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Abstract Electrocatalytic oxidation of more stable 2,5-furanedimethanol (FDM) for 2,5-furanediformic acid (FDCA) generation with concurrent hydrogen production is attractive but still nascent compared to 5-Hydroxymethyl-2-furaldehyde (HMF). The need for effective and stable bifunctional electrocatalysts that are efficient for the FDM cell is thus quite significant. Wood serves as an ideal matrix for boosting the performance of catalysts, since its hierarchical porous structures facilitate mass transport and provide abundant active sites. Unfortunately, it has never been demonstrated for electrochemically organic synthesis. Herein, the effectiveness of Fe-CoP in catalyzing FDM oxidation was demonstrated by density functional theory (DFT) calculations and experiments, and a renewable carbonized porous wood decorated with Fe-doped CoP nanoleaves (Fe-CoP/CW) was constructed for electrocatalytic FDCA and hydrogen generation. The obtained Fe-CoP/CW as an anode in FDM solution afforded a current density of 100 mA cm−2 with a yield of 90% FDCA at a potential no more than 1.50 V vs RHE, which was 90 mV and 350 mV lower than Fe-CoP/carbon cloth (CC) and IrO2. In addition, Fe-CoP/CW showed excellent long-term stability for 108-h FDM oxidation in strong alkaline solution. Remarkably, in stark contrast to Fe-CoP/CC and Pt, the hydrogen evolution performance of Fe-CoP/CW was not impacted by FDM at the cathode, and it required exceptionally low overpotentials of 0.19 V to achieve 100 mA cm−2. As a result, in terms of the overall cell, the hydrogen production rate was 0.756 mmol cm−2 h−1, which was 3.57 times higher than those of commonly used commercial Pt | IrO2 cell, presenting a Faraday efficiency of near 100%. This work will pave the way towards the implementation of highly suited bifunctional electrodes and the possibility of affordable, effective, and environmentally-friendly wood-derived electrocatalysts for electrochemically organic synthesis. Graphical Abstract

Abstract Predicting H3K27M mutation status in midline gliomas non-invasively is of considerable interest, particularly using deep learning with 11C-methionine (MET) and 18F-fluoroethyltyrosine (FET) positron emission tomography (PET). To optimise prediction efficiency, we derived an assistance training (AT) scheme to allow mutual benefits between MET and FET learning to boost the predictability but still only require either PET as inputs for predictions. Our method significantly surpassed conventional convolutional neural network (CNN), radiomics-based, and MR-based methods, achieved an area under the curve (AUC) of 0.9343 for MET, and an AUC of 0.8619 for FET during internal cross-validation (n = 90). The performance remained high in hold-out testing (n = 19) and consecutive testing cohorts (n = 21), with AUCs of 0.9205 and 0.7404. The clinical feasibility of the proposed method was confirmed by the agreements to multi-departmental decisions and outcomes in pathology-uncertain cases. The findings positions our method as a promising tool for aiding treatment decisions in midline glioma.

IntroductionGut microbiota (GM) has been implicated in gestational diabetes mellitus (GDM), yet longitudinal changes across trimesters remain insufficiently explored.MethodsThis nested cohort study aimed to investigate GM alterations before 24 weeks of gestation and their association with GDM. Ninety-three Chinese participants provided fecal samples during the first and second trimesters. Based on oral glucose tolerance tests, 11 participants were classified as GDM, and 82 as non-diabetic (ND). Using 16S rRNA sequencing, we analyzed both cross-sectional and longitudinal differences in GM structure between those two groups.ResultsIn the first trimester, GDM group exhibited lower levels of Bacteroides_H and Acetatifactor compared to ND group (p

