Metabolic characterization of sphere-derived prostate cancer stem cells reveals aberrant urea cycle in stemness maintenance.

Alteration of cell metabolism is one of the essential characteristics of tumor growth. Cancer stem cells (CSCs) are the initiating cells of tumorigenesis, proliferation, recurrence, and other processes, and play an important role in therapeutic resistance and metastasis. Thus, identification of the metabolic profiles in prostate cancer stem cells (PCSCs) is critical to understanding prostate cancer progression. Using untargeted metabolomics and lipidomics methods, we show distinct metabolic differences between prostate cancer cells and PCSCs. Urea cycle is the most significantly altered metabolic pathway in PCSCs, the key metabolites arginine and proline are evidently elevated. Proline promotes cancer stem-like characteristics via the JAK2/STAT3 signaling pathway. Meanwhile, the enzyme pyrroline-5-carboxylate reductase 1 (PYCR1), which catalyzes the conversion of pyrroline-5-carboxylic acid to proline, is highly expressed in PCSCs, and the inhibition of PYCR1 suppresses the stem-like characteristics of prostate cancer cells and tumor growth. In addition, carnitine and free fatty acid levels are significantly increased, indicating reprogramming of fatty acid metabolism in PCSCs. Reduced sphingolipid levels and increased triglyceride levels are also observed. Collectively, our data illustrate the comprehensive landscape of the metabolic reprogramming of PCSCs and provide potential therapeutic strategies for prostate cancer.

Improving Charge Storage of Biaxially-Oriented Polypropylene under Extreme Electric Fields by Excimer UV Irradiation.

Biaxially-oriented polypropylene (BOPP) is one of the most commonly used materials for film-based capacitors for power electronics and pulsed power systems. To address the pressing issue of performance-limiting loss under extreme electric-fields, here a one-step, high-throughput, and environment-friendly process based on very low-dose ultra-violet irradiation from KrCl (222 nm) and Xe2 (172 nm) excimer is demonstrated. The performance of commercial BOPP is boosted in terms of withstanding electric-field extremes (Weibull breakdown strength 694 to 811 V µm-1 by 17% at 25 °C and 428 to 651 V µm-1 by 52% at 120 °C), discharged energy density, and conduction losses. Importantly, the depth profile of space charge is precisely measured in situ with a high resolution of 500 nm by laser induced pressure pulse. Consequently, the space charge effect and electric-field distortion are reduced and related to the improved polymer films. It is demonstrated that energetic UV photons act as scissors for BOPP chains and dissociate oxygen molecules leading to the more thermally stable oxygen-containing structures, as deep traps to impede charge migration. This work provides a promising approach to produce polymers with customized microscopic characteristics that is compatible with the assembly lines of polymer-based capacitors.

Aurora kinase A regulates cancer-associated RNA aberrant splicing in breast cancer.

The contribution of oncogenes to tumor-associated RNA splicing and the relevant molecular mechanisms therein require further elaboration. Here, we show that oncogenic Aurora kinase A (AURKA) promotes breast cancer-related RNA aberrant splicing in a context-dependent manner. AURKA regulated pan-breast cancer-associated RNA splicing events including GOLGA4, RBM4 and UBQLN1. Aberrant splicing of GOLGA4 and RBM4 was closely related to breast cancer development. Mechanistically, AURKA interacted with the splicing factor YBX1 and promoted AURKA-YBX1 complex-mediated GOLGA4 exon inclusion. AURKA binding to the splicing factor hnRNPK promoted AURKA-hnRNPK complex-mediated RBM4 exon skipping. Analysis of clinical data identified an association between the AURKA-YBX1/hnRNPK complex and poor prognosis in breast cancer. Blocking AURKA nuclear translocation with small molecule drugs partially reversed the oncogenic splicing of RBM4 and GOLGA4 in breast cancer cells. In summary, oncogenic AURKA executes its function on modulating breast cancer-related RNA splicing, and nuclear AURKA is distinguished as a hopeful target in the case of treating breast cancer.

Pterostilbene Interferes With Lipopolysaccharide-Induced Myocardial Injury Through Oxidative Stress and Inflammasome Pathways.

