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1.
EMBO J ; 41(19): e110682, 2022 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-35950443

RESUMO

The plant defense hormone, salicylic acid (SA), plays essential roles in immunity and systemic acquired resistance. Salicylic acid induced by the pathogen is perceived by the receptor nonexpressor of pathogenesis-related genes 1 (NPR1), which is recruited by TGA transcription factors to induce the expression of pathogenesis-related (PR) genes. However, the mechanism by which post-translational modifications affect TGA's transcriptional activity by salicylic acid signaling/pathogen infection is not well-established. Here, we report that the loss-of-function mutant of brassinosteroid insensitive2 (BIN2) and its homologs, bin2-3 bil1 bil2, causes impaired pathogen resistance and insensitivity to SA-induced PR gene expression, whereas the gain-of-function mutant, bin2-1, exhibited enhanced SA signaling and immunity against the pathogen. Our results demonstrate that salicylic acid activates BIN2 kinase, which in turn phosphorylates TGA3 at Ser33 to enhance TGA3 DNA binding ability and NPR1-TGA3 complex formation, leading to the activation of PR gene expression. These findings implicate BIN2 as a new component of salicylic acid signaling, functioning as a key node in balancing brassinosteroid-mediated plant growth and SA-induced immunity.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica , Brassinosteroides/metabolismo , DNA/metabolismo , Resistência à Doença/genética , Expressão Gênica , Regulação da Expressão Gênica de Plantas , Hormônios/metabolismo , Fosforilação , Reguladores de Crescimento de Plantas/metabolismo , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Ácido Salicílico/metabolismo , Ácido Salicílico/farmacologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
2.
Proc Natl Acad Sci U S A ; 120(11): e2215250120, 2023 03 14.
Artigo em Inglês | MEDLINE | ID: mdl-36888655

RESUMO

Classical dynamins are best understood for their ability to generate vesicles by membrane fission. During clathrin-mediated endocytosis (CME), dynamin is recruited to the membrane through multivalent protein and lipid interactions between its proline-rich domain (PRD) with SRC Homology 3 (SH3) domains in endocytic proteins and its pleckstrin-homology domain (PHD) with membrane lipids. Variable loops (VL) in the PHD bind lipids and partially insert into the membrane thereby anchoring the PHD to the membrane. Recent molecular dynamics (MD) simulations reveal a novel VL4 that interacts with the membrane. Importantly, a missense mutation that reduces VL4 hydrophobicity is linked to an autosomal dominant form of Charcot-Marie-Tooth (CMT) neuropathy. We analyzed the orientation and function of the VL4 to mechanistically link data from simulations with the CMT neuropathy. Structural modeling of PHDs in the cryo-electron microscopy (cryo-EM) cryoEM map of the membrane-bound dynamin polymer confirms VL4 as a membrane-interacting loop. In assays that rely solely on lipid-based membrane recruitment, VL4 mutants with reduced hydrophobicity showed an acute membrane curvature-dependent binding and a catalytic defect in fission. Remarkably, in assays that mimic a physiological multivalent lipid- and protein-based recruitment, VL4 mutants were completely defective in fission across a range of membrane curvatures. Importantly, expression of these mutants in cells inhibited CME, consistent with the autosomal dominant phenotype associated with the CMT neuropathy. Together, our results emphasize the significance of finely tuned lipid and protein interactions for efficient dynamin function.


Assuntos
Proteínas Sanguíneas , Dinaminas , Microscopia Crioeletrônica , Dinaminas/metabolismo , Endocitose/fisiologia , Lipídeos , Dinamina I/metabolismo
3.
Annu Rev Physiol ; 84: 229-255, 2022 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-34780259

RESUMO

In mammalian cardiac myocytes, the plasma membrane includes the surface sarcolemma but also a network of membrane invaginations called transverse (t-) tubules. These structures carry the action potential deep into the cell interior, allowing efficient triggering of Ca2+ release and initiation of contraction. Once thought to serve as rather static enablers of excitation-contraction coupling, recent work has provided a newfound appreciation of the plasticity of the t-tubule network's structure and function. Indeed, t-tubules are now understood to support dynamic regulation of the heartbeat across a range of timescales, during all stages of life, in both health and disease. This review article aims to summarize these concepts, with consideration given to emerging t-tubule regulators and their targeting in future therapies.


