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Phosphatidylcholine (PC) is one of the major building blocks of biological membranes in mammalian cells. In PC, two fatty acyl chains are linked to the sn-1 and sn-2 positions of the glycerol backbone, and a head group containing a phosphate and choline is linked to the sn-3 position. The length and saturation of the fatty acyl chains of PC vary significantly and determine the biophysical and chemical properties of PC. In mammalian cells, active metabolism of PC takes place primarily through de novo synthesis, the remodelling cycle of deacylation and reacylation, and further modification of lysophosphatidylcholine. PC metabolism undergoes remarkable changes in obesity. In this mini review, we discuss PC metabolism and summarize recent advances about the important involvement of PC metabolism in obesity and obesity-related diseases.

Bovine milk exosomes (BMEs) are being explored in drug delivery despite their rapid elimination by macrophages. We aimed at identifying the BME transporter in murine bone marrow-derived macrophages (BMDMs). Fluorophore-labeled BMEs were used in transport studies in BMDMs from C57BL/6J and class A scavenger receptor type 1/2 (CASR-1/2) knockout mice and tissue accumulation in macrophage-depleted C57BL/6J mice. Parametric and nonparametric statistics tests for pairwise and multiple comparisons were used. Chemical inhibitors of phagocytosis by cytochalasin D led to a 69 ± 18% decrease in BME uptake compared with controls ( P < 0.05), whereas inhibitors of endocytic pathways other than phagocytosis had a modest effect on uptake ( P > 0.05). Inhibitors of class A scavenger receptors (CASRs) including CASR-1/2 caused a 70% decrease in BME uptake ( P < 0.05). The uptake of BMEs by BMDMs from CASR-1/2 knockout mice was smaller by 58 ± 23% compared with wild-type controls ( P < 0.05). Macrophage depletion by clodronate caused a more than 44% decrease in BME uptake in the spleen and lungs ( P < 0.05), whereas the decrease observed in liver was not statistically significant. In conclusion, CASR-1/2 facilitates the uptake of BMEs in BMDMs and C57BL/6J mice.

Aberrant activation of the nucleotide-binding domain and leucine-rich repeat related (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome drives the development of many complex inflammatory diseases, such as obesity, Alzheimer's disease, and atherosclerosis. However, no medications specifically targeting the NLRP3 inflammasome have become clinically available. Therefore, we aim to identify new inhibitors of the NLRP3 inflammasome in this study. Methods: Vesicle-like nanoparticles (VLNs) were extracted from garlic chives and other Allium vegetables and their effects on the NLRP3 inflammasome were evaluated in primary macrophages. After garlic chive-derived VLNs (GC-VLNs) were found to exhibit potent anti-NLRP3 inflammasome activity in cell culture, such function was further assessed in a murine acute liver injury disease model, as well as in diet-induced obesity. Finally, GC-VLNs were subjected to omics analysis to identify the active components with anti-NLRP3 inflammasome function. Results: GC-VLNs are membrane-enclosed nanoparticles containing lipids, proteins, and RNAs. They dose-dependently inhibit pathways downstream of NLRP3 inflammasome activation, including caspase-1 autocleavage, cytokine release, and pyroptotic cell death in primary macrophages. The inhibitory effects of GC-VLNs on the NLRP3 inflammasome are specific, considering their marginal impact on activation of other inflammasomes. Local administration of GC-VLNs in mice alleviates NLRP3 inflammasome-mediated inflammation in chemical-induced acute liver injury. When administered orally or intravenously, GC-VLNs accumulate in specific tissues and suppress activation of the NLRP3 inflammasome and chronic inflammation in diet-induced obese mice. The phospholipid 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC) in GC-VLNs has been identified to inhibit NLRP3 inflammasome activation. Conclusions: Identification of GC-VLNs and their active component DLPC as potent inflammasome inhibitors provides new therapeutic candidates in the treatment of NLRP3 inflammasome-driven diseases.

