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Abstract Architected Instability-based Metamaterials (AIMs) composed of curved beams can exhibit multi-stable geometric phase transformations. By tailoring geometry and topology, AIMs can accommodate large reversible deformations while dissipating energy beyond the capabilities of conventional materials. These exceptional mechanical properties are attractive for aerospace engineering applications, including energy-absorbing components in landing gears, impact-resistant protective structures, and vibration-damping systems. Nevertheless, this very mechanism that enables reversible energy dissipation also limits its capacity, because geometric phase transformations like snap-through buckling occur at low specific strength. Thus, the reversible energy dissipation capability cannot be easily leveraged in aerospace applications. In this work, we propose a theoretical and numerical modeling integrated approach to manipulate the out-of-plane stiffness distribution of curved beams. Compared to the conventional AIMs with uniform curved beams, the strength of the proposed beam configuration can be largely improved, while the local maximum strain remained relatively lower during phase transformations. Finite element analysis and experiments show this approach mitigates the local strain concentration effects of AIMs. Without inducing unreversible plastic deformation, the mechanical properties like the maximum peak strength, trapped energy during compression, and energy dissipation under cyclic loading can be increased by 62.1%, 82.5%, and 45.6%, respectively. 

Plant intracellular nucleotide-binding leucine-rich repeat (NLR) receptors with an N-terminal Toll/interleukin-1 receptor (TIR) domain sense pathogen effectors to initiate immune signaling. TIR domains across different kingdoms have NADase activities and can produce phosphoribosyl adenosine monophosphate/diphosphate (pRib-AMP/ADP) or cyclic ADPR (cADPR) isomers. The lipase-like proteins EDS1 and PAD4 transduce immune signals from sensor TIR-NLRs to a helper NLR called ADR1, which executes immune function. We report the structure and function of anArabidopsisEDS1-PAD4-ADR1 (EPA) heterotrimer in complex with pRib-AMP/ADP activated by plant or bacterial TIR signaling. 2′cADPR can be hydrolyzed into pRib-AMP and thus activate EPA signaling. Bacterial TIR domains producing 2′cADPR also activate EPA function. Our findings suggest that 2′cADPR may be the storage form of the unstable signaling molecule pRib-AMP. 

Plant intracellular nucleotide-binding leucine-rich repeat receptors (NLRs) analyzed to date oligomerize and form resistosomes upon activation to initiate immune responses. Some NLRs are encoded in tightly linked co-regulated head-to-head genes whose products function together as pairs. We uncover the oligomerization requirements for differentArabidopsispaired CHS3-CSA1 alleles. These pairs form resting-state heterodimers that oligomerize into complexes distinct from NLRs analyzed previously. Oligomerization requires both conserved and allele-specific features of the respective CHS3 and CSA1 Toll-like interleukin-1 receptor (TIR) domains. The receptor kinases BAK1 and BIRs inhibit CHS3-CSA1 pair oligomerization to maintain the CHS3-CSA1 heterodimer in an inactive state. Our study reveals that paired NLRs hetero-oligomerize and likely form a distinctive “dimer of heterodimers” and that structural heterogeneity is expected even among alleles of closely related paired NLRs. 

Intracellular plant immune receptors, termed NLRs (Nucleotide-binding Leucine-rich repeat Receptors), confer effector-triggered immunity. Sensor NLRs are responsible for pathogen effector recognition. Helper NLRs function downstream of sensor NLRs to transduce signaling and induce cell death and immunity. Activation of sensor NLRs that contain TIR (Toll/interleukin-1receptor) domains generates small molecules that induce an association between a downstream heterodimer signalosome of EDS1 (EnhancedDisease Susceptibility 1)/SAG101 (Senescence-AssociatedGene 101) and the helper NLR of NRG1 (NRequired Gene 1). Autoactive NRG1s oligomerize and form calcium signaling channels largely localized at the plasma membrane (PM). The molecular mechanisms of helper NLR PM association and effector-induced NRG1 oligomerization are not well characterized. We demonstrate that helper NLRs require positively charged residues in their N-terminal domains for phospholipid binding and PM association before and after activation, despite oligomerization and conformational changes that accompany activation. We demonstrate that effector activation of a TIR-containing sensor NLR induces NRG1 oligomerization at the PM and that the cytoplasmic pool of EDS1/SAG101 is critical for cell death function. EDS1/SAG101 cannot be detected in the oligomerized NRG1 resistosome, suggesting that additional unknown triggers might be required to induce the dissociation of EDS1/SAG101 from the previously described NRG1/EDS1/SAG101 heterotrimer before subsequent NRG1 oligomerization. Alternatively, the conformational changes resulting from NRG1 oligomerization abrogate the interface for EDS1/SAG101 association. Our data provide observations regarding dynamic PM association during helper NLR activation and underpin an updated model for effector-induced NRG1 resistosome formation. 

