BACKGROUND Metastasis associated in colon cancer 1 (MACC1) is a recently identified gene that plays a key role in regulating hepatocyte growth factor-MET signaling. In this study, we demonstrated the correlation and the clinical significance of MACC1 and MET expression in rectal cancer patients treated with chemoradiotherapy (CRT) followed by surgery. MATERIALS AND METHODS The expression of MACC1 and MET mRNA in residual cancer cells from 52 patients after CRT was determined by quantitative real-time polymerase chain reaction. Protein expression was also investigated by immunohistochemical staining. RESULTS MACC1 and MET expressions were positively correlated. Furthermore, these proteins were also co-expressed in immunohistochemical analyses. High expression of MACC1 or MET was associated with reduced relapse-free survival and the prognosis was worse when both genes were highly expressed. CONCLUSION Evaluation of MACC1 and MET expression may be useful for predicting prognosis in patients with rectal cancer treated with CRT followed by surgery.

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.

We have designed a new vector- and marker-free site-directed deletion system for gram-negative bacteria. In this system, a specific DNA fragment is amplified from a parental strain by using polymerase chain reaction (PCR), then circularized and introduced back into the parental strain for homologous recombination. The recombinant mutant is then detected and isolated by PCR-based sib selection. Unlike conventional methods, our Simple Deletion method requires no cloning procedures, and no foreign genes such as antibiotic-resistance genes are introduced as selection markers. The resulting mutant is, therefore, the same as the parental strain except for the lack of the target region. This method is categorized as a type of "self-cloning," and the resulting mutant can be used for laboratory research without restrictions. Using this method, we generated a mutant of a plant pathogenic bacterium, Xanthomonas campestris pv. campestris, in which the 20.4-kb hrp gene cluster involved in the type III secretion system and in pathogenicity was deleted. In addition, we proved that this method can also be used to delete smaller DNA regions of X. campestris pv. campestris and to generate deletion mutants of the bacterium Ralstonia solanacearum.

Orbital hypertelorism is defined as an abnormally wide bony interorbital distance. The aims of surgery are both correction of ocular dystopia and cosmetic reconstruction of the nasal crest. Marked improvement of visual function, especially binocular vision, by surgery is not expected. Here we report that surgical treatment unexpectedly resulted in a significant visual improvement for a 13-year-old boy with orbital hypertelorism who also had bilateral cleft lip and palate.

Actin filaments have a double-helix structure consisting of globular actin molecules. In many mechanical cellular activities, such as cell movement, division, and shape control, modulation of the extensional and torsional dynamics of the filament has been linked to regulatory actin-binding protein functions. Therefore, it is important to quantitatively evaluate extension-torsion coupling of filament to better understand the actin filament dynamics. In the present study, the extension-torsion coupling was investigated using molecular dynamics simulations. We constructed a model for the actin filament consisting of 14 actin subunits in an ionic solvent as a minimal functional unit, and analyzed longitudinal and twisting Brownian motions of the filament. We then derived the expected value of energy associated with extension and torsion at equilibrium, and evaluated the extension-torsion stiffness of the filament from the thermal fluctuations obtained from the MD simulations. The results demonstrated that as the analyzed sampling-window duration was increased, the extension-torsion coupling stiffness evaluated on a nanosecond scale tended to converge to a value of 7.6×10(-11)N. The results obtained from this study will contribute to the understanding of biomechanical events, under mechanical tension and torque, involving extension-torsion coupling of filaments.

BACKGROUND: The neurotrophic receptor tropomyosin related kinase (TrkB) is associated with tumor progression in neuroblastoma and certain human malignancies. Recent reports indicate TrkB may participate in the epithelial-mesenchymal transition (EMT). This study investigates whether TrkB expression is associated with the clinical outcome of colorectal cancer (CRC) patients and whether TrkB induces EMT in CRC cells. METHODS: TrkB and E-cadherin expression in surgical tissue samples and clinicopathological data from 102 CRC patients were analyzed by real-time polymerase chain reaction and immunohistochemistry. The biological role of TrkB in CRC was analyzed using RNA interference against TrkB in the CRC cell line SW480 to assess tumor progression and the correlation between TrkB and E-cadherin expression. RESULTS: Patients with high TrkB mRNA expression in clinical samples had a significantly poorer prognosis relative to those with low TrkB levels (p = 0.03). TrkB was inversely correlated with E-cadherin at both the mRNA and protein levels. In vitro, cell proliferation (p = 0.02), migration (p < 0.001), and invasion (p < 0.001) were significantly inhibited by TrkB knockdown while the anoikis rate increased in TrkB siRNA-transfected cells compared to control siRNA. Interestingly, E-cadherin expression in TrkB siRNA-transfected cells was higher than in control cells and vimentin was lower conversely. CONCLUSIONS: High TrkB expression is associated with poor prognosis in CRC patients and enhanced malignant potential in terms of proliferation, migration, invasion, and anoikis inhibition in CRC cells. These results indicate TrkB could induce EMT and play an important role in CRC progression to metastasis.

