Search Results (711)

The functional role of MAPKK genes in potato (Solanum tuberosum L.) under high-temperature stress remains unexplored, despite their critical importance in stress signaling and yield protection. We characterized StMAPKK1, a novel group D MAPKK localized to plasma membrane/cytoplasm. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed cultivar-specific upregulation in potato (‘Atlantic’ and ‘Desiree’) leaves under heat stress (25 °C, 30 °C, and 35 °C). Transgenic lines overexpressing (OE) StMAPKK1 exhibited elevated antioxidant enzyme activity, including ascorbate peroxidase (APX), catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD), mitigating oxidative damage. Increased proline and chlorophyll accumulation and reduced oxidative stress markers, hydrogen peroxide (H₂O₂) and malondialdehyde (MDA), indicate improved cellular redox homeostasis. The upregulation of key antioxidant and heat stress-responsive genes (StAPX, StCAT1/2, StPOD12/47, StFeSOD2/3, StMnSOD, StCuZnSOD1/2, StHSFA3 and StHSP20/70/90) strengthened the enzymatic defense system, enhanced thermotolerance, and improved photosynthetic efficiency, with significant improvements in net photosynthetic rate (Pn), transpiration rate (E), and stomatal conductance (Gs) under heat stress (35 °C) in StMAPKK1-OE plants. Superior growth and biomass (plant height, plant and its root fresh and dry weights, and tuber yield) accumulation, confirming the positive role of StMAPKK1 in thermotolerance. Conversely, RNA interference (RNAi)-mediated suppression of StMAPKK1 led to a reduction in enzymatic activity, proline content, and chlorophyll levels, exacerbating oxidative stress. Downregulation of antioxidant-related genes impaired ROS scavenging capacity and declines in photosynthetic efficiency, growth, and biomass, accompanied by elevated H₂O₂ and MDA accumulation, highlighting the essential role of StMAPKK1 in heat stress adaptation. These findings highlight StMAPKK1’s potential as a key genetic target for breeding heat-tolerant potato varieties, offering a foundation for improving crop resilience in warming climates. Full article

Recent insights into the p53-MDM2 nexus have advanced deeper understanding of their regulation and potent impact on cancer heterogeneity. The roles of nuclear phosphoinositide (PIP_ns) in modulating this pathway are emerging as a key mechanism. Here, we dissect the molecular mechanisms by which nuclear PIP_ns stabilize p53 through the recruitment of small heat shock proteins (sHSPs), activate the nuclear phosphatidylinositol 3-kinase (PI3K)-AKT signaling cascade, and modulate MDM2 function to regulate the p53-MDM2 interaction. We propose potential mechanisms by which nuclear PIP_ns coordinate signaling with nuclear p53, AKT, and MDM2. Ultimately, we highlight that nuclear PIP_ns serve as a ‘third messenger’ within the p53-MDM2 axis, expanding the current framework of non-canonical nuclear signaling in cancer biology. Full article

Water temperature significantly affects the physiological balance of aquatic organisms like crustaceans, and heat shock proteins (HSPs) are crucial for stress resistance and pathogen defense. This study conducted a genome-wide analysis to explore the functional characteristics of the Hsp70 gene family in Procambarus clarkii. Fifteen Hsp70 family members were identified, with several genes showing upregulation under non-lethal heat shock (NLHS) and pathogen challenges. RNA-Seq and qPCR analyses confirmed increased expression of certain PcHsp70s during NLHS, indicating NLHS activation of the Hsp70 family to enhance immune regulation. dsRNA-mediated silencing of Hsp70 led to downregulation of TLR pathway genes (e.g., TLR1, TLR6), suggesting Hsp70 regulates the TLR signaling pathway for immune responses. These findings reveal that NLHS-induced Hsp70 upregulation improves pathogen resistance, offering insights for addressing temperature fluctuations and disease outbreaks in aquaculture to optimize management practices. Full article