The nucleotide-binding domain and leucine-rich repeat-containing family, pyrin domain-containing 3 (NLRP3) inflammasome is a key regulator of innate immune responses, and its aberrant activation is implicated in the pathogenesis of many diseases such as Alzheimer's disease and type 2 diabetes. Targeting the NLRP3 inflammasome could hold promise to combat these complex diseases, but therapies specifically inhibiting the NLRP3 inflammasome have not been developed for patient treatment. The current study aimed to identify food-borne exosome-like nanoparticles (ELNs) that inhibit NLRP3 inflammasome activity. Nine vegetables or fruits were selected to extract ELNs, which were examined for their inhibitory effects on activation of the NLRP3 inflammasome in primary macrophages. Although most of the tested ELNs posed minimal impacts, the ELNs from ginger rhizomes (G-ELNs) strongly inhibited NLRP3 inflammasome activation. The G-ELNs contained lipids, proteins, and RNAs and were easily taken up by macrophages. G-ELN treatment suppressed pathways downstream of inflammasome activation including caspase1 autocleavage, interleukin (IL)-1β and IL-18 secretion, and pyroptotic cell death. Apoptotic speck protein containing a caspase recruitment domain (ASC) oligomerization and speck formation assays indicated that G-ELNs blocked assembly of the NLRP3 inflammasome. The lipids in G-ELNs, rather than the RNAs or proteins, were responsible for the inhibitory activity observed. Together, the data suggested G-ELNs as new potent agents that block NLRP3 inflammasome assembly and activation. The unique features of G-ELNs including biomolecule protection and tissue bioavailability should facilitate the development of G-ELN-based therapy to target the NLRP3 inflammasome in the disease settings.

OBJECTIVES: Bovine milk exosomes (BMEs) are promising candidates for delivering drugs to brain tumors because they are scalable, bioavailable after oral administration, and cross the blood-brain barrier. The use of BMEs in drug delivery is limited by their rapid elimination by macrophages. The objectives of this study were to identify the BME transporter and assess BME transport kinetics in murine bone marrow-derived macrophages (BMDMs) as a first step toward developing strategies that decrease the elimination of drug-loaded BMEs. METHODS: BMEs were isolated by differential centrifugation from skim milk. For transport studies, proteins and lipids on the BME surface were labeled with HiLyte™ Fluor 750 hydrazide and PKH26, respectively, and RNAs in BMEs were labeled with Exo-Red; unlabeled BMEs were used to assess background noise. Bone marrow cells were isolated from the femur and tibia of both C57BL/6J and scavenger receptor A-I/II knockout mice and differentiated ex vivo into BMDMs for use in transport studies. ANOVA, Dunnett's and Kruskal-Wallis test, Dunn's post-hoc test, unpaired t-test, and two-tailed Mann–Whitney U tests were used for statistical analysis; P 0.05). Treatment with inhibitors of class A scavenger receptor (CASR), fucoidan and dextran sulfate caused a 70 ± 8.1% and 70 ± 18% decrease (P ˂ 0.05), respectively, in BME uptake. The role of CASR in BME uptake was confirmed by using a genetics approach: the uptake of BMEs by BMDMs from scavenger receptor A-I/II knockout mice decreased by 58 ± 23% compared to BMDMs from wild-type mice (P ˂ 0.05). CONCLUSIONS: BME uptake is facilitated by CASR in BMDMs and uptake cannot be saturated under physiological conditions. FUNDING SOURCES: NIH 1P20GM104320, and NIFA 2016–67,001-25,301 and 2020–67,017-30,834, USDA Hatch-1,011,996, and USDA W4002 (all to J.Z.). J.Z is a consultant for PureTech Health, Inc.