Assuntos
Insuficiência Cardíaca , Sarcolema , Animais , Cálcio/metabolismo , Sinalização do Cálcio/fisiologia , Membrana Celular/metabolismo , Humanos , Mamíferos , Miócitos Cardíacos/fisiologia , Sarcolema/metabolismo
4.
Circ Res ; 132(11): e188-e205, 2023 05 26.
Artigo em Inglês | MEDLINE | ID: mdl-37139790

RESUMO

BACKGROUND: Transverse tubules (t-tubules) form gradually in the developing heart, critically enabling maturation of cardiomyocyte Ca2+ homeostasis. The membrane bending and scaffolding protein BIN1 (bridging integrator 1) has been implicated in this process. However, it is unclear which of the various reported BIN1 isoforms are involved, and whether BIN1 function is regulated by its putative binding partners MTM1 (myotubularin), a phosphoinositide 3'-phosphatase, and DNM2 (dynamin-2), a GTPase believed to mediate membrane fission. METHODS: We investigated the roles of BIN1, MTM1, and DNM2 in t-tubule formation in developing mouse cardiomyocytes, and in gene-modified HL-1 and human-induced pluripotent stem cell-derived cardiomyocytes. T-tubules and proteins of interest were imaged by confocal and Airyscan microscopy, and expression patterns were examined by RT-qPCR and Western blotting. Ca2+ release was recorded using Fluo-4. RESULTS: We observed that in the postnatal mouse heart, BIN1 localizes along Z-lines from early developmental stages, consistent with roles in initial budding and scaffolding of t-tubules. T-tubule proliferation and organization were linked to a progressive and parallel increase in 4 detected BIN1 isoforms. All isoforms were observed to induce tubulation in cardiomyocytes but produced t-tubules with differing geometries. BIN1-induced tubulations contained the L-type Ca2+ channel, were colocalized with caveolin-3 and the ryanodine receptor, and effectively triggered Ca2+ release. BIN1 upregulation during development was paralleled by increasing expression of MTM1. Despite no direct binding between MTM1 and murine cardiac BIN1 isoforms, which lack exon 11, high MTM1 levels were necessary for BIN1-induced tubulation, indicating a central role of phosphoinositide homeostasis. In contrast, the developing heart exhibited declining levels of DNM2. Indeed, we observed that high levels of DNM2 are inhibitory for t-tubule formation, although this protein colocalizes with BIN1 along Z-lines, and binds all 4 isoforms. CONCLUSIONS: These findings indicate that BIN1, MTM1, and DNM2 have balanced and collaborative roles in controlling t-tubule growth in cardiomyocytes.


Assuntos
Dinamina II , Miócitos Cardíacos , Animais , Humanos , Camundongos , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Dinamina II/genética , Dinamina II/metabolismo , Miócitos Cardíacos/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Proteínas Nucleares/metabolismo , Isoformas de Proteínas/metabolismo , Proteínas Tirosina Fosfatases não Receptoras/genética , Proteínas Tirosina Fosfatases não Receptoras/metabolismo , Proteínas Supressoras de Tumor/metabolismo
5.
Mol Cell ; 66(5): 648-657.e4, 2017 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-28575660

