Search Results (7,030)

Calmodulin-like proteins (CMLs) serve as core components in plant calcium signal transduction pathways, and they extensively modulate plant growth, development, and adaptive responses to various abiotic stresses. In this study, we cloned the StCML19 gene from potato and generated stable transgenic Arabidopsis thaliana lines constitutively expressing this gene to investigate its functional role under drought stress. Transcriptome analysis revealed that StCML19 was up-regulated under drought conditions. Phenotypic assays showed that overexpressing StCML19 notably increased the seed germination rate and root length of transgenic Arabidopsis under mannitol-induced osmotic stress, and greatly improved the plant survival rate under severe soil drought stress. Physiological analysis showed that when put under drought stress, transgenic plants had higher proline content, better SOD, CAT, and POD activities, and significantly less malondialdehyde (MDA) accumulation than wild-type plants. In addition, overexpression of StCML19 led to greater plant sensitivity to exogenous ABA, with inhibited root growth and delayed seed germination as indicators. Conclusively, this study is the first to make sense of the biological function of potato StCML19 in the drought stress response and views StCML19 as a promising candidate gene for the genetic improvement of drought-tolerant potato varieties. Full article

Bacterial wilt caused by Ralstonia solanacearum is a major constraint on tomato (Solanum lycopersicum L.) production, yet the molecular basis of quantitative resistance remains poorly understood. In this study, comparative transcriptome profiling was performed on resistant (‘ZM3’) and susceptible (‘ZM86’) tomato inbred lines following pathogen inoculation in roots, stems, and leaves. Differential expression analysis and weighted gene co-expression network analysis (WGCNA) were conducted to identify resistance-associated regulatory modules and hub genes. The results revealed distinct gene expression patterns between the two genotypes after infection. Several co-expression modules were significantly associated with resistance or susceptibility traits. Functional enrichment analysis showed that differentially expressed genes were mainly involved in plant hormone signal transduction, plant–pathogen interaction, phenylpropanoid biosynthesis, and cell wall modification. Genes related to ethylene and salicylic acid signaling were strongly induced following infection, whereas brassinosteroid-associated genes showed genotype-dependent expression patterns. Network analysis further identified several hub genes within defense-related modules, including ACO (Solyc04g007980), ERF1 (Solyc09g091950), MAPK9, receptor-like kinase RLK (Solyc07g006770), and a dirigent family gene (Solyc10g008900). Taken together, our results suggest that tomato resistance to Ralstonia solanacearum involves a coordinated defense network integrating hormone-mediated transcriptional regulation and structural reinforcement, and provides candidate genes for breeding bacterial wilt-resistant cultivars. Full article

Mechanical signals play key roles in the regulation of epidermal homeostasis and regeneration after injury. Integrins are key components of focal adhesions, and these complexes are major contributors to mechanotransduction. In keratinocytes, integrin-linked kinase (ILK) modulates essential processes for epidermal homeostasis and wound repair. However, its functions in the transduction of mechanical stimuli have remained virtually unexplored. In this study, we characterized epidermal tissues and primary keratinocytes from mice with epidermis-restricted inactivation of the Ilk gene (ILK-KO). ILK-deficient epidermis exhibits abnormalities in key components of mechanotransduction cascades, including disruptions in hemidesmosomal Collagen XVII immunoreactivity at the dermal–epidermal junction, and marked reduction in the nuclear localization of the mechanosensitive transcriptional regulator YAP. In wild-type (ILK+), but not in ILK-KO-cultured keratinocytes, exposure to cyclic bidirectional strain induced marked F-actin cytoskeletal rearrangements, characterized by the assembly of thick cortical actin bundles and stress fibers, as well as YAP nuclear translocation and transcriptional activity. Exposure to mechanical strain was additionally accompanied by differential changes in miRNA expression between ILK+ and ILK-KO cells. These findings reveal multiple and previously unappreciated key regulatory roles for ILK in epidermal keratinocyte responses to mechanical signals. Full article