OBJECTIVES: To investigate whether honey, as a medicinal food, contain exosome-like nanoparticles (ELNs). METHODS: Honey was diluted in phosphate-buffered saline and subjected to sequential centrifugation followed by ultracentrifugation. ELNs were obtained from honey. Such ELNs were characterized in detail by electron microscopy analysis and biomolecule omics analysis. The anti-inflammatory functions of these honey-derived ELNs (H-ELNs) were studied in primary macrophages and experimentally induced acute liver injury in mice. RESULTS: H-ELNs were membrane-enclosed nanoparticles composed of lipids, proteins, and RNAs. These nanoparticles specifically hindered assembly and activation of the nucleotide-binding domain and leucine-rich repeat related (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome, which is a critical pro-inflammatory enzyme complex in the innate immune system. H-ELNs protected mice from acute inflammation and liver damage in the experimentally induced acute liver injury. microRNA miR-4057 in H-ELNs was found to potently inhibit activation of the NLRP3 inflammasome. CONCLUSIONS: H-ELNs were identified as a new bioactive component in honey and these nanoparticles demonstrated potent anti-inflammatory function in vitro and in vivo. This finding opens a new avenue for studying honey, a medicinal food since ancient times. FUNDING SOURCES: This work was supported by the United States Department of Agriculture (USDA) National Institute of Food and Agriculture (NIFA) and the National Institutes of Health (NIH) National Institute for General Medical Science (NIGMS).

The nucleotide-binding domain and leucine-rich repeat related (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome is a multimeric protein complex that recognizes various danger or stress signals from pathogens, the host, and the environment, leading to activation of caspase-1 and inducing inflammatory responses. This pro-inflammatory protein complex plays critical roles in pathogenesis of a wide range of diseases including neurodegenerative diseases, autoinflammatory diseases, and metabolic disorders. Therefore, intensive efforts have been devoted to understanding its activation mechanisms and to searching for its specific inhibitors. Approximately forty natural compounds with anti-NLRP3 inflammasome properties have been identified. Here, we provide an update about new natural compounds that have been identified within the last three years to inhibit the NLRP3 inflammasome and offer an overview of the underlying molecular mechanisms of their anti-NLRP3 inflammasome activities.

Honey has been used as a nutrient, an ointment, and a medicine worldwide for many centuries. Modern research has demonstrated that honey has many medicinal properties, reflected in its anti‐microbial, anti‐oxidant, and anti‐inflammatory bioactivities. Honey is composed of sugars, water and a myriad of minor components, including minerals, vitamins, proteins and polyphenols. Here, we report a new bioactive component‒vesicle‐like nanoparticles‒in honey (H‐VLNs). These H‐VLNs are membrane‐bound nano‐scale particles that contain lipids, proteins and small‐sized RNAs. The presence of plant‐originated plasma transmembrane proteins and plasma membrane‐associated proteins suggests the potential vesicle‐like nature of these particles. H‐VLNs impede the formation and activation of the nucleotide‐binding domain and leucine‐rich repeat related (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome, which is a crucial inflammatory signalling platform in the innate immune system. Intraperitoneal administration of H‐VLNs in mice alleviates inflammation and liver damage in the experimentally induced acute liver injury. miR‐4057 in H‐VLNs was identified in inhibiting NLRP3 inflammasome activation. Together, our studies have identified anti‐inflammatory VLNs as a new bioactive agent in honey.

The research is based on the characteristics of instructional software of evaluation and exercise, taking a small examination system for driving license as an example, this study is devoted to exploring a feasible test method for instructional software of evaluation and exercise based on mobile platform. The test method consists of four testing phases of unit and integration testing, functionality testing, comprehensive testing and teaching evaluation which existed research of functionality testing for modern Android mobile Apps is fully used for reference. Furthermore, further work is put forward. Firstly, a special defect management database for bug definition and analysis needs to be established. Secondly, major test points in comprehensive testing phase should be converted to user stories in modern agile testing environment for software exploratory test methods positively.