RESUMO

The glycogen synthase kinase-3 (GSK3) family kinases are central cellular regulators highly conserved in all eukaryotes. In Arabidopsis, the GSK3-like kinase BIN2 phosphorylates a range of proteins to control broad developmental processes, and BIN2 is degraded through unknown mechanism upon receptor kinase-mediated brassinosteroid (BR) signaling. Here we identify KIB1 as an F-box E3 ubiquitin ligase that promotes the degradation of BIN2 while blocking its substrate access. Loss-of-function mutations of KIB1 and its homologs abolished BR-induced BIN2 degradation and caused severe BR-insensitive phenotypes. KIB1 directly interacted with BIN2 in a BR-dependent manner and promoted BIN2 ubiquitination in vitro. Expression of an F-box-truncated KIB1 caused BIN2 accumulation but dephosphorylation of its substrate BZR1 and activation of BR responses because KIB1 blocked BIN2 binding to BZR1. Our study demonstrates that KIB1 plays an essential role in BR signaling by inhibiting BIN2 through dual mechanisms of blocking substrate access and promoting degradation.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/efeitos dos fármacos , Brassinosteroides/farmacologia , Proteínas F-Box/metabolismo , Quinase 3 da Glicogênio Sintase/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Plantas Geneticamente Modificadas/efeitos dos fármacos , Proteínas Quinases/metabolismo , Esteroides Heterocíclicos/farmacologia , Ubiquitina-Proteína Ligases/metabolismo , Arabidopsis/enzimologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Sítios de Ligação , Domínio Catalítico , Proteínas de Ligação a DNA , Ativação Enzimática , Estabilidade Enzimática , Proteínas F-Box/genética , Genótipo , Quinase 3 da Glicogênio Sintase/genética , Mutação , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fenótipo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Ligação Proteica , Proteínas Quinases/genética , Proteólise , Transdução de Sinais/efeitos dos fármacos , Especificidade por Substrato , Ubiquitina-Proteína Ligases/genética , Ubiquitinação
6.
Plant Cell Physiol ; 2024 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-38372617

RESUMO

The polyhydroxylated steroid phytohormone brassinosteroids (BRs) control many aspects of plant growth, development and responses to environmental changes. Plasma membrane (PM) H+-ATPase, the well-known PM proton pump, is a central regulator in plant physiology, which mediates not only plant growth and development, but also adaptation to stresses. Recent studies highlight that PM H+-ATPase is at least partly regulated via the BR signaling. Firstly, the BR cell surface receptor BRASSINOSTEROID-INSENSITIVE 1 (BRI1) and multiple key components of BR signaling directly or indirectly influence PM H+-ATPase activity. Secondly, the SMALL AUXIN UP RNA (SAUR) gene family physically interacts with BRI1 to enhance organ development of Arabidopsis by activating PM H+-ATPase. Thirdly, RNA-sequencing (RNA-seq) assays showed that the expression of some SAUR genes is upregulated under the light or sucrose conditions, which is related to the phosphorylation state of the penultimate residue of PM H+-ATPase in a time-course manner. In this review, we describe the structural and functional features of PM H+-ATPase, and summarize recent progress toward understanding the regulatory mechanism of PM H+-ATPase by BRs, and briefly introduce how PM H+-ATPase activity is modulated by its own biterminal regions and the post-translational modifications.

7.
Plant Cell Physiol ; 2024 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-38957969

RESUMO

The INDUCER OF CBF EXPRESSION 1/C-REPEAT BINDING FACTOR (ICE1/CBF) pathway plays a crucial role in plant responses to cold stress, impacting growth and development. Here, we demonstrated that ATBS1-INTERACTING FACTOR 2 (AIF2), a non-DNA-binding basic helix-loop-helix transcription factor, positively regulates freezing tolerance through the ICE1/CBF-induced cold tolerance pathway in Arabidopsis. Cold stress transcriptionally upregulated AIF2 expression and induced AIF2 phosphorylation, thereby stabilizing the AIF2 protein during early stages of cold acclimation. The AIF2 loss-of-function mutant, aif2-1, exhibited heightened sensitivity to freezing before and after cold acclimation. In contrast, ectopic expression of AIF2, but not the C-terminal-deleted AIF2 variant, restored freezing tolerance. AIF2 enhanced ICE1 stability during cold acclimation and promoted the transcriptional expression of CBFs and downstream cold-responsive genes, ultimately enhancing plant tolerance to freezing stress. MITOGEN-ACTIVATED PROTEIN KINASES 3 and 6 (MPK3/6), known negative regulators of freezing tolerance, interacted with and phosphorylated AIF2, subjecting it to protein degradation. Furthermore, transient co-expression of MPK3/6 with AIF2 and ICE1 downregulated AIF2/ICE1-induced transactivation of CBF2 expression. AIF2 interacted preferentially with BIN2 and MPK3/6 during the early and later stages of cold acclimation, respectively, thereby differentially regulating AIF2 activity in a cold acclimation time-dependent manner. Moreover, AIF2 acted additively in a gain-of-function mutant of BRASSINAZOLE-RESISTANT 1 (BZR1; bzr1-1D) and a triple knockout mutant of BRASSINOSTEROID-INSENSITIVE 2 (BIN2) and its homologs (bin2bil1bil2) to induce CBFs-mediated freezing tolerance. This suggests that cold-induced AIF2 coordinates freezing tolerance along with BZR1 and BIN2, key positive and negative components, respectively, of brassinosteroid signaling pathways.

