Search Results (1,451)

This study examined the effects of dietary carbohydrate levels on growth performance, immune response, and antioxidant capacity of male giant freshwater prawn Macrobrachium rosenbergii. Prawns with an initial body weight of 15.04 ± 0.08 g were fed for 9 weeks on five isonitrogenous, isolipidic diets formulated to contain 10%, 15%, 20%, 25%, and 30% carbohydrate (designated CHO10–CHO30). Weight gain rate and specific growth rate rose from CHO10 to CHO20 and then declined (peak in CHO20); broken-line regression of weight gain rate estimated the optimal dietary carbohydrate level at approximately 19.36% under the present experimental conditions. Moderate carbohydrate inclusion (15–20%) enhanced hepatopancreatic total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and catalase (CAT) activities together with the transcription of cu/znsod, cat and glutathione peroxidase (gsh-px), whereas levels ≥ 25% lowered these defenses and raised hepatopancreatic malondialdehyde (MDA). High-carbohydrate diets also increased hemolymph glucose, triglyceride, total cholesterol and low-density lipoprotein cholesterol, progressively altered the mRNA expression of genes involved in the Toll-like receptor (TLR)/myeloid differentiation primary response 88 (MyD88) signaling pathway (upregulated tlr, myd88, relish and tumor necrosis factor; downregulated inhibitor of NF-κB alpha (IκB-α)), and altered the expression of apoptosis-related genes (downregulated B-cell lymphoma 2 (bcl-2), upregulated caspase-3). These results suggest that M. rosenbergii has an estimated optimal dietary carbohydrate level of approximately 19.36% under the present experimental conditions; intakes at or above 25% are associated with compromised hepatopancreatic health and growth, likely through the induction of oxidative stress, altered transcription of TLR/MyD88 pathway-related genes, and changes in apoptosis-related gene expression. Full article

The gut, brain, fat body, and various glands, although rich in “CSPs”, are not involved in chemosensory functions. We propose renaming the “CSPs” to “4CSPs” (4-Cysteine Soluble Proteins) to clarify their role, avoiding confusion with proteins also found in wings, hemolymph, and eggs. This report highlights studies related to insecticide resistance and lipid transport and expands on potential intracellular functions, strongly indicating non-chemosensory features of 4CSPs. Additionally, we show a significant correlation between 4CSPs and mucins, translation initiation factors, and actin complex proteins, reinforcing the proposal to rename “CSPs” and consider developmental pattern, tissue distribution, and intracellular localization. Full article

Vascular smooth muscle cells (VSMCs) exhibit remarkable phenotypic plasticity, dynamically transitioning from a quiescent contractile state to a dedifferentiated synthetic phenotype that constitutes the fundamental cytological driver of pathological vascular remodeling in cardiovascular diseases (CVDs) including atherosclerosis, vascular restenosis, aortic dissection (AD), and vascular calcification (VC). Signal transducer and activator of transcription 3 (STAT3) operates as a master transcriptional and functional convergence node, integrating diverse upstream biochemical stimuli, neurohumoral factors, and biomechanical stressors to govern downstream gene regulatory networks. Aberrant STAT3 activation orchestrates VSMC phenotypic modulation, excessive proliferation, directional migration, programmed cell death involving apoptosis resistance and pyroptosis initiation, extracellular matrix (ECM) reorganization, and glycolytic metabolic reprogramming via canonical nuclear transcription and non-canonical subcellular actions. Here, we systematically delineate the modular structural organization, post-translational modifications, and negative regulatory feedback mechanisms of STAT3 in shaping VSMC functionality. Furthermore, we synthesize recent advances in pharmacological interventions by comprehensively categorizing therapeutic modalities into direct structural-domain inhibitors, upstream kinase-targeted indirect agents, bioactive natural products, and emerging clinical-stage translational candidates. Finally, we critically address translational hurdles regarding on-target systemic toxicities, and highlight site-specific vascular delivery systems alongside localized drug-eluting vascular devices to advance precision STAT3-targeted cardiovascular therapeutics. Full article