Background/Objectives: Doxorubicin (DOX), a widely used anthracycline chemotherapeutic agent, is recognized for its efficacy in treating various malignancies. However, its clinical application is critically limited due to dose-dependent cardiotoxicity, predominantly induced by oxidative stress and compromised antioxidant defenses. Photobiomodulation (PBM), a non-invasive intervention that utilizes low-intensity light, has emerged as a promising therapeutic modality in regenerative medicine, demonstrating benefits such as enhanced tissue repair, reduced inflammation, and protection against oxidative damage. This investigation sought to evaluate the cardioprotective effects of PBM preconditioning in human-induced pluripotent stem cell-derived ventricular cardiomyocytes (hiPSC-vCMs) subjected to DOX-induced toxicity. Methods: Human iPSC-vCMs were allocated into three experimental groups: control cells (untreated), DOX-treated cells (exposed to 2 μM DOX for 24 h), and PBM+DOX-treated cells (preconditioned with PBM, utilizing 660 nm ±10 nm LED light at an intensity of 10 mW/cm² for 500 s, delivering an energy dose of 5 J/cm², followed by DOX exposure). Cell viability assessments were conducted in conjunction with evaluations of oxidative stress markers, including antioxidant enzyme activities and malondialdehyde (MDA) levels. Furthermore, transcriptional profiling of 40 genes implicated in cardiac dysfunction was performed using TaqMan quantitative polymerase chain reaction (qPCR), complemented by analyses of protein expression for markers of cardiac stress, inflammation, and apoptosis. Results: Exposure to DOX markedly reduced the viability of hiPSC-vCMs. The cells exhibited significant alterations in the expression of 32 out of 40 genes (80%) after DOX exposure, reflecting the upregulation of markers associated with apoptosis, inflammation, and adverse cardiac remodeling. PBM preconditioning partially restored the cell viability, modulating the expression of 20 genes (50%), effectively counteracting a substantial proportion of the dysregulation induced by DOX. Notably, PBM enhanced the expression of genes responsible for antioxidant defense, augmented antioxidant enzyme activity, and reduced oxidative stress indicators such as MDA levels. Additional benefits included downregulating stress-related mRNA markers (HSP1A1 and TNC) and apoptotic markers (BAX and TP53). PBM also demonstrated gene reprogramming effects in ventricular cells, encompassing regulatory changes in NPPA, NPPB, and MYH6. PBM reduced the protein expression levels of IL-6, TNF, and apoptotic markers in alignment with their corresponding mRNA expression profiles. Notably, PBM preconditioning showed a diminished expression of BNP, emphasizing its positive impact on mitigating cardiac stress. Conclusions: This study demonstrates that PBM preconditioning is an effective strategy for reducing DOX-induced chemotherapy-related cardiotoxicity by enhancing cell viability and modulating signaling pathways associated with oxidative stress, as well as inflammatory and hypertrophic markers. Full article

Biosynthesized ZnO nanostructures were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), ultraviolet–visible (UV-vis) spectroscopy, and Fourier transform–infrared (FT-IR) spectroscopy. XRD confirmed a hexagonal wurtzite phase with an average crystallite size of 36.44 nm, while UV-vis spectroscopy showed a distinct absorption peak at 321 nm. The Zeta potential of the ZnO nanostructures was −24.28 mV, indicating high stability in suspension, which is essential for their dispersion and functionality in biological and environmental applications. The nematicidal activity of ZnO was evaluated in vitro at concentrations of 150, 300, 450, and 600 ppm, with the highest concentration achieving 75.71% mortality of second-stage juveniles (J2s) after 72 h. The calculated LC₅₀ values for the treatments were 270.33 ppm at 72 h. Additionally, molecular docking studies indicated significant interactions between the ZnO nanostructures and nematode proteins, HSP-90 and ODR1, supporting their potential nematicidal mechanism. This research highlights the effectiveness of neem leaf extract-mediated ZnO nanostructures as an eco-friendly, sustainable alternative for nematode control, presenting a promising solution for agricultural pest management. Full article