8.
Brain ; 146(4): 1561-1579, 2023 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-36059072

RESUMO

Bridging integrator 1 (BIN1) is the second most prevalent genetic risk factor identified by genome-wide association studies (GWAS) for late-onset Alzheimer's disease. BIN1 encodes an adaptor protein that regulates membrane dynamics in the context of endocytosis and neurotransmitter vesicle release. In vitro evidence suggests that BIN1 can directly bind to tau in the cytosol. In addition, BIN1's function limits extracellular tau seed uptake by endocytosis and subsequent propagation as well as influences tau release through exosomes. However, the in vivo roles of BIN1 in tau pathogenesis and tauopathy-mediated neurodegeneration remain uncharacterized. We generated conditional knockout mice with a selective loss of Bin1 expression in the forebrain excitatory neurons and oligodendrocytes in P301S human tau transgenic background (line PS19). PS19 mice develop age-dependent tau neuropathology and motor deficits and are commonly used to study Alzheimer's disease tau pathophysiology. The severity of motor deficits and neuropathology was compared between experimental and control mice that differ with respect to forebrain BIN1 expression. BIN1's involvement in tau pathology and neuroinflammation was quantified by biochemical methods and immunostaining. Transcriptome changes were profiled by RNA-sequencing analysis to gain molecular insights. The loss of forebrain BIN1 expression in PS19 mice exacerbated tau pathology in the somatosensory cortex, thalamus, spinal cord and sciatic nerve, accelerated disease progression and caused early death. Intriguingly, the loss of BIN1 also mitigated tau neuropathology in select regions, including the hippocampus, entorhinal/piriform cortex, and amygdala, thus attenuating hippocampal synapse loss, neuronal death, neuroinflammation and brain atrophy. At the molecular level, the loss of forebrain BIN1 elicited complex neuronal and non-neuronal transcriptomic changes, including altered neuroinflammatory gene expression, concomitant with an impaired microglial transition towards the disease-associated microglial phenotype. These results provide crucial new information on in vivo BIN1 function in the context of tau pathogenesis. We conclude that forebrain neuronal BIN1 expression promotes hippocampal tau pathogenesis and neuroinflammation. Our findings highlight an exciting region specificity in neuronal BIN1 regulation of tau pathogenesis and reveal cell-autonomous and non-cell-autonomous mechanisms involved in BIN1 modulation of tau neuropathology.


Assuntos
Doença de Alzheimer , Tauopatias , Camundongos , Humanos , Animais , Doença de Alzheimer/patologia , Proteínas tau/metabolismo , Doenças Neuroinflamatórias , Camundongos Transgênicos , Estudo de Associação Genômica Ampla , Tauopatias/patologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Camundongos Knockout , Hipocampo/metabolismo , Modelos Animais de Doenças , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Proteínas do Tecido Nervoso/genética
9.
Sensors (Basel) ; 24(16)2024 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-39205064

RESUMO

This study proposes a method named Hybrid Heuristic Proximal Policy Optimization (HHPPO) to implement online 3D bin-packing tasks. Some heuristic algorithms for bin-packing and the Proximal Policy Optimization (PPO) algorithm of deep reinforcement learning are integrated to implement this method. In the heuristic algorithms for bin-packing, an extreme point priority sorting method is proposed to sort the generated extreme points according to their waste spaces to improve space utilization. In addition, a 3D grid representation of the space status of the container is used, and some partial support constraints are proposed to increase the possibilities for stacking objects and enhance overall space utilization. In the PPO algorithm, some heuristic algorithms are integrated, and the reward function and the action space of the policy network are designed so that the proposed method can effectively complete the online 3D bin-packing task. Some experimental results illustrate that the proposed method has good results in achieving online 3D bin-packing tasks in some simulation environments. In addition, an environment with image vision is constructed to show that the proposed method indeed enables an actual robot manipulator to successfully and effectively complete the bin-packing task in a real environment.

