2'-O-methylation at internal sites on mRNA promotes mRNA stability.

2'-O-methylation (Nm) is a prominent RNA modification well known in noncoding RNAs and more recently also found at many mRNA internal sites. However, their function and base-resolution stoichiometry remain underexplored. Here, we investigate the transcriptome-wide effect of internal site Nm on mRNA stability. Combining nanopore sequencing with our developed machine learning method, NanoNm, we identify thousands of Nm sites on mRNAs with a single-base resolution. We observe a positive effect of FBL-mediated Nm modification on mRNA stability and expression level. Elevated FBL expression in cancer cells is associated with increased expression levels for 2'-O-methylated mRNAs of cancer pathways, implying the role of FBL in post-transcriptional regulation. Lastly, we find that FBL-mediated 2'-O-methylation connects to widespread 3' UTR shortening, a mechanism that globally increases RNA stability. Collectively, we demonstrate that FBL-mediated Nm modifications at mRNA internal sites regulate gene expression by enhancing mRNA stability.

Epsin Nanotherapy Regulates Cholesterol Transport to Fortify Atheroma Regression.

BACKGROUND: Excess cholesterol accumulation in lesional macrophages elicits complex responses in atherosclerosis. Epsins, a family of endocytic adaptors, fuel the progression of atherosclerosis; however, the underlying mechanism and therapeutic potential of targeting Epsins remains unknown. In this study, we determined the role of Epsins in macrophage-mediated metabolic regulation. We then developed an innovative method to therapeutically target macrophage Epsins with specially designed S2P-conjugated lipid nanoparticles, which encapsulate small-interfering RNAs to suppress Epsins. METHODS: We used single-cell RNA sequencing with our newly developed algorithm MEBOCOST (Metabolite-mediated Cell Communication Modeling by Single Cell Transcriptome) to study cell-cell communications mediated by metabolites from sender cells and sensor proteins on receiver cells. Biomedical, cellular, and molecular approaches were utilized to investigate the role of macrophage Epsins in regulating lipid metabolism and transport. We performed this study using myeloid-specific Epsin double knockout (LysM-DKO) mice and mice with a genetic reduction of ABCG1 (ATP-binding cassette subfamily G member 1; LysM-DKO-ABCG1fl/+). The nanoparticles targeting lesional macrophages were developed to encapsulate interfering RNAs to treat atherosclerosis. RESULTS: We revealed that Epsins regulate lipid metabolism and transport in atherosclerotic macrophages. Inhibiting Epsins by nanotherapy halts inflammation and accelerates atheroma resolution. Harnessing lesional macrophage-specific nanoparticle delivery of Epsin small-interfering RNAs, we showed that silencing of macrophage Epsins diminished atherosclerotic plaque size and promoted plaque regression. Mechanistically, we demonstrated that Epsins bound to CD36 to facilitate lipid uptake by enhancing CD36 endocytosis and recycling. Conversely, Epsins promoted ABCG1 degradation via lysosomes and hampered ABCG1-mediated cholesterol efflux and reverse cholesterol transport. In a LysM-DKO-ABCG1fl/+ mouse model, enhanced cholesterol efflux and reverse transport due to Epsin deficiency was suppressed by the reduction of ABCG1. CONCLUSIONS: Our findings suggest that targeting Epsins in lesional macrophages may offer therapeutic benefits for advanced atherosclerosis by reducing CD36-mediated lipid uptake and increasing ABCG1-mediated cholesterol efflux.

Low RNA stability signifies strong expression regulatability of tumor suppressors.

RNA expression of a gene is determined by not only transcriptional regulation, but also post-transcriptional regulation of RNA decay. The precise regulation of RNA stability in the cell plays an important role in normal development. Dysregulation of RNA stability can lead to diseases such as cancer. Here we found tumor suppressor RNAs tended to decay fast in normal cell types when compared with other RNAs. Consistent with a negative effect of m6A modification on RNA stability, we observed preferential deposition of m6A on tumor suppressor RNAs. Moreover, abundant m6A and fast decay of tumor suppressor RNAs both tended to be further enhanced in prostate cancer cells relative to normal prostate epithelial cells. Further, knockdown of m6A methyltransferase METTL3 and reader YTHDF2 in prostate cancer cells both posed stronger effect on tumor suppressor RNAs than on other RNAs. These results indicated a strong post transcriptional expression regulatability mediated by abundant m6A modification on tumor suppressor RNAs.

