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SummaryVarious hormones, kinases, and stressors (fasting, heat shock) stimulate 26S proteasome activity. To understand how its capacity to degrade ubiquitylated protein can increase, we studied ZFAND5, which promotes protein degradation during muscle atrophy. Cryo-electron microscopy showed that ZFAND5 induces large conformational changes in the 19S regulatory particle. ZFAND5’s AN1 Zn finger interacts with the Rpt5 ATPase and its C-terminus with Rpt1 ATPase and Rpn1, a ubiquitin-binding subunit. Surprisingly, these C-terminal interactions are sufficient to activate proteolysis. With ZFAND5 bound, entry into the proteasome’s protein translocation channel is wider, and ZFAND5 dissociation causes opening of the 20S gate for substrate entry. Using single-molecular microscopy, we showed that ZFAND5 binds ubiquitylated substrates, prolongs their association with proteasomes, and increases the likelihood that bound substrates undergo degradation, even though ZFAND5 dissociates before substrate deubiquitylation. These changes in proteasome conformation and reaction cycle can explain the accelerated degradation and suggest how other proteasome activators may stimulate proteolysis.

Sonodynamic therapy (SDT) has aroused great interest for its potential in the treatment of glioblastoma (GBM). SDT relies on tumor-selective accumulation of a sonosensitizer that is activated by ultrasound irradiation (UI) to generate cytotoxic actions. The efficacy of GBM-SDT depends on sufficient sonosensitizer buildup in the tumor, which is, however, seriously hampered by the anatomical and biochemical barriers of the GBM. To overcome this difficulty, we herein propose a delivery strategy of ‘platelets with ultrasound-triggered release property’, which takes advantage of 1) the platelets’ ability to carry cargo and release cargo upon activation, and 2) the ROS-generating property of SDT. To provide proof of concept for the strategy, we first stably loaded platelets with IOPD-Ce6, a nano-formed sonosensitizer consisting of iron oxide nanoparticles coated with polyglycerol and doxorubicin and loaded with chlorine e6. UI of the IOPD-Ce6-loaded platelets (IOPD-Ce6@Plt) elicited ROS generation in the IOPD-Ce6@Plt, which were immediately activated to release IOPD-Ce6 into GBM cells in co-culture which, when subjected to a second time of UI, exhibited pronounced ROS production, DNA injury, viability loss, and cell death in the GBM cells. In the in vivo experiments, mice bearing intracranial GBM grafts exhibited substantial tumor distribution of IOPD-Ce6 following intravenous injection of IOPD-Ce6@Plt and subsequent UI at the tumor site. The GBM grafts then exhibited pronounced cell injury and death after another round of UI of the tumors. Finally, the growth of intra-cranial GBM grafts was significantly slowed when an SDT protocol consisting of an intravenous IOPD-Ce6@Plt injection followed by multiple times of tumor UI had been applied twice to the mice. Our results are strong evidence for the idea that platelets are sound and amenable carriers to deliver sonosensitizers in the GBM in an ultrasound-triggered manner and thus to produce highly targeted and effective SDT of GBM.

Targeting signaling pathways that drive cancer cell migration or proliferation is a common therapeutic approach. A popular experimental technique, the scratch assay, measures the migration and proliferation-driven cell monolayer formation. Scratch assay analyses do not differentiate between migration and proliferation effects and do not attempt to measure dynamic effects. To improve upon these methods, we combine high-throughput scratch assays, continuous video microscopy, and variational system identification (VSI) to infer partial differential equation (PDE) models of cell migration and proliferation. We capture the evolution of cell density fields over time using live cell microscopy and automated image processing. We employ VSI techniques to identify cell density dynamics modeled with first-order kinetics of advection-diffusion-reaction systems. We present a comparison of our methods to results obtained using traditional inference approaches on previously analyzed 1-dimensional scratch assay data. We demonstrate the application of this pipeline on high throughput 2-dimensional scratch assays and find that decreasing serum levels can decrease random cell migration by approximately 20%. Our integrated experimental and computational pipeline can be adapted for automatically quantifying the effect of biological perturbations on cell migration and proliferation in various cell lines.