10.
Sensors (Basel) ; 24(6)2024 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-38544045

RESUMO

Trafficability gives tracked vehicles adaptability, stability, and propulsion for various purposes, including deep-sea research in rough terrain. Terrain characteristics affect tracked vehicle mobility. This paper investigates the soil mechanical interaction dynamics between rubber-tracked vehicles and sedimental soils through controlled laboratory-simulated experiments. Focusing on Bentonite and Diatom sedimental soils, which possess distinct shear properties from typical land soils, the study employs innovative user-written subroutines to characterize mechanical models linked to the RecurDyn simulation environment. The experiment is centered around a dual-tracked crawler, which in itself represents a fully independent vehicle. A new three-dimensional multi-body dynamic simulation model of the tracked vehicle is developed, integrating the moist terrain's mechanical model. Simulations assess the vehicle's trafficability and performance, revealing optimal slip ratios for maximum traction force. Additionally, a mathematical model evaluates the vehicle's tractive trafficability based on slip ratio and primary design parameters. The study offers valuable insights and a practical simulation modeling approach for assessing trafficability, predicting locomotion, optimizing design, and controlling the motion of tracked vehicles across diverse moist terrain conditions. The focus is on the critical factors influencing the mobility of tracked vehicles, precisely the sinkage speed and its relationship with pressure. The study introduces a rubber-tracked vehicle, pressure, and moisture sensors to monitor pressure sinkage and moisture, evaluating cohesive soils (Bentonite/Diatom) in combination with sand and gravel mixtures. Findings reveal that higher moisture content in Bentonite correlates with increased track slippage and sinkage, contrasting with Diatom's notable compaction and sinkage characteristics. This research enhances precision in terrain assessment, improves tracked vehicle design, and advances terrain mechanics comprehension for off-road exploration, offering valuable insights for vehicle design practices and exploration endeavors.

11.
Alzheimers Dement ; 20(4): 2922-2942, 2024 04.
Artigo em Inglês | MEDLINE | ID: mdl-38460121

RESUMO

INTRODUCTION: The BIN1 coding variant rs138047593 (K358R) is linked to Late-Onset Alzheimer's Disease (LOAD) via targeted exome sequencing. METHODS: To elucidate the functional consequences of this rare coding variant on brain amyloidosis and neuroinflammation, we generated BIN1K358R knock-in mice using CRISPR/Cas9 technology. These mice were subsequently bred with 5xFAD transgenic mice, which serve as a model for Alzheimer's pathology. RESULTS: The presence of the BIN1K358R variant leads to increased cerebral amyloid deposition, with a dampened response of astrocytes and oligodendrocytes, but not microglia, at both the cellular and transcriptional levels. This correlates with decreased neurofilament light chain in both plasma and brain tissue. Synaptic densities are significantly increased in both wild-type and 5xFAD backgrounds homozygous for the BIN1K358R variant. DISCUSSION: The BIN1 K358R variant modulates amyloid pathology in 5xFAD mice, attenuates the astrocytic and oligodendrocytic responses to amyloid plaques, decreases damage markers, and elevates synaptic densities. HIGHLIGHTS: BIN1 rs138047593 (K358R) coding variant is associated with increased risk of LOAD. BIN1 K358R variant increases amyloid plaque load in 12-month-old 5xFAD mice. BIN1 K358R variant dampens astrocytic and oligodendrocytic response to plaques. BIN1 K358R variant decreases neuronal damage in 5xFAD mice. BIN1 K358R upregulates synaptic densities and modulates synaptic transmission.


