Efficient genome-wide first-generation phenotypic screening system in mice using the piggyBac transposon.

Genome-wide phenotypic screens provide an unbiased way to identify genes involved in particular biological traits, and have been widely used in lower model organisms. However, cost and time have limited the utility of such screens to address biological and disease questions in mammals. Here we report a highly efficient piggyBac (PB) transposon-based first-generation (F1) dominant screening system in mice that enables an individual investigator to conduct a genome-wide phenotypic screen within a year with fewer than 300 cages. The PB screening system uses visually trackable transposons to induce both gain- and loss-of-function mutations and generates genome-wide distributed new insertions in more than 55% of F1 progeny. Using this system, we successfully conducted a pilot F1 screen and identified 5 growth retardation mutations. One of these mutants, a Six1/4 PB/+ mutant, revealed a role in milk intake behavior. The mutant animals exhibit abnormalities in nipple recognition and milk ingestion, as well as developmental defects in cranial nerves V, IX, and X. This PB F1 screening system offers individual laboratories unprecedented opportunities to conduct affordable genome-wide phenotypic screens for deciphering the genetic basis of mammalian biology and disease pathogenesis.

SERINC2 increases the risk of bipolar disorder in the Chinese population.

BACKGROUND: Although common variants in a large collection of patients are associated with increased risk for bipolar disorder (BD), studies have only been able to predict 25%-45% of risks, suggesting that lots of variants that contribute to the risk for BD haven't been identified. Our study aims to identify novel BD risk genes. METHODS: We performed whole-exome sequencing of 27 individuals from 6 BD multi-affected Chinese families to identify candidate variants. Targeted sequencing of one of the novel risk genes, SERINC2, in additional sporadic 717 BD patients and 312 healthy controls (HC) validated the association. Magnetic resonance imaging (MRI) were performed to evaluate the effect of the variant to brain structures from 213 subjects (4 BD subjects from a multi-affected family, 130 sporadic BD subjects and 79 HC control). RESULTS: BD pedigrees had an increased burden of uncommon variants in extracellular matrix (ECM) and calcium ion binding. By large-scale sequencing we identified a novel recessive BD risk gene, SERINC2, which plays a role in synthesis of sphingolipid and phosphatidylserine (PS). MRI image results show the homozygous nonsense variant in SERINC2 affects the volume of white matter in cerebellum. CONCLUSIONS: Our study identified SERINC2 as a risk gene of BD in the Chinese population.

Modulation of immunosuppressive effect of rapamycin via microfluidic encapsulation within PEG-PLGA nanoparticles.

The high hydrophobicity and low oral availability of immunosuppressive drug, rapamycin, seriously limit its application. It was thus aimed to develop a PEG-PLGA based nano-loading system for rapamycin delivery to achieve improved bioavailability with sustained effects via a novel microfluidic chip and manipulation of the hydrophobic PLGA chain length. PDMS based microfluidic chip with Y shape was designed and PEG-PLGA polymers with different PLGA chain length were used to prepare rapamycin nano-delivery systems. Dendritic cells were selected to evaluate the immunosuppressive effect of the nanoparticles including cytotoxicity assay, dendritic cell activation, and cytokine levels. The effects of different PEG-PLGA nanoparticles on the immunomodulatory properties were finally compared. It was shown that PEG-PLGA could be successfully used for rapamycin encapsulation via microfluidics to obtain nano-delivery systems (Rapa&P-20 k, Rapa&P-50 k and Rapa&P-95 k) ranging from 100 nm to 116 nm. The encapsulation efficiency was ranged from 69.70% to 84.55% and drug loading from 10.45% to 12.68%. The Rapa&P-50 k (PLGA chain length: 50 k) could achieve the highest drug loading (DL) and encapsulation efficiency (EE) as 12.68% and 84.55%. The encapsulated rapamycin could be gradually released from three nanoparticles for more than 1 month without any noticeable burst release. The Rapa & P nanoparticles exhibited enhanced immunosuppressive effects over those of free rapamycin as shown by the expression of CD40 and CD80, and the secretion of IL-1ß, IL-12 and TGF-ß1. Rapa&P-50 k nanoparticles could be the optimal choice for rapamycin delivery as it also achieved the most effective immunosuppressive property. Hence, this study could provide an efficient technology with superior manipulation to offer a solution for rapamycin delivery and clinical application.

Enhanced Electron Delocalization within Coherent Nano-Heterocrystal Ensembles for Optimizing Polysulfide Conversion in High-Energy-Density Li-S Batteries.

