A 4D single-cell protein atlas of transcription factors delineates spatiotemporal patterning during embryogenesis.

Complex biological processes such as embryogenesis require precise coordination of cell differentiation programs across both space and time. Using protein-fusion fluorescent reporters and four-dimensional live imaging, we present a protein expression atlas of transcription factors (TFs) mapped onto developmental cell lineages during Caenorhabditis elegans embryogenesis, at single-cell resolution. This atlas reveals a spatiotemporal combinatorial code of TF expression, and a cascade of lineage-specific, tissue-specific and time-specific TFs that specify developmental states. The atlas uncovers regulators of embryogenesis, including an unexpected role of a skin specifier in neurogenesis and the critical function of an uncharacterized TF in convergent muscle differentiation. At the systems level, the atlas provides an opportunity to model cell state-fate relationships, revealing a lineage-dependent state diversity within functionally related cells and a winding trajectory of developmental state progression. Collectively, this single-cell protein atlas represents a valuable resource for elucidating metazoan embryogenesis at the molecular and systems levels.

Microbe-derived Antioxidants Enhance Lipid Synthesis by Regulating the Hepatic AMPKα-SREBP1c Pathway in Weanling Piglets.

BACKGROUND: Weaning usually causes low feed intake and weight loss in piglets, which mobilizes lipid to energize. The microbe-derived antioxidants (MAs) exhibit great potential in antioxidation, anti-inflammation, and metabolic regulation. OBJECTIVES: We aimed to investigate the changes of lipid metabolism postweaning and effects of MA on growth performance and hepatic lipid metabolism in weanling piglets. METHODS: In the first experiment, piglets weaned at 21 d of age were slaughtered on weaning day (d0), 4 (d4), and 14 (d14) postweaning (6 piglets per day). In the second experiment, piglets were divided into 2 groups, receiving MA (MA) and saline gavage (CON), respectively. All piglets were weaned at 21 d of age and 6 piglets from each group were slaughtered at 25 d of age. RESULTS: In experiment 1, the serum triglyceride, total cholesterol (TC), and LDL cholesterol on d4 and d14 declined significantly compared with d0 (P < 0.05). The serum leptin on d0 was higher than that on d4 and d14 (P < 0.05). The serum ghrelin kept increasing from d0 to d14 (P < 0.05). The hepatic hormone-sensitive lipase and adipose triglyceride lipase first increased from d0 to d4 and then decreased from d4 to d14 (P < 0.05). In experiment 2, the average daily gain and average daily feed intake from 21 to 25 d of age increased in the MA group compared with the CON group (P < 0.05). The serum TC, hepatic TC, and glucose of MA group showed a significant increase than that of the CON group (P < 0.05). The expression of SCD1, ACAT2, and PPARγ were upregulated in the MA group (P < 0.05). Contrary to the decreased expression of phosphorylation of adenosine 5'-monophosphate-activated protein kinase alfa subunit (Thr172), the nuclear sterol regulatory element-binding protein 1c, fatty acid synthase, and peroxisome proliferator-activated receptor gamma of MA group increased than that of CON group (P < 0.05). CONCLUSIONS: Weaning promoted hepatic lipolysis and MA could enhance lipid synthesis by regulating adenosine 5'-monophosphate-activated protein kinase alfa subunit-sterol regulatory element-binding protein 1c pathway, thus improving growth performance of weanling piglets.

A self-powered implantable and bioresorbable electrostimulation device for biofeedback bone fracture healing.

Electrostimulation has been recognized as a promising nonpharmacological treatment in orthopedics to promote bone fracture healing. However, clinical applications have been largely limited by the complexity of equipment operation and stimulation implementation. Here, we present a self-powered implantable and bioresorbable bone fracture electrostimulation device, which consists of a triboelectric nanogenerator for electricity generation and a pair of dressing electrodes for applying electrostimulations directly toward the fracture. The device can be attached to irregular tissue surfaces and provide biphasic electric pulses in response to nearby body movements. We demonstrated the operation of this device on rats and achieved effective bone fracture healing in as short as 6 wk versus the controls for more than 10 wk to reach the same healing result. The optimized electrical field could activate relevant growth factors to regulate bone microenvironment for promoting bone formation and bone remodeling to accelerate bone regeneration and maturation, with statistically significant 27% and 83% improvement over the control groups in mineral density and flexural strength, respectively. This work provided an effective implantable fracture therapy device that is self-responsive, battery free, and requires no surgical removal after fulfilling the biomedical intervention.