Assuntos
Doença de Alzheimer , Animais , Camundongos , Doença de Alzheimer/genética , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides , Modelos Animais de Doenças , Camundongos Transgênicos , Neuroglia/patologia , Placa Amiloide/patologia , Humanos
12.
Evol Comput ; : 1-41, 2024 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-38713741

RESUMO

Gathering sufficient instance data to either train algorithm-selection models or understand algorithm footprints within an instance space can be challenging. We propose an approach to generating synthetic instances that are tailored to perform well with respect to a target algorithm belonging to a predefined portfolio but are also diverse with respect to their features. Our approach uses a novelty search algorithm with a linearly weighted fitness function that balances novelty and performance to generate a large set of diverse and discriminatory instances in a single run of the algorithm. We consider two definitions of novelty: (1) with respect to discriminatory performance within a portfolio of solvers; (2) with respect to the features of the evolved instances. We evaluate the proposed method with respect to its ability to generate diverse and discriminatory instances in two domains (knapsack and bin-packing), comparing to another well-known quality diversity method, Multi-dimensional Archive of Phenotypic Elites (MAP-Elites) and an evolutionary algorithm that only evolves for discriminatory behaviour. The results demonstrate that the novelty search method outperforms its competitors in terms of coverage of the space and its ability to generate instances that are diverse regarding the relative size of the "performance gap" between the target solver and the remaining solvers in the portfolio. Moreover, for the Knapsack domain, we also show that we are able to generate novel instances in regions of an instance space not covered by existing benchmarks using a portfolio of state-of-the-art solvers. Finally, we demonstrate that the method is robust to different portfolios of solvers (stochastic approaches, deterministic heuristics and state-of-the-art methods), thereby providing further evidence of its generality.

13.
Mol Biol Evol ; 39(11)2022 11 03.
Artigo em Inglês | MEDLINE | ID: mdl-36256608

RESUMO

Type II DNA topoisomerases regulate topology by double-stranded DNA cleavage and ligation. The TopoVI family of DNA topoisomerase, first identified and biochemically characterized in Archaea, represents, with TopoVIII and mini-A, the type IIB family. TopoVI has several intriguing features in terms of function and evolution. TopoVI has been identified in some eukaryotes, and a global view is lacking to understand its evolutionary pattern. In addition, in eukaryotes, the two TopoVI subunits (TopoVIA and TopoVIB) have been duplicated and have evolved to give rise to Spo11 and TopoVIBL, forming TopoVI-like (TopoVIL), a complex essential for generating DNA breaks that initiate homologous recombination during meiosis. TopoVIL is essential for sexual reproduction. How the TopoVI subunits have evolved to ensure this meiotic function is unclear. Here, we investigated the phylogenetic conservation of TopoVI and TopoVIL. We demonstrate that BIN4 and RHL1, potentially interacting with TopoVIB, have co-evolved with TopoVI. Based on model structures, this observation supports the hypothesis for a role of TopoVI in decatenation of replicated chromatids and predicts that in eukaryotes the TopoVI catalytic complex includes BIN4 and RHL1. For TopoVIL, the phylogenetic analysis of Spo11, which is highly conserved among Eukarya, highlighted a eukaryal-specific N-terminal domain that may be important for its regulation. Conversely, TopoVIBL was poorly conserved, giving rise to ATP hydrolysis-mutated or -truncated protein variants, or was undetected in some species. This remarkable plasticity of TopoVIBL provides important information for the activity and function of TopoVIL during meiosis.


Assuntos
Proteínas Arqueais , DNA Topoisomerases Tipo II , Filogenia , Sequência de Aminoácidos , DNA Topoisomerases Tipo II/química , DNA Topoisomerases Tipo II/genética , DNA Topoisomerases Tipo II/metabolismo , Proteínas Arqueais/química , Meiose/genética , Eucariotos/genética , Eucariotos/metabolismo
14.
Biochem Biophys Res Commun ; 678: 17-23, 2023 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-37611348

