was reactivated only by 5-aza-2'-deoxycytidine (DAC) and not by TSA. However, the activation of the silenced sFRP2 gene could be achieved in all cells using a combination of DAC and TSA. This is the first report indicating that aberrant DNA methylation and histone modifications work together to silence the sFRP2 gene in RCC cells.

A disintegrin and metalloproteases (ADAM) are cell membrane-anchored proteins with potential implications for the metastasis of human cancer cells via cell adhesion and protease activities. In prostate cancer (PC), the ADAM-10 protein showed a nuclear localization whereas in benign prostate hypertrophy (BPH) it was predominantly bound to the cell membrane. We hypothesized that the pathogenesis and progression of PC are attributable to the nuclear translocation of ADAM-10. Immunoblotting revealed that after 5alpha-dihydrotestosterone treatment, a 60-kDa active form of ADAM-10 was increased in the nuclear fraction but decreased in the cell membrane and cytoplasmic fractions of human androgen-dependent PC cells. Immunocytochemistry revealed that after 5alpha-dihydrotestosterone treatment, the ADAM-10 protein was translocated from the cell membrane to the nucleus. Coimmunoprecipitation of androgen receptor and ADAM-10 was detected in the nuclear fraction but not in the cell membrane and cytoplasmic fractions. Immunohistochemical study of 64 PC and 20 BPH samples showed that the intensity of ADAM-10 staining was significantly higher in the nuclei of PC cells than in the nuclei of BPH cells (P < 0.0001). It was also significantly lower in the cell membrane of PC cells than in the cell membrane of BPH cells (P = 0.0017). Nuclear staining intensity was significantly correlated with the clinical T-factor (P = 0.004), the Gleason score (P < 0.0001) and preoperative prostate-specific antigen levels (P = 0.0061). ADAM-10 small interfering RNA transfectants showed a significant decrease in cell growth compared to the controls. Our results suggest that in human PC, the nuclear translocation of ADAM-10 coupled with the androgen receptor is involved in tumor growth and progression.

SKP2 and CKS1 promote aggressive tumor behavior via the regulation of p27 degradation. Our previous DNA microarray analysis of human urothelial carcinoma and normal urothelial epithelium showed that in urothelial carcinoma the 2 most highly up-regulated genes among SKP2-p27 interaction related genes are SKP2 (4.7-fold) and CKS1 (2.2-fold). We hypothesized that SKP2 and CKS1 gene expression is associated with urothelial carcinoma invasiveness and prognosis.A total of 84 urothelial carcinoma specimens from patients with bladder (71) and upper urinary tract (13) cancer were examined by real-time reverse transcriptase-polymerase chain reaction and immunohistochemical study.Real-time reverse transcriptase-polymerase chain reaction showed that the average mRNA expression level of SKP2 and CKS1 significantly correlated with tumor stage, that is superficial vs invasive urothelial carcinoma (SKP2 and CKS1, p0.001 and 0.006) and grade (p0.001 and 0.009, respectively). Of the superficial urothelial carcinomas examined the SKP2 and CKS1 expression level was significantly higher in pT1 than in pTa samples (p=0.005 and 0.017, respectively). Immunohistochemical expression patterns of SKP2 and CKS1 also significantly correlated with tumor stage (p0.001 and 0.048) and grade (p=0.003 and 0.025, respectively). In contrast, p27 expression inversely correlated with tumor stage and grade (p0.001 and 0.011, respectively). Logistic regression analysis revealed that while SKP2 mRNA expression was a significant dependent predictor of p27 expression (p=0.021), there was no correlation between CKS1 mRNA expression and p27 (p=0.748). Kaplan-Meier curves and log rank tests revealed that the high mRNA expression levels of SKP2 and CKS1 had a significant adverse effect on prognosis (p=0.043 and 0.003, respectively).Our results suggest that SKP2 has a major role in the regulation of p27 degradation and CKS1 has a supporting role for SKP2 function in human urothelial carcinoma.

It is important to have information on cavernous sinus extension and bony destruction in pituitary macroadenomas before surgery, but magnetic resonance (MR) imaging cannot always depict them. In the present study we sought to determine whether multidetector-row computed tomography (MDCT) could provide preoperative information in addition to that provided by MR imaging in pituitary macroadenoma.The subjects comprised 33 consecutive patients (15 women, 18 men; mean age 50 years) with surgically proven macroadenoma. For MDCT, using the soft-tissue window and bone window, three orthogonal multiplanar reconstruction images were generated from venous-phase contrast-enhanced 0.5-mm isotropic voxel data. MDCT and MR images were evaluated with regard to: (1) clarity of tumor margins; (2) identification of the normal pituitary gland; (3) identification of erosion or destruction of the sellar floor; and (4) visualization of the adjacent optic pathways.MDCT more clearly demonstrated the lateral tumor margin than MR imaging (P = 0.002). No significant differences in visualization of the normal pituitary gland were noted between MDCT and dynamic MR imaging (P = 0.7). MDCT more clearly demonstrated sellar floor erosion or destruction at the sphenoid sinus than MR imaging (P0.001). MR imaging was superior to MDCT for visualizing the adjacent optic pathways (P0.001).MDCT is superior to MR imaging for assessing lateral tumor margin and the sellar floor at the sphenoid sinus. MDCT offers useful preoperative information in addition to that obtained from MR imaging.

