Search Results (168)

Background/Objectives: The rapid progression of stroke often results in irreversible brain damage and poor outcomes when treatment is delayed. Prophylactic administration of FAD012 (3,5-dimethyl-4-hydroxycinnamic acid), a synthetic derivative of ferulic acid (FA), has demonstrated cerebroprotective effects in ischemic models through antioxidant and endothelial protective mechanisms. This study investigated the effects of FAD012 on cerebral infarction and blood–brain barrier (BBB) integrity using a photothrombotic stroke model in rats. Methods: Male Sprague Dawley rats received a single intraperitoneal injection of FAD012 or FA (100 or 300 mg/kg) 60 min prior to stroke induction. Under isoflurane anesthesia, the middle cerebral artery was exposed, and stroke was induced by intravenous administration of Rose Bengal followed by green laser irradiation. Cerebral blood flow (CBF) was monitored by laser Doppler flowmetry. BBB disruption was evaluated by Evans Blue extravasation and immunohistochemistry for tight junction (TJ) proteins. Results: Control rats exhibited extensive infarction, BBB disruption, and reduced expression of claudin-5, occludin, and ZO-1, along with fragmented collagen IV. In contrast, FAD012 (300 mg/kg) significantly attenuated CBF reduction, reduced infarct size, preserved BBB integrity, and maintained TJ protein expression, with greater efficacy than an equivalent dose of FA. FAD012 also preserved the expression and phosphorylation of endothelial nitric oxide synthase (eNOS), a key marker of vascular integrity. The CBF-preserving effect of FAD012 was completely abolished by N^G-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor. Conclusions: These findings suggest that FAD012 protects endothelial function, thereby contributing to the maintenance of CBF and BBB integrity, supporting its potential as a prophylactic therapeutic agent for ischemic stroke. Full article

Irregular rooting in vitro is a major problem in the micropropagation of culinary rhubarb (Rheum rhaponticum), a vegetable crop rich in bioactive compounds. To date, little is known about the factors and mechanisms underlying adventitious root (AR) formation in rhubarb under in vitro conditions. Here, we studied the effects of indole-3-butyric acid (IBA) and its interaction with ethylene (ET) on AR development in rhubarb ‘Raspberry’ selection. To evaluate the ET-effect, we applied a precursor of ET biosynthesis—1 aminocyclopropane-1-carboxylic acid (ACC); an inhibitor of ET synthesis—aminoethoxyvinylglycine (AVG); and an inhibitor of ET action—silver nitrate (AgNO₃). The best results (96.9% rooting frequency, 12.7 roots/shoot) were obtained after adding ACC to the IBA-containing medium. The positive effect of ET was linked to decreased levels of cytokinin and auxins in the rhubarb shoot bases at the initiation and expression stages of rooting. Moreover, the enhanced expression levels of genes involved in auxin signalling and homeostasis (IAA17, GH3.1) and ABA catabolism (CYP707A1) were observed. The blocking of ethylene synthesis significantly increased JA production, and the rooting frequency decreased to 29.8%. The presence of AgNO₃ in the auxin medium resulted in a significant reduction in root number, which was consistent with the enhanced levels of ABA and the expression of genes related to ABA biosynthesis and signalling (PP2C49 and CBF4), as well as ET synthesis (ACO5). Full article