RESUMO

Salt stress is a negative environmental factors to affecting plants. Salinity inhibits seed germination and root growth, which reduces the biomass of agricultural plants. BRASSINOSTEROID-INSENSITIVE2 (BIN2) functions as a signalling hub to integrate the perception and transduction of plant growth and stress tolerance by the phosphorylation of target proteins. However, only a small number of target molecules have been discovered thus far. In this study, we present evidence that BIN2 controls the post-transcriptional activity of AGL16. BIN2 interacts and phosphorylates AGL16, which increases AGL16 stability and transcriptional activity. Genetic testing showed that the agl16 mutant can restore the reduction in the seed germination rate and primary root growth of the bin2-1 mutant, while the overexpression of AGL16 in the bin2-3bil1bil2 mutant reduced the salt tolerance compared with bin2-3bil1bil2 in response to salt stress. Taken together, our data identify a BIN2-AGL16 core protein module that is mediates the inhibition of seed germination and primary root growth under salt stress.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Agricultura , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Brassinosteroides , Proteínas Quinases , Estresse Salino , Tolerância ao Sal/genética , Proteínas de Domínio MADS
15.
Basic Res Cardiol ; 118(1): 13, 2023 03 29.
Artigo em Inglês | MEDLINE | ID: mdl-36988697

RESUMO

The prospective use of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) for cardiac regenerative medicine strongly depends on the electro-mechanical properties of these cells, especially regarding the Ca2+-dependent excitation-contraction (EC) coupling mechanism. Currently, the immature structural and functional features of hiPSC-CM limit the progression towards clinical applications. Here, we show that a specific microarchitecture is essential for functional maturation of hiPSC-CM. Structural remodelling towards a cuboid cell shape and induction of BIN1, a facilitator of membrane invaginations, lead to transverse (t)-tubule-like structures. This transformation brings two Ca2+ channels critical for EC coupling in close proximity, the L-type Ca2+ channel at the sarcolemma and the ryanodine receptor at the sarcoplasmic reticulum. Consequently, the Ca2+-dependent functional interaction of these channels becomes more efficient, leading to improved spatio-temporal synchronisation of Ca2+ transients and higher EC coupling gain. Thus, functional maturation of hiPSC-cardiomyocytes by optimised cell microarchitecture needs to be considered for future cardiac regenerative approaches.


Assuntos
Células-Tronco Pluripotentes Induzidas , Miócitos Cardíacos , Humanos , Miócitos Cardíacos/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Acoplamento Excitação-Contração , Sinalização do Cálcio , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo , Cálcio/metabolismo
16.
Environ Res ; 231(Pt 3): 116202, 2023 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-37211183

RESUMO

Understanding the community assembly process is a central issue in microbial ecology. In this study, we analyzed the community assembly of particle-associated (PA) and free-living (FL) surface water microbiomes in 54 sites from the headstream to the river mouth of an urban river in Japan, the river basin of which has the highest human population density in the country. Analyses were conducted from two perspectives: (1) analysis of deterministic processes considering only environmental factors using a geo-multi-omics dataset and (2) analysis of deterministic and stochastic processes to estimate the contributions of heterogeneous selection (HeS), homogeneous selection (HoS), dispersal limitation (DL), homogenizing dispersal (HD), and drift (DR) as community assembly processes using a phylogenetic bin-based null model. The variation in microbiomes was successfully explained from a deterministic perspective by environmental factors, such as organic matter-related, nitrogen metabolism, and salinity-related parameters, using multivariate statistical analysis, network analysis, and habitat prediction. In addition, we demonstrated the dominance of stochastic processes (DL, HD, and DR) over deterministic processes (HeS and HoS) in community assembly from both deterministic and stochastic perspectives. Our analysis revealed that as the distance between two sites increased, the effect of HoS sharply decreased while the effect of HeS increased, particularly between upstream and estuary sites, indicating that the salinity gradient could potentially enhance the contribution of HeS to community assembly. Our study highlights the importance of both stochastic and deterministic processes in community assembly of PA and FL surface water microbiomes in urban riverine ecosystems.