Low RNA stability signifies increased post-transcriptional regulation of cell identity genes.

Cell identity genes are distinct from other genes with respect to the epigenetic mechanisms to activate their transcription, e.g. by super-enhancers and broad H3K4me3 domains. However, it remains unclear whether their post-transcriptional regulation is also unique. We performed a systematic analysis of transcriptome-wide RNA stability in nine cell types and found that unstable transcripts were enriched in cell identity-related pathways while stable transcripts were enriched in housekeeping pathways. Joint analyses of RNA stability and chromatin state revealed significant enrichment of super-enhancers and broad H3K4me3 domains at the gene loci of unstable transcripts. Intriguingly, the RNA m6A methyltransferase, METTL3, preferentially binds to chromatin at super-enhancers, broad H3K4me3 domains and their associated genes. METTL3 binding intensity is positively correlated with RNA m6A methylation and negatively correlated with RNA stability of cell identity genes, probably due to co-transcriptional m6A modifications promoting RNA decay. Nanopore direct RNA-sequencing showed that METTL3 knockdown has a stronger effect on RNA m6A and mRNA stability for cell identity genes. Our data suggest a run-and-brake model, where cell identity genes undergo both frequent transcription and fast RNA decay to achieve precise regulation of RNA expression.

CD45 Is Sufficient to Initiate Endothelial-to-Mesenchymal Transition in Human Endothelial Cells-Brief Report.

BACKGROUND: Endothelial-to-mesenchymal transition (EndMT) is a dynamic process in which endothelial cells acquire mesenchymal properties and in turn contribute to tissue remodeling and growth. Previously, we found EndMT associated with mitral valve adaptation after myocardial infarction. Furthermore, mitral valve endothelial cells collected at 6 months post-myocardial infarction expressed the pan-leukocyte marker CD45 and EndMT markers. Additionally, mitral valve endothelial cells induced to undergo EndMT with TGF (transforming growth factor)-ß1 strongly coexpressed CD45 but not CD11b or CD14. Pharmacologic inhibition of the CD45 PTPase (protein tyrosine phosphatase) domain in mitral valve endothelial cells blocked TGFß-induced EndMT. This prompted us to speculate that, downstream of TGFß, CD45 induces EndMT. METHODS: We activated the endogenous CD45 promoter in human endothelial colony forming cells (ECFCs) using CRISPR (cluster regularly interspaced short palindromic repeats)/inactive Cas9 (CRISPR-associated protein 9) transcriptional activation. Bulk RNA sequencing was performed on control ECFCs and CD45-positive ECFCs to identify transcriptomic changes. Three functional assays-cellular migration, collagen gel contraction, and transendothelial electrical resistance-were conducted to assess mesenchymal properties in CD45-positive ECFCs. RESULTS: Activation of the endogenous CD45 promoter in ECFC and 3 additional sources of endothelial cells induced expression of several genes implicated in EndMT. In addition, CD45-positive ECFCs showed increased migration, a hallmark of EndMT, increased collagen gel contraction, a hallmark of mesenchymal cells, and decreased cell-cell barrier integrity, indicating reduced endothelial function. CONCLUSIONS: CD45 is sufficient to incite an EndMT phenotype and acquisition of mesenchymal cell properties in normal human ECFCs. We speculate that CD45, through its C-terminal PTPase domain, initiates signaling events that drive EndMT.

Fe(II)-Modulated Microporous Electrocatalytic Membranes for Organic Microcontaminant Oxidation and Fouling Control: Mechanisms of Regulating Electron Transport toward Enhanced Reactive Oxygen Species Activation.