Introduction: Unilateral cleft lip and palate (CLP) is a severe orofacial birth defect characterized by improper fusion of facial parts and disturbed orofacial functions. The defect manifests as a gap in the orofacial tissues that is accompanied by defective healing patterns and chronic inflammation. The immune system’s defense factors modulate immunity, inflammation, and healing. Angiogenesis factors control blood-vessel formation. Therefore, these factors are vital in the immunological assessment and understanding of CLP morphopathogenesis. The aim of the study is to assess the distribution of vascular endothelial growth factor (VEGF), transforming growth factor beta 1 (TGF- β1), the total macrophage population and the M2 subtype, heat-shock proteins (HSP) 60 and 70, and nuclear factor kappa B (NF-κB) p50 and p65 subtypes in the affected tissue of children with CLP before and during primary dentition. Materials and Methods: Tissue samples were obtained from 15 patients aged from 3 to 8 months during veloplastic surgery. Five controls were used for comparison of data. Immunohistochemistry, light microscopy, semi-quantitative evaluation (from 0 to ++++), and statistics (Mann–Whitney U test and Spearman’s rank correlation) were used to evaluate the data for statistically significant differences and correlations between the groups. Results: Epithelial tissues affected by CLP presented with statistically significant increases in levels of VEGF (p = 0.007), total macrophages (p = 0.007), HSP60 (p = 0.001), NF-κB p65 (p = 0.000), and p50 (p = 0.045), but with a decrease in M2 macrophages (p = 0.025). Blood vessels in CLP-affected tissues showed a statistically significant increase in levels of NF-κB p65 (p = 0.003) and a statistically significant decrease in M2 numbers (p = 0.014). Connective tissue presented with no statistically significant differences. Spearman’s rank correlation revealed multiple statistically significant correlations—26 positive and 5 negative. Conclusions: Statistically significant changes in levels of VEGF and both NF-κB subtypes and numbers of total macrophages and M2 macrophages suggest a possible alteration of variable immune and inflammatory reactions and macrophage functions associated with the initiation and maintenance of the chronic process and the resulting damage. Full article

Parkinson’s disease (PD) is the second most prevalent neurodegenerative disease. It is characterized by mitochondrial dysfunction, increased reactive oxygen species (ROS), α-synuclein (α-syn) and phosphorylated-tau protein (p-tau) aggregation, and dopaminergic neuron cell death. Current drug therapies only provide temporary symptomatic relief and fail to stop or reverse disease progression due to the severe side effects or the blood–brain barrier. This study aimed to investigate the neuroprotective effects of an intermittent heating approach, thermal cycling-hyperthermia (TC-HT), in an in vitro PD model using rotenone (ROT)-induced human neural SH-SY5Y cells. Our results revealed that TC-HT pretreatment conferred neuroprotective effects in the ROT-induced in vitro PD model using human SH-SY5Y neuronal cells, including reducing ROT-induced mitochondrial apoptosis and ROS accumulation in SH-SY5Y cells. In addition, TC-HT also inhibited the expression of α-syn and p-tau through heat-activated pathways associated with sirtuin 1 (SIRT1) and heat-shock protein 70 (Hsp70), involved in protein chaperoning, and resulted in the phosphorylation of Akt and glycogen synthase kinase-3β (GSK-3β), which inhibit p-tau formation. These findings underscore the potential of TC-HT as an effective treatment for PD in vitro, supporting its further investigation in in vivo models with focused ultrasound (FUS) as a feasible heat-delivery approach. Full article