Este trabajo tuvo como objetivo comprender si la accountability es, o no, la razón de ser de la Contabilidad. Si la respuesta es positiva y correctamente comprendida por los maestros, investigadores y profesionales de la Contabilidad, podremos alimentar la esperanza de que nuestro estatus social, político y económica alcanzará brevemente el nivel deseado por todos nosotros, en la misma medida en la que el concepto de accountability sea aplicado en nuestro País, con due diligence. Las pesquisas que vienen siendo realizadas hace más de 15 (quince) años en la Fundación Instituto de Investigaciones Contables, Actuarias y Financieras (Fipecafi) vinculada a la FEA/USP, revelan que, en el mundo de los negocios públicos y privados, existen básicamente dos formas de accountability, de ellas es de naturaleza exógena y otra endógena. Las dos formas de accountability aportan para la comprensión del concepto de plena accountability a que se refiere el modelo de gobierno empresarial de la International Federation of Accountants (Ifac). La pesquisa de campo que, en el inicio de los años 90, fue objeto de una consulta de la directoria para América del Sur y Caribe del Banco Mundial a la Fipecafi, con el objetivo de medirse el nivel de accountability en Brasil, tanto en el sector público como privado, es algo que aún permanece en abierto, como desafío para los porvenires encuestadores de esa tema.

Cavitation bubble activity caused by ultrasonic irradiation in water was applied to the hard surface such as carburized steel and TiN-coated on WC-super hard alloy to investigate whether it is possible or not to introduce residual compressive stress on the hard surface. The main results obtained in this study are as follows.In the case of carburized steel with hardness of 700HV, the large residual compressive stress of approximately 820MPa was successfully introduced on the surface by the ultrasonic irradiation in water. Furthermore, it was revealed that the ultrasonic irradiation in water successfully introduced the large residual compressive stress of approximately 2400MPa on the surface of TiN-ceramic coated on WC-super hard alloy of which the initial surface residual compressive stress was about 1800MPa. The redundant application of the ultrasonic irradiation on the TiN ceramic surface after the residual stress reached the maximum caused not the variation in the surface roughness and micro-hardness but a little damage such as very small pits on the surface.

insulin signaling in the progression of a murine RCC model using metformin to treat hyperinsulinemia induced by a high-carbohydrate diet. METHODS: pT1-T4 tumor specimens from 99 patients with RCC were evaluated for IR expression by immunohistochemistry. Serum insulin and C-peptide levels were measured. The association between biomarker expression and serum concentration, prognostic factors, and clinical outcomes were assessed. C57BL/6 mice were randomized into four groups: low-carbohydrate diet, high-carbohydrate diet, low-carbohydrate diet þ metformin, and high-carbohydrate diet þ metformin groups. RENCA cells were injected subcutaneously. Body weight, glucose tolerance test, serum insulin level, and tumor volume were evaluated. Immunoblotting was performed to assess the insulin signaling pathway in the murine RCC model. RESULTS: The IR expression level was significantly lower in patients with tumor stage pT2-4 and with a preoperative serum Cpeptide level more than 5.14 ng/mL. High IR expression was an independent predictor of favorable progression-free survival after radical nephrectomy. In vivo progression of RENCA in the murine model was not significantly stimulated by hyperinsulinemia induced by a high-carbohydrate diet. However, in vivo progression of RENCA was significantly inhibited by metformin in both the lowand highcarbohydrate diet groups. IR expression was significantly higher in the tumors from the low-carbohydrate diet group and high-carbohydrate diet plus metformin group than that from the high-carbohydrate diet group. CONCLUSIONS: IR expression in RCC was inversely associated with cancer progression and serum insulin levels in both clinical and murine models. The anticancer effect of metformin in the murine RCC model might be derived from the direct effect of the agent on cancer cells rather than the effect of the lower insulin level. The lower expression of IR in the tumor from patients with a poor outcome might be the result of a higher serum insulin level, which is not strong cause of cancer progression.

and mean photon counts of bone metastases were measured using the IVIS imaging systemx (Xenogen, Alameda, CA, USA). RESULTS: Two weeks after castration, bone mineral density in the castration group was significantly lower than that in the noncastration group and the castration þ OPG group (p 1⁄4 0.034 and p 1⁄4 0.010, respectively). The number of mice with bone metastases in the castration group was significantly higher than that in the noncastration group and the castration þ OPG group (p 1⁄4 0.024 and p 1⁄4 0.028, respectively). The mean number of bone metastases and the mean photon counts of bone metastases in the castration group was significantly higher than those in the castration þ OPG group (p 1⁄4 0.020 and p 1⁄4 0.041, respectively). CONCLUSIONS: Castration reduced bone mineral density and accelerated bone metastasis in the murine CRPC model. OPG prevented both bone mineral loss and bone metastasis. The castration-induced acceleration of bone metastasis in CRPC could be caused by activation of osteoclasts due to RANK signaling. These results suggested that it is necessary to inhibit RANK signaling from the beginning of ADT to prevent bone metastasis in prostate cancer patients.