Cold stress severely limits legume productivity, threatening global food security, particularly in climate-vulnerable regions. This review synthesizes advances in understanding and enhancing cold tolerance in key legumes (chickpea, soybean, lentil, and cowpea), addressing three core questions: (1) molecular/physiological foundations of cold tolerance; (2) how emerging technologies accelerate stress dissection and breeding; and (3) integration strategies and deployment challenges. Legume cold tolerance involves conserved pathways (e.g., ICE-CBF-COR, Inducer of CBF Expression, C-repeat Binding Factor, Cold-Responsive genes) and species-specific mechanisms like soybean’s GmTCF1a-mediated pathway. Multi-omics have identified critical genes (e.g., CaDREB1E in chickpea, NFR5 in pea) underlying adaptive traits (membrane stabilization, osmolyte accumulation) that reduce yield losses by 30–50% in tolerant genotypes. Technologically, AI and high-throughput phenotyping achieve >95% accuracy in early cold detection (3–7 days pre-symptoms) via hyperspectral/thermal imaging; deep learning (e.g., CNN-LSTM hybrids) improves trait prediction by 23% over linear models. Genomic selection cuts breeding cycles by 30–50% (to 3–5 years) using GEBVs (Genomic estimated breeding values) from hundreds of thousands of SNPs (Single-nucleotide polymorphisms). Advanced sensors (LIG-based, LoRaWAN) enable real-time monitoring (±0.1 °C precision, <30 s response), supporting precision irrigation that saves 15–40% water while maintaining yields. Key barriers include multi-omics data standardization and cost constraints in resource-limited regions. Integrating molecular insights with AI-driven phenomics and multi-omics is revolutionizing cold-tolerance breeding, accelerating climate-resilient variety development, and offering a blueprint for sustainable agricultural adaptation. Full article

Histone methylation plays important roles in plant development and adaptation to multiple stresses. SET domain group (SDG) proteins are identified as plant histone lysine methyltransferases in Arabidopsis and other crops. However, the SDG gene family and its functional roles in tomato remain unknown. In this research, 48 tomato SDG (SlSDG) gene family members were identified, and their chromosomal locations and conserved motifs were determined. According to phylogenetic analysis, the SlSDGs are divided into seven groups, which is consistent with Arabidopsis and rice. Promoter analysis indicated that the SlSDGs may be associated with biotic and abiotic stress responses. The expression pattern of SlSDGs illustrates that heat and cold stress significantly influence the transcript abundance of SDG14/19/21/23/48. The results of a VIGS assay showed that silencing SlSDG19 and SlSDG48 decreases tomato heat tolerance, while silencing SlSDG14 improves the heat tolerance of tomato plants. The analysis of downstream regulating genes indicated that heat shock proteins (HSPs), especially HSP70 and HSP90, act as critical effectors. Similarly, the experimental assay and expression analysis suggest that SDG21 and SDG23 positively and negatively regulate tomato cold tolerance through the CBF-COR pathway, respectively. These findings clarify the function of tomato SDG proteins and provide insight for the genetic improvement of tomato for temperature stress tolerance. Full article

Low-temperature stress severely limits plant growth and reduces agricultural productivity. Calmodulin-like (CML) proteins are crucial calcium sensors in plant cold responses. Transcriptome analysis of cold-stressed Saussurea involucrata identified seven differentially expressed CML genes. qRT-PCR confirmed that SiCML6 was strongly induced at 4 °C and −2 °C. Bioinformatics analysis showed that SiCML6 encodes a transmembrane protein containing an EF-hand domain. This protein carries a signal peptide and shows the closest phylogenetic relationship to Helianthus annuus CML3. Its promoter contains ABA, methyl jasmonate (MeJA), and cold-response elements. Arabidopsis plants overexpressing SiCML6 showed significantly higher survival rates at −2 °C than wild-type plants. Under freezing stress, SiCML6-overexpressing lines exhibited reduced malondialdehyde content, relative electrolyte leakage, and ROS accumulation (H₂O₂ and O₂⁻), along with increased proline, soluble sugars, soluble proteins, and total antioxidant capacity (T-AOC). SiCML6 elevated the expression of cold-responsive genes CBF3 and COR15a under normal conditions and further upregulated CBF1/2/3 and COR15a at 4 °C. Thus, low temperatures induced SiCML6 expression, which was potentially regulated by ABA/MeJA. SiCML6 enhances freezing tolerance by mitigating oxidative damage through boosted T-AOC and osmoprotectant accumulation while activating the CBF-COR signaling pathway. This gene is a novel target for improving crop cold resistance. Full article