Immune pathways of plants and prokaryotes are activated by distinct TIR-generated metabolites. 

Developing new approaches for vascularizing synthetic tissue systems will have a tremendous impact in diverse areas. One area where this is particularly important is developing new skeletal muscle tissue systems, which could be utilized in physiological model studies and tissue regeneration. To develop vascularized approaches a microfluidic on-chip design for creating channels in polymer systems can be pursued. Current microfluidic tissue engineering methods include soft lithography, rapid prototyping, and cell printing; however, these have limitations such as having their scaffolding being inorganic, less desirable planar vasculature geometry, low fabrication efficiency, and limited resolution. Here we successfully developed a circular microfluidic channel embedded in a 3D extracellular matrix scaffolding with 3D myogenesis. We used a thermo-responsive polymer approach with micromilling-molding and designed a mixture of polyester wax and paraffin wax to fabricate the sacrificial template for microfluidic channel generation in the scaffolding. These findings will impact a number of fields including biomaterials, biomimetic structures, and personalized medicine in the future. 

Abstract Cell development and behavior are driven by internal genetic programming, but the external microenvironment is increasingly recognized as a significant factor in cell differentiation, migration, and in the case of cancer, metastatic progression. Yet it remains unclear how the microenvironment influences cell processes, especially when examining cell motility. One factor that affects cell motility is cell mechanics, which is known to be related to substrate stiffness. Examining how cells interact with each other in response to mechanically differential substrates would allow an increased understanding of their coordinated cell motility. In order to probe the effect of substrate stiffness on tumor related cells in greater detail, we created hard–soft–hard (HSH) polydimethylsiloxane (PDMS) substrates with alternating regions of different stiffness (200 and 800 kPa). We then cultured WI-38 fibroblasts and A549 epithelial cells to probe their motile response to the substrates. We found that when the 2 cell types were exposed simultaneously to the same substrate, fibroblasts moved at an increased speed over epithelial cells. Furthermore, the HSH substrate allowed us to physically guide and separate the different cell types based on their relative motile speed. We believe that this method and results will be important in a diversity of areas including mechanical microenvironment, cell motility, and cancer biology. 

Plant nucleotide-binding leucine-rich repeat (NLR) immune receptors activate cell death and confer disease resistance by unknown mechanisms. We demonstrate that plant Toll/interleukin-1 receptor (TIR) domains of NLRs are enzymes capable of degrading nicotinamide adenine dinucleotide in its oxidized form (NAD + ). Both cell death induction and NAD + cleavage activity of plant TIR domains require known self-association interfaces and a putative catalytic glutamic acid that is conserved in both bacterial TIR NAD + -cleaving enzymes (NADases) and the mammalian SARM1 (sterile alpha and TIR motif containing 1) NADase. We identify a variant of cyclic adenosine diphosphate ribose as a biomarker of TIR enzymatic activity. TIR enzymatic activity is induced by pathogen recognition and functions upstream of the genes enhanced disease susceptibility 1 ( EDS1 ) and N requirement gene 1 ( NRG1 ), which encode regulators required for TIR immune function. Thus, plant TIR-NLR receptors require NADase function to transduce recognition of pathogens into a cell death response.