INTRODUCTION AND OBJECTIVES: Generally in microRNA (miRNA) biogenesis, one strand (guide strand) of the duplex may potentially act as a functional miRNA and incorporated into the RNA-induced silencing complex and regulating target genes. In contrast, other strand (passenger strand: miRNA*) is disintegrated quickly. Our miRNA expression signature of bladder cancer (BC) by using deep-sequencing revealed that tumor-suppressive miRNA-145 and its passenger strand of miRNA-145* were significantly reduced in cancer tissues, suggesting these function as tumor suppressors in BC cells. The aim of the study was to investigate functional significance of miRNA-145* and its regulated molecular targets/ pathways in BC. METHODS: We evaluated the expression levels of miR-145/ miR-145* in 43 BC clinical specimens and 12 normal bladder epithelia by real-time PCR. The functional analysis of the miRNA and molecular targets/pathways searching strategies were described as our previous studies (J Urol,192:1822-1830,2014; FEBS Lett,588:3170-3179,2014; PLoS One,9:e84311,2014). RESULTS: Expression levels of miR-145 and miR-145* were significantly lower than that in normal bladder epithelia (P < 0.0001). Gain-of-function studies revealed that cell proliferation, migration, and invasion were significantly inhibited in both miR-145 and miRNA-145* transfection into BC cell lines (each, P < 0.001). Combination of gene expression analysis of miR-145*-transfectants and in silico analysis showed that several oncogenic genes (HK2, YES1, FOSL1 etc.) were identified as miR-145* regulation in BC cells. CONCLUSIONS: Our genome-wide miRNA expression signature showed that miR-145* as a tumor-suppressive miR-145 passenger strand frequently reduced in BC cells and acts as a tumorsuppressor. Elucidation of miR-145/145*-mediated novel cancer pathways provide new insights of the potential mechanisms of BC oncogenesis. Furthermore, tumor-suppressive miR-145/145* might be contributed to development of miRNA-based cancer therapeutics in the disease.

Here, we found that members of the microRNA-29 family (miR-29a/b/c; “miR-29s”) were significantly reduced in clear cell renal cell carcinoma (ccRCC) tissues, suggesting that they functioned as tumour suppressors. Restoration of all mature members of the miR-29 family inhibited cancer cell proliferation, migration and invasion. LOXL2 was a direct target gene of miR-29s, as shown by genome-wide gene expression analysis and luciferase reporter assay. Overexpressed LOXL2 was confirmed in ccRCC clinical specimens, and silencing of LOXL2 inhibited cancer cell migration and invasion in ccRCC cell lines. Our data demonstrated that the miR-29s-LOXL2 axis contributed to cancer cell migration and invasion in ccRCC.

In developed countries, endometrial cancer (EC) is the most common malignancy among women. Unopposed estrogen therapy, obesity, nulliparity, diabetes mellitus and arterial hypertension have been linked to an increased risk of EC. However, the molecular mechanisms of EC oncogenesis and metastasis have not yet been fully elucidated. Our recent studies of microRNA (miRNA) expression signatures revealed that the microRNA-1/133a (miR‑1/133a) cluster is frequently downregulated in various types of human cancers. However, the functional role of the miR‑1/133a cluster in EC cells is still unknown. Thus, the aim of this study was to investigate the functional significance of the miR‑1/133a cluster and its regulated molecular targets, with an emphasis on the contributions of miR‑1/133a to EC oncogenesis and metastasis. We found that the expression levels of miR‑1 and miR‑133a were significantly reduced in EC tissues. Moreover, restoration of mature miR‑1 or miR‑133a miRNAs significantly inhibited cancer cell migration and invasion, suggesting that these clustered miRNAs act as tumor suppressors. Prediction of miRNA targets revealed that phosphodiesterase 7A (PDE7A) was a potential target gene regulated by both miR‑1 and miR‑133a. PDE7A was confirmed to be overexpressed in EC clinical specimens and silencing of PDE7A significantly inhibited cancer cell migration and invasion. Our data demonstrated that downregulation of the miR‑1/133a cluster promoted cancer cell migration and invasion via overexpression of PDE7A in EC cells. Elucidation of the molecular networks regulated by tumor-suppressive miRNAs will provide insights into the molecular mechanisms of EC oncogenesis and metastasis.