Abstract Base editors (BEs) are a recent generation of genome editing tools that couple a cytidine or adenosine deaminase activity to a catalytically impaired Cas9 moiety (nCas9) to enable specific base conversions at the targeted genomic loci. Given their strong application potential, BEs are under active developments toward greater levels of efficiency and safety. Here, a previously overlooked nCas9‐centric strategy is explored for enhancement of BE. Based on a cytosine BE (CBE), 20 point mutations associated with nCas9‐target interaction are tested. Subsequently, from the initial positive X‐to‐arginine hits, combinatorial modifications are applied to establish further enhanced CBE variants (1.1–1.3). Parallel nCas9 modifications in other versions of CBEs including A3A‐Y130F‐BE4max, YEE‐BE4max, CGBE, and split‐AncBE4max, as well as in the context of two adenine BEs (ABE), likewise enhance their respective activities. The same strategy also substantially improves the efficiencies of high‐fidelity nCas9/BEs. Further evidence confirms that the stabilization of nCas9‐substrate interactions underlies the enhanced BE activities. In support of their translational potential, the engineered CBE and ABE variants respectively enable 82% and 25% higher rates of editing than the controls in primary human T‐cells. This study thus demonstrates a highly adaptable strategy for enhancing BE, and for optimizing other forms of Cas9‐derived tools.

Abstract It is important to monitor the intra-/extracellular concentration of hydrogen peroxide (H2O2) in biological processes. However, miniaturized devices that enable portable and accurate H2O2 measurement are still in their infancy because of the difficulty of developing facile sensing strategies and highly integrated sensing devices. In this work, portable H2O2 sensors based on Pt-Ni hydrogels with excellent peroxidase-like and electrocatalytic activities are demonstrated. Thus, simple and sensitive H2O2 sensing is achieved through both colorimetric and electrochemical strategies. The as-fabricated H2O2 sensing chips exhibit favorable performance, with low detection limits (0.030 μM & 0.15 μM), wide linearity ranges (0.10 μM–10.0 mM & 0.50 μM–5.0 mM), outstanding long-term stability (up to 60 days), and excellent selectivity. With the aid of an M5stack development board, portable visual and electrochemical H2O2 sensors are successfully constructed without complicated and expensive equipment or professional operators. When applied to the detection of H2O2 released from HeLa cells, the results obtained by the developed sensors are in good agreement with those from an ultraviolet‒visible spectrophotometer (UV‒vis) (1.97 μM vs. 2.08 μM) and electrochemical station (1.77 μM vs. 1.84 μM).

Abstract The characteristics of three‐phase imbalances are present in medium‐ and low‐voltage distribution networks. Integrating single‐phase distributed generation (DG) exacerbates network imbalances, resulting in increased power losses and potential safety hazards. To address these issues, a dynamic reconfiguration strategy (DNR) for three‐phase imbalanced distribution networks, considering soft open points (SOP), has been proposed. The objective is to alleviate the three‐phase imbalance and minimize the operational costs of the distribution network. Within the reconfiguration strategy, the constraints of DG current imbalance in practical system operations are considered. A three‐phase imbalanced DNR model that simultaneously considers the constraints of the SOP and DG current imbalances is introduced. This model aims to optimize the operation of all devices, including the SOP, to address the overall imbalance of the distribution network. This enables the authors to transform the non‐linear model into a mixed‐integer linear programming model, significantly improving the solution efficiency. To validate the proposed strategy, simulations of a modified IEEE 34‐node distribution system and an actual 78‐node distribution system were conducted. The results demonstrate that this strategy offers significant economic benefits and ensures the security of the distribution network.

