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Ultrasonographic studies have demonstrated transient reduction in spleen volume in relation to apnea diving. We measured spleen volume under various respiratory conditions by MR imaging to accurately determine the influence of ordinary breath holding on spleen volumetry. Twelve healthy adult volunteers were examined. Contiguous MR images of the spleen were acquired during free breathing and during respiratory manipulations, including breath holding at the end of normal expiration, breath holding at deep inspiration, and the valsalva maneuver, and spleen volume was measured from each image set based on the sum-of-areas method. Acquisition during free breathing was performed with respiratory triggering. The duration of each respiratory manipulation was 30 s, and five sets of MR images were acquired serially during each manipulation. Baseline spleen volume before respiratory manipulation was 173.0 ± 79.7 mL, and the coefficient of variance for two baseline measures was 1.4% ± 1.6%, suggesting excellent repeatability. Spleen volume decreased significantly just after the commencement of respiratory manipulation, remained constant during the manipulation, and returned to the control value 2 min after the cessation of the manipulation, irrespective of manipulation type. The percentages of volume reduction were 10.2% ± 2.9%, 10.2% ± 3.5%, and 13.3% ± 5.7% during expiration breath holding, deep-inspiration breath holding, and the valsalva maneuver, respectively, and these values did not differ significantly. Spleen volume is reduced during short breath-hold apnea in healthy adults. Physiological responses of the spleen to respiratory manipulations should be considered in the measurement and interpretation of spleen volume.

Purpose To investigate a method for optimizing the display conditions of brain magnetic resonance (MR) images. Materials and methods We retrospectively analyzed brain MR images of 120 adults classified into screening, acute cerebral infarction, and brain tumor groups ( n  = 40 each). Two observers independently displayed the images on a monitor and optimized the display conditions using the W/L and U/L methods. In the W/L method, the observers manipulated the width and level of the display window, while in the U/L method they manipulated the upper and lower levels of the window. The times required were compared between the two methods. Additionally, the appropriateness of the determined window setting was evaluated visually by the respective observer to exclude the possibility that rough, suboptimal adjustment shortened the adjustment time. Results For both observers and all groups, the time required for optimization was significantly shorter for the U/L method than for the W/L method. The appropriateness of the window setting for the U/L method was equal to or better than that for the W/L method. Conclusion Manipulating the upper and lower levels of the display window appears to improve the efficiency of interpreting brain MR images through rapid optimization of the display condition.

Background We investigated a practical method to measure peak filling rate (PFR) as an indicator of diastolic function of the left ventricle. Ten adult volunteers underwent cine MR imaging using steady-state free precession (SSFP) and phase contrast (PC) sequences to measure PFR. Two PC image sets were acquired at the mitral valve orifice, and PFR was determined from the set with high true temporal resolution (temporal PC method) or with high spatial resolution (spatial PC method). SSFP images covering the left ventricle were acquired, and a time–volume curve was generated around the peak filling phase. PFR was determined using parabolic curve fitting on the first-derivative curve of the LV time–volume curve. Findings PFR values estimated by the PC methods correlated well with those estimated by the SSFP method, despite apparent underestimation. The underestimation was smaller for the temporal PC method (12 %) than for the spatial PC method (28 %). Intra- and inter-observer repeatabilities were better for the PC methods than for the SSFP method. Conclusions PFR measurement by PC imaging with high true temporal resolution is convenient and offers excellent repeatability and acceptable accuracy, indicating suitability for clinical use.

Introduction. Display contrast can be changed nonlinearly by manipulating the gamma value of the grayscale. We investigated the contrast of the hepatobiliary-phase images acquired with different flip angles (FAs) and displayed with different gamma values in Gd-EOB-DTPA-enhanced magnetic resonance imaging. Material and Methods. Twenty patients with liver tumors were studied. Hepatobiliary-phase images were acquired at low (12°) and high (30°) FAs. Low-FA images were converted to simulate images displayed with different gamma values, using ImageJ software. To assess image contrast, the liver-to-muscle signal ratio (LMR), liver-to-spleen signal ratio (LSR), contrast ratio (CR), liver signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) were calculated. Results. The LMR, LSR, and CR were higher in the high-FA images than in the low-FA original images. Although the SNR was lower in the high-FA images, indicating an increase in noise, the CNR was higher. Raising the gamma value increased the LMR, LSR, and CR, notably decreased the SNR, and slightly decreased the CNR. Conclusion. Increasing the FA enhanced image contrast, supporting its usefulness for improving the delineation of focal liver lesions. Although the associated increase in noise may be problematic, raising the grayscale gamma value enhances the display contrast of low-FA images.

Objective Ultrasonographic studies have demonstrated transient reduction in spleen volume in relation to apnea diving. We measured spleen volume under various respiratory conditions by MR imaging to accurately determine the influence of ordinary breath holding on spleen volumetry. Materials and Methods Twelve healthy adult volunteers were examined. Contiguous MR images of the spleen were acquired during free breathing and during respiratory manipulations, including breath holding at the end of normal expiration, breath holding at deep inspiration, and the valsalva maneuver, and spleen volume was measured from each image set based on the sum-of-areas method. Acquisition during free breathing was performed with respiratory triggering. The duration of each respiratory manipulation was 30 s, and five sets of MR images were acquired serially during each manipulation. Results Baseline spleen volume before respiratory manipulation was 173.0 plus or minus 79.7 mL, and the coefficient of variance for two baseline measures was 1.4% plus or minus 1.6%, suggesting excellent repeatability. Spleen volume decreased significantly just after the commencement of respiratory manipulation, remained constant during the manipulation, and returned to the control value 2 min after the cessation of the manipulation, irrespective of manipulation type. The percentages of volume reduction were 10.2% plus or minus 2.9%, 10.2% plus or minus 3.5%, and 13.3% plus or minus 5.7% during expiration breath holding, deep-inspiration breath holding, and the valsalva maneuver, respectively, and these values did not differ significantly. Conclusions Spleen volume is reduced during short breath-hold apnea in healthy adults. Physiological responses of the spleen to respiratory manipulations should be considered in the measurement and interpretation of spleen volume.