A Highly Bright Near-Infrared Afterglow Luminophore for Activatable Ultrasensitive In Vivo Imaging.

Afterglow luminescence imaging probes, with long-lived emission after cessation of light excitation, have drawn increasing attention in biomedical imaging field owing to their elimination of autofluorescence. However, current afterglow agents always suffer from an unsatisfactory signal intensity and complex systems consisting of multiple ingredients. To address these issues, this study reports a near-infrared (NIR) afterglow luminophore (TPP-DO) by chemical conjugation of an afterglow substrate and a photosensitizer acting as both an afterglow initiator and an energy relay unit into a single molecule, resulting in an intramolecular energy transfer process to improve the afterglow brightness. The constructed TPP-DO NPs emit a strong NIR afterglow luminescence with a signal intensity of up to 108  p/s/cm2 /sr at a low concentration of 10 µM and a low irradiation power density of 0.05 W/cm2 , which is almost two orders of magnitude higher than most existing organic afterglow probes. The highly bright NIR afterglow luminescence with minimized background from TPP-DO NPs allows a deep tissue penetration depth ability. Moreover, we develop a GSH-activatable afterglow probe (Q-TPP-DO NPs) for ultrasensitive detection of subcutaneous tumor with the smallest tumor volume of 0.048 mm3 , demonstrating the high potential for early diagnosis and imaging-guided surgical resection of tumors.

Electrochemical Hydrophobic Tri-layer Interface Rendered Mechanically Graded Solid Electrolyte Interface for Stable Zinc Metal Anode.

The aqueous zinc-ion battery is promising as grid scale energy storage device, but hindered by the instable electrode/electrolyte interface. Herein, we report the lean-water ionic liquid electrolyte for aqueous zinc metal batteries. The lean-water ionic liquid electrolyte creates the hydrophobic tri-layer interface assembled by first two layers of hydrophobic OTF- and EMIM+ and third layer of loosely attached water, beyond the classical Gouy-Chapman-Stern theory based electrochemical double layer. By taking advantage of the hydrophobic tri-layer interface, the lean-water ionic liquid electrolyte enables a wide electrochemical working window (2.93 V) with relatively high zinc ion conductivity (17.3 mS/cm). Furthermore, the anion crowding interface facilitates the OTF- decomposition chemistry to create the mechanically graded solid electrolyte interface layer to simultaneously suppress the dendrite formation and maintain the mechanical stability. In this way, the lean-water based ionic liquid electrolyte realizes the ultralong cyclability of over 10000 cycles at 20 A/g and at practical condition of N/P ratio of 1.5, the cumulated areal capacity reach 1.8 Ah/cm2 , which outperforms the state-of-the-art zinc metal battery performance. Our work highlights the importance of the stable electrode/electrolyte interface stability, which would be practical for building high energy grid scale zinc-ion battery.

Microbe-Derived Antioxidants Alleviate Liver and Adipose Tissue Lipid Disorders and Metabolic Inflammation Induced by High Fat Diet in Mice.

Obesity induces lipodystrophy and metabolic inflammation. Microbe-derived antioxidants (MA) are novel small-molecule nutrients obtained from microbial fermentation, and have anti-oxidation, lipid-lowering and anti-inflammatory effects. Whether MA can regulate obesity-induced lipodystrophy and metabolic inflammation has not yet been investigated. The aim of this study was to investigate the effects of MA on oxidative stress, lipid disorders, and metabolic inflammation in liver and epididymal adipose tissues (EAT) of mice fed with a high-fat diet (HFD). Results showed that MA was able to reverse the HFD-induced increase in body weight, body fat rate and Lee's index in mice; reduce the fat content in serum, liver and EAT; and regulate the INS, LEP and resistin adipokines as well as free fatty acids to their normal levels. MA also reduced de novo synthesis of fat in the liver and EAT and promoted gene expression for lipolysis, fatty acid transport and ß-oxidation. MA decreased TNF-α and MCP1 content in serum, elevated SOD activity in liver and EAT, induced macrophage polarization toward the M2 type, inhibited the NLRP3 pathway, increased gene expression of the anti-inflammatory factors IL-4 and IL-13 and suppressed gene expression of the pro-inflammatory factors IL-6, TNF-α and MCP1, thereby attenuating oxidative stress and inflammation induced by HFD. In conclusion, MA can effectively reduce HFD-induced weight gain and alleviate obesity-induced oxidative stress, lipid disorders and metabolic inflammation in the liver and EAT, indicating that MA shows great promise as a functional food.