Abstract The balance among different CD4+ T cell subsets is crucial for repairing the injured spinal cord. Dendritic cell (DC)‐derived small extracellular vesicles (DsEVs) effectively activate T‐cell immunity. Altered peptide ligands (APLs), derived from myelin basic protein (MBP), have been shown to affect CD4+ T cell subsets and reduce neuroinflammation levels. However, the application of APLs is challenging because of their poor stability and associated side effects. Herein, it is demonstrate that DsEVs can act as carriers for APL MBP87‐99A91 (A91‐DsEVs) to induce the activation of 2 helper T (Th2) and regulatory T (Treg) cells for spinal cord injury (SCI) in mice. These stimulated CD4+ T cells can efficiently “home” to the lesion area and establish a beneficial microenvironment through inducing the activation of M2 macrophages/microglia, inhibiting the expression of inflammatory cytokines, and increasing the release of neurotrophic factors. The microenvironment mediated by A91‐DsEVs may enhance axon regrowth, protect neurons, and promote remyelination, which may support the recovery of motor function in the SCI model mice. In conclusion, using A91‐DsEVs as a therapeutic vaccine may help induce neuroprotective immunity in the treatment of SCI.

Abstract Neoantigens derived from somatic mutations in cancer cells can induce antigen‐specific T‐cell immune response for cancer immunotherapy. However, the 3D models for assessing neoepitope immunogenicity and efficacy of anti‐tumor T‐cell immune response to neoantigens are less than perfect. Here, a 3D tumor model based on recellularized liver matrix is leveraged with HepG2 cells to investigate T cell cytotoxic reactivity toward hepatocellular carcinoma (HCC) neoantigens. The whole exome sequencing (WES) data of 364 HCC patients in The Cancer Genome Atlas database are collected and 25 highly potential immunogenic neoantigens to human leukocyte antigen (HLA)‐A*02:01 molecule in silico are predicted. Six of the HCC neoantigen candidates are functionally validated with high immunogenicity by measuring cellular interferon‐γ secretion and cytotoxicity during neoantigen‐specific T‐cell immune responses in vitro. Then, the minigene of six functionally identified neoantigen peptides is constructed and the minigene‐modified GFP‐HepG2 cells are generated. Neoantigen‐specific immune response is observed with highly secreted Granzyme B, IFN‐γ, and PD‐1 when targeting the minigene‐modified GFP‐HepG2 cells in the 3D RLM HCC tumor model. Overall, the 3D RLM tumor model provides a novel strategy for preclinical assessment of the efficacy of neoantigen‐specific T cell immune response, which helps develop personalized cancer vaccines and immunotherapy treatments for HCC patients.

Abstract Purpose Ferroptosis plays a crucial role in the initiation and progression of melanoma. This study developed a robust signature with ferroptosis‐related genes (FRGs) and assessed the ability of this signature to predict OS in patients with skin cutaneous melanoma (SKCM). Methods RNA‐sequencing data and clinical information of melanoma patients were extracted from TCGA, GEO, and GTEx. Univariate, multivariate, and LASSO regression analyses were conducted to identify the gene signature. A 10 FRG signature was an independent and strong predictor of survival. The predictive performance was assessed using ROC curve. The functions of this gene signature were assessed by GO and KEGG analysis. The statuses of low‐risk and high‐risk groups according to the gene signature were compared by GSEA. In addition, we investigated the possible relationship of FRGs with immunotherapy efficacy. Results A prognostic signature with 10 FRGs (CYBB, IFNG, FBXW7, ARNTL, PROM2, GPX2, JDP2, SLC7A5, TUBE1, and HAMP) was identified by Cox regression analysis. This signature had a higher prediction efficiency than clinicopathological features (AUC = 0.70). The enrichment analyses of DEGs indicated that ferroptosis‐related immune pathways were largely enriched. Furthermore, GSEA showed that ferroptosis was associated with immunosuppression in the high‐risk group. Finally, immune checkpoints such as PDCD‐1 (PD‐1), CTLA4, CD274 (PD‐L1), and LAG3 were also differential expression in two risk groups. Conclusions The 10 FRGs signature were a strong predictor of OS in SKCM and could be used to predict therapeutic targets for melanoma.