Commercialization of high energy density Lithium-Sulfur (Li-S) batteries is impeded by challenges such as polysulfide shuttling, sluggish reaction kinetics, and limited Li+ transport. Herein, a jigsaw-inspired catalyst design strategy that involves in situ assembly of coherent nano-heterocrystal ensembles (CNEs) to stabilize high-activity crystal facets, enhance electron delocalization, and reduce associated energy barriers is proposed. On the catalyst surface, the stabilized high-activity facets induce polysulfide aggregation. Simultaneously, the surrounded surface facets with enhanced activity promote Li2S deposition and Li+ diffusion, synergistically facilitating continuous and efficient sulfur redox. Experimental and DFT computations results reveal that the dual-component hetero-facet design alters the coordination of Nb atoms, enabling the redistribution of 3D orbital electrons at the Nb center and promoting d-p hybridization with sulfur. The CNE, based on energy level gradient and lattice matching, endows maximum electron transfer to catalysts and establishes smooth pathways for ion diffusion. Encouragingly, the NbN-NbC-based pouch battery delivers a Weight energy density of 357 Wh kg-1, thereby demonstrating the practical application value of CNEs. This work unveils a novel paradigm for designing high-performance catalysts, which has the potential to shape future research on electrocatalysts for energy storage applications.

Mechanistic Insight into the Synergy between Platinum Single Atom and Cluster Dual Active Sites Boosting Photocatalytic Hydrogen Evolution.

In the energy transition context, the design and synthesis of high-performance Pt-based photocatalysts with low Pt content and ultrahigh atom-utilization efficiency for hydrogen production are essential. Herein, a facile approach for decorating atomically dispersed Pt cocatalysts having single-atom (SA) and atomic cluster (C) dual active sites on CdS nanorods (PtSA+C /CdS) via atomic layer deposition is reported. The size of the cocatalyst and the spatial intimacy of the cocatalyst active sites are precisely engineered at the atomic scale. The PtSA+C /CdS photocatalysts show the optimized photocatalytic hydrogen evolution activity, achieving a reaction rate of 80.4 mmol h-1 g-1 , which is 1.6- and 7.3-fold higher than those of the PtSA /CdS and PtNP /CdS photocatalysts, respectively. Thorough characterization and theoretical calculations reveal that the enhanced photocatalytic activity is due to a remarkable synergy between SAs and atomic clusters as dual active sites, which are responsible for water adsorption-dissociation and hydrogen desorption, respectively. A similar synergetic effect is found in a representative Pt/TiO2 system, indicating the generality of the strategy. This study demonstrates the significance of the synergy between active sites for enhancing the reaction efficiency, opening a new avenue for the rational design of atomically dispersed photocatalysts with high efficiency.

Vanadium as Auxiliary for Fe-V Dual-Atom Electrocatalyst in Lithium-Sulfur Batteries: "3D in 2D" Morphology Inducer and Coordination Structure Regulator.

The undesirable shuttling behavior, the sluggish redox kinetics of liquid-solid transformation, and the large energy barrier for decomposition of Li2S have been the recognized problems impeding the practical application of lithium-sulfur batteries. Herein, inspired by the spectacular catalytic activity of the Fe/V center in bioenzyme for nitrogen/sulfur fixation, we design an integrated electrocatalyst comprising N-bridged Fe-V dual-atom active sites (Fe/V-N7) dispersed on ingenious "3D in 2D" carbon nanosheets (denoted as DAC), in which vanadium induces the laminar structure and regulates the coordination configuration of active centers simultaneously, realizing the redistribution of the 3d-orbital electrons of Fe centers. The high coupling/conjunction between Fe/V 3d electrons and S 2p electrons shows strong affinity and enhanced reactivity of DAC-Li2Sn (1 ≤ n ≤ 8) systems. Thus, DAC presents strengthened chemisorption ability toward polysulfides and significantly boosts bidirectional sulfur redox reaction kinetics, which have been evidenced theoretically and experimentally. Besides, the well-designed "3D in 2D" morphology of DAC enables uniform sulfur distribution, facilitated electron transfer, and abundant active sites exposure. Therefore, the assembled Li-S cells present outstanding cycling stability (637.3 mAh g-1 after 1000 cycles at 1 C) and high rate capability (711 mAh g-1 at 4 C) under high sulfur content (70 wt %).

Clinical, genetic, and experimental research of hyperphenylalaninemia.