The research is based on the background of new engineering, taking the software engineering major of Anhui Sanlian University as an example, this study is devoted to exploring and implementing on a feasible mode of “one body-two wings” for course construction of software engineering major. Talents cultivation based on integration of industry and education is “one body”, curriculum design and case resource construction are “two wings” of the mode. Furthermore, further work is put forward to optimize for this mode from three aspects.

The research is mainly based on the concept of technologies of software exploratory testing, Questing Patterns (Q-Patterns) and reusable test case, taking a reusable test case as an example, this study is devoted to the development of a feasible reusable test case management system based on exploratory testing in rapid prototype development method, which the process of design, storage, reuse and management of test cases could be implemented positively in the first system version. Furthermore, further work for technical improvement in the next version of this system is put forward.

The research is based on the characteristics of mobile Applications (Apps), taking an Android mobile App of a small examination system as an application case, this study is devoted to exploring a feasible functionality testing method based on exploratory testing for Android mobile Apps, which includes fundamental functionality logic verification, Graphical User Interface (GUI) automated testing, and special testing. Furthermore, further work is put forward to apply this functionality testing method to IOS mobile Apps, adding new test items for mobile Apps and improving the method of depth-first search for control traversal in GUI automated testing.

The research is based on the idea of agile testing, taking the “order cancelled” module of an e-commerce website as an application case, this study is devoted to exploring a feasible and scientific agile testing framework of four quadrants to form an anticlockwise closed loop for guiding the software agile testing process. Furthermore, further work is put forward to establish of a special defect management database for bug definition and analysis, and build the particular test process models for each quadrant of this agile testing framework gradually.

Problem-Based Learning (PBL) is a teaching mode which is suitable for key courses of software engineering major in universities. Based on the concept of PBL, this study takes engineering of Anhui Sanlian University as an example, and is committed to building a feasible PBL teaching mode of software engineering course. Furthermore, further work is put forward to promote the development of the PBL teaching mode for local teaching environment.

The innovation of the curriculum system of software engineering major has become an important part under background of new engineering currently that cannot be ignored. As a case study of Anhui Sanlian University, from the three aspects of talent training, knowledge of vocation, curriculum setting to explore the construction of curriculum system for software engineering major under background of new engineering. Finally, further work is expected in the future. Firstly, sustainable agile teaching system for competitiveness should be developed. Secondly, a local platform for innovative practice on “Five Classes” based on Outcomes-Based Education (OBE) should be established.

Fulminant hepatic failure (FHF) is a rare, life-threatening liver disease with a poor prognosis. Administration of D-galactosamine (GalN) and lipopolysaccharide (LPS) triggers acute liver injury in mice, simulating many clinical features of FHF in humans, therefore, this disease model is often used to investigate potential therapeutic interventions to treat FHF. Recently, suppression of the nucleotide-binding domain and leucine-rich repeat related (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome, was shown to alleviate the severity of GalN/LPS-induced liver damage in mice. Therefore, the goal of this study was to find dietary exosome-like nanoparticles (ELNs) with therapeutic potential in curbing FHF by suppressing the NLRP3 inflammasome. Seven commonly consumed mushrooms were used to extract ELNs. These mushrooms were found to contain ELNs composed of RNAs, proteins, and lipids. Among these mushroom-derived ELNs, only shiitake mushroom-derived ELNs (S-ELNs) substantially inhibited NLRP3 inflammasome activation by preventing inflammasome formation in primary macrophages. S-ELNs also suppressed the secretion of interleukin (IL)-6, as well as both protein and mRNA levels of the Il1b gene. Remarkably, pre-treatment with S-ELNs protected mice from GalN/LPS-induced acute liver injury. Therefore, S-ELNs, identified as potent new inhibitors of the NLRP3 inflammasome, represent a promising class of agents with the potential to combat FHF.

