Inhibition of TMEM16A improves cisplatin-induced acute kidney injury via preventing DRP1-mediated mitochondrial fission.

Acute kidney injury (AKI) is associated with high morbidity and mortality. Our previous study has demonstrated that TMEM16A, a Ca2+-activated chloride channel, contributes to renal fibrosis progression in chronic kidney disease. However, whether TMEM16A is involved in AKI is still unknown. In this study, we established cisplatin AKI mice model and found that TMEM16A expression was upregulated in the injured kidney. In vivo knockdown of TMEM16A effectively prevented cisplatin-induced tubular cell apoptosis, inflammation and kidney function loss. Western blot and transmission electron microscopy (TEM) revealed that TMEM16A knockdown inhibited Drp1 translocation from the cytoplasm to mitochondria and prevented mitochondrial fission in tubular cells. Consistently, in cultured HK2 cells, knockdown or inhibition of TMEM16A by shRNA or its specific inhibitor suppressed cisplatin-induced mitochondrial fission and its associated energy dysfunction, ROS accumulation, and cell apoptosis via inhibiting Drp1 activation. Further investigation showed that genetic knockdown or pharmacological inhibition of TMEM16A inhibited cisplatin-induced Drp1 Ser-616 site phosphorylation through ERK1/2 signaling pathway, whereas overexpression of TMEM16A promoted this effect. Treatment with Drp1 or ERK1/2 inhibitor could efficiently prevent cisplatin-induced mitochondrial fission. Collectively, our data suggest that TMEM16A inhibition alleviated cisplatin-induced AKI by preventing tubular cell mitochondrial fission through the ERK1/2 / Drp1 pathway. Inhibition of TMEM16A may be a novel therapeutic strategy for AKI.

Non-destructive identification of Pseudostellaria heterophylla from different geographical origins by Vis/NIR and SWIR hyperspectral imaging techniques.

The composition of Pseudostellaria heterophylla (Tai-Zi-Shen, TZS) is greatly influenced by the growing area of the plants, making it significant to distinguish the origins of TZS. However, traditional methods for TZS origin identification are time-consuming, laborious, and destructive. To address this, two or three TZS accessions were selected from four different regions of China, with each of these resources including distinct quality grades of TZS samples. The visible near-infrared (Vis/NIR) and short-wave infrared (SWIR) hyperspectral information from these samples were then collected. Fast and high-precision methods to identify the origins of TZS were developed by combining various preprocessing algorithms, feature band extraction algorithms (CARS and SPA), traditional two-stage machine learning classifiers (PLS-DA, SVM, and RF), and an end-to-end deep learning classifier (DCNN). Specifically, SWIR hyperspectral information outperformed Vis/NIR hyperspectral information in detecting geographic origins of TZS. The SPA algorithm proved particularly effective in extracting SWIR information that was highly correlated with the origins of TZS. The corresponding FD-SPA-SVM model reduced the number of bands by 77.2% and improved the model accuracy from 97.6% to 98.1% compared to the full-band FD-SVM model. Overall, two sets of fast and high-precision models, SWIR-FD-SPA-SVM and SWIR-FD-DCNN, were established, achieving accuracies of 98.1% and 98.7% respectively. This work provides a potentially efficient alternative for rapidly detecting the origins of TZS during actual production.

Production of Mesoporous Magnetic Carbon Materials from Oily Sludge by Combining Thermal Activation and Post-Washing.

In this work, the oily sludge (OS) from a local waste oil recycling plant was reused as a precursor for producing porous magnetic carbon composites (CC) by pyrolysis, followed by carbon dioxide activation. Based on the thermogravimetric analysis (TGA) of the OS feedstock, the preparation experiments were performed at 800−900 °C. From the pore analysis of the CC products, it indicated an increasing trend, as the BET surface area greatly increased from about 1.0 to 44.30 m2/g. In addition, the enhancement effect on the pore properties can be consistently obtained from the acid-washed CC products because the existing and new pores were reformed due to the leaching-out of inorganic minerals. It showed an increase from 32.27 to 94.45 m2/g and 44.30 to 94.52 m2/g at 850 and 900 °C, respectively, showing their mesoporous features. These porous and iron-containing features were also observed by the scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS). In addition, the adsorption removal of total organic carbon (TOC) in the raw wastewater, by the CC product, showed its high performance (>80%).