Hyperphenylalaninemia (HPA) is the most common amino acid metabolism defect in humans. It is an autosomal-recessive disorder of the phenylalanine (Phe) metabolism, in which high Phe concentrations and low tyrosine (Tyr) concentrations in the blood cause phenylketonuria (PKU), brain dysfunction, light pigmentation and musty odor. Newborn screening data of HPA have revealed that the prevalence varies worldwide, with an average of 1:10,000. Most cases of HPA result from phenylalanine hydroxylase (PAH) deficiency, while a small number of HPA are caused by defects in the tetrahydrobiopterin (BH4) metabolism and DnaJ heat shock protein family (Hsp40) member C12 (DNAJC12) deficiency. Currently, the molecular pathophysiology of the neuropathology associated with HPA remains incompletely understood. Dietary restriction of Phe has been highly successful, although outcomes are still suboptimal and patients find it difficult to adhere to the treatment. Pharmacological treatments, such as BH4 and phenylalanine ammonia lyase, are available. Gene therapy for HPA is still in development.

A Lactoglobulin-Composite Self-Healing Coating for Mg Alloys.

Corrosion issue is one of the most crucial bottlenecks for extensive employment of Mg alloys, in particular under harsh engineering conditions. Differing from traditional approaches, a self-healing protective coating composed of lactoglobulin is proposed herein to offer sustainable protection to the underlying Mg parts. Corrosion resistance, evaluated by electrochemical measurements and hydrogen evolution tests, indicates that the lactoglobulin composite film exhibits nobler corrosion potential (-1.28 VSCE), smaller corrosion current density (8.4 × 10-6 A/cm2), and lower average corrosion rate (∼0.03 mm/y) than those of its bare counterparts. Moreover, the pre-made cracks in the film were evidently self-healed within 24 h of exposure to corrosive media. The proposed self-healing lactoglobulin composite film provides opportunities to tackle the corrosion challenges of Mg alloys.

Effect of different reconstruction algorithms on coronary artery calcium scores using the reduced radiation dose protocol: a clinical and phantom study.

BACKGROUND: This study aimed to evaluate the effects of different iterative reconstruction (IR) algorithms on coronary artery calcium (CAC) score quantification using the reduced radiation dose (RRD) protocol in an anthropomorphic phantom and in patients. METHODS: A thorax phantom, containing 9 calcification inserts with varying hydroxyapatite (HA) densities, was scanned with the reference protocol [120 kv, 80 mAs, filtered back projection (FBP)] and RRD protocol (120 kV, 20-80 mAs, 5 mAs interval) using a 256-slice computed tomography (CT) scanner. Raw data were reconstructed with different reconstruction algorithms [iDose4 levels 1-7 and iterative model reconstruction (IMR) levels 1-3]. Signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and Agatston score (AS) were calculated for each image series. The correction factor was derived from linear regression analysis between the reference image series and other image series with different parameters. Additionally, 40 patients were scanned with the RRD protocol (50 mAs) and reconstructed with FBP, iDose4 level 4, and IMR level 2. AS was calculated for the 3-group image series, and was corrected by applying a correction factor for the IMR group. The agreement of risk stratification with different reconstruction algorithms was also analyzed. RESULTS: For the phantom study, the iDose4 and IMR groups had significantly higher SNR and CNR than the FBP group (all P<0.05). There were no significant differences in the total AS after comparing image series reconstructed with iDose4 (level 1-7) and FBP (all P>0.05), while AS from the IMR (level 1-3) image series were lower than the FBP group (all P<0.05). The tube current of 50 mAs was determined for the clinical study, and the correction factor was 1.14. For the clinical study, the median AS from the iDose4 and IMR groups were both significantly lower compared to the FBP image series [(112.89 (63.01, 314.09), 113.22 (64.78, 364.95) vs. 118.59 (65.05, 374.48), both P<0.05]. After applying the correction factor, the adjusted AS from the IMR group was not significantly different from that of the FBP group [126.48 (69.62, 355.85) vs. 118.59 (65.05, 374.48), P=0.145]. Moreover, the agreement in risk stratification between FBP and IMR improved from 0.81 to 0.85. CONCLUSIONS: The RRD CAC scoring scan using the IMR reconstruction algorithm is clinically feasible, and a correction factor can help reduce the AS underestimation effect.

The accuracy of compressed sensing cardiovascular magnetic resonance imaging in heart failure classifications.

