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Eukaryotic cells share a basic scheme of internal organization featuring membrane-based organelles. The use of fluorescent proteins (FPs) greatly facilitated live-cell imaging of organelle dynamics and protein trafficking. One major limitation of this approach is that the fusion of an FP to a target protein can and often does compromise the function of the target protein and alter its subcellular localization. The optimization process to obtain a desirable fusion construct can be time-consuming or even unsuccessful. In this work, we set out to provide a validated set of FP-based markers for major organelles in the budding yeast ( Saccharomyces cerevisiae ). Out of over 160 plasmids constructed, we present a final set of 42 plasmids, the recommendations for which are backed up by meticulous evaluations. The tool set includes three colors (green, red, and blue) and covers the endoplasmic reticulum (ER), nucleus, Golgi apparatus, endosomes, vacuoles, mitochondria, peroxisomes, and lipid droplets. The fidelity of the markers was established by systematic cross-comparison and quantification. Functional assays were performed to examine the impact of marker expression on the secretory pathway, endocytic pathway, and metabolic activities of mitochondria and peroxisomes. Concomitantly, our work constitutes a reassessment of organelle identities in this model organism. Our data support the recognition that “late Golgi” and “early endosomes,” two seemingly distinct terms, denote the same compartment in yeast. Conversely, all other organelles can be visually separated from each other at the resolution of conventional light microscopy, and quantification results justify their classification as distinct entities. IMPORTANCE Cells contain elaborate internal structures. For eukaryotic cells, like those in our bodies, the internal space is compartmentalized into membrane-bound organelles, each tasked with specialized functions. Oftentimes, one needs to visualize organelles to understand a complex cellular process. Here, we provide a validated set of fluorescent protein-based markers for major organelles in budding yeast. Yeast is a commonly used model when investigating basic mechanisms shared among eukaryotes. Fluorescent proteins are produced by cells themselves, avoiding the need for expensive chemical dyes. Through extensive cross-comparison, we make sure that each of our markers labels and only labels the intended organelle. We also carefully examined if the presence of our markers has any negative impact on the functionality of the cells and found none. Our work also helps answer a related question: are the structures we see really what we think they are?

The formation of autophagosomes mediating the sequestration of cytoplasmic materials is the central step of autophagy. Several phosphoinositides, which are signaling molecules on the membrane, are involved in autophagy. However, it is not always clear whether these phosphoinositides act directly at the site of autophagosome formation, or indirectly via the regulation of other steps or pathways. To address this question, we used a set of phosphoinositide probes to systematically examine their potential presence on autophagosomal membranes in yeast (Saccharomyces cerevisiae). We verified the specificity of these probes using mutant cells deficient in the production of the corresponding phosphoinositides. We then examined starved yeast cells co-expressing a phosphoinositide probe together with an autophagosomal membrane marker, 2Katushka2S-Atg8. Our data revealed that PtdIns(4,5)P2 and PtdIns(3,5)P2 were mainly present on the plasma membrane and vacuolar membrane, respectively. We observed only occasional co-localization between the PtdIns(4)P probe and Atg8, some of which may represent the transient passage of a PtdIns(4)P-containing structure near the autophagosomal membrane. In contrast, substantial colocalization of the PtdIns(3)P probe with Atg8 was observed. Taken together, our data indicate that only PtdIns(3)P is present in a substantial amount on the autophagosomal membrane. For other phosphoinositides involved in autophagy, either their presence on the autophagosomal membrane is very transient, or they act on other cellular membranes to regulate autophagy.

With the considerations of the behaviors of shell deformation, mold flux film and air gap dynamic distribution in shell/mold gap, a two dimensional slice-travel transient thermo-mechanical coupled model of simulation shell solidification in wide and thick slab continuous casting mold was developed by using the commercial program ANSYS. The evolutions of strand-mold system thermal behaviors, including the air gap formation and the mold flux film dynamical distribution in shell/mold gap and shell temperature field, and the evolutions of shell deformation and stress distribution of peritectic steel solidified in a 2120 mm wide and 266 mm thick slab continuous casting mold were analyzed. The results show that the air gap formation and the thick mold flux film distribution mainly concentrate in the regions 0–21 mm and 0–7 mm, 0–120 mm and 0–100 mm off the shell wide and narrow faces corners, and thus the hot spots are given rise to form in the regions 15–55 mm and 15–50 mm off the shell wide and narrow face corners. The shell server deformation occurs in the off-corners in the middle and lower parts of the mold. The stress evolution in shell surface is tensile stress, while that in shell solidification front is compressive stress.