Background: Although POLD1 exhibits oncogenic properties in multiple malignancies, its precise role and regulatory mechanisms in cisplatin resistance of bladder cancer (BC) remain elusive. This study aims to elucidate the functional involvement and upstream regulatory axis of POLD1 in BC chemoresistance. Methods: We evaluated the impact of POLD1 on chemoresistance and DNA damage repair (DDR) using public clinical databases and cisplatin-resistant cell lines, employing CCK-8, colony formation, flow cytometry, immunofluorescence, and comet assays. Protein interactions were examined via Co-IP, molecular docking, and truncation mutant analysis. ChIP-PCR and dual-luciferase reporter assays were utilized to identify the upstream transcription factor. In vivo functionality was further validated in a nude mouse xenograft model. Results: POLD1 was markedly upregulated in BC tissues and cisplatin-resistant BC cells, and its elevated expression was tightly associated with advanced tumor stage, high pathological grade, and poor patient prognosis. Functional experiments verified that POLD1 knockdown aggravated cisplatin-induced DNA damage and drastically sensitized BC cells to cisplatin in vitro, and attenuated tumor growth under cisplatin treatment in vivo, indicating that POLD1 is a key driver of cisplatin resistance. Mechanistically, POLD1 physically interacted with and activated ATM, thereby initiating homologous recombination (HR) repair signaling. Moreover, transcription factor MAZ directly bound the promoter region of POLD1 to transcriptionally upregulate its expression. Rescue experiments in vitro further validated that the MAZ-POLD1 axis facilitates cisplatin resistance by modulating the DDR pathway in BC. Conclusions: Therapeutic targeting of the POLD1-regulated DDR pathway holds great promise as an effective strategy to reverse acquired cisplatin resistance in BC. Full article

The stress response is essential for cellular and organismal survival as it acts as a protective and adaptive mechanism to maintain homeostasis. At the organismal level, physical stressors induce responses in mammals that are mediated primarily by the hypothalamic–pituitary–adrenal (HPA) axis, which regulates glucocorticoid secretion. The HPA axis functions as a circadian-regulated, multi-oscillator system, in which the paraventricular nucleus, the pituitary, and the adrenal gland exhibit intrinsic rhythmicity while remaining coordinated by the output from the suprachiasmatic nucleus. Glucocorticoids act both as stress effectors and systemic zeitgebers that synchronize peripheral clocks. At the cellular level, cellular stressors are sensed by four protein kinases of eukaryotic translation initiation factor 2α (eIF2α) and activate the evolutionarily conserved integrated stress response (ISR), which converges on phosphorylation of Serine 51 on eIF2α. ISR signaling is temporally regulated by the circadian clock and controls time-of-day-dependent protein synthesis. In parallel, ISR pathways feed back onto the circadian clock through transcriptional, translational and epigenetic mechanisms, directly influencing core clock gene expression and stability of circadian oscillations. Physiological ISR activity supports circadian robustness and resetting, whereas excessive ISR activation dampens rhythmic gene expression and destabilizes behavioral rhythms. The current review summarizes recent advances in our understanding of the crosstalk mechanisms between the HPA axis, ISR, and the circadian clock to provide new insights into disease mechanisms and inform chronotherapeutic strategies to target dysregulated HPA and ISR activities and restore temporal homeostasis. Full article