Smart campus is a new stage in the development of educational informatization. Focusing on present construction situation of smart campus in universities in China and abroad, the paper firstly reviews of typical construction modes, types of design architecture and major functional modules of smart campus in universities. Secondly, from the perspective of application, service and management for smart education, some key technologies of construction of smart campus in universities are analyzed and summarized. Finally, further work of promoting the construction standards and evaluation of smart campus, training and improving the big data literacy for teachers and students in campus, and exploring effectively on sharing and constructing mechanism on smart campus for local application-oriented universities is expected in this paper.

A lightweight autonomous learning system for the course of software testing based on Android is developed according to the local teaching environment, and the system is fully applied on autonomous learning process outside class on campus. The system is designed on the architecture of SSH (Struts-Spring-Hibernate) which is based on J2EE for web server, and the functions of Course Information, Resource Management, Exercise, User Management and Online Discussion can be provided to client users by Android platform. Finally, further work such as how to add the function of online exam for subjective exam items in the system, how to add powerful function of micro video content editing for the management of course resource, and how to integrate this system into the local informational learning platform to complete the blended learning mode on SPOC (Small Private Online Course) are expected in the paper.

In recent years, the construction of MOOC (Massive Open Online Course) resources database has been paid more attention on MOOC research field. Based on the idea of integrable ware, MOOC elements are designed to form the corresponding MOOC learning knowledge points for some courses of software engineering major in a local university. They have the characteristics of openness, flexible and reusable. MOOC elements are fully applied in construction of the MOOC resources database on local campus for personalized learning effectively. The results show that teachers can select several different MOOC elements to form an important knowledge point for MOOC teaching process according to an actual course and reuse some existed MOOC elements to form another MOOC knowledge point for other courses. Different MOOC elements can be combined freely to form the related learning knowledge points for one course of software engineering major to stimulate and improve students’ interests in personalized learning. Finally, further work is put forward. Firstly, compatible supported conditions on software and hardware are to be created based on big data. Secondly, capacities on integrating and reusing of various types of MOOC resources by teachers should be improved. Thirdly, more reusable MOOC elements based on integrable ware for the courses of other majors should be designed and constructed for personalized learning.

Coding style is an important factor in measuring the program quality. The paper explores on the C program coding style of the program annotation writing, data declaration, program statement writing, and input/output design style from the perspective of software engineering. Good coding style plays an important role in improving the readability, testability and maintainability of the program.

OBJECTIVES: The NLRP3 inflammasome is a key regulator of innate immune responses, and its aberrant activation has been implicated in the pathogenesis of many complex diseases including multiple sclerosis, type 2 diabetes and atherosclerosis. Our study aimed to identify the food-borne exosome-like nanoparticles (ELNs) that inhibit NLRP3 inflammasome in macrophages. METHODS: ELNs from nine vegetables or fruits were purified using ultracentrifugation, incubated with bone-marrow derived macrophages, followed by the activation of NLRP3 inflammasome. After the dietary ELNs with strong inhibitory effects on inflammasome were identified, their roles in the inflammasome assembly and downstream pathways were further examined. RESULTS: We identified that ELNs from ginger rhizomes (G-ELNs) strongly inhibited NLRP3 inflammasome activation. We verified that G-ELNs were exosome-like nanoparticles containing biomolecules including RNAs, proteins, and lipids. The G-ELNs were taken up by macrophages. G-ELNs suppressed the downstream pathways of NLRP3 inflammasome activation including Casp1 auto-cleavage, interleukin (IL)-1 and IL-18 secretion, and pyroptotic cell death. G-ELNs blocked the assembling steps of NLRP3 inflammasome, but had no effects on protein levels of a key mediator of NLRP3 inflammasome NIMA-related kinase 7 (Nek7), or the inflammasome subunits NLRP3, ASC, and Casp1. CONCLUSIONS: Our findings identified G-ELNs as a new potent agent that blocks NLRP3 inflammasome assembly and activation. FUNDING SOURCES: This work was supported by the National Institutes of Health (NIH) 1P20GM104320 Nebraska Center Prevention of Obesity Diseases (NPOD) Seed Grant and Project Leader Grant, and the United States Department of Agriculture (USDA) National Institute of Food and Agriculture (NIFA) Hatch Project 1015948.