Purpose: We hypothesized that combined methylation analysis of Wnt antagonist genes could serve as a panel of biomarkers for diagnosis, staging, and prognosis in renal cell carcinoma (RCC). Experimental Design: Samples (n = 62) of RCC and corresponding normal renal tissue (NRT) were analyzed using methylation-specific PCR for methylation of six Wnt antagonist genes (sFRP-1, sFRP-2, sFRP-4, sFRP-5, Wif-1, and Dkk-3). To increase the sensitivity/specificity of RCC detection, the methylation score (M score) for multigene methylation analysis was developed. Receiver operator characteristic curve analysis was used to determine the optimal sensitivity/specificity of the M score. In addition, the M score was compared with the clinicopathologic outcome. Thirty-three serum DNA samples were also used to investigate the methylation status of Wnt antagonist genes. Results: The methylation levels of all Wnt antagonists were significantly higher in RCC than in NRT. In multivariate regression analysis, the methylation level of sFRP-1 was a significant independent predictor of RCC, whereas for sFRP-2 and sFRP-4 there was a trend toward significance as independent predictors. The M score of Wnt antagonist genes was significantly higher in RCC than in NRT. Overall, the M score had a sensitivity of 79.0% and a specificity of 75.8% (area under the curve, 0.808) as a diagnostic biomarker. In addition, the M score could significantly distinguish grade, pT category, M category, and overall survival of RCC patients. The M score was independent of age and gender in predicting overall survival by the Cox proportional hazards model. In RCC patients, 72.7% of the methylation-specific PCR results had identical methylation in samples of tumor and serum DNA. No serum DNA in normal controls showed aberrant methylation of the Wnt antagonist genes. In addition, the methylation status of Wnt antagonist genes in serum DNA was significantly correlated with tumor grade and stage. Conclusions: This is the first report showing that M score analysis of Wnt antagonist genes can serve as an excellent epigenetic biomarker panel for detection, staging, and prognosis of RCC using serum DNA.

Thymidine phosphorylase (TP) regulates intracellular thymidine metabolism and can enhance the anti-tumor effectiveness of 5′-deoxy-5-fluorouridine (5′-DFUR) by conversion of the pro-drug 5′-DFUR to 5-fluorouracil (5-FU) in tumor tissues. 5′-DFUR is an effective anti-tumor drug in cells expressing high levels of TP. 3′-Azido 3′-deoxythymidine (AZT) is a thymidine analog that has been proven useful in the treatment of acquired immunodefiency syndrome (AIDS). In this study, we found that AZT induces TP expression and enhances the sensitivity of human myeloid leukemia U937 cells to 5′-DFUR. Both the protein level and the activity of TP in U937 cells were elevated for 48 h after exposure to AZT (20, 100 or 300 μM). AZT enhanced TP promoter activity in a dose-dependent manner. AZT also increased TP mRNA levels in U937 cells as assayed by Real-time reverse-transcription PCR. AZT enhanced the cytotoxic effect of 5′-DFUR on U937 cells. A TP inhibitor, TPI, abrogated the cytotoxic activity of 5′-DFUR, and attenuated the combined cytotoxicity of AZT and 5′-DFUR. These results suggest that AZT enhances the cytotoxic effect of 5′-DFUR on U937 cells by upregulating TP activity in addition to its inhibition of thymidine kinase (TK) activity and reduction of intracellular dTTP pools.

Multidrug resistance protein 1 (MRP1) is a human ATP-binding cassette (ABC) transporter in the plasma membrane. It confers multidrug resistance to tumor cells by actively effluxing intracellular drugs. To examine the functional significance of intracellular loops (ICLs) in MRP1, we determined the effect of mutation of the amino acid sequence EXXXG, which is conserved in ICL5 and ICL7 of human MRP1, 2 and 3, sulfonylurea receptor (SUR) 1 and 2, and mouse MRP1 and 2. E and G in the ICLs of human MRP1 were mutated to L and P, respectively, and the N-terminal (including ICL5) and C-terminal (including ICL7) wild type or mutant halves of MRP1 were co-expressed in insect cells. The mutation of either ICL5 or ICL7 considerably decreased ATP-dependent LTC 4 uptake into vesicles of insect cells expressing mutated MRP1. GSH-dependent photolabeling of MRP1 with an 125 I-labeled photoaffinity analog of azido agosterol A (azido AG-A) was abolished by the mutations in ICL5 and ICL7. Mutations in ICL5 of MRP1 almost completely inhibited the labeling of NBD2, but not NBD1, by 8-azido-α-[ 32 P]ATP. In contrast, mutations in ICL7 of MRP1 abolished the labeling of both NBDs. Mutation of either ICL5 or ICL7 of MRP1 almost completely inhibited vanadate trapping with 8-azido-α-[ 32 P]ATP by both NBD1 and NBD2 domains. These findings indicate that the intramolecular signaling between NBD and ICL s in MRP1 is vital for MRP1 function.

BACKGROUND Aberrant CpG methylation profiles of gene promoters and their correlation with advanced pathologic features have been well investigated in prostate carcinoma (PC). Several case–control and prospective studies have revealed a positive association between current smoking and PC. The authors hypothesized that smoking influences both progression and prognosis of PC through CpG hypermethylation of related genes. METHODS A total of 164 PC patients (52 current, 30 former, and 82 never smokers) and 69 benign prostatic hyperplasia (BPH) patients were examined by methylation-specific PCR (MSP) for 3 genes: adenomatous polyposis coli (APC), glutathione S-transferase pi (GSTP1), and multidrug resistance one (MDR1). The methylation status of representative samples was confirmed by bisulfite DNA sequencing analysis. The newly defined methylation score (M-score) of each sample is the sum of the corresponding log hazard ratio (HR) coefficients derived from multivariate logistic regression analysis for pathology (BPH vs. PC), and was related to clinical and pathologic outcome including smoking status. RESULTS The M-score was significantly higher in the current smokers than in never smokers (P = 0.008). Spearman rank correlation test demonstrated a significant correlation between pack-years smoked and M-score in PCs (P = 0.039). Significant correlation of the M-score methylation was observed with high pT category (P < 0.001), high Gleason sum (P < 0.001), high preoperative prostate-specific antigen (PSA) (P = 0.041), and advanced pathologic features. In addition, Gleason sum was significantly associated with PSA failure-free probability as a poor outcome (P = 0.020). CONCLUSIONS This is the first study to demonstrate significant correlation of the methylation status of multigenes with smoking status in PC. Smoking status may influence both progression and prognosis of PC through CpG hypermethylation of related genes. Cancer 2006. © 2005 American Cancer Society.

