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PURPOSEThe exact pathogenesis of primary open-angle glaucoma (POAG) is poorly understood. Genome-wide association studies (GWAS) have recently uncovered many loci associated with variation in intraocular pressure (IOP); a crucial risk factor for POAG. Artificial intelligence (AI) can be used to interrogate the effect of specific genetic knockouts on the morphology of trabecular meshwork cells (TMCs), the regulatory cells of IOP.METHODSSixty-two genes at fifty-five loci associated with IOP variation were knocked out in primary TMC lines. All cells underwent high-throughput microscopy imaging after being stained with a five-channel fluorescent cell staining protocol. A convolutional neural network (CNN) was trained to distinguish between gene knockout and normal control cell images. The area under the receiver operator curve (AUC) metric was used to quantify morphological variation in gene knockouts to identify potential pathological perturbations.RESULTSCells whereRALGPS1had been perturbed demonstrated the greatest morphological variation from normal TMCs (AUC 0.851, SD 0.030), followed byLTBP2(AUC 0.846, SD 0.029) andBCAS3(AUC 0.845, SD 0.020). Of seven multi-gene loci, five had statistically significant differences in AUC (pCONCLUSIONSWe demonstrate a robust method for functionally interrogating genome-wide association signals using high-throughput microscopy and AI. Genetic variations inducing marked morphological variation can be readily identified, allowing for the gene-based dissection of loci associated with complex traits.

Exploring the structural and electrical properties of DNA origami nanowires is an important endeavor for the advancement of DNA nanotechnology and DNA nanoelectronics. Highly conductive DNA origami nanowires are a desirable target for creating low-cost self-assembled nanoelectronic devices and circuits. In this work, the structure-dependent electrical conductance of DNA origami nanowires is investigated. A silicon nitride (Si

Combining surface-initiated, TdT (terminal deoxynucleotidyl transferase) catalyzed enzymatic polymerization (SI-TcEP) with precisely engineered DNA origami nanostructures (DONs) presents an innovative pathway for the generation of stable, polynucleotide brush-functionalized origami nanostructures. We demonstrate that SI-TcEP can site-specifically pattern DONs with brushes containing both natural and non-natural nucleotides. The brush functionalization can be precisely controlled in terms of the location of initiation sites on the origami core and the brush height and composition. Coarse-grained simulations predict the conformation of the brush-functionalized DONs that agree well with the experimentally observed morphologies. We find that polynucleotide brush-functionalization increases the nuclease resistance of DONs significantly, and that this stability can be spatially programmed through the site-specific growth of polynucleotide brushes. The ability to site-specifically decorate DONs with brushes of natural and non-natural nucleotides provides access to a large range of functionalized DON architectures that would allow for further supramolecular assembly, and for potential applications in smart nanoscale delivery systems.

The human immune system displays substantial variation between individuals, leading to differences in susceptibility to autoimmune disease. We present single-cell RNA sequencing (scRNA-seq) data from 1,267,758 peripheral blood mononuclear cells from 982 healthy human subjects. For 14 cell types, we identified 26,597 independent cis-expression quantitative trait loci (eQTLs) and 990 trans-eQTLs, with most showing cell type–specific effects on gene expression. We subsequently show how eQTLs have dynamic allelic effects in B cells that are transitioning from naïve to memory states and demonstrate how commonly segregating alleles lead to interindividual variation in immune function. Finally, using a Mendelian randomization approach, we identify the causal route by which 305 risk loci contribute to autoimmune disease at the cellular level. This work brings together genetic epidemiology with scRNA-seq to uncover drivers of interindividual variation in the immune system.