The current study used an attentional blink (AB) paradigm to test how grouping influences the initial selection and the subsequent short-term memory consolidation of a target. On a given trial, two red Kanizsa-type targets (T1, T2) with varying grouping strength were embedded in a rapid serial visual presentation stream of irrelevant distractors.

Visual working memory (VWM) is typically considered to represent complete objects, that is, separate parts of an object are maintained as bound objects. Yet, it remains unclear whether and how the features of disparate parts are integrated into a whole-object memory representation. Using a change detection paradigm, the present study investigated whether VWM performance varies as a function of grouping strength for features that either determine the grouped object (orientation) or which are not directly grouping-relevant (color). Our results showed a large grouping benefit for grouping-relevant orientation features and additionally, a smaller, but still reliable benefit for grouping-irrelevant color features when both were potentially task-relevant. By contrast, when color was the only task-relevant feature, no grouping benefit from the orientation feature was revealed both under lower or relatively high demands for precision.

BackgroundHand, foot and mouth disease (HFMD) is an acute enterovirus-induced disease. Gut microbiota dysbiosis has been identified as a factor that plays an important role in enteral virus infection, but the gut microbiota profile in hand, foot and mouth disease has rarely been studied in a large population.MethodsA total of 749 children (HFMD: n = 262, healthy control: n = 487) aged 2 to 7 years were recruited from hospitals and communities in the period from May to July, 2017. Clinical and demographical information was collected by trained personnel, and fecal samples were collected and processed for 16S ribosomal RNA(rRNA) gene sequencing.ResultsWe observed a significant alteration in the microbiota profile of children with HFMD compared with that of control children. Patients with enteroviruses A71(EV71) positive had more dysbiotic gut microbiota than those with coxsackievirus A16 (CAV16) positive. We found that Prevotella and Streptococcus were enriched in children with HFMD, whereas beneficial bacteria, including Bifidobacterium and Faecalibacterium, were depleted. Children with synbiotics supplements had lower risk of HFMD and we observed that the gut microbiota of HFMD patients who were administered synbiotics exhibited potential resistance to the dysbiosis detected in HFMD.ConclusionsThis study suggested that the gut microbiota of patients with hand, foot and mouth disease exhibits dysbiosis and that synbiotics supplements potentially helps maintain the homeostasis of the gut flora.

Additional file 4: Table S4. Oligonucleotide sequences in this study.

Additional file 2: Table S2. Information of CRC patients collected in tissue microarray of metastasis.

Additional file 7: Table S7. Primer sequences for the miR-200 family members/CDH1 in ChIP assay.

Additional file 1: Table S1. Information of CRC patients collected for clinical studies.

Additional file 5: Table S5. Antibodies used for ChIP, western blot, IHC and IF assays in this study.

Additional file 3: Table S3. Information of CRC patients collected in tissue microarray of survival.

Additional file 10. Supplementary methods.

Additional file 9: Table S9. Primers used for construction of plasmids in CRISPR-Cas9 assay.

Additional file 6: Table S6. Primer sequences for the 36 miRNAs in qRT-PCR assay of CRC cells.

Additional file 8: Table S8. Primers used for qRT-PCR analysis in this study.

This work presented a new drone-based face detection dataset Drone LAMS in order to solve issues of low performance of drone-based face detection in scenarios such as large angles which was a predominant working condition when a drone flies high. The proposed dataset captured images from 261 videos with over 43k annotations and 4.0k images with pitch or yaw angle in the range of -90{\deg} to 90{\deg}. Drone LAMS showed significant improvement over currently available drone-based face detection datasets in terms of detection performance, especially with large pitch and yaw angle. Detailed analysis of how key factors, such as duplication rate, annotation method, etc., impact dataset performance was also provided to facilitate further usage of a drone on face detection., Comment: 10 pages, 6 figures,conference or other essential info