A global atmospheric general circulation model, with the horizontal grid size of about 20 km, has been developed, making use of the Earth Simulator, the fastest computer available at present for meteorological applications. We examine the model's performance of simulating the present-day climate from small scale through global scale by time integrations of over 10 years, using a climatological sea surface temperature. Global distributions of the seasonal mean precipitation, surface air temperature, geopotential height, zonal-mean wind and zonal-mean temperature agree well with the observations, except for an excessive amount of global precipitation, and warm bias in the tropical upper troposphere. This model improves the representation of regional-scale phenomena and local climate, by increasing horizontal resolution due to better representation of topographical effects and physical processes, with keeping the quality of representation of global climate. The model thus enables us to study global characteristics of small-scale phenomena and extreme events in unprecedented detail.

Promoter hypermethylation is one of the putative mechanisms underlying the inactivation of negative cell-cycle regulators. We examined whether the methylation status of p16(INK4a) and p14(ARF), genes located upstream of the RB and p53 pathway, is a useful biomarker for the staging, clinical outcome, and prognosis of human bladder cancer. Using methylation-specific PCR (MSP), we examined the methylation status of p16(INK4a) and p14(ARF) in 64 samples from 45 bladder cancer patients (34 males, 11 females). In 19 patients with recurrent bladder cancer, we examined paired tissue samples from their primary and recurrent tumors. The methylation status of representative samples was confirmed by bisulfite DNA sequencing analysis. The median follow-up duration was 34.3 months (range 27.0-100.1 months). The methylation rate for p16(INK4a) and p14(ARF) was 17.8% and 31.1%, respectively, in the 45 patients. The incidence of p16(INKa) and p14(ARF) methylation was significantly higher in patients with invasive (>or=pT2) than superficial bladder cancer (pT1) (p=0.006 and p=0.001, respectively). No MSP bands for p16(INK4a) and p14(ARF) were detected in the 8 patients with superficial, non-recurrent tumors. In 19 patients with tumor recurrence, the p16(INK4a) and p14(ARF) methylation status of the primary and recurrent tumors was similar. Of the 22 patients who had undergone cystectomy, 8 (36.4%) manifested p16(INKa) methylation; p16(INK4a) was not methylated in 23 patients without cystectomy (p=0.002). Kaplan-Meier analysis revealed that patients with p14(ARF) methylation had a significantly poorer prognosis than those without (p=0.029). This is the first study indicating that MSP analysis of p16(INK4a) and p14(ARF) genes is a useful biomarker for the pathological stage, clinical outcome, and prognosis of patients with bladder cancer.

MicroRNAs are small non-coding RNAs that negatively regulate gene expression post-transcriptionally. MicroRNA profiles have been studied in various cancers including prostate cancer since 2007. According to these results, microRNA signature in prostate cancer differed from their normal counterparts, suggesting that the differential expression of microRNAs in tumors functionally implicated in their pathogenesis. The up-regulation of oncogenic microRNA and down-regulation of tumor suppressor microRNA are demonstrated in various cancers. The target genes of microRNA identified include those of apoptosis avoidance, cell proliferation, epithelial mesenchymal transition, angiogenic signalling, and generation of androgen independence in prostate cancer. Elucidation of micro RNA function may provide further advance in diagnosis and treatment of prostate cancer.

The incidence and mortality of prostate cancer (PC) is approximately 2-fold higher among African-Americans as compared to Caucasians and very low in Asian. We hypothesize that inactivation of GSTP1 genes through CpG methylation plays a role in the pathogenesis of PC, and its ability to serve as a diagnostic marker that differs among ethnic groups. GSTP1 promoter hypermethylation and its correlation with clinico-pathological findings were evaluated in 291 PC (Asian = 170; African-American = 44; Caucasian = 77) and 172 benign prostate hypertrophy samples (BPH) (Asian = 96; African-American = 38; Caucasian = 38) using methylation-specific PCR. In PC cells, 5-aza-dC treatment increased expression of GSTP1 mRNA transcripts. The methylation of all CpG sites was found in 191 of 291 PC (65.6%), but only in 34 of 139 BPH (24.5%). The GSTP1 hypermethylation was significantly higher in PC as compared to BPH in each ethnic group (p < 0.0001). Logistic regression analysis (PC vs. BPH) showed that African-Americans had a higher hazard ratio (HR) (13.361) compared to Caucasians (3.829) and Asian (8.603). Chi-square analysis showed correlation of GSTP1 hypermethylation with pathological findings (pT categories and higher Gleason sum) in Asian PC (p < 0.0001) but not in African-Americans and Caucasian PC. Our results suggest that GSTP1 hypermethylation is a sensitive biomarker in African-Americans as compared to that in Caucasians or Asian, and that it strongly influences tumor progression in Asian PC. Ours is the first study investigating GSTP1 methylation differences in PC among African-American, Caucasian and Asian.