Regulation of the fast electron transport process for the generation and utilization of reactive oxygen species (ROS) by achieving fortified electron "nanofluidics" is effective for electrocatalytic oxidation of organic microcontaminants. However, limited available active sites and sluggish mass transfer impede oxidation efficiency. Herein, we fabricated a conductive electrocatalytic membrane decorated with hierarchical porous vertically aligned Fe(II)-modulated FeCo layered double hydroxide nanosheets (Fe(II)-FeCo LDHs) in an electro-Fenton system to maximize exposure of active sites and expedite mass transfer. The nanospaced interlayers of Fe(II)-FeCo LDHs within the microconfined porous structure formed by its vertical nanosheets highly boost the micro/nanofluidic distribution of target pollutants to active centers/species, achieving accelerated mass transferability. Aliovalent substitution by Fe(II) activates in-plane metallics to maximize the available active sites and makes each Fe(II)-FeCo LDH nanosheet a geometrical nanocarrier for constructing a fast electron "nanofluidic" to accelerate Fe(II) regeneration in Fe(III)/Fe(II) cycles. As a result, the Fe(II)-FeCo LDHs exhibited improved reactivity in catalyzing H2O2 to •OH and 1O2. Accordingly, the membrane exhibited a higher atrazine degradation kinetic (0.0441 min-1) and degradation rate (93.2%), which were 4.7 and 2.1 times more than those of the bare carbon nanotube membrane, respectively. Additionally, the enhanced hydrophilic and strongly oxidized reactivity synergistically mitigated the organic fouling occurring in the pores and surface of the membrane. These findings clarify the activation mechanism of ROS over an innovative electrocatalytic membrane reactor design for organic microcontaminant treatment.

Estimation of water footprint in seawater desalination with reverse osmosis process.

Seawater desalination is one of the most applied approaches for freshwater replenishment. However, the process not only generates freshwater but also consumes it. It is important to evaluate the balance of the production and consumption of freshwater in desalination, which is also called as water footprint. It will reveal the feasibility of seawater desalination in terms of water production, but related study has not been reported. In this study, the water footprint of reverse osmosis desalination process has been investigated based on a real reverse osmosis desalination plant data. According to the calculation, the freshwater utilization of the reverse osmosis desalination plant was about 8.16 × 10-3 m3 with 1 m3 freshwater production. The study reveals that RO desalination is freshwater gain process as the utilized freshwater amount was less than the one produced. The sensitivity study showed that the energy source used in the process was the most significant parameter affecting on the water footprint. The freshwater required in the reverse osmosis desalination with energy supplied by thermal and solar was 8.01 × 10-3 m3 and 9.90 × 10-3 m3 in 1 m3 freshwater generation, respectively. It suggests that energy source selection is important in RO desalination system.

A feasible approach for azo-dye methyl orange degradation in siderite/H2O2 assisted by persulfate: Optimization using response surface methodology and pathway.

Siderite was applied to the binary oxidant system of siderite-catalyzed hydrogen peroxide (H2O2) and enhanced with persulfate (PS). In the absence of PS, methyl orange (MO) almost could not be degraded by the siderite/H2O2 process. However, adding PS significantly improved the capacity of MO to oxidize azo-dye. The influence of individual and interaction of reaction factors have been explored with a simple response surface methodology (RSM) based on central composite design (CCD). The quadratic model with low probabilities (<0.0001) at a confidence level of 95% was satisfactory to predict MO degradation in siderite/H2O2/PS system, whose correlation coefficients of R2 and R2-adj were 0.9569 and 0.9264, respectively. Moreover, the optimum operation conditions of 21.20 mM, 2.75 g/L, 3.86 mM, and 4.69 for H2O2, siderite, PS and initial pH, respectively with the response of C/C0 around 0.047. Radical scavenging experiments and electron spin resonance (ESR) determined that ·OH was crucial for MO degradation, while the contribution of SO4·- was minor. The surface morphology and iron content of siderite before and after the oxidation process showed clear differences. Possible intermediates and a degradation pathway were proposed based on the results of UV-Vis spectral and GC-MS analysis. Moreover, the toxicity to Vibrio fischeri bioluminescent bacterium has increased in the earlier degradation stage due to the generated by-products and weaken with the continuous treatment. This study demonstrated that the siderite/H2O2/PS system was effective over a relatively wide pH range without producing secondary pollutants, making it a promising technology and potential environmentally benign approach to azo-dye wastewater treatment.