Blood–brain barrier (BBB) dysfunction is a hallmark of cerebral small vessel disease (cSVD). This study aimed to identify a mouse model that replicates BBB impairment and shares key cSVD risk factors. Transgenic db/db and LDLr^−/−.Leiden mice, both prone to obesity and hypertension, were compared to C57BL/6J controls. BBB leakage was assessed using DCE-MRI and sodium fluorescein (NaFl); cerebral blood flow (CBF) by MRI. Dyslipidemia and vascular inflammation were measured by plasma tests. Tight junction integrity, endothelial dysfunction (glucose transporter 1, GLUT-1) and neuroinflammation were evaluated with immunohistochemistry and PCR. Both transgenic models developed an obese phenotype with hyperinsulinemia, but only LDLr^−/−.Leiden mice showed human-like dyslipidemia. When fed a high-fat diet (HFD) or HFD plus cholesterol, LDLr^−/−.Leiden mice showed reduced CBF, endothelial dysfunction (lowered GLUT-1), elevated vascular inflammation (ICAM-1, VCAM-1, S-selectin), and BBB leakage, as evidenced by DCE-MRI and NaFl, together with reduced ZO-1 and claudin-5 expression. Contrastingly, db/db mice showed endothelial dysfunction without BBB leakage. Neuroinflammation (IBA-1, GFAP) was observed only in LDLr^−/−.Leiden groups, consistent with BBB disruption. These findings indicate that LDLr^−/−.Leiden mice, but not db/db mice, are a promising translational model for studying BBB dysfunction in cSVD, offering insights into disease mechanisms and a platform for therapeutic development. Full article

Cold stress adversely impacts tomato (Solanum lycopersicum) production, particularly in temperate regions, by impairing growth, development, and yield. Abscisic acid (ABA), a key phytohormone, plays a central role in mediating tomato’s response to cold stress through a complex crosstalk network with other hormones and signaling molecules. This review examines ABA’s interactions with hormones such as ethylene, jasmonates, auxin, gibberellins, salicylic acid, brassinosteroids, and strigolactones, as well as signaling molecules like hydrogen peroxide, nitric oxide, hydrogen sulfide, and calcium. These interactions regulate various physiological processes, including osmolyte accumulation, membrane stability, and oxidative stress mitigation, and influence the expression of cold-responsive genes, such as CBFs, COR, and LEA. Critical knowledge gaps remain, particularly in understanding ABA’s context-specific interactions with other hormones and the integration of calcium signaling with ABA pathways under cold stress. By synthesizing current research, this review enhances our understanding of tomato’s cold stress response and provides insights for genetically improving cold tolerance, supporting sustainable tomato production amid climate challenges. Full article

Drought stress is a devastating natural stress that threatens crop productivity and quality. Mitigating the adverse effects of drought stress on wheat is a key object in agriculture. C-repeat binding transcription factor/DROUGHT RESPONSE ELEMENT BINDING FACTOR 1 (CBF/DREB1) transcription factors are well known for their role in cold acclimation. However, the involvement of CBF genes in drought stress and the mechanisms underlying their function remain poorly understood. In this study, 81 CBFs were identified in wheat, which were further clustered into four distinct lineages based on phylogenetic analysis. Chromosomal localization indicated that most CBF genes were dispersed across chromosome 5. We identified three homoeologous genes (TaCBF14A, TaCBF14B, and TaCBF14D) that were simultaneously upregulated under drought stress based on RNA-seq analysis. According to the high expression after drought stress, TaCBF14B was selected for further functional analysis. Subcellular localization and transcriptional activation activity analysis indicated that TaCBF14B likely functions as a transcription factor involved in drought stress tolerance. Overexpression of TaCBF14B in Arabidopsis enhanced the primary root growth by 13.49% (OE1), 12.56% (OE2), and 19.53% (OE3) under 200 mM mannitol treatment, and 21.65% (OE1), 16.63% (OE2), and 28.13% (OE3) under 250 mM mannitol treatment compared to WT. Meanwhile, the water loss rate of transgenic lines was 56% in WT leaves, but only 44%, 50%, and 40% in OE1, OE2, and OE3 lines, respectively. Compared to the wild type, POD activities of OE1, OE2, and OE3 were significantly increased by 42.94%, 29.41%, and 62.52%, respectively. And the Pro activities in OE1, OE2, and OE3 were significantly increased by 16.33%, 5.18%, and 29.09%, respectively, compared to the wild type. Additionally, the MDA content in OE1, OE2, and OE3 was significantly reduced by 40.53%, 15.81%, and 54.36%, respectively. Further analysis showed that the transgenic lines were hypersensitive to abscisic acid (ABA), and exhibited increased expression of AtABI3. We speculate that TaCBF14B plays an important role in enhancing drought tolerance. In summary, our findings provide new insights into the functional roles of CBF genes in drought stress tolerance. Full article