The human androgen receptor (AR) gene contains a single nucleotide polymorphism (SNP) on codon 211 and two polymorphic short tandem repeats of CAG and GGC in the N-terminal domain that may influence transcription efficiency of AR gene. We previously reported that the lengths of the CAG and GGC repeats are inversely and linearly related to AR activity and associated with several cancers. However, little is known about this SNP on codon 211 of the AR gene in human renal cell cancer. The cause of renal cell cancer is not well understood although several factors such as chemical carcinogens and hormones have been implicated. AR has been identified in human and animal renal cell cancer. We hypothesize that the SNP on codon 211 is associated with human renal cell cancer. To test this hypothesis, the genetic distribution of the SNP on codon 211 of AR gene was investigated in renal cell cancer patients (211 cases) and healthy controls (200 cases). The allelic and genotypic distributions were determined by PCR-RFLP and direct DNA sequencing techniques. The χ2 test for these data revealed that the distribution of this SNP was not different between renal cell cancer patients as compared to normal healthy controls. The findings suggest that the SNP on codon 211 in the AR gene may not have an important role in the carcinogenesis of human renal cell cancers.

Objectives To evaluate the results of retroperitoneoscopic partial nephrectomy with transient occlusion of the renal artery for treatment of patients with small renal tumors. Methods From May 2002 to March 2003, 12 consecutive patients with T1N0M0 renal tumors with a median diameter of 3.0 cm underwent retroperitoneoscopic partial nephrectomy. The renal artery was clamped transiently without occlusion of the renal vein and the tumor was excised with scissors. Intracorporeal renal cooling was achieved by cold saline perfusion of the renal pelvis through a single- J ureteral catheter. Hemostasis was achieved by approximating the renal parenchyma over the perirenal fatty tissue using a retroperitoneoscopic suturing technique. Results Of the 12 patients, 11 successfully underwent partial nephrectomy retroperitoneoscopically. The median operative time, renal ischemic time, and amount of blood loss in the successful procedures was 220 minutes (range 132 to 340), 50 minutes (range 16 to 115), and 150 mL (range 50 to 800), respectively. The renal calix was opened and repaired in 10 patients. The surgical margin was negative in all patients. The postoperative course was uneventful in all patients, and the return to work was achieved in a median of 18 days (range 9 to 39). The renal function of the affected kidney was well preserved postoperatively, when examined by renal scintigraphy. Neither local recurrence nor distant metastasis was found during the median follow-up period of 10 months (range 1 to 18). Conclusions Retroperitoneoscopic partial nephrectomy with transient occlusion of the renal artery is a useful and less-invasive method for resection of small renal tumors. Additional technologic developments to reduce ischemic times and to cool the kidneys laparoscopically are required.

Purpose: CYP1B1 activates various environmental carcinogens in human tissues, including renal tissues. We hypothesize that certain polymorphisms of the CYP1B1 gene are risk factors for renal cell cancer. The rationale for this hypothesis is that chemical procarcinogenic compounds require metabolic activation by oxidative enzymes such as CYP1B1 to be transformed into potentially carcinogenic forms. To test this hypothesis, we investigated the genotypic distributions of six different loci on the CYP1B1 gene and their association with renal cell cancer. Experimental Design: DNA from 211 cases of human renal cell cancer and 200 healthy controls was analyzed by sequence-specific PCR and direct DNA sequencing to determine the genotypic frequencies of six different polymorphic loci on the CYP1B1 gene. Results: The results of this study demonstrate that the frequencies of genotype 119T/T and genotype 432G/G were significantly higher in renal cell cancer patients compared with healthy normal controls. The relative risks were calculated as 3.01 and 2.17 for genotypes 119T/T and 432G/G, respectively, in renal cell carcinoma patients. These genotypic distributions were also significantly different between male and female patients. The relative risks of genotype 119T/T were calculated as 3.95 in males and 1.92 in females, and the relative risks of genotype 432G/G were calculated as 2.81 in males and 1.35 in females. Conclusions: The present study demonstrates for the first time that the polymorphisms at codons 119 and 432 may be risk factors for renal cancer, especially in the male population.

