The Mechanism of Tubular Recycling Endosome Biogenesis

Endocytic trafficking is a critical process for cellular homeostasis, and multiple ailments that include cardiovascular disease and cancer are related to the dysregulation of endocytic transport. As vesicles and target membranes are key to endocytic transport, lipids are essential for the regulation of endocytic trafficking pathways. We have shown that the tubular recycling endosomes (TRE) are essential for the regulation of endocytic recycling pathways. However, the mechanisms by which TRE are biosynthesized and carry out their functions remain unsolved. Studies from our lab have shown that phosphatidic acid (PA) recruits Molecule Interacting with Casl-Like protein 1 (MICAL-L1) as well as Syndapin2, and subsequently Eps15 homology domain containing protein EHD1 to the membrane of TREs as a complex, and this complex is essential for the biogenesis of TRE and efficient recycling of internalized receptors back to the plasma membrane. However, the involvement of PA in endocytic trafficking has not been well characterized. Diacylglycerol kinase (DGK) α is one of ten DGK isoforms that converts diacylglycerol (DAG) to PA. We showed that depletion of DGKα, a kinase devoid of a clathrin-dependent adaptor protein complex 2 binding site, results in an impaired biogenesis of TRE. As a consequence, we observed a delay in MHC I recycling to the plasma membrane. On the other hand, the rate of MHC I internalization remained unaffected. By Co-immunoprecipitation assay, we showed that DGKα forms a complex with the TRE hub protein, MICAL-L1. Given that MICAL-L1 and the F-BAR-containing membrane-tubulating protein Syndapin2 associate selectively with PA, whose generation is majorly mediated by DGKα, we propose a positive feedback loop in which DGKα generates PA to drive its own recruitment to TRE via its interaction with MICAL-L1. Our data support a novel role for the involvement of DGKα in TRE biogenesis and MHC I recycling. While the molecular mechanism of MICAL-L1 decorated TRE biogenesis is further revealed by our studies of the involvement of DGKα, major questions with regards to their function in endocytic recycling still remained unsolved, such as what cargos travel through these tubular membrane structures, where is the destination of these cargos, etc. We showed that TRE preferentially traffic cargos internalized via clathrin-independent endocytosis (CIE), and may originate from the sorting endosomes (SE). Since MICAL-L1 TRE is a major component of the endocytic recycling compartment (ERC), the understanding of the composition and cargo distribution within the ERC would further add to our knowledge of TRE functions. We used 3D Structured Illumination Microscopy, dual-channel and 3D direct Stochastic Optical Reconstruction Microscopy (dSTORM) to obtain new information about ERC morphology and cargo segregation. For the first time, we discovered that cargo internalized either via clathrin-mediated endocytosis (CME) or CIE remains segregated in the ERC, likely on distinct carriers. This suggests that no further sorting occurs upon cargo exit from SE. Moreover, 3D dSTORM data support a model in which some, but not all ERC vesicles are tethered by contiguous ‘membrane bridges’. These findings support a significantly altered model for endocytic recycling in mammalian cells, in which cargos are sorted in the peripheral endosomes and carried by MICAL-L1 decorated TRE to the ERC, while segregation is maintained at the ERC

Utjecaj dodatka ekstrakta maslačka, saharoze i starter kulture na viskoznost, sposobnost zadržavanja vode i pH jogurta