About 47% of pathogenic point mutations could be corrected by ABE-induced A·T-to-G·C conversions. However, the applications of ABEs are still hindered by undesired editing efficiency, limited editing scopes, and off-targeting effects. Here, we develop a new adenine base editor, by embedding TadA-8e monomer into SpRY-nCas9, named as CE-8e-SpRY, which exhibits higher activity at NRN than NYN PAMs favored by SpRY nuclease. CE-8e-SpRY could target nearly all genomic sites in principle and induces the highest targeting efficiency among tested SpRY-based ABEs. In addition, CE-8e-SpRY also shows reduced RNA and DNA off-targeting activities. With optimized sgRNAs, CE-8e-SpRY induces efficient or desired target editing at some disease-relevant loci where conventional ABEs were unable to induce precise and satisfied editing. Taken together, our CE-8e-SpRY could broaden the applicability of ABEs in correcting or introducing pathogenic point mutations.

Inborn errors of immunity (IEIs) are a group of inherited disorders caused by mutations in the protein-coding genes involved in innate and/or adaptive immunity. Hematopoietic stem cell transplantation (HSCT) is a mainstay definitive therapy for many severe IEIs. However, the lack of HLA-matched donors increases the risk of developing severe immunological complications. Gene therapy provides long-term clinical benefits and could be an attractive therapeutic strategy for IEIs. In this review, we describe the development and evolution of clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated proteins (Cas) gene-editing systems, including double-strand break (DSB)-based gene editing and DSB-free base editing or prime editing systems. Here, we discuss the advances in and issues associated with CRISPR/Cas gene editing tools and their potential as therapeutic alternatives for IEIs. We also highlight the progress of preclinical studies for the treatment of human genetic diseases, including IEIs, using CRISR/Cas and ongoing clinical trials based on this versatile technology.

Objectives: Osteoporosis and Parkinson's disease (PD) are both aging-related diseases. PD patients with comorbid osteoporosis are vulnerable to the risk of fracture, which leads to a serious public health burden to the whole society. Therefore, this study sought to reveal the clinical and genetic correlations between PD and osteoporosis based on a cross-sectional study and bioinformatics analysis. Methods: A cross-sectional study of 95 PD patients and 99 healthy controls was conducted. Ordinal logistic regression analysis was utilized to investigate the clinical correlations between PD and osteoporosis. Two microarray datasets (GSE20292, GSE35958) including PD, osteoporosis and normal control samples were retrieved from the GEO database for GO analysis, KEGG pathway analysis and PPI network. Results: PD patients had lower 25(OH)VitD, FN BMD, BMD and T-score of the LS and TH, as well as poorer bone mass diagnosis, yet higher PINP compared to healthy controls. Both age and UPDRS II score of PD patients were adversely correlated with BMD of LS and TH. PD diagnosis acted as an independent risk factor of osteoporosis, and PD patients had approximately double risk for osteoporosis. Bioinformatics analysis further revealed that SNAP25, AQP4, SV2B, KCND3, and ABCA2 had important diagnostic value and risk prediction value for both PD and osteoporosis. Conclusions: PD diagnosis can be used as an independent risk factor for osteoporosis. Moreover, SNAP25, AQP4, SV2B, KCND3 and ABCA2 as the top 5 hub genes have important diagnostic and risk predictive value for both PD and osteoporosis.

Multi-nucleotide variants (MNVs) represent an important type of genetic variation and have biological and clinical significance. To simulate MNVs, we designed four dual-mutation base editors combining hA3A(Y130F), TadA8e(V106W), and protospacer adjacent motif (PAM)-flexible SpRY and selected cytosine and adenine base editor-SpRY (CABE-RY), which had the best editing performance, for further study. Characterization and comparison showed that CABE-RY had a smaller DNA editing window and lower RNA off-target edits than the corresponding single base editors. Thus, we have established a versatile tool to efficiently simulate MNVs over the genome, which could be very useful for functional studies on MNVs in humans.