Dry eye disease (DED) is an ocular surface disorder characterized by tear film instability, inflammation, epithelial damage, and neurosensory abnormalities. Due to its multifactorial etiology and pathophysiology, conventional therapies that focus on lubrication and immunosuppression often fall short in addressing the neuropathic component of ocular pain experienced by a growing subset of patients. Recent developments in sensory neuroscience have highlighted the pivotal role of ion channels in mediating ocular surface homeostasis, pain signaling, and inflammation. This review examines the role of the following major ion channel families in the pathophysiology of DED and neuropathic ocular pain: transient receptor potential (TRP) channels, voltage-gated sodium (Nav) channels, and purinergic P2X receptors. The review details their anatomical distribution, molecular function, and responses to environmental stimuli such as heat, cold, osmolarity, and injury. Current treatments, such as artificial tears, anti-inflammatory drops, and systemic neuromodulators, are also reviewed in relation to their effects on ion channel modulation. Additionally, emerging therapies that directly target sensory transduction pathways are introduced. This review highlights the therapeutic potential of ion channel modulation in personalizing treatment for patients with ocular surface pain, particularly those with neuropathic features unresponsive to standard care. Full article

Background/Objectives: Protein kinases play crucial roles in signal transduction pathways that regulate growth and differentiation in Trypanosoma cruzi. These protein kinases are attractive targets to develop new drugs to treat Chagas disease. Methods: We used several protein kinase inhibitors targeting the p38 MAPK, MEK, and ERK pathways to evaluate their effects on the in vivo phosphorylation status of T. cruzi proteins, particularly DNA polymerase beta (TcPolβ). We also used Genistein, a protein tyrosine kinase inhibitor, to assess its effects on global protein phosphorylation and TcPolβ phosphorylation. Also, we investigated the effect of oxidative stress on global tyrosine phosphorylation. Finally, we determined the phosphorylation sites on TcPolβ by the protein kinases TcPKC2 and TcWee570 in vitro. Results: p38 MAPK and MEK protein kinase inhibitors inhibited approximately 50% of the Ser/Thr phosphorylation of TcPolβ. Genistein inhibited both Ser/Thr and Tyr phosphorylation of several polypeptides in epimastigotes. Oxidative stress increases global Tyr phosphorylation by about twofold and also TcPolβ phosphorylation. TcPKC2 and TcWee570 were able to phosphorylate TcPolβ at both Ser/Thr and Tyr residues. Conclusions: Small-molecule protein kinase inhibitors can affect the phosphorylation status of TcPolβ in vivo. Since Genistein can inhibit both Ser/Thr and Tyr protein phosphorylation, and TcPKC2 and TcWee570 can phosphorylate both Ser/Thr and Tyr residues, it suggests the existence of dual protein kinases in T. cruzi. However, this possibility must be further studied. Full article

Diabetic cognitive impairment (DCI) is a serious and growing public health concern. The role of N6-methyladenosine (m⁶A), the predominant mRNA modification in the mammalian brain, in DCI pathogenesis remains not fully elucidated. Here, GEO-derived diabetes datasets were combined with in vivo and in vitro models to reveal aberrant expression of m⁶A-related genes. The results showed that the overall level of m⁶A RNA methylation in both the diabetic group and the high-glucose group was significantly decreased compared to the normal group. In addition, the expression of methyltransferase METTL3, which is involved in the regulation of m⁶A RNA methylation, was downregulated in both diabetic and hyperglycemic groups, and was positively correlated with the downregulation of the overall m⁶A level. Neuronal models with stable METTL3 knockdown were generated using lentiviral transduction. Subsequent ¹H-NMR metabolomic and MeRIP-qPCR analyses demonstrated that METTL3 deficiency disrupts key metabolic pathways, including phosphatidylethanolamine and phosphatidylcholine biosynthesis and glucose–alanine metabolism, and identified Fgf15 (the mouse ortholog of human FGF19) and H6PD as candidate downstream targets. Collectively, these data suggest that METTL3-dependent m⁶A RNA methylation alterations may contribute to DCI through metabolic dysregulation, positioning METTL3 as a promising therapeutic target for DCI. Full article