The use of gait analysis to differentiate among paediatric populations with neurological and developmental conditions such as idiopathic toe walking (ITW), cerebral palsy (CP), and hereditary spastic paraplegia (HSP) remains challenging due to the insufficient precision of current diagnostic approaches, leading in some cases to misdiagnosis. Existing methods often isolate the analysis of gait variables, overlooking the whole complexity of biomechanical patterns and variations in motor control strategies. While previous studies have explored the use of statistical physics principles for the analysis of impaired gait patterns, gaps remain in integrating both kinematic and kinetic information or benchmarking these approaches against Deep Learning models. This study evaluates the robustness of statistical physics metrics in differentiating between normal and abnormal gait patterns and quantifies how the data source affects model performance. The analysis was conducted using gait data sets from two research institutions in Madrid and Dublin, with a total of 81 children with ITW, 300 with CP, 20 with HSP, and 127 typically developing children as controls. From each kinematic and kinetic time series, Shannon’s entropy, permutation entropy, weighted permutation entropy, and time irreversibility metrics were derived and used with Random Forest models. The classification accuracy of these features was compared to a ResNet Deep Learning model. Further analyses explored the effects of inter-laboratory comparisons and the spatiotemporal resolution of time series on classification performance and evaluated the impact of age and walking speed with linear mixed models. The results revealed that statistical physics metrics were able to differentiate among impaired gait patterns, achieving classification scores comparable to ResNet. The effects of walking speed and age on gait predictability and temporal organisation were observed as disease-specific patterns. However, performance differences across laboratories limit the generalisation of the trained models. These findings highlight the value of statistical physics metrics in the classification of children with different toe walking conditions and point towards the need of multimetric integration to improve diagnostic accuracy and gain a more comprehensive understanding of gait disorders. Full article

Background: Exploring new strategies to improve type 2 diabetes mellitus (T2DM) is one of the frontier hotspots in the field of healthy food. Flavonoid–metal complexes have become one of the research hotspots in the field of health foods due to their unique structural and functional properties. Methods: In this study, the effect of hesperetin–copper(II) complex [Hsp–Cu(II)] on the gut microbiota of mice with T2DM was investigated by the 16S rRNA high-throughput sequencing. Results: The analyses of α and β diversity indicated that the richness and diversity of gut microbiota in the T2DM mice decreased and the community structure was significantly different from the normal mice. Hsp–Cu(II) increased the abundances of the beneficial bacteria (Lactobacillus, Ligilactobacillus, Romboutsia, Faecalibaculum, and Dubosiella), and decreased the amounts of the harmful bacteria (Desulfobacterota, Corynebacterium, and Desulfovibrio) and the ratio of Firmicutes/Bacteroidetes (from 44.5 to 5.8) in the T2DM mice, which was beneficial for regulating the composition of intestinal microbiota. The linear discriminant analysis effect size analysis showed that the intervention of Hsp–Cu(II) made the short-chain fatty acid (SCFA) producers (o_Lachnospirales, f_Lachnospiraceae, g_Faecalibaculum, g_Romboutsia, and g_Turicibacter) and the lactic acid bacteria producers (f_Lactobacillaceae and o_Lactobacillales) highly enriched, and the production of its metabolite SCFAs (acetic acid, propionic acid, butyric acid, and valeric acid) were increased in a dose-dependent manner, promoting the SCFA metabolism. Conclusions: Hsp–Cu(II) may improve glucose metabolic disorders and alleviate T2DM by modulating gut microbiota composition, promoting probiotics proliferation and SCFAs production, restoring intestinal barrier integrity, and suppressing local inflammation. These research findings may provide a theoretical basis for developing Hsp–Cu(II) as a new hypoglycemic nutritional supplement, and offer new ideas for the dietary food nutritional regulation to alleviate T2DM. Full article

The APETALA2/ethylene response factor (AP2/ERF) is a class of plant-specific transcription factors, among which the dehydration-responsive element-binding protein (DREB) subfamily has been widely reported to enhance plant resistance to abiotic stresses. A high-temperature-related gene, Apium graveolens DREBA6b (AgDREBA6b; accession number: OR727346), was previously cloned from a heat-tolerant celery variety. In this study, we transformed this gene into Arabidopsis thaliana using an Agrobacterium rhizogenes-mediated method to explore its function. The results showed that overexpressing AgDREBA6b in Arabidopsis thaliana significantly improved plant growth under high-temperature stress (38 °C) compared to the dreb mutant and wild-type (WT) plants. The anatomical structure of the leaves revealed that the number and degree of stomatal openings in the overexpressed plants were significantly higher than those in the WT and dreb plants, suggesting that AgDREBA6b enhances stomatal opening. Additionally, the chlorophyll content, chlorophyll fluorescence properties, proline (Pro), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities were higher in the transgenic plants, indicating better stress tolerance. qPCR analysis showed that four heat tolerance-related genes (AtHSP98.7, AtHSP70-1, AtAPX1, and AtGOLS1) were upregulated in the transgenic plants, with higher expression levels than in WT and mutant plants. This study provides valuable genetic resources for understanding the molecular mechanisms of celery’s heat tolerance and offers insights for breeding heat-tolerant celery varieties. Full article