Alfalfa (Medicago sativa) is a globally distributed economic legume crop used for forage and ecological restoration. We aimed to explore the mechanisms underlying the cold acclimation observed in this species. Our results for fall plant growth showed that non-dormant alfalfa (SD) maintained a vigorous growth rate compared to that of fall-dormant alfalfa (ZD); however, the winter survival rate of ZD was higher than that of SD. Among the ZD samples, the starch content first accumulated and then decreased; the sucrose content was consumed first along with simultaneous raffinose accumulation, which was followed by sucrose content accumulation, with consistent changes in the corresponding related synthase and hydrolase activity. SD exhibited the opposite trend. The transcriptome data showed that most of the differentially expressed genes were involved in carbon metabolism (ko01200), amino acid biosynthesis (ko01230), and starch and sucrose metabolism (ko00500). Our data clearly show that alfalfa’s cold acclimation mechanism is a complex process, with the establishment of stable carbon homeostasis; sucrose is first converted into starch and raffinose, and then, starch is converted into sucrose, which enables alfalfa’s cold resistance. The process is accompanied by CBF/DREB1A TF regulation. This study provides important insights into the cold acclimation mechanisms of alfalfa. Full article

Low temperature significantly contributes to the medicinal quality of Conyza blinii. The CBF/DREB1-dependent cold-responsive signaling pathway is a major contributor to plant cold stress resistance. However, whether the CBF/DREB1 signaling pathway affects the terpenoid metabolism of C. blinii under cold stress remains to be explored. Here, we systematically identified and analyzed the impact of CbCBFs on the terpenoid metabolism of C. blinii. The results showed that three CbCBFs and CbICE1 were identified based on the transcriptome. The functions significantly correlated with CbCBFs encompass plant hormones and stress responses. Co-expression analysis revealed that key genes in BR and JA signaling pathways were correlated with CbCBFs. Among them, CbCBF2 is the predominant factor under low-temperature conditions and is significantly positively correlated with oleanolic acid. Overexpression of CbCBF2 significantly upregulated the catalase gene CbβAS and increased oleanolic acid content in the leaves. These results indicate that CbCBF2 can act as a major regulatory factor to promote the synthesis of oleanolic acid by integrating JA and BR signaling under low temperature conditions. Full article