Energy balance assessment on chicken manure for biogas production in Rabat-Salé-Zemmour-Zaïr of Morocco.

Chicken manure management has grabbed significant attention in Morocco due to the increasing demand on chicken and eggs. Bioconversion of chicken manure to biogas could reduce the chicken manure amount as well as generate clean energy. To evaluate the feasibility of converting chicken manure to biogas in terms of energy gain, the energy balance of the chicken manure for biogas production in the region of Rabat-Salé-Zemmour-Zaïr of Morocco has been investigated. The result showed that there was an energy gain of 1350 MJ for per tonne of dry chicken manure was converted to biogas. The energy gain increased to 3996 MJ/tonne of dry chicken manure when the wheat straw was added to co-digest with chicken manure. With consideration of converting the obtained biogas for electricity generation, there was extra electricity for sale after subtracting the electricity consumed inside the poultry industry. Comparing with co-firing, pyrolysis, and gasification, chicken manure anaerobic digestion was superior in terms of energy gain.

Identification of genes underlying phenotypic plasticity of wing size via insulin signaling pathway by network-based analysis in Sogatella furcifera.

BACKGROUND: Phenotypic plasticity is a common and highly adaptive phenomenon where the same genotype produces different phenotypes in response to environmental cues. Sogatella furcifera, a migratory pest of rice exhibits wing dimorphism, is a model insect for studying phenotypic plasticity of wing size. The Insullin-PI3K-Akt-FOXO signaling pathway plays a crucial role in the manipulation of wing size in the migratory insects. However, the regulatory mechanism via the pathway involved in wing dimorphism are still unexplored. RESULTS: Accompanied by special alternative splicing, genes involved in muscle contraction and energy metabolism were highly expressed in the wing hinges of macropters, demonstrating their adaptation for energy-demanding long-distance flights. Based on FOXO ChIP-Seq analysis, a total of 1259 putative target genes were observed in the wing hinges, including wing morph development, flight muscle and energy metabolism genes. An integrated gene interaction network was built by combining four heterogeneous datasets, and the IIS-PI3K-Akt-FOXO pathway was clustered in a divided functional module. In total, 45 genes in the module directly interacting with the IIS-PI3K-Akt-FOXO pathway showed differential expression levels between the two wing hinges, thus are regarded as potential Insulin pathway mediated wing dimorphism related genes (IWDRGs). Of the 45 IWDRGs, 5 were selected for verification by gene knockdown experiments, and played significant roles in the insect wing size regulation. CONCLUSIONS: We provided valuable insights on the genetic basis of wing dimorphism, and also demonstrated that network analysis is a powerful approach to identify new genes regulating wing dimorphic development via insulin signaling pathway in the migratory insect.

Structures of an all-α protein running along the DNA major groove.

Despite over 3300 protein-DNA complex structures have been reported in the past decades, there remain some unknown recognition patterns between protein and target DNA. The silkgland-specific transcription factor FMBP-1 from the silkworm Bombyx mori contains a unique DNA-binding domain of four tandem STPRs, namely the score and three amino acid peptide repeats. Here we report three structures of this STPR domain (termed BmSTPR) in complex with DNA of various lengths. In the presence of target DNA, BmSTPR adopts a zig-zag structure of three or four tandem α-helices that run along the major groove of DNA. Structural analyses combined with binding assays indicate BmSTPR prefers the AT-rich sequences, with each α-helix covering a DNA sequence of 4 bp. The successive AT-rich DNAs adopt a wider major groove, which is in complementary in shape and size to the tandem α-helices of BmSTPR. Substitutions of DNA sequences and affinity comparison further prove that BmSTPR recognizes the major groove mainly via shape readout. Multiple-sequence alignment suggests this unique DNA-binding pattern should be highly conserved for the STPR domain containing proteins which are widespread in animals. Together, our findings provide structural insights into the specific interactions between a novel DNA-binding protein and a unique deformed B-DNA.