Background: The peony (Paeonia suffruticosa Andr.), a globally valued woody ornamental species, suffers severe heat-induced floral damage that compromises its horticultural value. While the HSP20 proteins are critical for plant thermotolerance, their genomic organization and regulatory dynamics remain uncharacterized in the peony. This study aims to systematically identify the PsHSP20 genes, resolve their molecular features, and elucidate their heat-responsive expression patterns to enable targeted thermotolerance breeding. Methods: The genome-wide identification employed HMMER and BLASTP searches against the peony genome. The physicochemical properties and protein structures of the gene family were analyzed using online websites, such as Expasy, Plant-mPLoc, and SOPMA. The cis-regulatory elements were predicted using PlantCARE. Expression profiles under different times of 40 °C heat stress were validated by qRT-PCR (p < 0.05). Results: We identified 58 PsHSP20 genes, classified into 11 subfamilies. All members retain the conserved α-crystallin domain, and exhibit predominant nuclear/cytoplasmic localization. Chromosomal mapping revealed uneven distribution without lineage-specific duplications. The promoters were enriched in stress-responsive elements (e.g., HSE, ABRE) and in 24 TF families. The protein networks linked 13 PsHSP20s to co-expressed partners in heat response (GO:0009408) and ER protein processing (KEGG:04141). Transcriptomics demonstrated rapid upregulation of 48 PsHSP20s within 2 h of heat exposure, with PsHSP20-12, -34, and -51 showing the highest induction (>15-fold) at 6 h/24 h. Conclusions: This first genome-wide study resolves the architecture and heat-responsive dynamics of the PsHSP20 family. The discovery of early-induced genes (PsHSP20-12/-34/-51) provides candidates for thermotolerance enhancement. These findings establish a foundation for molecular breeding in the peony. Full article

Marine heatwaves (MHWs) have recently become more frequent, intense, and prolonged, posing significant threats to marine life and fisheries. In this study, transcriptomic analysis was employed to investigate the genes and pathways in Seriola dumerili that respond to MHW-induced stress at 28 °C (T28) and 32 °C (T32), using 24 °C (T24) as the control. Transcriptome sequencing revealed that 17 differentially expressed genes (DEGs) belonging to the heat shock protein (HSP) families—HSP30, HSP40, HSP70, and HSP90—were significantly upregulated under short-lasting MHW stress in the T24-4d vs. T32-4d comparison. Additionally, genes related to oxidative stress (e.g., protein disulfide isomerase family A member 6 [pdia6]), immune responses (e.g., interferon regulatory factor 5 [irf5]), and energy metabolism (e.g., hexokinase-1 [hk1]) were also identified. Enrichment analysis of DEGs in the T24-4d vs. T32-4d group revealed that S. dumerili exhibited adaptive responses to MHWs through the upregulation of HSPs and the activation of antioxidant, energy metabolism, and immune response pathways. However, in the T24-13d vs. T32-13d group, DEGs associated with these pathways were either not significantly expressed or were downregulated. These findings indicate that S. dumerili is unable to sustain its adaptive responses under repeated, intense MHW exposure, resulting in the disorder of its antioxidant defense system, immune suppression, and metabolic dysfunction. This study provides valuable insights into the molecular responses of S. dumerili to MHWs and supports the selection for thermal resistance in this species. Full article