Tomato (Solanum lycopersicum L.) is one of the major vegetable crops worldwide. Research on the Janus kinase–signal transducer and activator of transcription (JAK–STAT) signaling pathway in tomatoes and other plant systems is extremely limited. In this study, the roles of STAT, a crucial element of the JAK–STAT signaling pathway in tomato seed germination and low-temperature stress responses are examined, employing gene family analysis and genetic transformation. The results indicate that the S. lycopersicum genome contains only one member of the STAT gene family, SlSTAT. Subcellular localization experiments reveal that SlSTAT is found in both the cytoplasm and nucleus, suggesting its potential involvement in biological functions within these cellular compartments. Among the 26 different tomato tissue/organs tested, SlSTAT exhibited higher expression levels in hypocotyl (8 days past germination; 8 DPG), and low expression of SlSTAT significantly reduced the germination rate and impacted biomass at 8 DPG. In addition, the SlSTAT gene was significantly downregulated during low-temperature treatment. Compared with the wild-type (WT) tomatoes, the SlSTAT-overexpressing plants showed more resistance to low-temperature conditions, whereas the downexpressing tomatoes exhibited increased sensitivity. The expressions of low-temperature marker genes (SlCBF1-3) and N⁶-methyladenosine (m⁶A)-modification-related genes (m⁶A writer, reader, and eraser genes) were detected to explore possible molecular mechanisms by which SlSTAT causes changes in tomato low-temperature stress resistance. The expression changes of SlCBF1-3 in transgenic plants do not merely follow a straightforward linear relationship with the changes in SlSTAT expression, suggesting a more complex molecular mechanism and a non-direct interaction between SlSTAT and the promoters of SlCBFs. On the other hand, SlSTAT also changes the expression levels of RNA m⁶A-modification-related genes, especially SlFIP37 (writer gene), SlYTP8/9 (reader genes), and SlALKBH8 (eraser gene), ultimately leading to changes in the levels of m⁶A modification. These research findings lay the groundwork for exploring functions of JAK–STAT pathway in tomato development and stress responses, expanding the scope of JAK–STAT signaling studies in plant systems. Full article

Background/Objectives: In vitro models play a crucial role in preclinical respiratory research, enabling the testing and screening of mucosal formulations, dosage forms, and inhaled drugs. Mucociliary clearance (MCC) is an essential defense mechanism in mucosal drug delivery but is often impaired in respiratory diseases. Despite its importance, standardized in vitro MCC assays are rarely reported. Furthermore, many published methods primarily measure cilia beat frequency (CBF), which requires high-speed cameras that are not accessible to all laboratories. Therefore, this study aimed to develop a physiologically relevant, differentiated in vitro model of the respiratory epithelium that incorporates both beating cilia and functional MCC. We chose porcine airway mucosa as an alternative to human tissue due to ethical considerations and limited availability. The established model is designed to provide a reproducible and accessible method for a broad range of research laboratories. Methods: The previously published tracheal mucosal primary cell (TMPC DS) model, derived from porcine tissue, lacked the presence of beating cilia, which are crucial for effective MCC analysis. For accurate MCC assessment, beating cilia are essential as they play a key role in mucus clearance. To address this limitation, the here-described ciliated tracheal mucosal primary cell (cTMPC) model was developed. cTMPCs were isolated from porcine tissue and cultured under air–liquid interface (ALI) conditions for 21 days to promote differentiation. This model was evaluated for cell morphology, tight junction formation, ciliated and mucus-producing cells, barrier function, gene expression, and tracer/IgG transport. MCC and the model’s suitability for standardized MCC assays were assessed using an inverted microscope. In contrast to the TMPC DS model, which lacked beating cilia and thus could not support MCC analysis, the cTMPC model allows for comprehensive MCC studies. Results: The developed differentiated in vitro model demonstrated key structural and functional features of the respiratory epithelium, including well-differentiated cell morphology, tight junction integrity, ciliated and mucus-producing cells, and effective barrier function. Functional MCC was observed, confirming the model’s potential for standardized clearance assays. Conclusions: This differentiated in vitro model closely replicates the structural and functional characteristics of in vivo airways. It provides a valuable platform for studying mucociliary clearance, toxicology, drug uptake, and evaluating mucosal formulations and dosage forms in respiratory research. Full article