Preparation and Characterization of Porous Materials from Pineapple Peel at Elevated Pyrolysis Temperatures.

In this work, pineapple peel (PP) was reused as a precursor in biochar (BC) production at elevated temperatures (i.e., 500−900 °C) for residence times of 0−60 min. The findings showed that pyrolysis temperature and residence time played a vital role in pore development. As pyrolysis temperature increased from 800 to 900 °C for residence times of 20 and 60 min, the data on the Brunauer−Emmett−Teller (BET) surface area of the resulting biochar products significantly jumped from 11.98−32.34 to 119.43−133.40 m2/g. In addition, there was a significant increase in the BET surface area from 1.02 to 133.40 m2/g with the residence time of 0 to 20 min at 900 °C. From the data of the nitrogen adsorption−desorption isotherms and the pore size distribution, both micropores (pore diameters of <2.0 nm) and mesopores (pore diameters of 2.0−50.0 nm) are present in the PP-based biochar products. Due to its good fittings in the pseudo-second-order model and its hydrophilic nature, as seen in the Fourier transform infrared spectroscopy (FTIR), the resulting biochar could be a porous material to be used for the effective removal of cationic compounds (i.e., methylene blue (MB)) from liquid phases.

Genome-wide CRISPR screen identifies CDK6 as a therapeutic target in adult T-cell leukemia/lymphoma.

Adult T-cell leukemia/lymphoma (ATLL) is an aggressive T-cell malignancy with a poor prognosis with current therapy. Here we report genome-wide CRISPR-Cas9 screening of ATLL models, which identified CDK6, CCND2, BATF3, JUNB, STAT3, and IL10RB as genes that are essential for the proliferation and/or survival of ATLL cells. As a single agent, the CDK6 inhibitor palbociclib induced cell cycle arrest and apoptosis in ATLL models with wild-type TP53. ATLL models that had inactivated TP53 genetically were relatively resistant to palbociclib owing to compensatory CDK2 activity, and this resistance could be reversed by APR-246, a small molecule activator of mutant TP53. The CRISPR-Cas9 screen further highlighted the dependence of ATLL cells on mTORC1 signaling. Treatment of ATLL cells with palbociclib in combination with mTORC1 inhibitors was synergistically toxic irrespective of the TP53 status. This work defines CDK6 as a novel therapeutic target for ATLL and supports the clinical evaluation of palbociclib in combination with mTORC1 inhibitors in this recalcitrant malignancy.

Enhanced efficacy of JAK1 inhibitor with mTORC1/C2 targeting in smoldering/chronic adult T cell leukemia.

Adult T-cell leukemia (ATL) is an aggressive T-cell lymphoproliferative malignancy of regulatory T lymphocytes (Tregs), caused by human T-cell lymphotropic virus 1 (HTLV-1). Interleukin 2 receptor alpha (IL-2Rα) is expressed in the leukemic cells of smoldering/chronic ATL patients, leading to constitutive activation of the JAK/STAT pathway and spontaneous proliferation. The PI3K/AKT/mTOR pathway also plays a critical role in ATL cell survival and proliferation. We previously performed a high-throughput screen that demonstrated additive/synergistic activity of Ruxolitinib, a JAK1/2 inhibitor, with AZD8055, an mTORC1/C2 inhibitor. However, effects of unintended JAK2 inhibition with Ruxolitinib limits it therapeutic potential for ATL patients, which lead us to evaluate a JAK1-specific inhibitor. Here, we demonstrated that Upadacitinib, a JAK-1 inhibitor, inhibited the proliferation of cytokine-dependent ATL cell lines and the expression of p-STAT5. Combinations of Upadacitinib with either AZD8055 or Sapanisertib, mTORC1/C2 inhibitors, showed anti-proliferative effects against cytokine-dependent ATL cell lines and synergistic effect with reducing tumor growth in NSG mice bearing IL-2 transgenic tumors. Importantly, the combination of these two agents inhibited ex vivo spontaneous proliferation of ATL cells from patients with smoldering/chronic ATL. Combined targeting of JAK/STAT and PI3K/AKT/mTOR pathways represents a promising therapeutic intervention for patients with smoldering/chronic ATL.