To evaluate the accuracy of compressed sensing (CS) cardiovascular magnetic resonance imaging (CMR) in the classifications of heart failure (HF). One hundred and fifty-four patients underwent 3 T CMR using CS single-breath-hold cine (SBH-cine) after a standard multiple-breath-hold cine (MBH-cine). The image quality of the two methods was compared. Cardiac function parameters were quantitatively analyzed. The patients were divided into different HF types based on diagnostic criteria using two sequences. The scan time for CS SBH-cine was reduced by 88% compared with that of standard MBH-cine. In the non-atrial-fibrillation (NAF) group (n = 121), the image quality score of CS SBH-cine was slightly decreased compared with standard MBH-cine (4.5 ± 0.6 for the CS SBH-cine vs. 4.7 ± 0.5 for the standard MBH-cine, T = 5.038, p < 0.05). In the atrial fibrillation (AF) group (n = 33), the CS SBH-cine image quality score was slightly higher than that of the standard MBH-cine (3.8 ± 0.7 for the CS SBH-cine vs. 3.3 ± 0.5 for the standard MBH-cine, T = - 4.503, p < 0.05). The left ventricular (LV) end diastolic volume, LV end systolic volume, LV ejection fraction, and LV mass index calculated by CS SBH-cine had no significant differences from those calculated by standard MBH-cine. The agreement on HF classifications of both the standard MBH-cine and the CS SBH-cine was excellent (kappa = 0.934, p < 0.05). The result showed that HF could be classified accurately using CS SBH-cine.

Early and Quantitative Assessment of Myocardial Deformation in Essential Hypertension Patients by Using Cardiovascular Magnetic Resonance Feature Tracking.

The aims of the study were to identify subclinical global systolic function abnormalities and evaluate influencing factors associated with left ventricular (LV) strain parameters in hypertensive subjects using cardiovascular magnetic resonance imaging feature tracking (CMR-FT). The study enrolled 57 patients with essential hypertension (mean age: 43.04 ± 10.90 years; 35 males) and 26 healthy volunteers (mean age: 38.69 ± 10.44 years; 11 males) who underwent clinical evaluation and CMR examination. Compared with controls, hypertensive patients had significantly impaired myocardial strain values while ejection fraction (EF) did not differ. After multivariate regression analyses adjustment for confounders, the global radial strains (GRS) was independently associated with the mean arterial pressure (MAP) and left ventricular mass index (LVMI) (ß = -0.219, p = 0.009 and ß = -0.224, p = 0.015, respectively; Adjusted R2 = 0.4); the global circumferential strains (GCS) was also independently associated with the MAP and LVMI (ß = 0.084, p = 0.002 and ß = 0.073, p = 0.01, respectively; Adjusted R2 = 0.439); the global longitudinal strains (GLS) was independently associated with the Age and MAP (ß = 0.065, p = 0.021 and ß = 0.077, p = 0.009, respectively; Adjusted R2 = 0.289). Myocardial strain can early detect the myocardial damage and may be an appropriate target for preventive strategies before abnormalities of EF.

TTRAP is a novel PML nuclear bodies-associated protein.

PML nuclear body (PML NB) is an important macromolecular nuclear structure that is involved in many essential aspects of cellular function. Tens of proteins have been found in PML NBs, and promyelocytic leukemia protein (PML) has been proven to be essential for the formation of this structure. Here, we showed that TRAF and TNF receptor-associated protein (TTRAP) was a novel PML NBs-associated protein. TTRAP colocalized with three important PML NBs-associated proteins, PML, DAXX and Sp100 in the typical fashion of PML NBs. By yeast mating assay, TTRAP was identified to interact with these PML NBs-associated proteins. The transcription and expression of TTRAP could be induced by IFN-gamma, representing another common feature of PML NBs-associated proteins. These results would not only be important for understanding PML NBs but also be helpful in studying the TTRAP function in the future.

piggyBac insertional mutagenesis screen identifies a role for nuclear RHOA in human ES cell differentiation.

The mechanisms regulating human embryonic stem (ES) cell self-renewal and differentiation are not well defined in part due to the lack of tools for forward genetic analysis. We present a piggyBac transposon gain of function screen in human ES cells that identifies DENND2C, which genetically cooperates with NANOG to maintain self-renewal in the presence of retinoic acid. We show that DENND2C negatively regulates RHOA activity, which cooperates with NANOG to block differentiation. It has been recently shown that RHOA exists in the nucleus and is activated by DNA damage; however, its nuclear function remains unknown. We discovered that RHOA associates with DNA and that DENND2C affects nuclear RHOA localization, activity, and DNA association. Our study illustrates the power of piggyBac as a cost-effective, efficient, and easy to use tool for forward genetic screens in human ES cells and provides insight into the role of RHOA in the nucleus.