Flooding, as a major abiotic stress, significantly impacts the growth and survival of poplar plantations in the floodplains of the middle and lower reaches of the Yangtze River. Elucidating the molecular mechanisms underlying flooding responses in poplar is crucial for enhancing plantation productivity. In this study, two important eastern cottonwood cultivars, Populus deltoides ‘Jianghan 1’ (HBI) and P. deltoides Bartr. CL (CL), were investigated. By integrating long-term growth surveys and transcriptome sequencing, we analyzed their phenotypic traits and molecular responses to flooding stress. After 7 years of seasonal flooding, HBI exhibited a survival rate of 73.91%, along with superior height (23.1 m) and diameter at breast height (DBH, 26.3 cm), compared with CL, indicating HBI as a flooding-tolerant cultivar. Transcriptome analysis identified 1098 shared differentially expressed genes (DEGs) in the leaves of flooded HBI and CL, which were mainly enriched in stress signal perception, oxidative stress regulation, energy metabolism and circadian rhythm. Cultivar-specific DEG analysis revealed that CL mainly activated pathways related to oxidative stress and damage repair pathways, whereas HBI-specific genes were significantly enriched in hormone signal transduction, growth regulation, flavonoid synthesis and photosynthesis. Based on this distinct enrichment pattern in the tolerant cultivar HBI, we propose that it possesses adaptive advantages under flooding stress. Specifically, HBI likely coordinates multiple physiological processes by activating ethylene and other hormone-related genes, thereby regulating hypoxia adaptation, reoxygenation-induced oxidative stress, photosynthetic recovery, and flavonoid-mediated antioxidant defense. This coordinated regulation collectively sustains growth vigor and enhances survival under seasonal inundation. Our findings demonstrate clear transcriptomic divergence underlying flooding tolerance among poplar cultivars, laying a theoretical foundation for the selection of flooding-tolerant varieties and the sustainable development of forestry in flood-prone regions. Furthermore, these results broaden the current knowledge of flooding stress biology in woody plants. Full article

Background/Objectives: Human epidermal growth factor receptor 2 (HER2)-positive breast cancer is linked to poorer overall survival and a higher risk of brain metastases compared to HER2-negative breast cancer. Current preclinical studies lack robust HER2+ metastatic syngeneic mouse models for investigating targeted and immunomodulatory therapies. This study aims to develop effective HER2+ mouse models to investigate response dynamics to HER2-targeted therapy and immunotherapy. Methods: The human HER2 gene (WT or mutant p.A775_G776insYVMA, GFP-tagged at the C-terminus) was introduced into triple-negative breast cancer (TNBC) mouse mammary carcinoma cells with known metastatic potential (4T1 and EO771) via lentiviral transduction. HER2 expression and phosphorylation were analyzed using Western blotting and immunohistochemistry. Tumors were treated with HER2-targeted therapy (trastuzumab and tucatinib), immune checkpoint blockade (anti-PD-1 and anti-CTLA-4), and anti-HER2 antibody–drug conjugate (ADC) to evaluate treatment efficacy. Metastatic potential was assessed with brain fluorescence imaging. Statistical analysis included ANOVA and Kaplan–Meier tests. Results: Newly established lines demonstrated expression of HER2+, with HER2^YVMA lines showing higher phosphorylation than HER2^WT lines. Cells were tumorigenic, demonstrating in vivo tumor take rates at 100% for 4T1-HER2 and 15–30% for EO771-HER2. HER2 overexpression led to a 30% increase in spontaneous brain metastasis in the 4T1-HER2 models. Trastuzumab alone did not reduce primary tumor size but significantly reduced brain GFP signal by 17% ± 8% and 26% ± 7% in the 4T1-HER2^WT and 4T1-HER2^YVMA models, respectively. Combinational therapies with anti-HER2 therapy and immune checkpoint blockade effectively suppressed primary tumor growth and prolonged survival in EO771-HER2^YVMA model. T-Dxd, but not T-DM1, demonstrated partial treatment response in the EO771-HER2^WT model. Conclusions: HER2+ syngeneic tumor models were developed that spontaneously metastasize to the brain and demonstrate variable responses to immunotherapies and ADCs. These models are valuable for advancing molecular imaging modalities for HER2+ brain metastasis, studying blood–brain barrier penetration of HER2-targeted drugs, and exploring the combination of therapies, including immunotherapy. Full article