The objective of this study was to investigate the effect of temperature changes during heat–moisture treatment (HMT) on the appearance, structure and digestibility of sweet potato starch (SPS). The results showed that after HMT, there were depressions, cavities and fragments on the surface of SPS particles. The polarized crosses of SPS were irregular and partially blurred. The relative crystallinity and short-range order of SPS decreased, while rearrangement and reorientation of the starch molecules occurred and the thermal stability increased. The resistant starch content of SPS reached the highest (24.77%) after 4 h treatment at 110 °C and 25% moisture. The obtained results can provide a reference for the modification of SPS.

Using theoretical analysis combined with numerical simulation, the inclination composite force chain arch structure is proposed, and the characteristics of the force chain arch under the influence of surface length are analyzed. The research result shows that the inclination collapse rock formation presents in a “trapezoidal” collapse form that is gradually narrowed from bottom to top. When the length of the working face increases, the collapse height also increases. The strong chain is distributed in the form of flat arches or half arches, forming a composite force chain arch structure. The composite force chain-arch structure forms a carrier structure above the stope with coal pillars at two sides; the working face tends to be sparse with cracks in the middle, and the porosity is low, while dense cracks on both sides increase the porosity, the fracture field and porosity as a whole show the characteristics of “O” ring; the composite force chain arch structure transfers the overburden stress to the coal pillars on both sides, and when the length of the working face increases, the cracks of the coal pillars are more developed, and the stress concentrate coefficient increases.

Rice can accumulate more organic and inorganic arsenic (iAs) than other crop plants. In this study, the localization of As in rice grains was investigated using High Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry (HPLC-ICP-MS) based on 26 rice varieties collected from two provinces. In all the samples, the total As contents in polished rice were 0.03–0.37 mg/kg, with average values of 0.28 and 0.21 mg/kg for two sample sets. The results of the determination of arsenic speciation in different components of rice grain showed that in the polished and brown rice the mean value of arsenite (As(III)) was nearly twice than that of arsenate (As(V)). The regional difference was observed in both total As contents and As speciation. The reason may be that As(III) is more mobile than As(V) in a dissociated form and because of soil properties, rice varieties, and the growing environment. The proportion of iAs and the total As in rice bran was higher than that in polished rice, and this is because As tends accumulate between the husk and the endosperm. In our study, selenium could alleviate the risk of arsenic toxicity at the primary stage of rice growth. Co-exposure to As and Se in germinated rice indicated that the reduction in As accumulation in polished rice reached 73.8%, 76.8%, and 78.3% for total As, As(III), and As(V) when compared with rice treated with As alone. The addition of Se (0.3 mg/kg) along with As significantly reduced the As amount in different parts of germinated rice. Our results indicated that Se biofortification could alleviate the As accumulation and toxicity in rice crops.

Abstract Background Site-specific C>T DNA base editing has been achieved by recruiting cytidine deaminases to the target C using catalytically impaired Cas proteins; the target C is typically located within 5-nt editing window specified by the guide RNAs. The prototypical cytidine base editor BE3, comprising rat APOBEC1 (rA1) fused to nCas9, can indiscriminately deaminate multiple C’s within the editing window and also create substantial off-target edits on the transcriptome. A powerful countermeasure for the DNA off-target editing is to replace rA1 with APOBEC proteins which selectively edit C’s in the context of specific motifs, as illustrated in eA3A-BE3 which targets TC. However, analogous editors selective for other motifs have not been described. In particular, it has been challenging to target a particular C in C-rich sequences. Here, we sought to confront this challenge and also to overcome the RNA off-target effects seen in BE3. Results By replacing rA1 with an optimized human A3G (oA3G), we developed oA3G-BE3, which selectively targets CC and CCC and is also free of global off-target effects on the transcriptome. Furthermore, we created oA3G-BE4max, an upgraded version of oA3G-BE3 with robust on-target editing. Finally, we showed that oA3G-BE4max has negligible Cas9-independent off-target effects at the genome. Conclusions oA3G-BE4max can edit C(C)C with high efficiency and selectivity, which complements eA3A-editors to broaden the collective editing scope of motif selective editors, thus filling a void in the base editing tool box.