HIKESHI-related hypomyelinating leukodystrophy (HHL) is a life-threatening disorder caused by homozygous pathogenic variants in HIKESHI. Symptoms include infantile onset progressive spastic dystonic quadriplegia, nystagmus, failure to thrive, diffused hypomyelination, and severe morbidity or death following febrile illness. V54L variants in HIKESHI are particularly prevalent within the Ashkenazi Jewish population. Here, we identified a novel P78S disease-causing variant in HIKESHI in a patient of Christian Arab origin, presenting with clinical and radiologic features characteristic of HHL. In silico analysis suggests that the mutated residue may affect the HIKESHI protein’s dimerization domain. We generated a comprehensive set of induced pluripotent stem cells (iPSCs) from the index case and two additional HHL patients. To investigate mechanisms potentially linked to febrile illness in HHL, we used these cells to study the heat shock (HS) response. HHL-iPSCs showed dramatically decreased levels of HIKESHI compared with healthy controls following HS. In addition, they exhibited increased HSP70 mRNA levels in response to HS, suggesting an increased sensitivity. HHL-iPSCs had impaired HSP70 translocation to the nucleus. Our results provide a human-relevant model for HHL. Full article

Small heat shock proteins (sHsps) play crucial roles in organismal adaptation to stress tolerance. Sitodiplosis mosellana, a devastating insect wheat pest, undergoes long obligatory larval diapause to survive temperature extremes during summer and winter. To elucidate the function of sHsps in this process, two sHsp-encoding genes (SmHsp22.2 and SmHsp26.7) were characterized from S. mosellana, and their responsiveness to diapause and thermal stress, as well as their roles in cold stress, was analyzed. Both SmHsp22.2 and SmHsp26.7 possessed the canonical α-crystallin domain and lacked introns. Quantitative PCR indicated significant upregulation of SmHsp22.2 and SmHsp26.7 during diapause, especially in summer and winter. Notably, SmHsp22.2 exhibited higher expression in summer relative to winter, whereas SmHsp26.7 showed the opposite profile. Moreover, short-term heat shock (≥35 °C) in over-summering larvae or cold shock (≤−10 °C) in over-wintering larvae was found to trigger transcriptional upregulation of both genes, while prolonged temperature extremes (i.e., 45–50 °C or −15 °C) did not elicit a comparable response. RNA interference-mediated knockdown of both genes significantly increased the mortality of S. mosellana larvae under cold stress. These findings indicate the importance of both SmHsps in diapause and environmental adaptation in S. mosellana. Full article

Background: BCR-ABL inhibitors such as imatinib and nilotinib exhibit multi-kinase activity that extends beyond oncology, offering significant potential for drug repurposing. Objectives: This study aims to systematically evaluate and prioritize the repurposing potential of BCR-ABL inhibitors, particularly imatinib and nilotinib. Methods: An integrated pharmacoinformatics framework was applied to analyze seven BCR-ABL inhibitors. Structural clustering, cheminformatics analysis, and transcriptomic profiling using the Connectivity Map were employed to evaluate structural relationships, target profiles, and gene expression signatures associated with non-oncology indications. Results: Structurally, imatinib and nilotinib clustered closely, while HY-11007 exhibited distinct features. Nilotinib’s high selectivity correlated with strong transcriptional effects in neurodegeneration-related pathways (e.g., HSP90 and LYN), whereas imatinib’s broader kinase profile (PDGFR and c-KIT) was linked to fibrosis and metabolic regulation. Connectivity Map analysis identified more than 30 non-cancer indications, including known off-target uses (e.g., imatinib for pulmonary hypertension) and novel hypotheses (e.g., nilotinib for Alzheimer’s via HSPA5 modulation). A substantial portion of these predictions aligned with the existing literature, underscoring the translational relevance of the approach. Conclusions: These findings highlight the importance of integrating structure–activity relationships and transcriptomic signatures to guide rational repurposing. We propose prioritizing nilotinib for CNS disorders and imatinib for systemic fibrotic diseases, supporting their advancement into preclinical and clinical evaluation. More broadly, this framework offers a versatile platform for uncovering hidden therapeutic potential across other drug classes with complex polypharmacology. Full article