Dandelion extract is a traditional Chinese medicine and contains significant nutritional value. The aim of this study was to research the optimum fermentation conditions for dandelion addition to plain yogurt using a single factor experiments and orthogonal experiment. The results of the present study demonstrated that the addition of dandelion extract affected the viscosity, water-holding capacity and pH of yogurt. Optimized conditions for dandelion addition to plain yogurt based on viscosity, incubation time, pH and sensory score were 10 % sucrose, 0.3 % of the starter cultures, incubation time of 6.5 hours and 3 % dandelion extract. A new kind of dandelion yogurt with high viscosity, good water-holding capacity and good taste was prepared in this study.Ekstrakt maslačka je tradicionalni kineski lijek, a također ima i značajnu nutritivnu vrijednost. Cilj ovog istraživanja bio je ispitati optimalne uvjete fermentacije za dodavanje maslačka u jogurt korištenjem pojedinih faktorskih eksperimenata i ortogonalnog eksperimenta. Rezultati ove studije pokazali su da dodavanje ekstrakta maslačaka utječe na viskoznost, sposobnost zadržavanja vode i pH jogurta. Optimalni uvjeti za dodavanje maslačka u jogurt na temelju viskoznosti, vremena inkubacije, pH i senzorskog rezultata bili su 10 % saharoze, 0,3 % starter kulture, vrijeme inkubacije od 6,5 sati i dodatak 3 % ekstrakta maslačka. U ovoj studiji pripremljena je nova vrsta jogurta s dodatkom maslačka karakteriziranog visokom viskoznošću, dobrom sposobnošću zadržavanja vode i dobrim okusom

Rethinking Alignment in Video Super-Resolution Transformers

The alignment of adjacent frames is considered an essential operation in video super-resolution (VSR). Advanced VSR models, including the latest VSR Transformers, are generally equipped with well-designed alignment modules. However, the progress of the self-attention mechanism may violate this common sense. In this paper, we rethink the role of alignment in VSR Transformers and make several counter-intuitive observations. Our experiments show that: (i) VSR Transformers can directly utilize multi-frame information from unaligned videos, and (ii) existing alignment methods are sometimes harmful to VSR Transformers. These observations indicate that we can further improve the performance of VSR Transformers simply by removing the alignment module and adopting a larger attention window. Nevertheless, such designs will dramatically increase the computational burden, and cannot deal with large motions. Therefore, we propose a new and efficient alignment method called patch alignment, which aligns image patches instead of pixels. VSR Transformers equipped with patch alignment could demonstrate state-of-the-art performance on multiple benchmarks. Our work provides valuable insights on how multi-frame information is used in VSR and how to select alignment methods for different networks/datasets. Codes and models will be released at https://github.com/XPixelGroup/RethinkVSRAlignment.Comment: This paper has been accepted for NeurIPS 202

Extreme long-lifetime self-assembled monolayer for air-stable molecular junctions

The molecular electronic devices based on self-assembled monolayer (SAM) on metal surfaces demonstrate novel electronic functions for device minimization yet are unable to realize in practical applications, due to their instability against oxidation of the sulfur-metal bond. This paper describes an alternative to the thiolate anchoring group to form stable SAMs on gold by selenides anchoring group. Because of the formation of strong selenium-gold bonds, these stable SAMs allow us to incorporate them in molecular tunnel junctions to yield extremely stable junctions for over 200 days. A detailed structural characterization supported by spectroscopy and first-principles modeling shows that the oxidation process is much slower with the selenium-gold bond than the sulfur-gold bond, and the selenium-gold bond is strong enough to avoid bond breaking even when it is eventually oxidized. This proof of concept demonstrates that the extraordinarily stable SAMs derived from sel-enides are useful for long-lived molecular electronic devices and can possibly become important in many air-stable applications involving SAMs.</p

Site-specific incorporation of phosphotyrosine using an expanded genetic code.

Access to phosphoproteins with stoichiometric and site-specific phosphorylation status is key to understanding the role of protein phosphorylation. Here we report an efficient method to generate pure, active phosphotyrosine-containing proteins by genetically encoding a stable phosphotyrosine analog that is convertible to native phosphotyrosine. We demonstrate its general compatibility with proteins of various sizes, phosphotyrosine sites and functions, and reveal a possible role of tyrosine phosphorylation in negative regulation of ubiquitination

Investigation of the Structure Requirement for 5-HT6 Binding Affinity of Arylsulfonyl Derivatives: A Computational Study