Synthesis, Characterization, and Performance Evaluation of Sulfur-Containing Diphenylamines Based on Intramolecular Synergism.

To obtain novel structural antioxidants that have different antioxidant mechanisms, four 2-(alkylthio)-N-(4-(phenylamino)phenyl)acetamides 2a-d as dual functional antioxidants are designed, synthesized, and confirmed by ¹H-NMR, FTIR, MS, and elemental analysis. The antioxidant behavior of compounds 2a-d as additives of base oil triisodecyl trimellitate (TIDTM) is evaluated by non-isothermal and isothermal DSC analyses. The results showed all compounds can greatly increase the incipient oxidation temperature (IOT) and oxidation induction time (OIT) of TIDTM, especially, compound 2c exhibited an OIT value of 72.5 min at 230 °C, which is almost 28 times the length of TIDTM. Moreover, compounds 2a-d do not affect the tribological performance of TIDTM. The mechanism of antioxidants involved an intramolecular synergism are proposed. This work demonstrates compound 2c can be used as a novel potential antioxidant additive of TIDTM; in addition, it would inspire the emergence of highly potent antioxidants with different antioxidant mechanisms.

DIP1 plays an antiviral role against DCV infection in Drosophila melanogaster.

Disconnected Interacting Protein 1 (DIP1) is a dsRNA-binding protein that participates in a wide range of cellular processes. Whether DIP1 is involved in innate immunity remains unclear. Here, DIP1 was found to play an antiviral role in S2 cells. Its antiviral action is specific for DCV infection and not for DXV infection. dip1 mutant flies are hypersensitive to DCV infection. The increased mortality in dip1 mutant flies is associated with the accumulation of DCV positive-stranded RNAs in vivo. This study demonstrated that dip1 is a novel antiviral gene that restricts DCV replication in vitro and in vivo.

Operation performance and microbial community dynamics of phosphorus removal sludge with different electron acceptors.

Operation performances of phosphorus removal sludge with different electron acceptors in three parallel SBRs were firstly compared in the present study, and the effect of post-aeration on denitrifying phosphorus removal was also studied. Moreover, community dynamics of different phosphorus removal sludge was systematically investigated with high-throughput sequencing for the first time. TP removal rates for nitrate-, nitrite-, and oxygen-based phosphorus removal sludge were 84.8, 78.5, and 87.4 %, with an average effluent TP concentration of 0.758, 0.931, and 0.632 mg/l. The average specific phosphorus release and uptake rates were 20.3, 10.8, and 21.5, and 9.43, 8.68, and 10.8 mgP/(gVSS h), respectively. Moreover, electron utilization efficiency of denitrifying phosphorus removal sludge with nitrate as electron acceptor was higher than nitrite, with P/e(-) were 2.21 and 1.51 mol-P/mol-e(-), respectively. With the assistance of post-aeration for nitrate-based denitrifying phosphorus removal sludge, settling ability could be improved, with SVI decreased from 120 to 80 and 72 ml/g when post-aeration time was 0, 10, and 30 min, respectively. Moreover, further phosphorus removal could be achieved during post-aeration with increased aeration time. However, the anoxic phosphorus uptake was deteriorated, which was likely a result of shifted microbial community structure. Post-aeration of approximately 10 min was proposed for denitrifying phosphorus removal. Nitrate- and nitrite-based denitrifying phosphorus removal sludge exhibited similar community structure. More phosphorus accumulating organisms were enriched under anaerobic-aerobic conditions, while anaerobic-anoxic conditions were favorable for suppressing glycogen-accumulating organisms. Significant differences in pathogenic bacterial community profiles revealed in the current study indicated the potential public health hazards of non-aeration activated sludge system.

Embryonic alcohol exposure in zebrafish predisposes adults to cardiomyopathy and diastolic dysfunction.

