It is well known that oestrogens play important roles in both the pathogenesis and development of invasive ductal carcinoma (IDC) of human breast. However, molecular features of oestrogen actions have remained largely unclear in pure ductal carcinoma in situ (pDCIS), regarded as a precursor lesion of many IDCs. This is partly due to the fact that gene expression profiles of oestrogen-responsive genes have not been examined in pDCIS. Therefore, we first examined the profiles of oestrogen-induced genes in oestrogen receptor (ER)-positive pDCIS and DCIS (DCIS-c) and IDC (IDC-c) components of IDC cases (n = 4, respectively) by microarray analysis. Oestrogen-induced genes identified in this study were tentatively classified into three different groups in the hierarchical clustering analysis, and 33% of the genes were predominantly expressed in pDCIS rather than DCIS-c or IDC-c cases. Among these genes, the status of MYB (c-MYB), RBBP7 (RbAp46) and BIRC5 (survivin) expression in carcinoma cells was significantly higher in ER-positive pDCIS (n = 53) than that in ER-positive DCIS-c (n = 27) or IDC-c (n = 27) by subsequent immunohistochemical analysis of the corresponding genes (P < 0.0001, P = 0.03 and P = 0.0003, respectively). In particular, the status of c-MYB immunoreactivity was inversely (P = 0.006) correlated with Ki-67 in the pDCIS cases. These results suggest that expression profiles of oestrogen-induced genes in pDCIS may be different from those in IDC, and c-MYB, RbAp46 and survivin may play particularly important roles among oestrogen induced genes in ER-positive pDCIS.

A new automated cleanup system for the analysis of dioxins (PCDDs, PCDFs and DL-PCBs) has been developed. It was controlled by PLC through the touch-panel. This automated cleanup system can simultaneously treat six samples in 2h, using only about 30mL of solvent. In this study, the recovery rates of the internal standard added as cleanup spiked were between 70% and 120% in the fly ash sample. The RSDs (relative standard deviations) were below 15%. The shortest analysis time from cleanup to calculation of concentration was approximately 6h. Moreover, this automated cleanup system eliminates personal error in sample preparation and training time for the analyst, and improves the accuracy of the experiment. Additionally, this automated cleanup system allowed rapid analysis and less consumption of organic solvent.

Apoptotic cells must be recognized early for phagocytosis to ensure that their toxic contents do not damage neighboring cells. In some cases this is achieved via CD43-capped membrane glycoproteins, the sialylpolylactosaminyl chains of which serve as ligands for phagocytosis by macrophages. However, because many additional changes occur during apoptosis, determining exactly which events are responsible for signaling macrophages to initiate phagocytosis remains a challenge. Here, we examined one clearance mechanism in detail and determined that capping of CD43 alone is sufficient to initiate phagocytosis. We induced macrophage-mediated phagocytosis by using cytochalasin B to artificially cap CD43 on healthy (non-apoptotic) Jurkat cells. Additional experiments confirmed that sialylpolylactosaminyl chains formed through this capping method are a prerequisite for removal, and that nucleolin is the macrophage receptor responsible for their detection. These findings strongly suggest that capping of CD43 presents a sufficient signal for phagocytosis without any additional membrane changes.

A gender difference has been reported in the morbidity of esophageal squamous cell carcinoma (ESCC). Estrogens have been proposed to play some roles in this difference　but its details have not been clarified yet. Therefore, in this study, we first examined the status of estrogen receptor (ER)α and ERβ in 90 Japanese ESCC patients. ERα and ERβ immunoreactivity was detected in the nuclei of ESCC cells (41.1% and 97.8%, respectively). There was a significant positive association between the ERβ H score and histological differentiation (P = 0.0403), TNM-pM (LYM)(P=0.00164), and Ki67 ⁄MIB1 LI of carcinoma cells (P = 0.0497, r = 0.207). In addition, the ERβ status of carcinoma cells was significantly correlated with unfavorable clinical outcome of the patients. Multivariate analysis further revealed the ERβ status in carcinoma cells as an independent unfavorable prognostic factor of these patients. We further examined the effects of estrogen treatment on ESCC cell line (ECGI-10) transfected with ERα or ERβ in vitro. The number of ECGI-10 transfected with ERβ was increased by estradiol or ERβ specific agonist but estradiol did not exert any effects upon the cell number of ECGI-10 transfected with ERα. In summary, results of our present study clearly demonstrated that the status of ERβ in ESCC was closely associated with the unfavorable prognosis possibly through altering cell proliferation of carcinoma cells. © 2012 Japanese Cancer Association.