Enhancing the biosynthesis of nicotinamide mononucleotide in Lactococcus lactis by heterologous expression of FtnadE.

BACKGROUND: Nicotinamide mononucleotide (NMN), a key intermediate of nicotinamide adenine dinucleotide, plays an important in anti-aging and disease. Lactococcus lactis, an important probiotic lactic acid bacteria (LAB), has shown great potential for the biosynthesis of NMN, which will significantly affect the probiotic effects of the dairy products. RESULTS: We used the CRISPR/nCas9 technique to knockout nadR gene of L. lactis NZ9000 to enhance the accumulation of NMN by 61%. The nadE* gene from Francisella tularensis with codon optimization was heterologous in L. lactis NZ9000ΔnadR and has a positive effect on NMN production. Combined with optimization of the concentration of substrate nicotinamide, a final intracellular NMN titer was 2289 µmol L-1  mg-1 with 10 g L-1 nicotinamide supplement, which was 5.7-fold higher than that of the control. The transcription levels of key genes (pncA, nadD and prs1) involved in NMN biosynthesis were up-regulated by more than two-fold, indicating that the increase of NMN titer was attributed to FtnadE* heterologous expression. CONCLUSION: Our study provides a better understanding of the NMN biosynthesis pathway in L. lactis, and can facilitate NMN production in LAB via synthetic biology approaches. © 2022 Society of Chemical Industry.

Single-cell dynamics of chromatin activity during cell lineage differentiation in Caenorhabditis elegans embryos.

Elucidating the chromatin dynamics that orchestrate embryogenesis is a fundamental question in developmental biology. Here, we exploit position effects on expression as an indicator of chromatin activity and infer the chromatin activity landscape in every lineaged cell during Caenorhabditis elegans early embryogenesis. Systems-level analyses reveal that chromatin activity distinguishes cellular states and correlates with fate patterning in the early embryos. As cell lineage unfolds, chromatin activity diversifies in a lineage-dependent manner, with switch-like changes accompanying anterior-posterior fate asymmetry and characteristic landscapes being established in different cell lineages. Upon tissue differentiation, cellular chromatin from distinct lineages converges according to tissue types but retains stable memories of lineage history, contributing to intra-tissue cell heterogeneity. However, the chromatin landscapes of cells organized in a left-right symmetric pattern are predetermined to be analogous in early progenitors so as to pre-set equivalent states. Finally, genome-wide analysis identifies many regions exhibiting concordant chromatin activity changes that mediate the co-regulation of functionally related genes during differentiation. Collectively, our study reveals the developmental and genomic dynamics of chromatin activity at the single-cell level.

First-principles determination of the crystallography of the low-temperature phase for NbRu and TaRu shape memory alloys.

The crystallography of the low-temperature phases (ß'') for shape memory alloys (NbRu and TaRu) has been debated for decades. Though a P2/m monoclinic structure has been proposed for the ß'' phase, the proposed structure is not able to completely represent the measured diffraction data. In this work, the crystallography of the ß'' phase was investigated by first-principles calculations. We showed that the previously reported P2/m monoclinic structure was lattice unstable due to the presence of the soft phonon mode. A P21/m monoclinic structure was derived from the P2/m monoclinic structure by displacing its atoms according to the eigenvector of the soft phonon mode at the Γ point. The P21/m and the P2/m monoclinic structures are structurally similar, but the former one is energetically and structurally more favorable than the latter one. We concluded that the ß'' phase preferred to crystallize in the P21/m monoclinic structure rather than the previously reported P2/m monoclinic structure. Our results offer guidance for the experimental determination of the crystallography of the ß'' phase for NbRu and TaRu.

Microbe-derived antioxidants attenuate cobalt chloride-induced mitochondrial function, autophagy and BNIP3-dependent mitophagy pathways in BRL3A cells.

Environmental excessive cobalt (Co) exposure increases risks of public health. This study aimed to evaluate the potential mechanism of microbe-derived antioxidants (MA) blend fermented by probiotics in attenuating cobalt chloride (CoCl2)-induced toxicology in buffalo rat liver (BRL3A) cells. Herein, results showed that some phenolic acids increased in MA compared with the samples before fermentation through UHPLC-QTOF-MS analysis. Also, the contents of essential and non-essential amino acids, their derivatives and minerals were rich in MA. The DPPH, O2-, OH- and ABTS+ scavenging ability of MA is comparable to those of vitamin C and better than mitoquinone mesylate (mitoQ). In vitro cell experiments showed that CoCl2 treatment increased the percentage of apoptosis cells, lactate dehydrogenase and genes involved in glycolysis, increased ATP production and decreased mitochondrial membrane potential, increased genes involved in canonical autophagy process (including initiation, autophagosomes maturation and fusion with lysosome) and BNIP3-dependent mitophagy pathways in BRL3A cells, while MA attenuated CoCl2-induced reactive oxygen species (ROS) production, apoptosis, mitochondrial protein expression and dysfunction, and BNIP3-dependent mitophagy. Collectively, these results provide insights into the role of MA in reversing CoCl2-induced toxicology in BRL3A cells, providing the promising constituents for decreasing Co-induced toxicology in functional foods.