Recently, the incidence of thyroid cancer (THCA) has been on the rise. RNA binding proteins (RBPs) and their abnormal expression are closely related to the emergence and pathogenesis of tumor diseases. In this study, we obtained gene expression data and corresponding clinical information from the TCGA database. A total of 162 aberrantly expressed RBPs were obtained, comprising 92 up-regulated and 70 down-regulated RBPs. Then, we performed a functional enrichment analysis and constructed a PPI network. Through univariate Cox regression analysis of key genes and found that NOLC1 ( p = 0.036), RPS27L ( p = 0.011), TDRD9 ( p = 0.016), TDRD6 ( p = 0.002), IFIT2 ( p = 0.037), and IFIT3 ( p = 0.02) were significantly related to the prognosis. Through the online website Kaplan-Meier plotter and multivariate Cox analysis, we identified 2 RBP-coding genes ( RPS27L and IFIT3 ) to construct a predictive model in the entire TCGA dataset and then validate in two subsets. In-depth analysis revealed that the data gave by this model, the patient’s high-risk score is very closely related to the overall survival rate difference ( p = 0.038). Further, we investigated the correlation between the model and the clinic, and the results indicated that the high-risk was in the male group ( p = 0.011) and the T3-4 group ( p = 0.046) was associated with a poor prognosis. On the whole, the conclusions of our research this time can make it possible to find more insights into the research on the pathogenesis of THCA, this could be beneficial for individualized treatment and medical decision making.

Shot peen forming is an important method for preparing aircraft wall panels, but it introduces residual compressive stress fields on a material′s surface, as well as surface roughness and defects. The corrosion behavior of a shot peen-formed aluminum alloy sheet (FPA) in a salt spray environment was studied. The results demonstrated that the corrosion rates of FPA samples were faster than those without shot peen forming, especially during the initial stage. Both FPA-treated and untreated samples experienced pitting, but the pits in the FPA-treated sample were larger and deeper. The cross-sectional analysis of the corrosion layer showed that the corrosion layer of the shot-peened sheet was thicker, and the anodic oxidation protective film was easily broken. Intergranular corrosion of the FPA-treated sample occurred in a salt spray environment after only 10 min, and the corrosion pits were 35.04 μm deep. For samples without FPA treatment, intergranular corrosion occurred after 5 h of salt spray corrosion. The cracks that formed after shot peen forming acted as corrosion channels that increased the corrosion susceptibility of the aluminum alloy. After shot peen forming, the impedance value and the radius of capacitive resistance arc of the samples decreased in the process of salt spray corrosion, indicating that the corrosion resistance of the alloys was reduced.

Trophic Level Index (TLI) is often used to assess the general eutrophication state of inland lakes in water science, technology, and engineering. In this paper, a data-driven inland-lake eutrophication assessment method was proposed by using an artificial neural network (ANN) to build relationships from remote sensing data and in-situ TLI sampling. In order to train the net, Moderate Resolution Imaging Spectroradiometer (MODIS, which has a revisit cycle of 4 times per day) data were combined with in-situ observations. Results demonstrate that the TLI obtained directly from remote-sensing images using the data-driven method is more accurate than the TLI calculated from the water quality factors retrieved from remote-sensing images using a multivariate regression method. Spatially continuous and quasi-real time results were retrieved by using MODIS data. This method provides an efficient way to map the TLI spatial distribution in inland lakes, and provides a scheme for increased automation in TLI estimation.