Human papillomavirus (HPV) type 16 is the most common type implicated as the etiological agent that causes cervical cancer. The marketed prophylactic vaccines against HPV infection are composed of virus-like particles (VLPs) assembled from the recombinant major capsid protein L1. Elicitation of functional and neutralizing antibodies by vaccination is the mode of action by which the vaccines prevent the viral infection. In this study, a panel of murine mAbs against HPV16 L1 were generated and comprehensively characterized with respect to their mapping to the epitope spectrum on the viral capsid. These mAbs were categorized into five epitope bins by two different methods based on the pairwise cross-inhibition and competition with human polyclonal antibodies. In addition, a preliminary demonstration of the spatial relationship of the epitopes recognized by these mAbs was performed using a cross-blocking assay with a well-characterized human mAb, 26D1. Interestingly, two mAbs recognizing different epitopes were found to act synergistically in the pseudovirion-based neutralization assay (PBNA). To facilitate cross-lab and cross-study comparison, the international standard (IS) serum 05/134 was used to calibrate the mAbs as well as the human serum samples from the HPV16/18 vaccine recipients. The neutralizing mAbs, particularly those that recognizing immunodominant epitopes, would be useful in developing epitope-specific assays for monitoring the vaccine production process and for serological assessment.

Thimerosal has been widely used as a preservative in human vaccines for decades. Thimerosal, a thiol capping agent with ethyl mercury being the active degradant, could have impacts on the vaccine potency due to potential thiol modification. The effects on the antigenicity and immunogenicity of human papillomavirus (HPV) virus-like particles (VLPs) in the presence of thimerosal was studied. In general, reduced binding activity was observed between HPV antigens and monoclonal antibodies (mAbs) upon thimerosal treatment, accompanied by reduced protein conformational stability. The immunogenicity of a pentavalent vaccine formulation (HPV6, HPV11, HPV16, HPV18 and hepatitis E virus) with or without thimerosal was studied in mice. The functional antibody titres, as well as the binding titres, were determined, showing a substantial decrease for vaccine formulations containing thimerosal for HPV16/18. Similarly, epitope-specific competition assays using specific and functional mAbs as tracers also showed a significant reduction in immunogenicity for HPV16/18 in the presence of thimerosal. Structural alterations in the capsid protein for HPV18 were observed with cryo-electron microscopy and 3-dimensional reconstruction in the comparative structural analysis. The results should alert scientists in formulation development field on the choice for vaccine preservatives, in particular for thiol-containing antigens.

Ultra-dense networking is widely accepted as a promising enabling technology to realize high power and spectrum efficient communications in future 5G communication systems. Although joint resource allocation schemes promise huge performance improvement at the cost of cooperation among base stations, the large numbers of user equipment and base station make jointly optimizing the available resource very challenging and even prohibitive. How to decompose the resource allocation problem is a critical issue. In this paper, we exploit factor graphs to design a distributed resource allocation algorithm for ultra dense networks, which consists of power allocation, subcarrier allocation and cell association. The proposed factor graph based distributed algorithm can decompose the joint optimization problem of resource allocation into a series of low complexity subproblems with much lower dimensionality, and the original optimization problem can be efficiently solved via solving these subproblems iteratively. In addition, based on the proposed algorithm the amounts of exchanging information overhead between the resulting subprob-lems are also reduced. The proposed distributed algorithm can be understood as solving largely dimensional optimization problem in a soft manner, which is much preferred in practical scenarios. Finally,the performance of the proposed low complexity distributed algorithm is evaluated by several numerical results.

The factors affecting the economy efficiency of rail grinding and their interrelation are researched. Based on the management theory of life-cycle cost, the model of analyzing rail grinding parameters is established, the calculation method is determined, and minimizing the rail life-cycle cost is regarded as optimization objective for the optimal analysis of grinding factor. The results prove that with certain parameters (grinding interval and mental removal), the total present value of rail cost tends to decrease firstly and then increase with the increase of rail service life. With the increase of the grinding interval, the total present value of rail cost also tends to decrease firstly and then increase, and there is an extreme point in the variation tendency. In the economic cost of rail service life, rail replacement and waste of railway taking-up cost are two of the biggest parts. Therefore, reasonable rail maintenance strategy plays a significant role in reducing life-cycle cost.