Myocardial contractile dysfunction caused by sepsis is a serious threat to human health, and its pathogenesis is not completely clear. It is generally believed that excessive inflammation and oxidative stress are the main causes of myocardial damage caused by sepsis. Pterostilbene (PTS) has a variety of biological activities, including anti-oxidant, anti-inflammatory, and anti-aging. Whether PTS protect myocardial function in rats with sepsis through anti-inflammatory and anti-oxidant effects has not been reported. In this study, we investigated the role of PTS in septic mice induced by lipopolysaccharide (LPS). Mice were injected intraperitoneally with LPS (20 mg/kg) to simulate sepsis. Use Echocardiography, Masson, DHE, H&E, IHC, IF and other experimental methods to explore the effects of PTS on LPS. The results showed that PTS was indicated to significantly increase the cardiac function of mice with sepsis. PTS treatment also reduced the mRNA expression of IL-1α, IL-6, MCP-1, and IL-1ß and the protein expression of NLRP3 in vivo and in vitro, and inhibited the migration of inflammatory cells. PTS treatment also reduced the mRNA expression of collagen I, collagen III and α-SMA, and inhibited fibrosis. PTS treatment reduced the mRNA expression of NOX1, NOX2, and NOX4 and inhibited DHE levels in vivo and in vitro. In summary, our data indicated that PTS played a crucial role in LPS-induced myocardial injured and might be a key target for the prevention and treatment of sepsis-induced myocardial dysfunction.

Artificial Intelligence of Manufacturing Robotics Health Monitoring System by Semantic Modeling.

Robotics is widely used in nearly all sorts of manufacturing. Steady performance and accurate movement of robotics are vital in quality control. Along with the coming of the Industry 4.0 era, oceans of sensor data from robotics are available, within which the health condition and faults are enclosed. Considering the growing complexity of the manufacturing system, an automatic and intelligent health-monitoring system is required to detect abnormalities of robotics in real-time to promote quality and reduce safety risks. Therefore, in this study, we designed a novel semantic-based modeling method for multistage robotic systems. Experiments show that sole modeling is not sufficient for multiple stages. We propose a descriptor to conclude the stages of robotic systems by learning from operational data. The descriptors are akin to a vocabulary of the systems; hence, semantic checking can be carried out to monitor the correctness of operations. Furthermore, the stage classification and its semantics were used to apply various regression models to each stage to monitor the quality of each operation. The proposed method was applied to a photovoltaic manufacturing system. Benchmarks on production datasets from actual factories show the effectiveness of the proposed method to realize an AI-enabled real-time health-monitoring system of robotics.

Spectrum Analysis Enabled Periodic Feature Reconstruction Based Automatic Defect Detection System for Electroluminescence Images of Photovoltaic Modules.

Electroluminescence (EL) imaging is a widely adopted method in quality assurance of the photovoltaic (PV) manufacturing industry. With the growing demand for high-quality PV products, automatic inspection methods based on machine vision have become an emerging area concern to replace manual inspectors. Therefore, this paper presents an automatic defect-inspection method for multi-cell monocrystalline PV modules with EL images. A processing routine is designed to extract the defect features of the PV module, eliminating the influence of the intrinsic structural features. Spectrum domain analysis is applied to effectively reconstruct an improved PV layout from a defective one by spectrum filtering in a certain direction. The reconstructed image is used to segment the PV module into cells and slices. Based on the segmentation, defect detection is carried out on individual cells or slices to detect cracks, breaks, and speckles. Robust performance has been achieved from experiments on many samples with varying illumination conditions and defect shapes/sizes, which shows the proposed method can efficiently distinguish intrinsic structural features from the defect features, enabling precise and speedy defect detections on multi-cell PV modules.

Nuclear Aurora kinase A switches m6A reader YTHDC1 to enhance an oncogenic RNA splicing of tumor suppressor RBM4.