To enhance the resolution timeliness and accuracy of inverse kinematics of industrial robots, a robot inverse kinematics method based on ELM-SSA-SCA was proposed. The positive kinematic model of the mechanical arm with Six Degrees of Freedom was established using D-H method, and ELM (extreme learning machine) with fast training speed was used to predict the initial resolution of robot inverse kinematics. The blended optimization method of SSA (Sparrow Search Algorithm) and SCA (Sine Cosine Algorithm) was applied to optimize the obtained initial inverse solution. The position under the optimum fitness was used as the output, so as to obtain the optimum solution of inverse kinematics. It is shown from the experimental results that the algorithm based on SSA-SCA-ELM achieved inverse solutions with faster convergence rate, higher precision and better timeliness compared to the solution method based on PSA-BP neural network.

An experiment was conducted to investigate the influences of supplemental lysophospholipids (LPL) on the growth performance, nutrient digestibility, and fecal bacterial profile, and short-chain fatty acids (SCFAs) of beef cattle. Thirty-six Angus beef cattle [565 ± 10.25 kg body weight (BW)] were grouped by BW and age, and randomly allocated to 1 of 3 treatment groups: (1) control (CON, basal diet); (2) LLPL [CON supplemented with 0.5 g/kg LPL, dry matter (DM) basis]; and (3) HLPL (CON supplemented with 0.75 g/kg, DM basis). The Angus cattle were fed a total mixed ration that consisted of 25% roughage and 75% concentrate (dry matter [DM] basis). The results reveal that LPL inclusion linearly increased the average daily gain (P = 0.02) and the feed efficiency (ADG/feed intake, P = 0.02), while quadratically increasing the final weight (P = 0.02) of the beef cattle. Compared with CON, the total tract digestibilities of DM (P < 0.01), ether extract (P = 0.04) and crude protein (P < 0.01) were increased with LPL supplementation. At the phylum-level, the relative abundance of Firmicutes (P = 0.05) and ratio of Firmicutes: Bacteroidetes (P = 0.04) were linearly increased, while the relative abundances of Bacteroidetes (P = 0.04) and Proteobacteria (P < 0.01) were linearly decreased with increasing LPL inclusion. At the genus-level, the relative abundances of Clostridium (P < 0.01) and Roseburia (P < 0.01) were quadratically increased, and the relative abundances of Ruminococcus was linearly increased (P < 0.01) with LPL supplementation. Additionally, increasing the dose of LPL in diets linearly increased the molar proportion of butyrate (P < 0.01) and total SCFAs (P = 0.01) concentrations. A conclusion was drawn that, as a promising feed additive, LPL promoted growth performance and nutrient digestibility, which may be associated with the change of fecal microbiome and SCFAs.

Abstract Colostrum quality is a vital factor in mortality and growth performance for piglets. Lactoferrin is an immuno-active milk protein that contributes to the formation of a protective layer above intestinal mucosa, possesses the antibacterial and antiviral activities that are favorable for piglet development. However, there is a notable reduction in lactoferrin in sow milk during lactation after the first few days, which causes many piglets to fail to ingest enough colostrum thereby leading to an increase in piglet mortality. In this study, we successfully constructed genome-edited Large-White pigs with marker-free site-specific knock-in of lactoferrin gene in the 3′-end of Casein alpha-s1 via CRISPR/Cas9 mediated homologous recombination. Thus, the lactoferrin protein can be expressed in the mammary gland in the control of Casein alpha-s1 promoter. As expected, the lactoferrin protein in genetically modified pigs sustained high expression in both colostrum and milk when compared with wild-type pigs. Moreover, the bacterial plate assay indicated that the milk from genetically modified pigs showed bacteriostatic effects when compared with control pigs. Taken together, our study demonstrated that the milk from genetically modified pigs had antibacterial activity which may reduce the costs of veterinary drug and improve the surviving rate of piglets, which is promising for pig breeding.