Microbe-Derived Antioxidants Reduce Lipopolysaccharide-Induced Inflammatory Responses by Activating the Nrf2 Pathway to Inhibit the ROS/NLRP3/IL-1ß Signaling Pathway.

Inflammation plays an important role in the innate immune response, yet overproduction of inflammation can lead to a variety of chronic diseases associated with the innate immune system; therefore, modulation of the excessive inflammatory response has been considered a major strategy in the treatment of inflammatory diseases. Activation of the ROS/NLRP3/IL-1ß signaling axis has been suggested to be a key initiating phase of inflammation. Our previous study found that microbe-derived antioxidants (MA) are shown to have excellent antioxidant and anti-inflammatory properties; however, the mechanism of action of MA remains unclear. The current study aims to investigate whether MA could protect cells from LPS-induced oxidative stress and inflammatory responses by modulating the Nrf2-ROS-NLRP3-IL-1ß signaling pathway. In this study, we find that MA treatment significantly alleviates LPS-induced oxidative stress and inflammatory responses in RAW264.7 cells. MA significantly reduce the accumulation of ROS in RAW264.7 cells, down-regulate the levels of pro-inflammatory factors (TNF-α and IL-6), inhibit NLRP3, ASC, caspase-1 mRNA, and protein levels, and reduce the mRNA, protein levels, and content of inflammatory factors (IL-1ß and IL-18). The protective effect of MA is significantly reduced after the siRNA knockdown of the NLRP3 gene, presumably related to the ability of MA to inhibit the ROS-NLRP3-IL-1ß signaling pathway. MA is able to reduce the accumulation of ROS and alleviate oxidative stress by increasing the content of antioxidant enzymes, such as SOD, GSH-Px, and CAT. The protective effect of MA may be due to its ability of MA to induce Nrf2 to enter the nucleus and initiate the expression of antioxidant enzymes. The antioxidant properties of MA are further enhanced in the presence of the Nrf2 activator SFN. After the siRNA knockdown of the Nrf2 gene, the antioxidant and anti-inflammatory properties of MA are significantly affected. These findings suggest that MA may inhibit the LPS-stimulated ROS/NLRP3/IL-1ß signaling axis by activating Nrf2-antioxidant signaling in RAW264.7 cells. As a result of this study, MA has been found to alleviate inflammatory responses and holds promise as a therapeutic agent for inflammation-related diseases.

Berberine improved intestinal barrier function by modulating the intestinal microbiota in blunt snout bream (Megalobrama amblycephala) under dietary high-fat and high-carbohydrate stress.

The study investigated whether dietary berberine supplementation could improve intestinal barrier against inflammation induced by high-fat and high-carbohydrate diets in blunt snout bream. Fish (average initial weight 44.83 ± 0.06 g) were fed with six kinds of diets (control, high-fat diet (10% lipid) and high-carbohydrate (43% nitrogen-free extract) diet, control/berberine, high-fat/berberine or high-carbohydrate/berberine) for 8 weeks, respectively. Feeding mode of berberine (50 mg/kg diet) was adopted to two-week interval. After feeding trial, fish growth performance and intestinal barrier function were estimated. The result showed that no significant interactions between diet and berberine in growth performance, whole body composition or protein utilization were observed (P > 0.05). Specific growth rate (SGR) and feed conversion ratio (FCR) were significantly affected by berberine (P < 0.05). Protein efficiency ratio (PER), nitrogen retention (NRE), fish whole-body lipid contents increased greatly in high-fat or high-carbohydrate diets (P < 0.05). Significant interactions between diet and berberine were observed in fish intestinal barrier (physical, chemical, immunological and microbiological barriers) (P < 0.05). High-fat and high-carbohydrate diets could increase significantly intestinal permeability and inflammatory response, decrease intestinal mucins gene expression levels, and make the intestinal microbiota out of balance (P < 0.05). Berberine significantly inhibited inflammation response and modulated intestinal microflora profile (P < 0.05). Taken together, berberine could alleviate intestinal barrier damage injured by high-fat or high-carbohydrate diet and improve the growth performance of blunt snout bream.