ObjectiveTo investigate the differences in ocular surface characteristics, tear film quality, and the incidence of dry eye disease (DED) between Systemic Lupus Erythematosus (SLE) patients and healthy populations.MethodsThis age and gender-matched cross-sectional study included 96 SLE patients without secondary Sjögren's syndrome (SS) and 72 healthy subjects. The Ocular Surface Disease Index (OSDI), tear meniscus height (TMH), non-invasive tear film breakup time (NIKBUT), meibography, and tear film lipid layer grade were assessed. A receiver operative characteristic (ROC) curve was constructed to evaluate the predictive value of risk factors.ResultsCompared with the control subjects, a significantly greater proportion of SLE patients met the TFOS DEWS II DED diagnostic criteria (34.3 vs. 18.1%, P = 0.019). SLE patients without SS had higher OSDI scores [10.0 (4.5,18.0) vs. 5.0 (2.5,11.9), P < 0.001], and shorter NIKBUT [9.6 (6.6,15.0) vs. 12.3 (8.4, 15.8), P = 0.035]. Furthermore, TMH, Tear film lipid layer grade, and Meibomian gland (MG) dropout in SLE patients were worse than those in control subjects (all P < 0.05). For ROC analysis, the area under curve (AUC), sensitivity and specificity of prediction were 0.915, 75.8 and 92.1% for the combination of SLE disease activity index (SLEDAI), age and NIKBUT.ConclusionsSLE patients without SS exhibited a higher risk for DED than healthy subjects, and the poorer Meibomian gland function in SLE patients may potentially contribute to the development of DED. The combined parameters of SLEDAI, age and NIKBUT showed a high efficiency for the diagnosis of DED in SLE patients, with practical clinical applications.

Interactive teaching is very important for students to learn English. In the junior English classroom, language is not only a tool for teachers to organize teaching, but also the main source of students’ target language input. Therefore, more and more researchers at home and abroad pay attention to it. Classroom interactive language mainly includes questions and feedback. Among them, classroom questions are divided into display questions and reference questions. Junior high school is a very important stage of English learning. This paper will elaborate on interactive teaching and interactive classroom, and analyze the characteristics of junior high school English interactive classroom in detail, so as to provide a theoretical and methodological perspective for studying the relationship between second language acquisition and classroom teaching.

Reading is a dynamic process in which the reader and the text interact and construct meaning together. More and more junior high school English teachers and students have begun to pay attention to the importance of English reading and put forward higher requirements for students' English reading ability. Studying the reading habits and reading strategies of junior high school students and cultivating students to master reading strategies is a very important issue in the current junior high school English teaching, which has very important practical significance. This paper discusses reading strategies for junior English teaching, designs reading training methods for different teaching stages, and discusses the functions and characteristics of three learning modes to improve students' English reading ability and self-learning ability. Through the use of different learning modes, according to the students' personal abilities and personality characteristics, we can mobilize their enthusiasm and initiative, stimulate their interest in English reading, and achieve practical quality-oriented teaching.

OBJECTIVES: In the GI tract, Pomegranates (Pom) ellagitannins undergo hydrolysis to ellagic acid (EA) and are further metabolized into urolithins by the gut microbiota. Both EA and UA have significant health benefits. EA is also available as dietary supplement. We aimed to compare the bioavailability of EA and its biotransformation to UA between pure EA and Pom juice (PomJ). METHODS: 17 healthy male participants were randomly assigned to take a single dose of 8 oz of PomJ or 500 mg of EA, followed by 2-week washout, and crossover to the other intervention. We collected blood prior and 1 and 2 h after consumption of EA or PomJ and urine for 24hours. Plasma concentration of EA was measured using HPLC. Urinary EA metabolites (DMEAG and UAG) were determined by LC/MS and normalized to urine creatinine levels. Circulating MCP1, insulin and leptin were analyzed using the multiplex human cytokine panel (Millipore). RESULTS: There was no difference in plasma EA concentration after administration of PomJ versus 500 mg EA. Urinary DMEAG, normalized to creatinine, was significantly higher after PomJ compared to EA intake (PomJ: 0.11 + 0.02; EA: 0.05 + 0.01 µg/mg; P