Phosphorylated corn starch derivatives show improved pasting properties compared to native corn starch. In this study, starch phosphate carbamides (SPC) were prepared through a dry process reaction of starch with urea and phosphate salts. The effects of major factors on SPC pasting properties were studied by response surface methodology. Validation results showed that the polynomial quadratic models were adequate for predicting SPC pasting properties. Fourier transform infrared spectroscopy (FTIR) and a Rapid Visco Analyzer (RVA) were used to characterize SPC. RVA results showed SPC had higher peak viscosity, trough, breakdown, setback and final viscosities and lower gelatinization temperature compared to native starch. FTIR showed the characteristic absorption of the ester phosphate group in SPC at 1244/cm. SPC with high viscosity stability were applied to replace part of wheat flour used for making noodles. The best quality noodles were obtained with 12% SPC substitution.

The development of CRISPR/Cas9-mediated base editors (BEs) provided a versatile tool for precise genome editing. The recently developed xCas9-derived base editors (xBEs) that recognize the NG PAM substantially expand the targeting scope in the genome, while their editing efficiency needs to be improved. Here, we described an improved version of xBEs by fusing the BPNLS and Gam to the N terminus of xBEs (BPNLS-Gam-xBE3 and BPNLS-xABE), and this version of base editor displayed higher targeting efficiency for the majority of detected sites. By using this improved version of xBEs, we successfully created and corrected pathogenic mutations at genomic sites with the NGN protospacer-adjacent motif in human cells. Lastly, we used BPNLS-Gam-xBE3 to model pathogenic mutations in discarded human tripronuclear (3PN) zygotes, and no obvious off-targets and indels were detected. Taken together, the data in our study offer an efficient tool for precise genome editing and, thus, an enriched base editing toolkit. Keywords: CRISPR/Cas, base editor, xCas9, base editing efficiency, pathogenic mutation, Wilson disease

Base editing systems show their power in modeling and correcting the pathogenic mutations of genetic diseases. Previous studies have already demonstrated the editing efficiency of BE3-mediated C-to-T conversion in human embryos. However, the precision and efficiency of a recently developed adenine base editor (ABE), which converts A-to-G editing in human embryos, remain to be addressed. Here we selected reported pathogenic mutations to characterize the ABE in human tripronuclear embryos. We found effective A-to-G editing occurred at the desirable sites using the ABE system. Furthermore, ABE-mediated A-to-G editing in the single blastomere of the edited embryos exhibited high product purity. By deep sequencing and whole-genome sequencing, A or T mutations didn’t increase significantly, and no off-target or insertion or deletion (indel) mutations were detected in these edited embryos, indicating the ABE-mediated base editing in human embryos is precise and controllable. For some sites, since a different editing pattern was obtained from the cells and the embryos targeted with the same single guide RNA (sgRNA), it suggests that ABE-mediated editing might have different specificity in vivo. Taken together, we efficiently generated pathogenic A-to-G mutations in human tripronuclear embryos via ABE-mediated base editing.

Summary: RNA splicing is related to many human diseases; however, lack of efficient genetic approaches to modulate splicing has prevented us from dissecting their functions in human diseases. Recently developed base editors (BEs) offer a new strategy to modulate RNA splicing by converting conservative splice sites, but it is limited by the editing precision and scope. To overcome the limitations of currently available BE-based tools, we combined SpCas9-NG with ABEmax to generate a new BE, ABEmax-NG. We demonstrated that ABEmax-NG performed precise A⋅T to G⋅C conversion with an expanded scope, thus covering many more splicing sites. Taking advantage of this tool, we precisely achieved A⋅T to G⋅C conversion exactly at the splice sites. We further modeled pathogenic RNA splicing in vitro and in vivo. Taken together, we successfully generated a versatile tool suitable for precise and broad editing at the splice sites. : Biological Sciences; Molecular Biology; Biological Sciences Tools Subject Areas: Biological Sciences, Molecular Biology, Biological Sciences Tools