Objectives: The purpose of this study was to evaluate the MRI characteristics of venous thrombus over set time thresholds with histopathological correlation in a porcine model. Methods: Inferior vena cava thrombi were induced in 12 pigs. MRI was performed in three pigs 2 h, 1 day, 3 days and 2 weeks after thrombus induction. Results: The MRI characteristics were analysed in correlation with histopathological findings. The thrombi after 2 hours, which consisted of red blood cells (RBCs), showed isointensity on T(1 )weighted images (T(1)WIs) and hyperintensity on both T(2 )weighted images (T(2)WIs) and diffusion-weighted images (DWIs). The mean apparent diffusion coefficient (ADC) value was 1.93×10(-3) mm(2) s(-1). The thrombi after Day 1, which consisted of RBCs and migrating neutrophils at the periphery, showed isointensity on T(1)WIs, slight hyperintensity on T(2)WIs and hypointensity on DWIs. The mean ADC value was 1.62×10(-3) mm/s(-2). The thrombi after Day 3, which consisted of RBCs and peripheral inflammatory cells including macrophages, showed isointensity with peripheral hyperintense regions on T(1)WIs and hypointensity on both T(2)WIs and DWIs. The mean ADC value was 1.67×10(-3) mm(2) s(-1). After 2 weeks, the thrombi, which revealed RBC lysis surrounded by granulation tissues, showed isointensity on T(1)WIs and hyperintensity on T(2)WIs and DWIs. The mean ADC value was 2.48×10(-3) mm(2) s(-1). Conclusion: The temporal MRI characteristics seemed to be related to chemical and physical changes in RBC and organisation of granulation tissues. Free radicals generated by macrophages might also be related to some extent.

A bacterial strain that assimilates fucoidan from Cladosiphon okamuranus as sole carbon source was isolated as Luteolibacter algae H-18. It was found that it degraded fucoidan by intracellular enzymes, and that the degradation reactions were catalyzed by multiple enzymes. One enzyme, designated fraction B, was established to exhibit the deacetylation reaction of fucoidan. Other enzyme(s), designated fraction A, catalyzed the reaction(s) lowering the molecular weight of fucoidan.

Objectives: To evaluate an implanted thermal ablation device that can be heated with high efficiency using a resonant circuit as the implant.Methods: 16 rats were used. The implants, adjusted at a resonance frequency of 4 MHz, were fixed on the surface of the liver of rats under laparotomy. In 14 of 16 rats, an alternating magnetic field (AMF) was applied for 6 min with an output of 300 W from outside the body using a ferrite core applicator. The implant temperature during AMF exposure was measured. The 14 rats were divided into five groups, depending on time from AMF application until they were sacrificed (1 h, 1 day, 3 days, 7 days and 1 month after application). Two rats not exposed to AMF were used as controls. Livers were removed and evaluated; the cross-sectional area and width of the ablated region were measured.Results: During AMF exposure, the implant temperature rose to 127.8±39.3 °C (mean ±standard deviation). The cross-sectional area of the ablated region was largest after 1 day and tended to decrease with time. The widths of the ablated region were 4.87±0.22 mm, 4.15±0.36 mm, 3.67±0.58 mm and 3.24±0.16 mm in the 1 day, 3 day, 7 day and 1 month groups, respectively. No significant differences (p<0.05) were seen in either cross-sectional area or width of the ablated region.Conclusion: Sufficient heat for ablation was obtained in vivo using a newly developed implanted thermal ablation device. This device may be a new option for thermal ablation therapy.