Technologies of the architecture on SSH(Struts-Spring-Hibernate), database interface access, and web data mining are used to analyse and design of a lightweight course website for software testing which is based on SPOC(Small Private Online Course) for blended teaching on campus. Functions of learning topics releasing on the course, organizing and coordinating learners' discussion to promote the learning process can be achieved through the website for teachers, and the website can also provide learners with selecting some social learning tools(BBS, QQ, WeChat) to complete online communication for the course learning. Finally, further work is expected in the paper. Firstly, the targeted and distinctive mobile learning resources and smart education mode should be constructed which is based on SPOC. Secondly, the visualized, diversified third-party platforms and tools in the course website should be integrated effectively which are enable learners to complete the process of online exercising or examining for some types of subjective testing items. Thirdly, how to ensure the reliability, toughness, running performance, safety, stability for the course website which is formed by cutting out some non-essential functions from existed MOOC(Massive Open Online Course) platforms, and improve the functions of multiple data statistics with web data mining technology and motivational observation for the website development.

As an important measure on quality assurance for web applications of website, web testing has been paid more attention in software testing recently. Based on the analysis of website and its application characteristics, from the view of affecting the quality of web applications, testing activities from the five aspects on functionality testing, performance testing, usability testing, compatibility testing and security testing are explored to assure the quality on web applications of website. Finally, further researches on comprehensive and adequate testing of the web service composition with some test techniques under the background of service-oriented technology and cloud computing technology, and how to combine exploratory testing with testing activities of functionality testing and usability testing effectively in web testing are expected in the paper.

Instructional software is a kind of software program or system, which reflects the teaching strategy according to the teaching objectives and teaching contents for the purpose on promoting teaching and learning by both computer technology and network technology. Firstly, the concept of instructional software, features and functions are introduced in the article. Secondly, making a detailed description on the current types of instructional software features for total eleven types, and discussing on main points on designing of instructional software for each type fully. Finally, an initial expectation on researches on directions of instructional software designing is given, which from three aspects on designing of teaching platform for SPOC(Small Private Online Course), designing of mobile learning resources, and the intelligence on designing of instructional software in the future.

On the basis of the feature analysis on development of online course from the perspective of software engineering, a testing framework of online course based on agile is designed firstly. The framework consists of four test cycles with iteration by test stages on preparing for testing, pre-testing, functionality testing and regression testing, and with a test process of acceptance testing at the end of four test cycles before product delivery. The evaluation on organization of the contents of chapter-section structure is also embodied in this framework. Secondly, the framework is fully applied to the agile testing project for an actual online course of "Software Engineering", experimental result shows that more bugs for types of structure of contents, usability, compatibility, installing, security, and navigability of the online course are detected in time and test result is noticeable. Finally, putting forward that some features and aspects on non-functionality testing through test automation together with effective exploratory testing method, and constructing an automated testing environment on cloud computing for big data processing will be the future research.

Macrophage activation/polarization to distinct functional states is critically supported by metabolic shifts. How polarizing signals coordinate metabolic and functional reprogramming, and the potential implications for control of macrophage activation, remains poorly understood. Here we show that IL-4 signaling co-opts the Akt-mTORC1 pathway to regulate Acly, a key enzyme in Ac-CoA synthesis, leading to increased histone acetylation and M2 gene induction. Only a subset of M2 genes is controlled in this way, including those regulating cellular proliferation and chemokine production. Moreover, metabolic signals impinge on the Akt-mTORC1 axis for such control of M2 activation. We propose that Akt-mTORC1 signaling calibrates metabolic state to energetically demanding aspects of M2 activation, which may define a new role for metabolism in supporting macrophage activation.