A20 and RBX1 Regulate Brentuximab Vedotin Sensitivity in Hodgkin Lymphoma Models.

PURPOSE: For patients with refractory/relapsed Hodgkin lymphoma (roughly 20% of total cases), few effective therapeutic options exist. Currently, brentuximab vedotin (BV), a drug-conjugated anti-CD30 antibody, is one of the most effective approved therapy agents for these patients. However, many patients do not achieve complete remission and ultimately develop BV-resistant disease, necessitating a more detailed understanding of the molecular circuitry that drives BV sensitivity and the mechanism of BV resistance. EXPERIMENTAL DESIGN: Here, we established a ubiquitin regulator-focused CRISPR library screening platform in Hodgkin lymphoma and carried out a drug sensitization screen against BV to identify genes regulating BV treatment sensitivity. RESULTS: Our CRISPR library screens revealed the ubiquitin-editing enzymes A20 and RBX1 as key molecule effectors that regulate BV sensitivity in Hodgkin lymphoma line L428. A20 negatively regulates NF-κB activity which is required to prevent BV cytotoxicity. In line with these results, the RNA-seq analysis of the BV-resistant single-cell clones demonstrated a consistent upregulation of NF-κB signature genes, as well as the ABC transporter gene ABCB1. Mechanically, NF-κB regulates BV treatment sensitivity through mediating ABCB1 expression. Targeting NF-κB activity synergized well with BV in killing Hodgkin lymphoma cell lines, augmented BV sensitivity, and overcame BV resistance in vitro and in Hodgkin lymphoma xenograft mouse models. CONCLUSIONS: Our identification of this previously unrecognized mechanism provides novel knowledge of possible BV responsiveness and resistance mechanisms in Hodgkin lymphoma, as well as leads to promising hypotheses for the development of therapeutic strategies to overcome BV resistance in this disease.

A Hybrid Method Applied to Improve the Efficiency of Full-Waveform Inversion for Pavement Characterization.

Ground penetrating radar (GPR), as a nondestructive testing tool, is suitable for estimating the thickness and permittivity of layers within the pavement. However, it would become problematic when the layer is thin with respect to the probing pulse width, in which case overlapping between the reflected pulses occurs. In order to deal with this problem, a hybrid method based on multilayer perceptrons (MLPs) and a local optimization algorithm is proposed. This method can be divided into two stages. In the first stage, the MLPs roughly estimate the thickness and the permittivity of the GPR signal. In the second stage, these roughly estimated values are used as the initial solution of the full-waveform inversion algorithm. The hybrid method and the conventional global optimization algorithm are respectively used to perform the full-waveform inversion of the simulated GPR data. Under the same inversion precision, the objective function needs to be calculated for 450 times and 30 times for the conventional method and the hybrid method, respectively. The hybrid method is also applied to a measured data, and the thickness estimation error is 1.2 mm. The results show the high efficiency and accuracy of such hybrid method to resolve the problem of estimating the thickness and permittivity of a "thin layer".

Characterization of biochar prepared from biogas digestate.