Aberrant RNA splicing produces alternative isoforms of genes to facilitate tumor progression, yet how this process is regulated by oncogenic signal remains largely unknown. Here, we unveil that non-canonical activation of nuclear AURKA promotes an oncogenic RNA splicing of tumor suppressor RBM4 directed by m6A reader YTHDC1 in lung cancer. Nuclear translocation of AURKA is a prerequisite for RNA aberrant splicing, specifically triggering RBM4 splicing from the full isoform (RBM4-FL) to the short isoform (RBM4-S) in a kinase-independent manner. RBM4-S functions as a tumor promoter by abolishing RBM4-FL-mediated inhibition of the activity of the SRSF1-mTORC1 signaling pathway. Mechanistically, AURKA disrupts the binding of SRSF3 to YTHDC1, resulting in the inhibition of RBM4-FL production induced by the m6A-YTHDC1-SRSF3 complex. In turn, AURKA recruits hnRNP K to YTHDC1, leading to an m6A-YTHDC1-hnRNP K-dependent exon skipping to produce RBM4-S. Importantly, the small molecules that block AURKA nuclear translocation, reverse the oncogenic splicing of RBM4 and significantly suppress lung tumor progression. Together, our study unveils a previously unappreciated role of nuclear AURKA in m6A reader YTHDC1-dependent oncogenic RNA splicing switch, providing a novel therapeutic route to target nuclear oncogenic events.

Scorpion Venom peptide, AGAP inhibits TRPV1 and potentiates the analgesic effect of lidocaine.

The current study was designed to test the hypothesis that BmK AGAP (AGAP) potentiates the analgesic effect of lidocaine. The chronic constrictive injury was performed on 72 rats to induce a rapid onset and long-lasting pain. The rats were randomly assigned to one of six groups; Group A (n = 12) received an intrathecal administration of saline, Group B (n = 12) received an intrathecal injection of lidocaine, Group C (n = 12) received an intrathecal administration of AGAP, Group D, E, and F (n = 12 each) received an intrathecal administration of lidocaine 0.005 mg/ml + AGAP 25, 50, 100 µg/kg respectively. The von Frey filaments were used to assess mechanical allodynia. Nav1.7 and TRPV1 currents were recorded by the whole-cell aspiration patch-clamp technique, and KCNQ2/3 currents were recorded by the whole-cell drilling patch-clamp technique. The whole-cell aspiration patch-clamp technique showed that AGAP inhibited TRPV1and KCNQ2/3 currents and increased the analgesic effect of lidocaine. AGAP may have a synergistic effect with lidocaine which demonstrates a potential therapeutic approach for optimizing post-operative analgesia.

Nalbuphine suppresses breast cancer stem-like properties and epithelial-mesenchymal transition via the AKT-NFκB signaling pathway.

BACKGROUND: Cancer pain is a debilitating disorder of human breast cancer and a primary determinant of the poor quality of life, and relieving pain is fundamental strategy in the cancer treatment. However, opioid analgesics, like morphine and fentanyl, which are widely used in cancer pain treatment have been reported to enhance stem-like traits and epithelial-mesenchymal transition (EMT) of breast cancer cells. As such, it is vital to make the best choice of analgesic for breast cancer management. METHODS: MTT assays and colony formation assays were performed to examine tumor cell proliferation upon nalbuphine treatment. RT-PCR, western blot, flow cytometry, sphere formation, immunohistochemistry, transwell assays, wound healing assays and mouse xenograft were used to assess the biological effects of nalbuphine treatment. RESULTS: Nalbuphine inhibited breast cancer cell growth and tumorigenesis, with little effect on noncancerous breast cell lines. Nalbuphine suppressed cancer stem-like traits and EMT in both breast cancer cells and mouse xenograft tumor tissues. Additionally, activation of AKT reversed the nalbuphine-induced inhibition of cancer stem-like properties, tumorigenesis and EMT. CONCLUSIONS: Our results demonstrate a new role of nalbuphine in inhibiting cancer stem-like properties and EMT in addition to relieving pain, which suggests that nalbuphine may be effective in breast cancer treatment.

Stress-induced epinephrine enhances lactate dehydrogenase A and promotes breast cancer stem-like cells.

Chronic stress triggers activation of the sympathetic nervous system and drives malignancy. Using an immunodeficient murine system, we showed that chronic stress-induced epinephrine promoted breast cancer stem-like properties via lactate dehydrogenase A-dependent (LDHA-dependent) metabolic rewiring. Chronic stress-induced epinephrine activated LDHA to generate lactate, and the adjusted pH directed USP28-mediated deubiquitination and stabilization of MYC. The SLUG promoter was then activated by MYC, which promoted development of breast cancer stem-like traits. Using a drug screen that targeted LDHA, we found that a chronic stress-induced cancer stem-like phenotype could be reversed by vitamin C. These findings demonstrated the critical importance of psychological factors in promoting stem-like properties in breast cancer cells. Thus, the LDHA-lowering agent vitamin C can be a potential approach for combating stress-associated breast cancer.