The accelerating impact of climate change on giant panda (Ailuropoda melanoleuca) habitats have become an international research topic. Recently, many studies have also focused on medium-sized mountain ranges or entire giant panda habitats to predict how habitats will change as the climate warms, but few say in detail what to do or where to focus efforts. To fill this gap, this paper presents a new method to take comprehensive, fine-scale evaluations incorporating climate change, human disturbance, and current conservation networks and translate them into practical countermeasures in order to help decision-makers set priority regions for conservation. This study looked at the core area of the Sichuan Giant Panda Sanctuaries United Nations Educational, Scientific and Cultural Organisation (UNESCO) World Natural Heritage site, namely Ya’an Prefecture, as a case study. The research employs the Maximum Entropy (MaxEnt) modeling algorithm to analyze how climate change will affect the habitats by 2050 under two scenarios: only considering the influence of climate change, and thinking about the coupled influence of climate change and human disturbance together. The results showed the following: (1) only considering climate change, the overall habitat that can be used by giant pandas in this region will increase, which differs from most of the previous results showing a decrease; (2) the new suitable habitat will shift westward, northward and eastward in this region; (3) conversely, the suitable habitat will be significantly reduced (about 58.56%) and fragmentized when taking into account human disturbance factors; (4) at present, the three small nature reserves are far from each other and cannot cover the present habitat well nor protect the potentially suitable habitats. Based on the comprehensive analysis of habitat shifts and our two field investigations, we suggest two regions that can be expanded into the conservation network to contain more potentially suitable habitats in the future. Furthermore, we used a geographical information system to incorporate high-resolution remote-sensing images from the GF-1 satellite, land-cover maps, and a digital elevation model (DEM) to verify the possibility of our two suggested regions.

At the end of March 2013, the first case of human infection with avian influenza A（H7N9） virus was confirmed in Shanghai. From April to May 2013, 18 patients with avian influenza A（H7N9） virus infection were hospitalized in Shanghai Public Health Clinical Center, and finally, 12 of them survived. The short-term prognosis of these patients had been described previously, but the long-term prognosis remained unclear.

Angiogenesis is a key process in regenerative medicine generally, as well as in the specific field of nerve regeneration. However, no convenient and objective method for evaluating the angiogenesis of tissue-engineered nerves has been reported. In this study, tissue-engineered nerves were constructed in vitro using Schwann cells differentiated from rat skin-derived precursors as supporting cells and chitosan nerve conduits combined with silk fibroin fibers as scaffolds to bridge 10-mm sciatic nerve defects in rats. Four weeks after surgery, three-dimensional blood vessel reconstructions were made through MICROFIL perfusion and micro-CT scanning, and parameter analysis of the tissue-engineered nerves was performed. New blood vessels grew into the tissue-engineered nerves from three main directions： the proximal end, the distal end, and the middle. The parameter analysis of the three-dimensional blood vessel images yielded several parameters, including the number, diameter, connection, and spatial distribution of blood vessels. The new blood vessels were mainly capillaries and microvessels, with diameters ranging from 9 to 301 μm. The blood vessels with diameters from 27 to 155 μm accounted for 82.84% of the new vessels. The microvessels in the tissue-engineered nerves implanted in vivo were relatively well-identified using the MICROFIL perfusion and micro-CT scanning method, which allows the evaluation and comparison of differences and changes of angiogenesis in tissue-engineered nerves implanted in vivo.

Improving charge transfer is the key to the performance of non-noble metal semiconductor-based surface enhanced Raman scattering (SERS) substrates. In this work, the O-incorporated 1T-MoS2 nanosheets with rich sulfur defects (ID-MoS2) are obtained by simple calcination of 1T-MoS2 nanosheets in air atmosphere. Using rhodamine 6G (R6G) as typical probe molecules, ID-MoS2 nanosheets show ultrahigh Raman enhancement effects with an enhancement factor of 1.24 × 107 due to sulfur defects and O incorporation in the 1T-MoS2 lattice. First-principle density functional theory calculations suggest that the existence of sulfur defects and O incorporation significantly increase the Fermi energy level (Ef) and electronic density of states of ID-MoS2. Moreover, O incorporation can enhance the interactions between the substrate and the adsorbed molecules through electrostatic and hydrogen bonding. All these improve the charge transfer resonance and result in the remarkable SERS activity of ID-MoS2 nanosheets. This is the first study on the increasing SERS performance of semiconductor substrates by simultaneously employing defect and dopant incorporation. This study provides an approach to optimize the performance of semiconductor-based SERS substrates.