Unliganded thyroid hormone receptor (TR) actively represses transcription via the nuclear receptor corepressor (N-CoR)/histone deacetylase 3 (HDAC3) complex. Although transcriptional activation by liganded receptors involves chromatin remodeling, the role of ATP-dependent remodeling in receptor-mediated repression is unknown. Here we report that SNF2H, the mammalian ISWI chromatin remodeling ATPase, is critical for repression of a genomically integrated, TR-regulated reporter gene. N-CoR and HDAC3 are both required for recruitment of SNF2H to the repressed gene. SNF2H does not interact directly with the N-CoR/HDAC3 complex, but binds to unacetylated histone H4 tails, suggesting that deacetylase activity of the corepressor complex is critical to SNF2H function. Indeed, HDAC3 as well as SNF2H are required for nucleosomal organization on the TR target gene. Consistent with these findings, reduction of SNF2H induces expression of an endogenous TR-regulated gene, dio1, in liver cells. Thus, although not apparent from studies of transiently transfected reporter genes, gene repression by TR involves the targeting of chromatin remodeling factors to repressed genes by the HDAC activity of nuclear receptor corepressors.

Nuclear receptor corepressors SMRT (silencing mediator of retinoid and thyroid receptors) and N-CoR (nuclear receptor corepressor) recruit histone deacetylase (HDAC) activity to targeted regions of chromatin. These corepressors contain a closely spaced pair of SANT motifs whose sequence and organization is highly conserved. The N-terminal SANT is a critical component of a deacetylase activation domain (DAD) that binds and activates HDAC3. Here, we show that the second SANT motif functions as part of a histone interaction domain (HID). The HID enhances repression by increasing the affinity of the DAD-HDAC3 enzyme for histone substrate. The two SANT motifs synergistically promote histone deacetylation and repression through unique functions. The HID contribution to repression is magnified by its ability to inhibit histone acetyltransferase enzyme activity. Remarkably, the SANT-containing HID preferentially binds to unacetylated histone tails. This implies that the SMRT HID participates in interpreting the histone code in a feed-forward mechanism that promotes and maintains histone deacetylation at genomic sites of SMRT recruitment.

Drosophila melanogaster display overt circadian rhythms in rest:activity behavior and eclosion. These rhythms have an endogenous period of approximately 24 hr and can adjust or "entrain" to environmental inputs such as light. Circadian rhythms depend upon a functioning molecular clock that includes the core clock genes period and timeless (reviewed in and ). Although we know that a clock in the lateral neurons (LNs) of the brain controls rest:activity rhythms, the cellular basis of eclosion rhythms is less well understood. We show that the LN clock is insufficient to drive eclosion rhythms. We establish that the prothoracic gland (PG), a tissue required for fly development, contains a functional clock at the time of eclosion. This clock is required for normal eclosion rhythms. However, both the PG clock function and eclosion rhythms require the presence of LNs. In addition, we demonstrate that pigment-dispersing factor (PDF), a neuropeptide secreted from LNs, is necessary for the PG clock and eclosion rhythms. Unlike other clocks in the fly periphery, the PG is similar to mammalian peripheral oscillators because it depends upon input, including PDF, from central pacemaker cells. This is the first report of a peripheral clock necessary for a circadian event.

As the derivation and innovation of MOOC (Massive Open Online Course), the development of SPOC (Small Private Online Course) platform which is based on MOOC idea can be better applied to the blended teaching of various courses in local universities in post-MOOC era. Based on the analysis of the current application of SPOC platform, from the view of technology in implementation, we explore the key technologies of identification on face recognition based on emotion calculation,course creating, editing and sharing, analysis of online learning behaviour and exercising or examining on subjective items for developing of a SPOC platform. Finally, further researches on how to construct on the targeted and distinctive mobile learning resources and smart education mode which is based on SPOC, how to provide formative assessment data and consciousness assessment data for learners accurately and function of data statistics and motivational observation that could be completed by teachers, how to cut some non-essential auxiliary functions of an existed MOOC platform for a certain local user group to form a flexible and usable SPOC platform with good expansibility are expected in the paper.

