Long term in vitro expansion of epithelial stem cells enabled by pharmacological inhibition of PAK1-ROCK-Myosin II and TGF-β signaling

Summary: Despite substantial self-renewal capability in vivo, epithelial stem and progenitor cells located in various tissues expand for a few passages in vitro in feeder-free condition before they succumb to growth arrest. Here, we describe the EpiX method, which utilizes small molecules that inhibit PAK1-ROCK-Myosin II and TGF-β signaling to achieve over one trillion-fold expansion of human epithelial stem and progenitor cells from skin, airway, mammary, and prostate glands in the absence of feeder cells. Transcriptomic and epigenomic studies show that this condition helps epithelial cells to overcome stresses for continuous proliferation. EpiX-expanded basal epithelial cells differentiate into mature epithelial cells consistent with their tissue origins. Whole-genome sequencing reveals that the cells retain remarkable genome integrity after extensive in vitro expansion without acquiring tumorigenicity. EpiX technology provides a solution to exploit the potential of tissue-resident epithelial stem and progenitor cells for regenerative medicine. : Zhang et al. screen a small-molecule collection and find that pharmacologic inhibition of TGF-β and PAK1-ROCK-Myosin II, in low calcium conditions, supports extended expansion of epithelial stem cells in 2D format. This approach enhances the potential of tissue-resident epithelial stem cells for cell therapy. Keywords: epithelial stem and progenitor cells, cell culture method, TGF-β, PAK1/ROCK/Myosin II, feeder-free, regenerative medicine, cell therap

A salient edges detection algorithm of multi-sensor images and its rapid calculation based on PFCM kernel clustering

AbstractMulti-sensor image matching based on salient edges has broad prospect in applications, but it is difficult to extract salient edges of real multi-sensor images with noises fast and accurately by using common algorithms. According to the analysis of the features of salient edges, a novel salient edges detection algorithm and its rapid calculation are proposed based on possibility fuzzy C-means (PFCM) kernel clustering using two-dimensional vectors composed of the values of gray and texture. PFCM clustering can overcome the shortcomings that fuzzy C-means (FCM) clustering is sensitive to noises and possibility C-means (PCM) clustering tends to find identical clusters. On this basis, a method is proposed to improve real-time performance by compressing data sets based on the idea of data reduction in the field of mathematical analysis. In addition, the idea that kernel-space is linearly separable is used to enhance robustness further. Experimental results show that this method extracts salient edges for real multi-sensor images with noises more accurately than the algorithm based on force fields and the FCM algorithm; and the proposed method is on average about 56 times faster than the PFCM algorithm in real time and has better robustness

Negative Regulation of p53 Functions by Daxx and the Involvement of MDM2

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Optimize Individualized Energy Delivery for Septic Patients Using Predictive Deep Learning Models: A Real World Study

Background and Objectives: We aim to establish deep learning models to optimize the individualized energy delivery for septic patients. Methods and Study Design: We conducted a study of adult septic patients in Intensive Care Unit (ICU), collecting 47 indicators for 14 days. After data cleaning and preprocessing, we used stats to explore energy delivery in deceased and surviving patients. We filtered out nutrition-related features and divided the data into three metabolic phases: acute early, acute late, and rehabilitation. Models were built using data before September 2020 and validated on the rest. We then established optimal energy target models for each phase using deep learning. Results: A total of 277 patients and 3115 data were included in this study. The models indicated that the optimal energy targets in the three phases were 900kcal/d, 2300kcal/d, and 2000kcal/d, respectively. Excessive energy intake increased mortality rapidly in the early period of the acute phase. Insufficient energy in the late period of the acute phase significantly raised the mortality of septic patients. For the rehabilitation phase, too much or too little energy delivery both associated with high mortality. Conclusion: Our study established time-series prediction models for septic patients to optimize energy delivery in the ICU. This approach indicated the feasibility of developing nutritional tools for critically ill patients. We recommended permissive underfeeding only in the early acute phase. Later, increased energy intake may improve survival and settle energy debts caused by underfeeding

Vertical Stress and Deformation Characteristics of Roadside Backfilling Body in Gob-Side Entry for Thick Coal Seams with Different Pre-Split Angles

Retained gob-side entry (RGE) is a significant improvement for fully-mechanized longwall mining. The environment of surrounding rock directly affects its stability. Roadside backfilling body (RBB), a man-made structure in RGE plays the most important role in successful application of the technology. In the field, however, the vertical deformation of RBB is large during the panel extraction, which leads to malfunction of the RGE. In order to solve the problem, roof pre-split is employed. According to geological conditions as well as the physical modeling of roof behavior and deformation of surrounding rock, the support resistance of RBB is calculated. The environment of surrounding rock, vertical stress and vertical deformation of the RBB in the RGE with different roof pre-split angles are analyzed using FLAC3D software. With the increase of roof pre-split angle, the vertical stresses both in the coal wall and RBB are minimum, and the vertical deformation of RBB also decreases from 110.51 mm to 6.1 mm. Therefore, based on the results of numerical modeling and field observation, roof pre-split angle of 90° is more beneficial to the maintenance of the RGE