Dogs represent a promising animal model for analyzing human neurodegenerative diseases, owing to their similarities to humans in nervous system architecture and behavioral phenotypes. Neural stem cells (NSCs) serve as a highly valuable in vitro experimental model for investigating neurogenesis, neurodegenerative disease pathogenesis, and neural molecular biology; however, studies on immortalized canine neural stem cell lines remain scarce. Herein, we successfully established an immortalized canine hippocampal neural stem cell line that can be continuously passaged in vitro via SV40 large T antigen (SV40LT) viral infection and subsequent cellular transformation. Both the immortalized NSCs and their normal parental counterparts differentiated into neuronal and glial lineages under induced differentiation conditions. Normal canine hippocampal NSCs can be passaged for no more than 10 generations, whereas the immortalized line can be passaged indefinitely while maintaining a normal karyotype. This immortalized canine hippocampal NSC line can act as a critical experimental tool for future research into neural differentiation mechanisms and stem cell-derived therapeutic strategies for neurological disorders in dogs. Full article

Heat stress (HS), induced by global climate warming, is one of the major limiting factors in edible fungi production. HS suppresses mycelial growth and fruiting body formation by causing excessive accumulation of intracellular reactive oxygen species (ROS), disrupting the integrity of cell membranes and cell walls, and impairing cellular metabolism. Increasing evidence suggests that the application of exogenous substances (ESs) effectively mitigates HS in edible fungi. Based on the recent literature, this review categorizes ESs into three groups—core signaling molecules, plant growth regulators, and cytoprotective agents—and summarizes their beneficial effects against HS in edible fungi. The underlying mechanisms of ES-mediated alleviation of heat-induced damage primarily involve four pathways: (1) regulation of antioxidant systems; (2) preservation of cell wall and membrane structural integrity; (3) modulation of defense-related gene expression; and (4) regulation of carbon metabolic flux. Current challenges and corresponding strategies are discussed to provide a reference for elucidating the mechanisms by which ESs alleviate HS and to promote their practical application in edible fungi production. Full article

Gastrointestinal malignant tumors account for approximately one-third of global cancer-related deaths, primarily including colorectal, gastric, pancreatic ductal adenocarcinoma, and hepatocellular carcinomas. These tumors have a high incidence, are often asymptomatic, and are prone to metastasis and recurrence, posing a significant public health burden. Although traditional methods such as radiotherapy and chemotherapy can delay disease progression, their nonspecific effects often lead to severe side effects and drug resistance, resulting in limited efficacy. Therefore, developing novel treatment strategies with high target specificity and favorable biological safety is a critical scientific issue in this field. Peptide drugs offer advantages such as good biocompatibility, low immunogenicity, diverse structures, and ease of modification, collectively demonstrating unique potential for tumor treatment. They can not only achieve precise delivery by specifically recognizing tumor receptors but can also directly interfere with signal transduction, metabolism, and immune regulation, producing multi-target antitumor effects. This article systematically reviews the research progress of peptide drugs in gastrointestinal tumors, focusing on their molecular mechanisms, delivery modification strategies, and the latest applications. It also summarizes the challenges and future directions for clinical translation, providing a theoretical foundation and future perspectives for the precise treatment of gastrointestinal tumors and the design of new drugs. Full article

Salt stress is an injurious concern of global climate change that negatively impacts the growth and yield of rice plants. Identifying salt tolerance genes is essential to understanding the molecular mechanism regulating salt tolerance in rice. In this study, we treated two rice varieties, Xiangxiuzhan (XXZ) and Changxiang (CXG), with 100 mM NaCl to examine the effect on the germination and growth stages. Transcriptome analysis was investigated for changes in gene expression between the two varieties. During the germination stage, the CXG variety had higher germination potential than the XXZ variety, whereas in the growth stage, the XXZ variety showed higher survival efficiency than the CXG variety. Transcriptome analysis showed that the XXZ variety had more DEGs in grains, while CXG displayed greater DEGs in leaves and roots. Gene Ontology (GO) and KEGG pathway showed that beta-alanine metabolism, cutin biosynthesis, and plant hormone signal transduction were over-represented, whereas heatmap analysis showed cellular and environmental signal transduction. This study focuses on the molecular pathways of the salt stress tolerance mechanism of Xiangxiuzhan and Changxiang varieties. Full article