AIMS: Fetal alcohol spectrum disorders (FASDs) impact up to 0.8% of the global population. However, cardiovascular health outcomes in adult patients, along with predictive biomarkers for cardiac risk stratification, remain unknown. Our aim was to utilize a longitudinal cohort study in an animal model to evaluate the impact of embryonic alcohol exposure (EAE) on cardiac structure, function, and transcriptional profile across the lifespan. METHODS AND RESULTS: Using zebrafish, we characterized the aftereffects of embryonic alcohol exposure (EAE) in adults binned by congenital heart defect (CHD) severity. Chamber sizes were quantified on dissected adult hearts to identify structural changes indicative of cardiomyopathy. Using echocardiography, we quantified systolic function based on ejection fraction and longitudinal strain, and diastolic function based on ventricular filling dynamics, ventricular wall movement, and estimated atrial pressures. Finally, we performed RNA sequencing on EAE ventricles and assessed how differentially expressed genes (DEGs) correlated with cardiac function. Here, we demonstrate that embryonic alcohol exposure (EAE) causes cardiomyopathy and diastolic dysfunction through persistent alterations to ventricular wall structure and gene expression. Following abnormal ventricular morphogenesis, >30% of all EAE adults developed increased atrial-to-ventricular size ratios, abnormal ventricular filling dynamics, and reduced myocardial wall relaxation during early diastole despite preserved systolic function. RNA sequencing of the EAE ventricle revealed novel and heart failure-associated genes (slc25a33, ankrd9, dusp2, dusp4, spry4, eya4, and edn1) whose expression levels were altered across the animal's lifespan or correlated with the degree of diastolic dysfunction detected in adulthood. CONCLUSIONS: Our study identifies EAE as a risk factor for adult-onset cardiomyopathy and diastolic dysfunction, regardless of CHD status, and suggests novel molecular indicators of adult EAE-induced heart disease.

A review of the most economically important poisonous plants to the livestock industry on temperate grasslands of China.

The majority of the literature on poisonous plant species in China is published in Chinese and not available to the majority of interested researchers and grassland managers in other countries. Therefore, a review of the Chinese literature was conducted to summarize the occurrence of poisonous plant species on temperate grasslands in China. We reviewed the literature to obtain general information on poisonous species but focus on locoweeds (Astragalus and Oxytropis spp.), drunken horse grass [Achnatherum inebrians (Hance) Keng ex Tzvelev] and langdu (Stellera chamaejasme L.) for information on their toxins, distribution and ecology, control methods and alternate uses. Of the almost 1300 poisonous species found on grasslands in China, these species are responsible for an estimated 80% of all livestock losses. This includes loss of performance as well as mortality. The locoweeds are a complex made up of Oxytropis and Astragalus species. The toxic principle in this complex, as well as in drunken horse grass, is the result of an endophyte fungus whereas in langdu it is produced by the plant. All these species are native to the grasslands, which suggest they have been a problem ever since herding began. Over that period of at least several millennia, herders would have learned and adapted to the presence of poisonous species. Strategies were developed and therapies employed to allow the animals to cope before and after poisoning. Nevertheless, grazing management could still be refined that would allow the use of the toxic legumes, while preventing poisonous symptoms, as has been tested elsewhere.

Transcriptional inhibition after irradiation occurs preferentially at highly expressed genes in a manner dependent on cell cycle progression.

In response to DNA double strand damage, ongoing transcription is inhibited to facilitate accurate DNA repair while transcriptional recovery occurs after DNA repair is complete. However, the mechanisms at play and identity of the transcripts being regulated in this manner are unclear. In contrast to the situation following UV damage, we found that transcriptional recovery after ionizing radiation (IR) occurs in a manner independent of the HIRA histone chaperone. Sequencing of the nascent transcripts identified a programmed transcriptional response, where certain transcripts and pathways are rapidly downregulated after IR, while other transcripts and pathways are upregulated. Specifically, most of the loss of nascent transcripts occurring after IR is due to inhibition of transcriptional initiation of the highly transcribed histone genes and the rDNA. To identify factors responsible for transcriptional inhibition after IR in an unbiased manner, we performed a whole genome gRNA library CRISPR / Cas9 screen. Many of the top hits in our screen were factors required for protein neddylation. However, at short times after inhibition of neddylation, transcriptional inhibition still occurred after IR, even though neddylation was effectively inhibited. Persistent inhibition of neddylation blocked transcriptional inhibition after IR, and it also leads to cell cycle arrest. Indeed, we uncovered that many inhibitors and conditions that lead to cell cycle arrest in G1 or G2 phase also prevent transcriptional inhibition after IR. As such, it appears that transcriptional inhibition after IR occurs preferentially at highly expressed genes in cycling cells.