The aim of this study was to assess how tinnitus retraining therapy (TRT) employing monaural noise generators (NGs) affects tinnitus patients. Ninety-five patients with chronic tinnitus were included in this study. All received directive counseling and monaural NGs without any other combination treatment. Effects were evaluated with the Tinnitus Handicap Inventory (THI) 6, 12, and 24 months after the start of treatment. Multiple regression analysis was used to evaluate factors associated with improvement of THI scores. We observed a significant improvement in the average THI scores, which ranged from 59 at baseline to 36 after 6 months and were stable up to 24 months. Severely distressed (SD) patients experienced more benefits from the therapy than moderately distressed (MD) patients. Multiple regression analysis revealed that the presence of transient anxiety symptoms, measured by the state section of the State Trait Anxiety Inventory at the beginning of treatment, were associated the most to a decline in THI scores in SD patients. Pure-tone average at three mid frequencies was a negatively contributing factor. Analysis of MD patients did not provide a good model. In conclusion, TRT employing monaural NGs was an effective intervention especially for SD patients, although plateau of the effects after 6 months may be indicative of limitation of applying NGs monoaurally. Reducing the patients' hearing handicap may reinforce the effects of TRT in SD patients. Additional evaluation and interventions for both SD and MD patients might be needed.

BACKGROUND: E-cadherin expression has been associated with an outcome in patients with colorectal cancer, and serum levels of soluble E-cadherin (sE-cadherin) are significantly elevated in patients with malignant disease. However, the prognostic value of serum sE-cadherin level has not been demonstrated in colorectal cancer. METHODS: Serum samples were collected from 186 patients with colorectal cancer and 21 normal volunteers. Serum sE-cadherin levels were measured using an enzyme-linked immunosorbent assay kit. We investigated the relationship between serum sE-cadherin level and clinicopathologic findings. RESULTS: Mean serum sE-cadherin levels were significantly higher in CRC patients than in controls. Mean sE-cadherin levels were significantly correlated with hepatic metastasis, UICC classification, and poor prognosis. Elevated serum sE-cadherin level was an independent risk factor for predicting poor prognosis, and was an independent marker for predicting hepatic metastasis. Among patients with synchronous hepatic metastases, the prognosis was significantly worse in patients with elevated serum sE-cadherin levels than in those with lower levels. CONCLUSIONS: Preoperative elevated sE-cadherin level is associated with poor prognosis in colorectal cancer. Measuring serum sE-cadherin may provide valuable information for predicting prognosis in patients with hepatic metastasis.

The aberrant regulation of B-cell receptor (BCR) signaling allows unwanted B cells to persist, thereby potentially leading to autoimmunity and B-cell malignancies. Casitas B-lineage lymphoma (Cbl) proteins suppress BCR signaling; however, the molecular mechanisms that control Cbl function in human B cells remain unclear. Here, we demonstrate that CIN85 (c-Cbl interacting protein of 85 kDa) is constitutively associated with c-Cbl, Cbl-b, and B-cell linker in B cells. Experiments using CIN85-overexpressing and CIN85-knockdown B-cell lines revealed that CIN85 increased c-Cbl phosphorylation and inhibited BCR-induced calcium flux and phosphorylation of Syk and PLCγ2, whereas it did not affect BCR internalization. The Syk phosphorylation in CIN85-overexpressing and CIN85-knockdown cells was inversely correlated with the ubiquitination and degradation of Syk. Moreover, CIN85 knockdown in primary B cells enhanced BCR-induced survival and growth, and increased the expression of BcLxL, A1, cyclin D2, and myc. Following the stimulation of BCR and Toll-like receptor 9, B-cell differentiation- associated molecules were up-regulated in CIN85-knockdown cells. Together, these results suggest that CIN85 is required for Cbl-mediated regulation of BCR signaling and for downstream events such as survival, growth, and differentiation of human B cells.