Ab Initio Studies on Interactions in K3_3C60_{60} under High Pressure

Fullerene solids doped with alkali metals (A$_3$C$_{60}$, A = K, Rb, Cs) exhibit a superconducting transition temperature ($T_c$) as high as 40 K, and their unconventional superconducting properties have been a subject of debate. With application of high pressure on K$_3$C$_{60}$ and Rb$_3$C$_{60}$, the experiments demonstrate the decrease of $T_c$. In this paper, we focus on K$_3$C$_{60}$ and derive the structure of K$_3$C$_{60}$ under different pressures based on first-principles calculations, exploring the trends of Coulomb interactions at various pressures. By utilizing the Maximally Localized Wannier function approach, Constrained Density Functional Perturbation Theory (cDFPT), and Constrained Random Phase Approximation (cRPA), we construct a microscopic low-energy model near the Fermi level. Our results strongly indicate that, in the K$_3$C$_{60}$ system, as pressure increases, the effect of phonons is the key to intraorbital electron pairing. There is a dominance of the phonon-driven superconducting mechanism at high pressure

Rapamycin enhances LPS induction of tissue factor and tumor necrosis factor-α expression in macrophages by reducing IL-10 expression

Bacterial lipopolysaccharide (LPS) induces monocytes/macrophages to express proinflammatory cytokines and tissue factor (TF), the primary activator of the coagulation cascade. Anti-inflammatory signaling pathways including the phosphatidylinositol-3-kinase (PI3K)-Akt pathway inhibit proinflammatory and TF gene expression in macrophages. We determined the role of Akt, the mammalian target of rapamycin (mTOR) and interleukin-10 in the inhibition of LPS-induced proinflammatory cytokine and TF gene expression in peritoneal macrophages (PMs). We used wild type (WT) peritoneal macrophages (PMs), and PMs from PTENflox/flox/LysMCre mice (PTEN−/− PMs), which have increased Akt activity. Pharmacologic inhibition of mTOR with rapamycin inhibited LPS induction of IL-10 mRNA and protein, and enhanced the expression of TF and the proinflammatory cytokine TNFα in WT PMs. Furthermore, neutralizing IL-10 with anti-IL-10 antibody enhanced LPS induction of TNFα and TF expression in WT PMs. The addition of recombinant IL-10 abolished rapamycin enhancement of LPS-induced TNFα and TF expression in WT PMs. Consistent with enhanced Akt activation, LPS-induced IL-10 expression was increased in PTEN−/− PMs compared to WT PMs. In contrast, LPS-induced TNFα and TF expression was significantly reduced in PTEN−/− PMs compared to WT PMs. However, the neutralizing IL-10 antibody did not completely prevent inhibition of LPS-induced TNFα and TF expression in PTEN−/− PMs. The results indicate that mTOR-dependent IL-10 expression leads to inhibition of LPS induction of TF and the proinflammatory cytokine TNFα in WT macrophages. In contrast, the decrease in LPS-induced TNFα and TF expression in PTEN−/− PMs also requires an IL-10-independent pathway

Macrophages Phenotype Regulated by IL-6 Are Associated with the Prognosis of Platinum-Resistant Serous Ovarian Cancer: Integrated Analysis of Clinical Trial and Omics

Background. The treatment of platinum-resistant recurrent ovarian cancer (PROC) is a clinical challenge and a hot topic. Tumor microenvironment (TME) as a key factor promoting ovarian cancer progression. Macrophage is a component of TME, and it has been reported that macrophage phenotype is related to the development of PROC. However, the mechanism underlying macrophage polarization and whether macrophage phenotype can be used as a prognostic indicator of PROC remains unclear. Methods. We used ESTIMATE to calculate the number of immune and stromal components in high-grade serous ovarian cancer (HGSOC) cases from The Cancer Genome Atlas database. The differential expression genes (DEGs) were analyzed via protein–protein interaction network, Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene ontology (GO) analysis to reveal major pathways of DEGs. CD80 was selected for survival analysis. IL-6 was selected for gene set enrichment analysis (GSEA). A subsequent cohort study was performed to confirm the correlation of IL-6 expression with macrophage phenotype in peripheral blood and to explore the clinical utility of macrophage phenotype for the prognosis of PROC patients. Results. A total of 993 intersecting genes were identified as candidates for further survival analysis. Further analysis revealed that CD80 expression was positively correlated with the survival of HGSOC patients. The results of GO and KEGG analysis suggested that macrophage polarization could be regulated via chemokine pathway and cytokine–cytokine receptor interaction. GSEA showed that the genes were mainly enriched in IL-6-STAT-3. Correlation analysis for the proportion of tumor infiltration macrophages revealed that M2 was correlated with IL-6. The results of a cohort study demonstrated that the regulation of macrophage phenotype by IL-6 is bidirectional. The high M1% was a protective factor for progression-free survival. Conclusion. Thus, the macrophage phenotype is a prognostic indicator in PROC patients, possibly via a hyperactive IL-6-related pathway, providing an additional clue for the therapeutic intervention of PROC