Salt stress severely constrains wheat growth and yield by inducing osmotic imbalance, ion toxicity, and excessive accumulation of reactive oxygen species (ROS). Although salt-tolerant cultivars can adapt through rapid signaling transduction and maintenance of cellular homeostasis, the underlying dynamic regulatory networks remain insufficiently characterized. In this study, we reanalyzed publicly available time-series RNA-seq data (0, 1, 3, 6, 12, and 24 h) from the salt-tolerant wheat cultivar Xiaoyan22 under salt stress and constructed a time-series-based co-expression network using weighted gene co-expression network analysis (WGCNA). Multiple gene modules were identified, among which the black module showed significant positive correlations with both salt treatment (treatment_bin) and stress duration (time_h). This module displayed a progressively increasing eigengene expression pattern throughout the stress period. Gene significance (GS) was positively correlated with module membership (MM), facilitating the identification of highly connected hub genes within this module. Functional enrichment analysis indicated that genes in the black module were primarily associated with DNA replication and genome stability maintenance, RNA metabolic regulation, phenylpropanoid metabolism, and cuticle/suberin/wax biosynthesis. Physiological analysis further revealed enhanced activities of superoxide (SOD), peroxide (POD), and catalase (CAT), enhanced accumulation of proline and soluble sugars, and a time-dependent increase in MDA under salt stress. qRT-PCR confirmed significant induction of candidate genes, including a ZAR1-like receptor kinase, Remorin, and NETWORKED 1D. Collectively, these findings integrate co-expression network inference with physiological and molecular validation, providing candidate regulators and pathways for understanding salt tolerance and supporting future molecular breeding efforts. Full article

The mitogen-activated protein kinase (MAPK) cascade constitutes a core component of signal transduction pathways in eukaryotic organisms. With its precise, efficient, and specific mechanism of action, this cascade pathway integrates, amplifies, and rapidly transmits signals. Among them, the specificity and functional diversity of the MPK3 cascade depend on the phosphorylation interaction between MKK and MPK3, as well as the specific interaction between MPK3 and its substrates. MPK3 targets an extremely diverse array of substrates, including transcription factors, RNA-binding proteins, enzymes, and transporters. The summary of the regulatory role of the MPK3 signal mainly focuses on three functional mechanisms: The most well-known regulatory mechanism is to recognize and phosphorylate substrate proteins or transcription factors, thereby affecting the stability and transcriptional activity of downstream substrates, and thus regulating the transcriptional regulatory activity and expression of downstream genes. MPK3 can also participate in downstream functional regulation by triggering the MAPKKK-MKK4/5-MPK3/6 signaling pathways or feedback mechanisms. MPK3 can exert regulatory effects independently or together with MPK6. The redundancy of the MPK3/6 function is related to the synergistic effect of the component cascade reaction, as well as the dose-dependent activation effect. This article presents a comprehensive synthesis of the latest research progress on the regulatory role of MPK3, in plant growth, development, and stress adaptation and defence. Moreover, it provides critical evaluations and forward-looking perspectives on the future investigation of the underlying molecular mechanisms governing MPK3-mediated regulation. Full article

We describe AI agents as stochastic dynamical systems and frame the problem of learning to reason as in transductive inference: Rather than approximating the distribution of past data as in classical induction, the objective is to capture its algorithmic structure so as to reduce the time needed to solve new tasks. In this view, information from past experience serves not only to reduce a model’s uncertainty, as in Shannon’s classical theory, but to reduce the computational effort required to find solutions to unforeseen tasks. Working in the verifiable setting, where a checker or reward function is available, we establish three main results. First, we show that the optimal speed-up for a new task is tightly related to the algorithmic information it shares with the training data, yielding a theoretical justification for the power-law scaling empirically observed in reasoning models. Second, while the compression view of learning, rooted in Occam’s Razor, favors simplicity, we show that transductive inference yields its greatest benefits precisely when the data-generating mechanism is most complex. Third, we identify a possible failure mode of naïve scaling: in the limit of unbounded model size and computing, models with access to a reward signal can behave as savants, brute-forcing solutions without acquiring transferable reasoning strategies. Accordingly, we argue that a critical quantity to optimize when scaling reasoning models is time, the role of which in learning has remained largely unexplored. Full article