Longitudinal monitoring of cerebrospinal fluid-derived circulating tumor cells: a pilot study.

Breast cancer-associated leptomeningeal metastasis (LM) is a particularly aggressive syndrome with an abysmal prognosis. Here we applied the SE-i•FISH platform for surveillance and function analysis of cerebrospinal fluid-derived circulating tumor cells (CSFTCs) from five breast cancer patients with LM. We observed a negative correlation of CK18 expression on CSFTCs with clinical symptom remission, and confirmed that at least six intrathecal chemotherapies were necessary. We also present the case of a breast cancer patient with bladder and leptomeningeal metastasis. The bladder is an extremely unusual site for metastasis from primary breast cancer, with only 14 cases sporadically reported worldwide. Our platform could help monitor CSFTC numbers and treatment responses for patients with LM.

Three-dimensional coculture of primary hepatocytes and stellate cells in silk scaffold improves hepatic morphology and functionality in vitro.

A vigorous in vitro model of liver that could recapitulate hepatic phenotype and functionality in vivo would exclusively improve the efficiency of bioartificial liver, drug discovery, or even transplantation therapy. Owing to the indispensable role of three-dimensional (3D) microenvironment in supporting viability and function of hepatocytes in vitro, much effort recently has been focused on improving reproducibility and standardization of primary hepatocyte cultures with a paradigm shift to 3D culture system, In the present study, an improved 3D coculture system of hepatocytes was established in which rat primary hepatocytes were cocultured with hepatic stellate cells in silk porous scaffolds. Silk scaffolds with incorporated extracellular matrix provided a suitable microenvironment for maintaining the viability, morphology and gene expression of the primary hepatocyte in vitro. The presence of stromal cells promoted primary hepatocyte to generate cellular aggregates with well-organized 3D architecture after 3 days of coculture in vitro. These aggregates exhibited proper morphology similar to liver tissue in vivo. Consistent with their phenotypic appearance, well-maintained functionality of hepatocytes was also observed in the cocultures, where albumin secretion/expression, urea synthesis as well as messenger ribonucleic acid expression of multiple cytochrome Ps (CYPs) enzymes increased significantly compared to either the 3D monocultures or monolayer cultures. Additionally, this 3D multicellular coculture model displayed an improved metabolic activity of CYPs enzymes to the probe drugs treatment. Thus, this culture system would not only contribute to the construction of micro-organoid tissue of liver but also potentially provide a robust tool for drug metabolism evaluation in vitro. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2171-2180, 2018.

Cisatracurium-induced proliferation impairment and death of colorectal cancer cells, HCT116 is mediated by p53 dependent intrinsic apoptotic pathway in vitro.

Activation of oncogenes and suppression of repressor genes are believed to play crucial roles in the pathogenesis of human colorectal carcinoma. Cisatracurium, a nondepolarizing neuromuscular blocking agent, has been reported to inhibit cell proliferation while promoting apoptosis. However, the underlining mechanism, of these growth setbacks are not well understood. We assessed the growth of human colorectal carcinoma (HCT116) and its cell cycle distribution upon cisatracurium exposure. Significant cell growth inhibition and accumulation of cells in G1 phase of the cell cycle was observed in treated cells compared with untreated cells (control). In furtherance to these observations, FITC Annexin V and propidium iodide apoptosis assay demonstrated concentration and time dependent percentage increase in apoptosis of cells treated with cisatracurium compared with untreated cells. qRT-PCR analysis showed concentration-dependent alterations in CD1, E2F, CE1, p53 and p21 mRNA expression. Western blot analysis indicated remarkable concentration dependent alterations in the expression of proliferation and survival proteins CD1, E2F, CE1, p53, p21, BAX, BCL-2, cytochrome C and cleaved PARP in cisatracurium-treated groups as compared with the untreated group. Cisatracurium also significantly promoted caspase-9 and caspase-3 activities in cells treated with cisatracurium compared with untreated cells. Thus, cisatracurium effectively inhibited proliferation and induced apoptosis of HCT116 cells in vitro at least via alteration of p53-dependent apoptotic pathway.