Osteocalcin (OC), a major noncollagenous bone matrix protein, is expressed prevalently in prostate cancer epithelial cells, adjacent fibromuscular stromal cells, and osteoblasts in locally recurrent prostate cancer and prostate cancer bone metastasis [Matsubara, S., Wada, Y., Gardner, T.A., Egawa, M., Park, M.S., Hsieh, C.L., Zhau, H.E., Kao, C., Kamidono, S., Gillenwater, J.Y., and Chung, L.W. (2001). Cancer Res. 61, 6012-6019]. We constructed an adenovirus vector carrying osteocalcin promoter-driven herpes simplex virus thymidine kinase (Ad-OC-hsv-TK) to cotarget prostate cancer cells and their surrounding stromal cells. A phase I dose escalation clinical trial of the intralesional administration of Ad-OC-hsv-TK followed by oral valacyclovir was conducted at the University of Virginia (Charlottesville, VA) in 11 men with localized recurrent and metastatic hormone-refractory prostate cancer (2 local recurrent, 5 osseous metastasis, and 4 lymph node metastasis) in order to determine the usefulness of this vector for the palliation of androgen-independent prostate cancer metastasis. This is the first clinical trial in which therapeutic adenoviruses are injected directly into prostate cancer lymph node and bone metastasis. Results show that (1). all patients tolerated this therapy with no serious adverse events; (2). local cell death was observed in treated lesions in seven patients (63.6%) as assessed by TUNEL assay, and histomorphological change (mediation of fibrosis) was detected in all posttreated specimens; (3). one patient showed stabilization of the treated lesion for 317 days with no alternative therapy. Of the two patients who complained of tumor-associated symptoms before the treatment, one patient with bone pain had resolution of pain, although significant remission of treated lesions was not observed by image examination; (4). CD8-positive T cells were predominant compared with CD4-positive T cells, B cells (L26 positive), and natural killer cells (CD56 positive) in posttreated tissue specimens; (5). levels of HSV TK gene transduction correlated well with coxsackie-adenovirus receptor expression but less well with the titers of adenovirus injected; and (6). intrinsic OC expression and the efficiency of HSV TK gene transduction affected the levels of HSV TK protein expression in clinical specimens. Our data suggest that this form of gene therapy requires further development for the treatment of androgen-independent prostate cancer metastasis although histopathological and immunohistochemical evidence of apoptosis was observed in the specimens treated. Further studies including the development of viral delivery will enhance the efficacy of Ad-OC-hsv-TK.

症例は糖尿病を有する58歳,男性,会陰部の疼痛・腫脹を主訴に入院となった. CTにて会陰部から陰嚢の結合組織内にガスの貯留を認め,切開を行うと膿汁とガスが噴出した.切開後も炎症は下腹部,鼠径部へ拡大し,切開を追加したが消退しなかった.入院20日目にARDSを発症し,患者は呼吸不全となった.下腹部,鼠径部,会陰部の皮膚が壊死に陥り,デブリドマンを繰り返すことにより,大きな皮膚欠損となったがこれにより炎症は消退した.呼吸状態も改善し29日目に人工呼吸器を離脱した. Fournier症候群は会陰部を中心とする壊疽性筋膜炎で,しばしば治療に抵抗し重篤化する.こうした例では壊死組織の積極的な広範囲切除が必要と考えられる.

We isolated a paclitaxel-resistant cell line (KK47/TX30) from a human bladder cancer cell line (KK47/WT) in order to investigate the mechanism of and reversal agents for paclitaxel resistance. KK47/TX30 cells exhibited 700-fold resistance to paclitaxel and cross-resistance to vinca alkaloids and topoisomerase II inhibitors. Tubulin polymerization assay showed no significant difference in the ratio of polymerized alpha- and beta-tubulin between KK47/WT and KK47/TX30 cells. Western blot analysis demonstrated overexpression of P-glycoprotein (P-gp) and lung resistance-related protein (LRP) in KK47/TX30 cells. Drug accumulation and efflux studies showed that the decreased paclitaxel accumulation in KK47/TX30 cells was due to enhanced paclitaxel efflux. Cell survival assay revealed that verapamil and cepharanthine, conventional P-gp modulators, could completely overcome paclitaxel resistance. To investigate whether new synthetic isoprenoids could overcome paclitaxel resistance, we synthesized 31 isoprenoids based on the structure of N-solanesyl-N,N'-bis(3,4-dimethoxybenzyl)ethylenediamine (SDB), which could reverse multidrug resistance (MDR), as shown previously. Among those examined, trans-N,N'-bis(3,4-dimethoxybenzyl)-N-solanesyl-1,2-diaminocyclohexane (N-5228) could completely reverse paclitaxel resistance in KK47/TX30 cells. N-5228 inhibited photoaffinity labeling of P-gp by [(3)H]azidopine, suggesting that N-5228 could bind to P-gp directly and could be a substrate of P-gp. Next, we investigated structural features of these 31 isoprenoids in order to determine the structural requirements for the reversal of P-gp-mediated paclitaxel resistance, suggesting that the following structural features are important for overcoming paclitaxel resistance: (1) a basic structure of 8 to 10 isoprene units, (2) a cyclohexane ring or benzene ring within the framework, (3) two cationic sites in close proximity to each other, and (4) a benzyl group with 3,4-dimethoxy functionalities, which have moderate electron-donating ability. These findings may provide valuable information for the development of P-gp-mediated MDR-reversing agents.