Background: Opioid use during pregnancy is linked to preterm birth, fetal growth restriction, and Neonatal Opioid Withdrawal Syndrome. While the placenta is the primary regulator of the uterine environment, the precise molecular mechanisms by which opioids dismantle placental function remain poorly defined. Methods: The transcriptomic landscape of opioid-exposed human placentas from a unique Hawai’i-based cohort was characterized. Results: Transcriptional signatures were defined by genes involved in translational inhibition and metabolic attenuation. Integrative network modeling predicted a prominent stress-signaling hub centered on p38 Mitogen-Activated Protein Kinase and Extracellular Signal-Related Kinase 1/2, paralleling a broad suppression of ribosomal protein transcripts (Ribosomal Protein 27S, -29, and -39). This response was further characterized by the inhibition of the Myelecytomatosis Proto-Oncogene regulatory hub, predicting a loss of mitochondrial phosphate transport through associated genes (via Solute Carrier Family 25A3) and cell-cycle progression (via S-Phase Kinase-Associated Protein 1). Additionally, the genes in this model suggested upregulation of antisense regulatory brakes, such as RAP2C Antisense RNA 1, which might further reinforce this inhibitory state. Conclusion: The findings suggest that prenatal opioid exposure is associated with the disruption of placental homeostasis. This may be coordinated by protein synthesis and bioenergetic flux, as inferred from the modeled eukaryotic Translation Initiation Factor-mediated Integrated Stress Response. These findings establish a localized, clinical baseline of human placental stress, highlighting candidate molecular regulatory nodes to guide future mechanistic studies and biomarker discovery in exposed pregnancies. Full article

In this study, Alfalfa (Medicago sativa L.) was used as the experimental material to systematically identify members of the AP2/ERF transcription factor family in its genome and to elucidate the response patterns and potential regulatory networks of this family under drought stress. A total of 756 AP2/ERF genes were identified and classified into four subfamilies: AP2, ERF, DREB, and RAV, among which the DREB and ERF subfamilies exhibited significant expansion. Evolutionary analysis indicated that whole-genome duplication (WGD)/segmental duplication was the primary driving force underlying the expansion of this family. Under graded drought treatments, multiple AP2/ERF genes were significantly upregulated, with MsERF210 showing continuously increasing expression as drought intensity escalated. WGCNA revealed that MsERF210 was highly positively correlated with antioxidant-related modules, including “Glutathione metabolism,” “Peroxisome,” and “Ascorbate and aldarate metabolism.” Further analysis showed that several antioxidant genes, such as SOD, GST, ALDH, and IDH, were significantly upregulated under drought stress, and their promoter regions were predicted to be bound by MsERF210. Physiological indicator measurements demonstrated that drought stress led to increased accumulation of H₂O₂ and MDA, along with elevated contents of AsA, DHA, GSH, and GSSG, while SOD activity initially increased and then decreased. qRT-PCR validated the expression trends of key genes, confirming the reliability of the transcriptome data. In summary, this study reveals the evolutionary characteristics of the AP2/ERF family in alfalfa and identifies MsERF210 as a key regulator that may enhance alfalfa’s adaptability to drought stress by modulating the antioxidant defense system and redox homeostasis. These findings provide important candidate gene resources and a theoretical basis for breeding new drought-resistant alfalfa varieties. Full article

Diosgenin is an important steroidal sapogenin widely used for the industrial production of steroidal drugs. Phytosulfokine-α (PSK-α) is a sulfated pentapeptide hormone known to regulate plant growth, development, and stress responses. However, its role in the regulation of diosgenin biosynthesis remains unclear. Fenugreek seedlings were treated with PSK-α to investigate its effects on diosgenin biosynthesis through metabolite profiling and comparative transcriptomic analyses. PSK-α induced a dynamic response in diosgenin accumulation, characterized by an initial decrease during the early stage of treatment followed by a gradual increase, ultimately resulting in 61.4% and 72.1% increases in diosgenin accumulation after 10 days and 15 days, respectively, compared with the corresponding random pentapeptide control. Comparative transcriptomic analysis between PSK-α-treated seedlings and the corresponding controls at 5 and 15 days identified key genes or transcript isoforms involved in diosgenin biosynthesis, PSK signaling and MAPK signaling pathways. Meanwhile, five candidate genes involved in diosgenin biosynthesis and PSK signaling exhibited expression patterns consistent with the dynamic accumulation pattern of diosgenin. To further dissect the regulatory network, co-expression network analysis between these 28 candidate genes involved in diosgenin biosynthesis and PSK signaling and 1407 transcription factor-encoding genes identified candidate transcription factors potentially involved in the coordinated regulation of diosgenin biosynthesis and PSK signaling. This study suggests that PSK-α may contribute to enhanced diosgenin accumulation through coordinated transcriptional regulation of diosgenin biosynthesis and PSK signaling pathways, providing new insights into the regulation of plant secondary metabolism by peptide hormones. Full article