Aromatase inhibitors (AIs) are considered the gold standard of endocrine therapy for estrogen receptor positive postmenopausal breast cancer patients. AI treatment was reported to result in marked alterations of genetic profiles in cancer tissues but its detailed molecular mechanisms have not been elucidated. Therefore, we profiled miRNA expression before and after the treatment with letrozole in MCF-7 co-cultured with primary breast cancer stromal cells. Letrozole significantly altered the expression profiles of cancer miRNAs in vitro. Among the elevated miRNAs following letrozole treatment, computational analysis identified let-7f, a tumor-suppressor miRNA which targeted the aromatase gene (CYP19A1) expression. Quantitative real time PCR assay using MCF-7 and SK-BR-3 cells as well as clinical specimens of neoadjuvant study demonstrated a significant inverse correlation between aromatase mRNA and let-7f expression. In addition, high let-7f expression was significantly correlated with low aromatase protein levels evaluated by both immunohistochemistry and western blotting method in breast cancer cases. Results of 3'UTR luciferase assay also demonstrated the actual let-7f binding sites in CYP19A1, indicating that let-7f directly targets the aromatase gene. Subsequent WST-8 and migration assays performed in let-7f transfected MCF-7 and SK-BR-3 cells revealed significant decrement of their proliferation and migration. These findings all demonstrated that let-7f, a tumor suppressor miRNA in breast cancer, directly targeted the gene and were restored by AI treatment. Therefore, AI may exert tumor-suppressing effects upon breast cancer cells by suppressing aromatase gene expression via restoration of let-7f. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Runt-related transcription factor 2 (RUNX2) belongs to the RUNX family of heterodimeric transcription factors, and is mainly associated with osteogenesis. Previous in vitro studies demonstrated that RUNX2 increased the cell proliferation of mouse and rat colon carcinoma cells but the status of RUNX2 has remained unknown in human colon carcinoma. Therefore, we examined clinical significance and biological functions of RUNX2 in colon carcinoma. RUNX2 immunoreactivity was examined in 157 colon carcinoma tissues using immunohistochemistry. RUNX2 immunoreactivity was evaluated as % of positive carcinoma cells (i.e., labeling index (LI)). We used SW480 and DLD-1 human colon carcinoma cells, expressing estrogen receptor (ER)-β in subsequent in vitro studies. RUNX2 immunoreactivity was detected in colon carcinoma cells, and the median value of RUNX2 LI was 67%. RUNX2 LI was significantly associated with Dukes' stage, liver metastasis and ERβ status. In addition, RUNX2 LI was significantly associated with adverse clinical outcome of the colon carcinoma patients, and turned out an independent prognostic factor following multivariate analysis. Results of in vitro studies demonstrated that both SW480 and DLD-1 cells transfected with small interfering RNA (siRNA) against RUNX2 significantly decreased their cell proliferation, migration and invasive properties. In addition, RUNX2 mRNA level was significantly decreased by ER antagonist in these two cells. These findings all suggest that RUNX2 is a potent prognostic factor in human colon carcinoma patients through the promotion of cell proliferation and invasion properties, and is at least partly up-regulated by estrogen signals through ERβ of carcinoma cells. © 2012 Wiley-Liss, Inc.

Notch is a transmembrane receptor functioning in the determination of cell fate. Abnormal Notch signaling promotes tumor development, showing either oncogenic or tumor suppressive activity. The uncertainty about the exact role of Notch signaling, partially, stems from inconsistencies in descriptions of Notch expression in human cancers. Here, we clarified basal-cell dominant expression of NOTCH1 in squamous epithelium. NOTCH1 was downregulated in squamous neoplasms of oral mucosa, esophagus and uterine cervix, compared with the normal basal cells, although the expression tended to be retained in cervical lesions. NOTCH1 downregulation was observed even in precancers, and there was little difference between cancers and high-grade precancerous lesions, suggesting its minor contribution to cancer-specific events such as invasion. In culture experiments, reduction of NOTCH1 expression resulted in downregulation of keratin 13 and keratin 15, and upregulation of keratin 17, and NOTCH1 knockdown cells formed a dysplastic stratified epithelium mimicking a precancerous lesion. The NOTCH1 downregulation and the concomitant alterations of those keratin expressions were confirmed in the squamous neoplasms both by immunohistochemical and cDNA microarray analyses. Our data indicate that reduction of NOTCH1 expression directs the basal cells to cease terminal differentiation and to form an immature epithelium, thereby playing a major role in the histopathogenesis of epithelial dysplasia. Furthermore, downregulation of NOTCH1 expression seems to be an inherent mechanism for switching the epithelium from a normal and mature state to an activated and immature state, suggesting its essential role in maintaining the epithelial integrity.