In order to solve the problems of low accuracy and low efficiency of point cloud registration for stereo camera systems, we propose a binocular stereo camera point cloud registration method based on IWOA and Improved ICP. We propose the following approaches in this paper—the registration process is divided into two steps to complete the initial coarse registration and the exact registration. In the initial registration stage, an improved Whale Optimization Algorithm (IWOA) based on nonlinear convergence factor and adaptive weight coefficients was proposed to realize the initial registration in combination with the RANSAC algorithm, and the obtained transformation matrix was used as the initial estimate of the subsequent exact registration algorithm. In the second step of the exact registration stage, an IICP algorithm with the introduction of normal vector weighting constraints at key points was proposed for achieving point cloud exact registration. This algorithm was verified by using Stanford point clouds (bunnies and monkeys) and our own point clouds algorithm, and the proposed algorithm in this paper has high registration accuracy, improved registration speed, and convergence speed.

Abstract Using T‐cell chimeric antigen receptors (CAR‐T) to activate and redirect T cells to tumors expressing the cognate antigen represents a powerful approach in cancer therapy. However, normal tissues with low expression of tumor‐associated antigens (TAAs) can be mistargeted, resulting in severe side effects. An approach using a collection of T cells expressing a diverse, 106‐member combinatorial cellular library of CARs, in which members can be specifically enriched based on avidity for cell membrane antigens, is reported. Using CD38 as the target antigen, an efficient and effective selection of CARs specifically recognizing CD38+ tumor cells is demonstrated. These selected CAR‐T's produce cytokines known to be associated with T cell activation in a CD38 expression‐dependent manner. This avidity‐based selection endows the engineered T cells with minimal off‐tumor effects, while retaining robust antitumor efficacy both in vitro and in vivo. The described method may facilitate the application of CAR‐T therapy to TAAs previously considered undruggable.

This study was conducted to evaluate the effect of dietary supplementation with lysophospholipids (LPLs) on the growth performance, nutrient digestibility, nitrogen utilization, and blood metabolites of finishing beef cattle. In total, 40 Angus beef bulls were blocked for body weight (447 ± 9.64 kg) and age (420 ± 6.1 days) and randomly assigned to one of four treatments (10 beef cattle per treatment): (1) control (CON; basal diet); (2) LLPL (CON supplemented with 0.012% dietary LPL, dry matter (DM) basis); (3) MLPL (CON supplemented with 0.024% dietary LPL, DM basis); and (4) HLPL (CON supplemented with 0.048% dietary LPLs, DM basis). The results showed that dietary supplementation with LPLs linearly increased the average daily gain (p < 0.01), digestibility of DM (p < 0.01), crude protein (p < 0.01), and ether extract (p < 0.01) and decreased the feed conversion ratio (p < 0.01). A linear increase in N retention (p = 0.01) and a decrease in urinary (p = 0.04) and fecal N (p = 0.02) levels were observed with increasing the supplemental doses of LPLs. Bulls fed LPLs showed a linear increase in glutathione peroxidase (p = 0.04) and hepatic lipase (p < 0.01) activity and a decrease in cholesterol (p < 0.01), triglyceride (p < 0.01), and malondialdehyde (p < 0.01) levels. In conclusion, supplementation with LPLs has the potential to improve the growth performance, nutrient digestibility, and antioxidant status of beef cattle.

CRISPR-based base editors allow for single nucleotide genome editing in a range of organisms. Here the authors demonstrate the in vivo generation of mouse models carrying clinically relevant mutations using C→T and A→G editors.