ROS-induced autophagy regulates porcine trophectoderm cell apoptosis, proliferation, and differentiation.

Significant embryo loss remains a serious problem in pig production. Reactive oxygen species (ROS) play a critical role in embryonic implantation and placentation. However, the potential mechanism of ROS on porcine trophectoderm (pTr) cell fate during the peri-implantation period has not been investigated. This study aimed to elucidate the effects of ROS on pTr cell phenotypes and the regulatory role in cell attachment and differentiation. Herein, results showed that exogenous H2O2 inhibited pTr cell viability, arrested the cell cycle at S and G2/M phases, and increased cell apoptosis and autophagy protein light chain 3B and Beclin-1, whereas these effects were reversed by different concentrations of N-acetyl-l-cysteine (NAC) posttreatment. In addition, NAC abolished H2O2-induced autophagic flux, inhibited intracellular and mitochondrial ROS, and restored expression of genes important for mitochondrial DNA and biogenesis, cell attachment, and differentiation. NAC reversed H2O2-activated MAPK and Akt/mammalian target of rapamycin pathways in dose-dependent manners. Furthermore, analyses with pharmacological and RNA interference approaches suggested that autophagy regulated cell apoptosis and gene expression of caudal-related homeobox 2 and IL-1ß. Collectively, these results provide new insights into the role of the ROS-induced autophagy in pTr cell apoptosis, attachment, and differentiation, indicating a promising target for decreasing porcine conceptus loss during the peri-implantation period.

Berberine attenuates sodium palmitate-induced lipid accumulation, oxidative stress and apoptosis in grass carp(Ctenopharyngodon idella)hepatocyte in vitro.

The objective of this work was to investigate the effect of berberine (BBR) on the Cell viability, lipid accumulation, apoptosis, cytochrome c, caspase-9 and caspase-3 in lipid accumulation-hepatocytes induced by sodium palmitate in vitro. The lipid accumulation-hepatocytes (induced by 0.5 mM sodium palmitate for 24 h) were treated with 5 µM berberine for 12 h. Then, the Cell viability, intracellular triglyceride (TG) content, lipid peroxide (LPO), malonaldehyde (MDA) content, cytochrome c, caspase-9, caspase-3 and apoptosis were detected. Sodium palmitate decreased Cell viability and increased intracellular TG content, lipid droplet accumulation, LPO and MDA concentrations, caused caspase-3 and caspase-9 activation, then led to apoptosis accompanied by cytochrome c release from mitochondria into the cytoplasm. Beberine could improve intracellular lipid droplet accumulation and oxidative stress, while reduce apoptosis induced by sodium palmitate.

Molecular dynamics simulations reveal how graphene oxide stabilizes and activates lipase in an anhydrous gas.

The interaction between Candida antarctica lipase B (CALB) and graphene oxide (GO) in an anhydrous gas was studied using molecular dynamics (MD) simulations augmented with a simulated annealing procedure to accelerate relaxation toward equilibrium. Three kinds of GO sheets with different oxygen contents were constructed to elucidate their effectiveness for stabilizing the active CALB conformation. It was shown that electrostatic forces are pivotal for the formation of CALB/GO complexes, and that a GO sheet with a higher oxygen content leads to stronger association with the protein. The simulation results suggest replacement of protein-binding water molecules by the GO surface, which was confirmed by thermogravimetric analysis. The CALB/GO assembly stabilizes the active enzyme conformation at elevated temperatures and, moreover, increases the protein flexibility near its active sites. The molecular details of GO interaction with CALB and the consequential effects on CALB stability and functionality are important for the development of unprecedented applications of gaseous enzymatic catalysis.

Molecular cloning, tissue distribution, and pharmacological characterization of blunt snout bream (Megalobrama amblycephala) melanocortin-5 receptor.

