PMEPA1/TMEPAI Is a Unique Tumorigenic Activator of AKT Promoting Proteasomal Degradation of PHLPP1 in Triple-Negative Breast Cancer Cells.

Transmembrane prostate androgen-induced protein (TMEPAI), also known as PMEPA1, is highly expressed in many types of cancer and promotes oncogenic abilities. However, the mechanisms whereby TMEPAI facilitates tumorigenesis are not fully understood. We previously established TMEPAI-knockout (KO) cells from human triple-negative breast cancer (TNBC) cell lines and found that TMEPAI-KO cells showed reduced tumorigenic abilities. Here, we report that TMEPAI-KO cells upregulated the expression of pleckstrin homology (PH) domain and leucine-rich repeat protein phosphatase 1 (PHLPP1) and suppressed AKT Ser473 phosphorylation, which was consistent with TCGA dataset analysis. Additionally, the knockdown (KD) of PHLPP1 in TMEPAI-KO cells partially but significantly rescued AKT Ser473 phosphorylation, as well as in vitro and in vivo tumorigenic activities, thus showing that TMEPAI functions as an oncogenic protein through the regulation of PHLPP1 subsequent to AKT activation. Furthermore, we demonstrated that TMEPAI PPxY (PY) motifs are essential for binding to NEDD4-2, an E3 ubiquitin ligase, and PHLPP1-downregulatory ability. Moreover, TMEPAI enhanced the complex formation of PHLPP1 with NEDD4-2 and PHLPP1 polyubiquitination, which leads to its proteasomal degradation. These findings indicate that the PY motifs of TMEPAI suppress the amount of PHLPP1 and maintain AKT Ser473 phosphorylation at high levels to enhance the tumorigenic potentiality of TNBC.

Heterologous Biosynthesis of Fungal Indole Sesquiterpene Sespendole.

Indole sesquiterpene sespendole, which has been isolated from the filamentous fungus Pseudobotrytis terrestris FKA-25, is a specific inhibitor of lipid droplet synthesis in mouse macrophages. The biosynthetic pathway that involves genes encoding six enzymes (spdEMBQHJ) was elucidated through heterologous expression of spd genes in Aspergillus oryzae, biotransformation experiments, and in vitro enzymatic reactions with a recombinant protein, thereby revealing the mechanism underlying the characteristic modification on the indole ring, catalyzed by a set of prenyltransferase (SpdE)/cytochrome P450 (SpdJ) enzymes. Functional analysis of the homologous genes encoding these enzymes involved in the biosynthesis of lolitrem allowed a biosynthetic pathway for the bicyclic ring skeleton fused to the indole ring to be proposed.

Computer-aided diagnosis of prostate cancer on magnetic resonance imaging using a convolutional neural network algorithm.

OBJECTIVE: To develop a computer-aided diagnosis (CAD) algorithm with a deep learning architecture for detecting prostate cancer on magnetic resonance imaging (MRI) to promote global standardisation and diminish variation in the interpretation of prostate MRI. PATIENTS AND METHODS: We retrospectively reviewed data from 335 patients with a prostate-specific antigen level of <20 ng/mL who underwent MRI and extended systematic prostate biopsy with or without MRI-targeted biopsy. The data were divided into a training data set (n = 301), which was used to develop the CAD algorithm, and two evaluation data sets (n = 34). A deep convolutional neural network (CNN) was trained using MR images labelled as 'cancer' or 'no cancer' confirmed by the above-mentioned biopsy. Using the CAD algorithm that showed the best diagnostic accuracy with the two evaluation data sets, the data set not used for evaluation was analysed, and receiver operating curve analysis was performed. RESULTS: Graphics processing unit computing required 5.5 h to learn to analyse 2 million images. The time required for the CAD algorithm to evaluate a new image was 30 ms/image. The two algorithms showed area under the curve values of 0.645 and 0.636, respectively, in the validation data sets. The number of patients mistakenly diagnosed as having cancer was 16/17 patients and seven of 17 patients in the two validation data sets, respectively. Zero and two oversights were found in the two validation data sets, respectively. CONCLUSION: We developed a CAD system using a CNN algorithm for the fully automated detection of prostate cancer using MRI, which has the potential to provide reproducible interpretation and a greater level of standardisation and consistency.

Focused Genome Mining of Structurally Related Sesterterpenes: Enzymatic Formation of Enantiomeric and Diastereomeric Products.

Heterologous expression of four clade-A bifunctional terpene synthases (BFTSs), giving di/sesterterpenes with unique polycyclic carbon skeletons such as sesterfisherol, enabled the isolation of the sesterterpenes Bm1, Bm2, Bm3, and Pb1. Their structures suggested that formation of the products occurs via various diastereomeric carbocation intermediates, allowing the proposal that clade-A BFTSs catalyze three-step cyclizations using several stereofacial combinations of allylic cation-olefin pairs in the corresponding intermediates to generate various stereoisomers.

Preoperative RFA simulation for liver cancer using a CT virtual ultrasound system.

We developed a computed tomography (CT) virtual ultrasound system (CVUS) as an imaging system to support treatment under percutaneous ultrasound (US) guidance. This prototype clinical system, produced in collaboration with Tokyo Medical University, uses display software developed by Toshiba Medical Systems. We examined the utility of this system by scheduling treatment plans preoperatively and simulating puncture and radiofrequency ablation (RFA) for liver cancer. The study enrolled 51 liver cancer patients with 66 nodules 0.8-8cm in diameter in which RFA was performed between June 2004 and December 2004. Virtual US and multiplanar reconstruction (MPR) images were constructed on the basis of DICOM CT data and puncture and ablation of liver cancer were simulated. The following were evaluated: (1) how to avoid complications and determine an appropriate puncture route by simulating puncture with C-mode MPR images; (2) determination of the three-dimensional location of the tumor for ablation, as well as the adjacent organs and vessels, by MPR rotation 360 degrees around the center of the tumor (center lock); and (3) how to determine the center and volume of ablation and avoid injuries to nearby organs and vessels by simulating ablation procedures. C -mode MPR images were effective for (1) determining and modifying the puncture route in 35 of 51 cases (69.6%) and (2) determining the spatial location of vessels and nearby organs in 50 of 51 cases (98.0%) by the center lock; and (3) simulating the ablation helped determine the center and volume of ablation by avoiding injuries to vessels and nearby organs in 45 or 51 cases (88.2%). Taken together, the CVUS allowed easy simulation of local treatment of liver cancer under US guidance using CT data alone and the preoperative simulation predicted an improvement in the safety of local therapy of liver cancer.

Quantification of hepatic parenchymal blood flow by contrast ultrasonography with flash-replenishment imaging.

Flash-replenishment (FR) utilizes destruction of microbubbles in the scan volume by high-power ultrasound and enables to observe reperfusion at a low acoustic power. In this paper, we introduced theoretic equation between probability density function (PDF) of the transit time in the scan volume and time intensity curve (TIC) measured by FR method. From the equations, it was explained that the mean transit time (MTT) through the scan volume was calculated from the plateau level and tangent of the initial slope. Animal experiments were also performed to measure TIC in the parenchymal region of the liver using FR method. From the result of the TIC, the variant of the PDF for the transit time was found to be small and the average MTT was 11.1 s. Hepatic blood flow by an ultrasonic transit time flowmeter was also measured in the same experiment, and adequate correlation was obtained from between the two methods. The results suggested that the FR method, which is a noninvasive measurement, can predict the blood flow of the liver.