Objective – To clarify the relationship between mitochondrial DNA (mtDNA) sequence variations and phenotypes in patients with A3243G mutation. Materials and methods – We studied whole mtDNA sequences in two families with A3243G mutation and characteristic clinical features. Two brothers in Family 1 had shown thiamine deficiency and mitochondrial myopathy without central nervous system involvement. In Family 2, a 16-year-old woman showed the symptoms of mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Her mother had had diabetes mellitus and died at the age of 42. The proportion of A3243G mtDNA in blood was 87 and 89% in the patients of Family 1, and 25% in the patient and less than 5% in the mother of Family 2. Results – The mtDNA analysis revealed the following homoplasmic substitutions: T1520C and C12153T found only in Family 1, and A15954G found only in Family 2. These substitutions were not detected in seven other MELAS patients or in 50 controls. Conclusion – These substitutions might be specific to these families and could be one of the factors that modulate their clinical features together with the A3243G mutation.

Background: Adrenomedullin (AM) has pluripotent activities and is involved in the regulation of vasomotor tone, cell differentiation and embryogenesis. However, the expression and pathophysiological role of AM has not been determined in human renal cell carcinoma (RCC). Methods: Twenty-six RCC specimens and three cultured human RCC cell lines (A498, SN12C and KPK-13) were analyzed. Expression of AM was determined by immunohistochemistry and reverse transcription–polymerase chain reaction (RT-PCR) analysis. The correlation between AM expression and microvessel count (MVC) in RCC specimens was examined to determine if AM plays a role in tumor angiogenesis. The correlation between the expression of AM and vascular endothelial growth factor (VEGF) was also investigated. Lastly, the effect of hypoxia upon the mRNA expression of AM, VEGF and hypoxia inducible factor-1 (HIF-1) by RCC cell lines was determined. Results: Immunohistochemistry indicated that AM and VEGF were primarily localized in the cytosol of RCC cells. AM and VEGF mRNA were detected in all RCC specimens and cultured RCC cell lines analyzed by RT-PCR. There was a positive correlation between AM mRNA expression and MVC (r = 0.516, P = 0.0062), and between VEGF mRNA expression and MVC (r = 0.485, P = 0.0111). We also observed a positive correlation between AM mRNA expression and VEGF mRNA expression (r = 0.552, P = 0.0029). Hypoxia significantly induced AM and VEGF mRNA expression, although the increase of the AM mRNA level (10.6–26.7 fold) was markedly greater than that of the VEGF mRNA level (1.5–1.9 fold). Conclusion: These results suggest that hypoxia-induced AM plays a part in tumor angiogenesis in conjunction with VEGF and facilitates human RCC growth under hypoxic conditions.

A two-phase fluid loop (TPFL), which enables removal of a large amount of heat and long-distance heat transportation utilizing latent heat of the pumped working fluid, is expected to be an essential thermal control system for the future spacecraft and orbital infrastructure. Two-Phase Fluid Loop experiment (TPFLEX) on Space Shuttle (STS-85) was conducted in order to demonstrate the operational principle, DP(=vapor pressure in the evaporator minus liquid pressure in the evaporator)>0, of TPFL under micro-gravity. As a result of this experiment we obtained the DP data as a function of heat load on the evaporator for a fixed flow rate.

透析患者の多嚢胞化萎縮腎 (acquired cystic disease of the kidney: ACDK) は高率に腎癌を合併する. 1991年6月から2001年12月までの約11年間に当施設で経験した透析患者のACDKに合併した腎癌症例14例16腎について, 臨床的・病理組織学的検討を行い, 診断・治療上の問題点を中心に考察を行った. 対象の年齢は39歳から75歳, 平均54.5歳, 性別は男性12人, 女性が2人であった. 透析に至った原疾患は, 慢性糸球体腎炎が13例, IgA腎症が1例であり, 糖尿病性腎症はいなかった. 透析方法は, 血液透析13例, 腹膜透析が1例, 透析期間は, 28か月から最長300か月, 平均143か月であった. 診断の手がかりとなったのは, CTが8例, 超音波検査が6例であった. 診断のきっかけになるような自覚症状を伴ったものは4例であり, 血尿が3例, 微熱, 側腹部痛が各々1例であった. 病的骨折部の生検で腺癌が認められ, 原発巣の検索の結果, 右腎癌が発見された症例を1例, 右腎癌に左副腎褐色細胞腫を合併した症例を1例, 両側の同時性および異時性ACDK合併腎癌を各1例経験した. また, 献腎移植希望登録の更新手続きのための受診時に行った超音波検査で偶然発見された症例を2例経験し, 献腎移植希望患者におけるスクリーニングも重要であると思われた. 患側は, 両側性が2例, 右腎が10例, 左腎が2例であった. 手術はいずれも開腹手術で行い, 14腎は経腰的アプローチ, 2腎は経腹的アプローチであった. 手術時間は100分から390分, 平均220分, 出血量は44-1305mL, 平均396mLであった. 症例11は両側性を疑い, 異時的に両側腎摘を行ったが, 後に摘出した左腎には腫瘍は認めず, 術前診断の困難さを改めて認識させられた. 術後合併症はpT4 (膵への直接浸潤) の1例で, 術後にDIC, 心不全を起こし, 術後21日目に死亡した症例を経験したが, 他には特に重篤な合併症はみられなかった.