Assuntos
Microbiota , Rios , Humanos , Filogenia , Multiômica , Processos Estocásticos
17.
Int J Biometeorol ; 67(4): 587-596, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36749414

RESUMO

In order to explore the influence of outdoor microclimate on the cooling effect of constant temperature community bin, the temperature prediction model was predicted. The temperature and microclimate data sets of the community bin were collected in summer from May 2021 to September 2021. The climatic characteristics included cloudy and sunny conditions, and the environmental factors included outdoor temperature, air speed, air relative humidity, and solar radiation intensity. Stepwise regression method was used to test the significance of environmental factors, and the corresponding regression equation was obtained. BP neural network was used to establish temperature prediction models under cloudy and sunny conditions, respectively. The results showed that the coefficient of determination (R2) of the two models was above 0.8, and the environmental factors with significant influence were screened out. The root mean square error (RMSE) between the training value and the actual value established by BP neural network was 0.83 °C, and the determination coefficient (R2) was 0.968. Under sunny conditions, the root mean square error (RMSE) of predicted value and measured value was 0.65 °C, and the determination coefficient (R2) was 0.982. According to the analysis of the sample data, it showed that the BP neural network was more accurate than stepwise regression, and could be used to predict the temperature of community bin, which provided model basis for the practical application of intelligent temperature control community bin in summer.


Assuntos
Temperatura Baixa , Redes Neurais de Computação , Temperatura , Estações do Ano , Microclima
18.
Sensors (Basel) ; 23(5)2023 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-36904992

RESUMO

In this paper, two of the most common calibration methods of synchronous TDCs, which are the bin-by-bin calibration and the average-bin-width calibration, are first presented and compared. Then, an innovative new robust calibration method for asynchronous TDCs is proposed and evaluated. Simulation results showed that: (i) For a synchronous TDC, the bin-by-bin calibration, applied to a histogram, does not improve the TDC's differential non-linearity (DNL); nevertheless, it improves its Integral Non-Linearity (INL), whereas the average-bin-width calibration significantly improves both the DNL and the INL. (ii) For an asynchronous TDC, the DNL can be improved up to 10 times by applying the bin-by-bin calibration, whereas the proposed method is almost independent of the non-linearity of the TDC and can improve the DNL up to 100 times. The simulation results were confirmed by experiments carried out using real TDCs implemented on a Cyclone V SoC-FPGA. For an asynchronous TDC, the proposed calibration method is 10 times better than the bin-by-bin method in terms of the DNL improvement.

19.
Sensors (Basel) ; 23(15)2023 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-37571710

RESUMO

Manufacturing systems need to be resilient and self-organizing to adapt to unexpected disruptions, such as product changes or rapid order, in supply chain changes while increasing the automation level of robotized logistics processes to cope with the lack of human experts. Deep Reinforcement Learning is a potential solution to solve more complex problems by introducing artificial neural networks in Reinforcement Learning. In this paper, a game engine was used for Deep Reinforcement Learning training, which allows visualization of view learning and result processes more intuitively than other tools, as well as a physical engine for a more realistic problem-solving environment. The present research demonstrates that a Deep Reinforcement Learning model can effectively address the real-time sequential 3D bin packing problem by utilizing a game engine to visualize the environment. The results indicate that this approach holds promise for tackling complex logistical challenges in dynamic settings.

20.
Sensors (Basel) ; 23(7)2023 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-37050435

RESUMO

We present an integrated single-photon detection device custom designed for quantum key distribution (QKD) with time-bin encoded single photons. We implemented and demonstrated a prototype photon-to-digital converter (PDC) that integrates an 8 × 8 single-photon avalanche diode (SPAD) array with on-chip digital signal processing built in TSMC 65 nm CMOS. The prototype SPADs are used to validate the QKD functionalities with an array of time-to-digital converters (TDCs) to timestamp and process the photon detection events. The PDC uses window gating to reject noise counts and on-chip processing to sort the photon detections into respective time-bins. The PDC prototype achieved a 22.7 ps RMS timing resolution and demonstrated operation in a time-bin setup with 158 ps time-bins at an optical wavelength of 410 nm. This PDC can therefore be an important building block for a QKD receiver and enables compact and robust time-bin QKD systems with imaging detectors.

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