5-HT6 receptor has been implicated in a series of diseases including anxiety, depression, schizophrenia and cognitive dysfunctions. 5-HT6 ligands have been reported to play a significant role in the treatment for central nervous system (CNS) diseases. Presently, a large series of 223 5-HT6 ligands were studied using a combinational method by 3D-QSAR, molecular docking and molecular dynamics calculations for further improvement of potency. The optimal 3D models exhibit satisfying statistical results with r2ncv, q2 values of 0.85 and 0.50 for CoMFA, 0.81 and 0.53 for CoMSIA, respectively. Their predictive powers were validated by external test set, showing r2pred of 0.71 and 0.76. The contour maps also provide a visual representation of contributions of steric, electrostatic, hydrophobic and hydrogen bond fields as well as the prospective binding models. In addition, the agreement between 3D-QSAR, molecular docking and molecular dynamics simulation proves the rationality of the developed models. These results, we hope, may be helpful in designing novel and potential 5-HT6 ligands

3D printed nanocellulose-based label for fruit freshness keeping and visual monitoring

Food packaging systems with a single function of freshness keeping or monitoring may not be able to meet all practical needs. Herein, cellulose nanofibers (CNF)-based labels with dual functions of fruit freshness keeping and visual monitoring were prepared by coaxial 3D printing. CNF-based ink with blueberry anthocyanin was used to create the shell of fibers, exhibiting high formability and print fidelity as well as sensitive visual pH-responsiveness for freshness monitoring. Chitosan containing 1-methylcyclopropene (1-MCP) was loaded into the hollow microchannels of fibers, in which 1-MCP was trapped by the electrostatic effect of chitosan and CNF and exhibited a sustained release behavior. The 3D printed labels prolonged the shelf life of litchis for 6 days, meanwhile, they sensitively indicated the changes in freshness and the accuracy was confirmed by Headspace-Gas Chromatography-Ion Mobility Spectrometry. The CNF-based integrated labels developed in this work provided a new idea for the development of food intelligent packaging

Mitigating Artifacts in Real-World Video Super-resolution Models

The recurrent structure is a prevalent framework for the task of video super-resolution, which models the temporal dependency between frames via hidden states. When applied to real-world scenarios with unknown and complex degradations, hidden states tend to contain unpleasant artifacts and propagate them to restored frames. In this circumstance, our analyses show that such artifacts can be largely alleviated when the hidden state is replaced with a cleaner counterpart. Based on the observations, we propose a Hidden State Attention (HSA) module to mitigate artifacts in real-world video super-resolution. Specifically, we first adopt various cheap filters to produce a hidden state pool. For example, Gaussian blur filters are for smoothing artifacts while sharpening filters are for enhancing details. To aggregate a new hidden state that contains fewer artifacts from the hidden state pool, we devise a Selective Cross Attention (SCA) module, in which the attention between input features and each hidden state is calculated. Equipped with HSA, our proposed method, namely FastRealVSR, is able to achieve 2x speedup while obtaining better performance than Real-BasicVSR. Codes will be available at https://github.com/TencentARC/FastRealVSR

Sucrose non-fermenting protein kinase gene UvSnf1 is required for virulence in Ustilaginoidea virens

ABSTRACTRice false smut caused by Ustilaginoidea virens is becoming one of the most devastating diseases in rice production areas in the world. Revealing U. virens potential pathogenic mechanisms provides ideas for formulating more effective prevention and control strategies. Sucrose non-fermenting 1 (Snf1) protein kinase plays a critical role in activating transcription and suppressing gene expression, as well as in cellular response to various stresses, such as nutrient limitation. In our study, we identified the Snf1 homolog UvSnf1 and analyzed its biological functions in U. virens. The expression level of UvSnf1 was dramatically up-regulated during invasion, indicating that UvSnf1 may participate in infection. Phenotypic analyses of UvSnf1 deletion mutants revealed that UvSnf1 is necessary for hyphae growth, spore production, and virulence in U. virens. Moreover, UvSnf1 promotes U. virens to use unfavorable carbon sources when the sucrose is insufficient. In addition, deletion of UvSnf1 down-regulates the expression of the cell wall-degrading enzymes (CWDEs) genes under sucrose limitation conditions in U. virens. Further analyses showed that CWDEs (UvCut1 and UvXyp1) are not only involved in growth, spore production, and virulence but are also required for the utilization of carbon sources. In conclusion, this study demonstrates that UvSnf1 plays vital roles in virulence and carbon source utilization in U. virens, and one of the possible mechanisms is playing a role in regulating the expression of CWDE genes