DNA methyl groups are selectively removed at target loci using inactive Cas9 fused to a SunTag-based peptide repeat that recruits the enzymatic activity. Despite the importance of DNA methylation in health and disease, technologies to readily manipulate methylation of specific sequences for functional analysis and therapeutic purposes are lacking. Here we adapt the previously described dCas9–SunTag for efficient, targeted demethylation of specific DNA loci. The original SunTag consists of ten copies of the GCN4 peptide separated by 5-amino-acid linkers. To achieve efficient recruitment of an anti-GCN4 scFv fused to the ten-eleven (TET) 1 hydroxylase, which induces demethylation, we changed the linker length to 22 amino acids. The system attains demethylation efficiencies >50% in seven out of nine loci tested. Four of these seven loci showed demethylation of >90%. We demonstrate targeted demethylation of CpGs in regulatory regions and demethylation-dependent 1.7- to 50-fold upregulation of associated genes both in cell culture (embryonic stem cells, cancer cell lines, primary neural precursor cells) and in vivo in mouse fetuses.

Background It is important to establish cancer stem cell (CSC)-targeted therapies to eradicate cancer. As it is a CSC marker, we focused on Kruppel-like factor 5 ( KLF5 ) in this study. Methods We searched for candidate microRNAs (miRNAs) that inhibited KLF5 expression by in silico analyses and screened them in colon cancer cell lines. Results We identified one promising miRNA, miR-4711-5p, that downregulated KLF5 expression by direct binding. This miRNA suppressed cell proliferation, migration and invasion ability, as well as stemness, including decreased stem cell marker expression, reactive oxygen species activity and sphere formation ability. MiR-4711-5p inhibited the growth of DLD-1 xenografts in nude mice with no adverse effects. We found that miR-4711-5p provoked G1 arrest, which could be attributed to direct binding of miR-4711-5p to TFDP1 (a heterodimeric partner of the E2F family). Our findings also suggested that direct binding of miR-4711-5p to MDM2 could upregulate wild-type p53, leading to strong induction of apoptosis. Finally, we found that miR-4711-5p had a potent tumour-suppressive effect compared with a putative anti-oncomiR, miR-34a, in tumour cell cultures derived from five patients with colorectal cancer. Conclusions Our data suggest that miR-4711-5p could be a promising target for CSC therapy.

The BRAF V600E mutation occurs in approximately 10% of patients with metastatic colorectal cancer (CRC) and constitutes a distinct subtype of the disease with extremely poor prognosis. To address this refractory disease, we investigated the unique metabolic gene profile of BRAF V600E‐mutated tumors via in silico analysis using a large‐scale clinical database. We found that BRAF V600E‐mutated tumors exhibited a specific metabolic gene expression signature, including some genes that are associated with poor prognosis in CRC. We discovered that BRAF V600E‐mutated tumors expressed high levels of glycolytic enzyme enolase 2 (ENO2), which is mainly expressed in neuronal tissues under physiological conditions. In vitro experiments using CRC cells demonstrated that BRAF V600E‐mutated cells exhibited enhanced dependency on ENO2 compared to BRAF wild‐type cancer cells and that knockdown of ENO2 led to the inhibition of proliferation and migration of BRAF V600E‐mutated cancer cells. Moreover, inhibition of ENO2 resulted in enhanced sensitivity to vemurafenib, a selective inhibitor of BRAF V600E. We identified AP‐1 transcription factor subunit (FOSL1) as being involved in the transcription of ENO2 in CRC cells. In addition, both MAPK and PI3K/Akt signaling were suppressed upon inhibition of ENO2, implying an additional oncogenic role of ENO2. These results suggest the crucial role of ENO2 in the progression of BRAF V600E‐mutated CRC and indicate the therapeutic implications of targeting this gene. There was no significant difference in the expression of ENO1 in BRAF V600E‐mutated CRC and other types of CRC, but ENO2 levels were significantly higher in BRAF V600E‐mutated CRC.

We developed a novel clamping device for laparoscopic surgery, free from conventional pinch structure, capable of uniformly occluding any ductal organ. This study aimed to evaluate performance of the new clamper compared to the pinch-type clamper. The new clamper consists of two metal bars with ties at each end, which enables parallel clamping. A resected porcine stomach was used, with an infusion tube at the anal end to increase intra-luminal pressure. The oral side of the stomach was clamped with either the new clamper or the pinch-type clamper, and their performances were evaluated in qualitative and semi-quantitative manner. Qualitative evaluation involved imaging the clamping site at intra-gastric pressures from 0 to 15 mmHg using microfocus computed tomography. The new clamper showed no gap even under increased intra-luminal pressure, while the pinch-type clamper showed a gap on the distal side. Quantitative evaluation measured bursting pressure under continuous air insufflation. Air leakages were observed in the new clamper at higher intra-luminal pressures than in the pinch-type clamper (46.1 mmHg vs. 13.6 mmHg, P  < 0.01). Our new clamping device showed superior performance in preclinical setting compared to the conventional pinch-type clamper. We are currently working on its design freezing and aiming for early commercialization.