The melanocortin-5 receptor (MC5R) plays an important role in the regulation of exocrine secretion in mammals. Its function in fish is not well established. In this study, we reported the molecular cloning, tissue expression, and pharmacological characterization of Megalobrama amblycephala MC5R (MamMC5R), as well as the effect of catching stress on its expression. The full-length cDNA of Mammc5r gene was 1237 bp, consisted of a 990-bp open reading frame encoding 329 amino acids. Sequence analyses revealed that the nucleotide and amino acid sequences of Mammc5r were highly homologous (> 90%) with MC5Rs of zebrafish, common carp, and goldfish. Tissue expression profile analysis showed that Mammc5r was widely expressed in both central and peripheral tissues, with the highest expression in the brain. Catching stress significantly changed the expression of Mammc5r in the skin, brain, and eye. In the skin, the expression level of Mammc5r was significantly reduced at 1 h and 4 h and increased at 24 h after catching stress. The Mammc5r expression level was rapidly upregulated in the brain and eye at 1 h and then decreased to the level before stress at 4 h and 24 h. With human MC5R (HsaMC5R) as a control, several agonists including α-melanocyte-stimulating hormone (α-MSH), and ß-MSH in addition to an analogue [Nle4, D-Phe7]-α-MSH (NDP-MSH), were used to investigate the binding and signaling properties of MamMC5R. The results revealed that MamMC5R had the highest affinity for NDP-MSH, followed by α- and ß-MSH. Taken together, the data suggested that MamMC5R might play a role in stress response in M. amblycephala.

ROS mediated EGFR/MEK/ERK/HIF-1α Loop Regulates Glucose metabolism in pancreatic cancer.

To investigate the glycometabolism associated mechanism in invasion and metastasis of pancreatic cancer, We screened out genes involved in anaerobic glycolysis headed by HIF-1α,using pre-established a pair of pancreatic cancer cell lines. In this study, we further detected the glucose metabolism state not only in the cells but all also in two groups of patients with different SUVmax on 18F-FDG PET/CT. The data suggests that ROS mediated EGFR/MEK/ERK/HIF-1α loop is activated in high glucose metabolic samples both in vitro and in vivo: The increasing of HIF-1α expression is controlled by activation of EGFR/MEK/ERK pathway in hypoxia condition, HIF-1α inhibits excessive release of ROS, the reduction of ROS further activates EGFR to form a positive feedback loop. This difference is closely related to invasion and metastasis capacity of pancreatic cancer, and can be rescued by separate or combined inhibition of EGFR or HIF-1α in various degree. These results indicate a new clue to develop therapy of pancreatic cancer by regulating the glucose metabolism.

Functional Conservation and Divergence among Homoeologs of TaSPL20 and TaSPL21, Two SBP-Box Genes Governing Yield-Related Traits in Hexaploid Wheat.

Maintaining high and stable yields has become an increasing challenge in wheat breeding due to climate change. Although Squamosa-promoter binding protein (SBP)-box genes have important roles in plant development, very little is known about the actual biological functions of wheat SBP-box family members. Here, we dissect the functional conservation, divergence, and exploitation of homoeologs of two paralogous TaSPL wheat loci during domestication and breeding. TaSPL20 and TaSPL21 were highly expressed in the lemma and palea. Ectopic expressions of TaSPL20/21 in rice exhibited similar functions in terms of promoting panicle branching but had different functions during seed development. We characterized all six TaSPL20/21 genes located across the three homoeologous (A, B, and D) genomes. According to the functional analysis of naturally occurring variants in 20 environments, four favorable haplotypes were identified. Together, they reduced plant height by up to 27.5%, and TaSPL21-6D-HapII increased 1000-grain weight by 9.73%. Our study suggests that TaSPL20 and TaSPL21 homoeologs underwent diversification in function with each evolving its own distinctive characteristics. During domestication and breeding of wheat in China, favorable haplotypes of each set were selected and exploited to varying degrees due to their large effects on plant height and 1000-grain weight.

Markov-state model for CO2 binding with carbonic anhydrase under confinement.

Enzyme immobilization with a nanostructure material can enhance its stability and facilitate reusability. However, the apparent activity is often compromised due to additional diffusion barriers and complex interactions with the substrates and solvent molecules. The present study elucidates the effects of the surface hydrophobicity of nano-confinement on CO2 diffusion to the active site of human carbonic anhydrase II (CA), an enzyme that is able to catalyze CO2 hydration at extremely high turnover rates. Using the Markov-state model in combination with coarse-grained molecular dynamics simulations, we demonstrate that a hydrophobic cage increases CO2 local density but hinders its diffusion towards the active site of CA under confinement. By contrast, a hydrophilic cage hinders CO2 adsorption but promotes its binding with CA. An optimal surface hydrophobicity can be identified to maximize both the CO2 occupation probability and the diffusion rate. The simulation results offer insight into understanding enzyme performance under nano-confinement and help us to advance broader applications of CA for CO2 absorption and recovery.