The acetylation of histone tails is a primary determinant of gene activity. Histone deacetylase 3 (HDAC3) requires the nuclear receptor corepressor SMRT for HDAC enzyme activity. Here we report that HDAC3 interacts with SMRT only after priming by cellular chaperones including the TCP-1 ring complex (TRiC), which is required for proper folding of HDAC3 in an ATP-dependent process. SMRT displaces TRiC from HDAC3, yielding an active HDAC enzyme. The SMRT–HDAC3 repression complex thus joins the VHL–elongin BC tumor suppression complex and the cyclin E–Cdk2 cell cycle regulation complex as critical cellular machines requiring TRiC for proper assembly and function. The strict control of HDAC3 activity underscores the cellular imperative that histone deacetylation occur only in targeted regions of the genome.

Inflammasomes are intracellular sensors that couple detection of pathogens and cellular stress to activation of Caspase-1, and consequent IL-1β and IL-18 maturation and pyroptotic cell death. Here, we show that the absent in melanoma 2 (AIM2) and nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasomes trigger Caspase-1-dependent mitochondrial damage. Caspase-1 activates multiple pathways to precipitate mitochondrial disassembly, resulting in mitochondrial reactive oxygen species (ROS) production, dissipation of mitochondrial membrane potential, mitochondrial permeabilization, and fragmentation of the mitochondrial network. Moreover, Caspase-1 inhibits mitophagy to amplify mitochondrial damage, mediated in part by cleavage of the key mitophagy regulator Parkin. In the absence of Parkin activity, increased mitochondrial damage augments pyroptosis, as indicated by enhanced plasma membrane permeabilization and release of danger-associated molecular patterns (DAMPs). Therefore, like other initiator caspases, Caspase-1 activation by inflammasomes results in mitochondrial damage.

A chlL-deletion mutant of Synechocystis sp. PCC 6803 designated as chlL- was unable to make significant amounts of chlorophyll in darkness. However, an apparent pseudorevertant has been generated spontaneously that can synthesize an increased amount of chlorophyll under light-activated heterotrophic growth conditions. Under these conditions, the chlorophyll content in this pseudorevertant was about 20% of that in the wild-type strain and about 4 times more than that in the original and in the recently recreated chlL-deletion mutant. This is paralleled by increased performance of dark-grown cells in terms of chlorophyll fluorescence induction and oxygen evolution rates in the pseudorevertant versus in the original mutant. PCR analysis confirmed that the chlL- -pseudorevertant mutant still lacked the chlL gene. These results imply that the light-independent chlorophyll biosynthesis pathway was partly recovered.

Sirt5, localized in the mitochondria, is a member of sirtuin family of NAD(+)-dependent deacetylases. Sirt5 was shown to deacetylate and activate carbamoyl phosphate synthase 1. Most recently, Sirt5 was reported to be the predominant protein desuccinylase and demalonylase in the mitochondria because the ablation of Sirt5 enhanced the global succinylation and malonylation of mitochondrial proteins, including many metabolic enzymes. In order to determine the physiological role of Sirt5 in metabolic homeostasis, we generated a germline Sirt5 deficient (Sirt5(-/-)) mouse model and performed a thorough metabolic characterization of this mouse line. Although a global protein hypersuccinylation and elevated serum ammonia during fasting were observed in our Sirt5(-/-) mouse model, Sirt5 deficiency did not lead to any overt metabolic abnormalities under either chow or high fat diet conditions. These observations suggest that Sirt5 is likely to be dispensable for the metabolic homeostasis under the basal conditions.