Design of Superlubricity System Using Si3N4/Polyimide as the Friction Pair and Nematic Liquid Crystals as the Lubricant.

Polyimide (PI) is a high-performance engineering plastic used as a bearing material. A superlubricity system using Si3N4/PI as the friction pair and nematic liquid crystals (LCs) as the lubricant was designed. The superlubricity performance was studied by simulating the start-stop condition of the machine, and it was found that the superlubricity system had good reproducibility and stability. In the superlubricity system, friction aligned with the PI molecules, and this alignment was less relevant compared to which substance was rubbing on the PI. Oriented PI molecules induced LC molecule alignment when the pretilt angle was very small, and the LC molecules were almost parallel to the PI molecules due to the one-dimensional ordered arrangement of LC molecules and low viscosity, which is conducive to the occurrence of the superlubricity phenomenon.

Autoimmune receptor encephalitis in ApoE­/­ mice induced by active immunization with NMDA1.

Subacute progressive neuropsychiatric symptoms with cognitive and motor impairment and autoimmune seizures are some of the typical symptoms of anti­N­methyl­D­aspartate receptor (anti­NMDAR) encephalitis. The mechanisms underlying this disease are yet to be elucidated, which could be partly attributed to the lack of appropriate animal models. The present study aimed to establish an active immune mouse model of anti­NMDAR encephalitis. Mice were immunized with the extracellular segment of the NMDA1 protein, then subjected to open­field and novel object recognition experiments. Plasma was collected after euthanasia on day 30 after immunization and anti­NMDA1 antibodies were detected using ELISA. Furthermore, brain slices were analyzed to measure postsynaptic density protein 95 (PSD­95) and NMDA1 expression. Western blot analysis of NMDA1 and PSD­95 protein expression levels in the hippocampus was also performed. In addition, protein expression levels of PSD­95 and NMDA1 in mouse neuronal HT­22 cells were evaluated. Compared with controls, mice immunized with NMDA1 exhibited anxiety, depression and memory impairment. Moreover, high anti­NMDA1 antibody titers were detected with ELISA and the levels of anti­NMDA1 antibody reduced postsynaptic NMDA1 protein density in the mouse hippocampus. These findings demonstrated the successful construction of a novel mouse model of anti­NMDAR encephalitis by actively immunizing the mice with the extracellular segment of the NMDA1 protein. This model may be useful for studying the pathogenesis and drug treatment of anti­NMDAR encephalitis in the future.

A newly predicted stable calcium argon compound byab initiocalculations under high pressure.

High pressure can change the valence electron arrangement of the elements, and it can be as a new method for the emergence of unexpected new compounds. In this paper, the Ca-Ar compounds at 0-200 GPa are systematically investigated by using CALYPSO structure prediction methods combined with first principles calculations. The study of the Ca-Ar system can provide theoretical guidance for the exploration of new structures of inert elemental Ar compounds under high pressure. A stable structure:P63/mmc-CaAr and six metastable structures:Rm-CaAr2,P4/mmm-CaAr2,Pm1-CaAr3,P4/mmm-CaAr3,P21/m-CaAr4andPm1-CaAr5were obtained. Our calculations show that the only stable phaseP63/mmc-CaAr can be synthesized at high pressure of 90 GPa. All the structures are ionic compounds of metallic nature, and surprisingly all Ar atoms attract electrons and act as an oxidant under high pressure conditions. The calculation results ofab initiomolecular dynamics show thatP63/mmc-CaAr compound maintains significant thermodynamic stability at high temperatures up to 1000 K. The high-pressure structures and electronic behaviors of the Ca-Ar system are expected to expand the understanding of the high-pressure chemical reactivity of compounds containing inert elements, and provide important theoretical support for the search of novel anomalous alkaline-earth metal inert element compounds.