In the study, the biogas digestate was evaluated as a potential feedstock for preparing biochars at a broad temperature range of 300-900°C. The physico-chemical and pore properties of the resulting biochars (denoted as SDBC, solid digestate biochar), including calorific value (higher heating value), surface area/pore volume/pore size distribution, true density, scanning electron microscopy - energy dispersive X-ray spectroscopy (SEM-EDS) and X-ray powder diffraction (XRD), were studied. It was found that the higher heating values of the SDBC products were on a decreasing trend as pyrolysis temperature increased, but they indicated an increase in true density. The results are probably associated with the active pyrolysis of the lignocellulosic fragments and the calcination (or shrinkage) processes, thus resulting in the increased contents of aromatic carbon clusters and main mineral constituents remained. Based on the pore properties, pyrolysis temperature at around 800°C seemed to be the optimal condition for producing SDBC, where its Brunauer-Emmet-Teller (BET) surface area (>100m2/g) largely increased as compared to that of the digestate feedstock (<1m2/g). Furthermore, the main compositions of mineral ash in the resulting biochar could exist as phosphates, carbonates, or oxides of calcium and other alkali/alkaline earth elements. According to the data on EDS and XRD, more pores could be significantly generated under severe pyrolysis (>700°C) due to the high aromaticity via the thermal decomposition of lignocelluloses and the volatilization of inorganic minerals.

Therapeutic effects of a novel tylophorine analog, NK-007, on collagen-induced arthritis through suppressing tumor necrosis factor α production and Th17 cell differentiation.

OBJECTIVE: To analyze the effects of a novel compound, NK-007, on the prevention and treatment of collagen-induced arthritis (CIA) and the underlying mechanisms. METHODS: We determined the effect of NK-007 on lipopolysaccharide (LPS)-triggered tumor necrosis factor α (TNFα) production by murine splenocytes and a macrophage cell line (RAW 264.7) by enzyme-linked immunosorbent assay, intracellular cytokine staining, and Western blotting. The LPS-boosted CIA model was adopted, and NK-007 or vehicle was administered at different time points after immunization. Mice were monitored for clinical severity of arthritis, and joint tissues were used for histologic examination, cytokine detection, and immunohistochemical staining. Finally, stability of TNFα production and Th17 cell differentiation were studied using quantitative polymerase chain reaction and flow cytometry. RESULTS: NK-007 significantly suppressed LPS-induced TNFα production in vitro. Administration of NK-007 completely blocked CIA development and delayed its progression. Furthermore, treatment with NK-007 at the onset of arthritis significantly inhibited the progress of joint inflammation. Administration of NK-007 also suppressed production of TNFα, interleukin-6 (IL-6), and IL-17A in the joint and reduced percentages of IL-17+ cells among CD4+ and γ/δ T cells in draining lymph nodes. We further demonstrated that NK-007 acted on the stability of TNFα messenger RNA and reduced Th17 cell differentiation. In addition, it significantly inhibited levels of IL-6 and IL-17A in human coculture assay. CONCLUSION: For its effects on the development and progression of CIA and for its therapeutic effect on CIA, NK-007 has great potential to be a therapeutic agent for human rheumatoid arthritis.

The mouse Mageb18 gene encodes a ubiquitously expressed type I MAGE protein and regulates cell proliferation and apoptosis in melanoma B16-F0 cells.

Although many cancer vaccines have been developed against type I MAGE (melanoma antigen) genes owing to their shared tumour-specific expression properties, studies about their expression and functions are relatively limited. In the present study, we first identify a non-testis-specific type I MAGE gene, Mageb18 (melanoma antigen family B 18). Mouse Mageb18 is also expressed in digestion- and immune-related tissues as well as testis, and its expression in testis is age-dependent. Mageb18 is expressed in many mouse-derived cell lines, and DNA demethylation and histone acetylation mediate the reactivation of Mageb18 in Mageb18-negtive H22 and C6 cells. We also show that mouse Mageb18 encodes a 46 kDa protein which is predominantly localized in the cytoplasm. In testis, the endogenous MAGEB18 protein is mainly expressed in proliferative spermatogonia and primary and secondary spermatocytes, but less so in spermatids. Finally, we demonstrate that knockdown of MAGEB18 inhibits the growth of B16-F0 cells and induces apoptosis, which correlates with increased levels of TP53 (tumour protein 53), p21, Bax and caspase 3. The results of the present study thus uncover an important phenomenon that the expression of certain type I MAGE genes, at least for Mageb18, is non-testis-specific. Although they can regulate various malignant phenotypes of cancer cells, it is necessary to study further their expression pattern in normal tissues before using them to develop more effective and safer cancer vaccines.