Dysphagia is a serious complication of stroke, yet effective pharmacological treatments remain limited. This study investigated the effects of FAD012 (3,5-dimethyl-4-hydroxy cinnamic acid), a synthetic derivative of ferulic acid (FA), on cerebral damage and swallowing dysfunction in a rat model of bilateral common carotid artery occlusion (2VO). Sprague–Dawley rats were orally administered FAD012 (3 or 10 mg/kg), FA (10 mg/kg), or 0.5% carboxymethyl cellulose (CMC, suspension vehicle) starting one week before 2VO. Two weeks after 2VO surgery, which was performed under isoflurane anesthesia, reflex swallowing was assessed by electromyographic recordings of the mylohyoid muscle under urethane anesthesia. Two weeks after 2VO, cerebral blood flow (CBF) declined to approximately 40% of baseline, and the number of reflex swallowing responses was significantly reduced in the CMC group. Additionally, 2VO induced O₂⁻ production, apoptotic cell death in the striatum, and a reduction in tyrosine hydroxylase expression. Substance P (SP) levels in the laryngopharyngeal mucosa, positively regulated by dopaminergic signaling in the basal ganglia, also decreased. FAD012 (10 mg/kg) effectively prevented the 2VO-induced reduction in CBF, enhanced the reflex swallowing, and preserved the dopamine-SP system. Notably, FAD012 exerted significantly stronger effects than FA at the same dose. These findings suggest that FAD012 maintains CBF under cerebral hypoperfusion and enhances the swallowing reflex by maintaining neuronal function in the striatal and laryngopharyngeal regions of 2VO rats. Full article

Cold stress is a significant factor limiting plant growth and development. Pomegranate is particularly susceptible to low temperatures. Calmodulin-binding transcriptional activators (CAMTAs) are key regulators of cold stress tolerance in plants. In this study, we conducted a comprehensive analysis of the CAMTA family proteins across 12 species, including Punica granatum (pomegranate), using bioinformatic methods. Pomegranate CAMTA3 (PgCAMTA3) was isolated and characterized, and it demonstrated enhanced cold tolerance when expressed in Arabidopsis thaliana. Quantitative real-time PCR (qRT-PCR) analysis showed that the expression of PgCAMTA3 was up-regulated under cold and ABA treatments in pomegranates. Two A. thaliana transgenic lines, OE1 and OE2, which overexpress PgCAMTA3, were generated through genetic transformation. The overexpression of PgCAMTA3 enhanced the cold stress tolerance in transgenic A. thaliana. OE1 and OE2 exhibited higher survival rates under cold stress. Furthermore, enzymatic activity assays revealed enhanced peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) in OE lines. These antioxidant enzymatic activities collectively contribute to better cold stress tolerance by providing more effective reactive oxygen species (ROS) scavenging and cellular protection mechanisms, which was confirmed by lower levels of malondialdehyde (MDA) and ROS production. In addition, the overexpression of PgCAMTA3 led to the upregulation of the expression levels of AtCBF2, AtNCED3, and AtWRKY22, which were modulated by CAMTA3. In summary, we report the significant role of PgCAMTA3 in plant cold tolerance. Our findings provide valuable insights into the CAMATA family in plants and offer new perspectives on the molecular mechanisms underlying cold tolerance in pomegranates. Full article

Background/Objectives: Acute myeloid leukemia (AML) is a rare hematological malignancy with a poor prognosis. Activating c-Kit (CD117) mutations occur in 5% of de novo AML and 30% of core-binding factor (CBF) AML, leading to worse clinical outcomes. Posttranslational modifications, particularly with myristic and palmitic acid, are crucial for various cellular processes, including membrane organization, signal transduction, and apoptosis regulation. However, most research has focused on solid tumors, with limited understanding of these mechanisms in AML. Fatty acid synthase (FASN), a key palmitoyl-acyltransferase, regulates the subcellular localization, trafficking, and degradation of target proteins, such as H-Ras, N-Ras, and FLT3-ITDmut receptors in AML. Methods: In this study, we investigated the role of FASN in two c-Kit-N822K-mutated AML cell lines using FASN knockdown via shRNA and the FASN inhibitor TVB-3166. Functional implications, including cell proliferation, were assessed through Western blotting, mass spectrometry, and PamGene. Results: FASN inhibition led to an increased phosphorylation of c-Kit (p-c-Kit), Lyn kinase (pLyn), MAP kinase (pMAPK), and S6 kinase (pS6). Furthermore, we observed sustained high expression of Gli1 in Kasumi1 cells following FASN inhibition, which is well known to be mediated by the upregulation of pS6. Conclusions: The combination of TVB-3166 and the Gli inhibitor GANT61 resulted in a significant reduction in the survival of Kasumi1 cells. Full article