In vitro 3D cell culture systems provide a physiologically relevant microenvironment that mimics native tissue architecture. However, establishing functional 3D testicular aggregates in teleosts remains challenging. In this study, we established a novel 3D co-culture aggregate system for olive flounder (Paralichthys olivaceus) primary testicular cells by incorporating the established olive flounder testicular cell (OFT) line. Primary testicular cells alone failed to form 3D aggregates within the first 24 h in ultra-low attachment plates. However, co-culturing with OFT cells rapidly initiated 3D aggregate assembly within 24 h. Furthermore, supplementation with a cocktail of sex hormones and growth factors maintained high cell viability without cytotoxic effects. Notably, quantitative real-time PCR analysis revealed that hormone and growth factor supplementation significantly upregulated the transcript levels of both plzf (2.15-fold) and scp3 (2.34-fold). These results demonstrate that the OFT cell line is essential for facilitating the assembly and incorporation of primary testicular cells into 3D aggregates, while sex hormones and growth factors promote spermatogonial stem cell maintenance and meiotic progression within the aggregates. Overall, this 3D co-culture platform serves as a valuable in vitro tool for fish spermatogonial stem cell research and reproductive biotechnology in aquaculture. Full article

Pain is a complex process involving dynamic transcriptional changes in cells of the PNS and CNS following injury or inflammation. Among stress-inducible transcription factors, ATF3 has emerged as one of the most robust molecular markers of neuronal injury, particularly in sensory neurons of the DRG. Although ATF3 is widely used as an indicator of axonal damage in experimental pain models, its functional contribution to the initiation, maintenance, and resolution of pain remains poorly understood. Recent transcriptomic and functional studies suggest that ATF3 not only reflects neuronal stress but also orchestrates gene expression programs involved in axonal regeneration, neuroimmune communication, ion channel remodeling, and nociceptor plasticity. Moreover, ATF3 expression has been identified in non-neuronal cell populations, including Schwann cells and satellite glial cells, indicating broader roles in peripheral nerve repair and neuroinflammation. Despite the growing body of experimental evidence, the literature remains fragmented, and no consensus has yet been reached as to whether ATF3 primarily promotes adaptive regeneration or directly contributes to maladaptive pain signaling. This review aims to provide a comprehensive and critical overview of the current understanding of ATF3 biology in pain, building on evidence from transcriptomic and molecular analyses, experimental models of neuropathic, inflammatory, and cancer-associated pain, and emerging mechanistic insights into its role in pain-related neuronal plasticity. This review examines the regulation of ATF3 expression, its downstream transcriptional targets, its interactions with inflammatory signaling pathways, and its potential value as a therapeutic target. By consolidating current evidence and highlighting existing knowledge gaps, this review seeks to clarify the multifaceted role of ATF3 in pain pathophysiology. Full article

Seed coat color is an important quality trait in mung bean (Vigna radiata) and is closely associated with seed appearance, commercial value, and phytochemical composition. To investigate the genetic basis of black seed coat formation, six F₂ populations were derived from reciprocal crosses between the black-seeded accession LZL115 and the green-seeded accession LZL156. Among 755 F₂ plants, 559 produced black-coated seeds and 196 produced green-coated seeds, conforming to a 3:1 segregation ratio (χ² = 0.37, p = 0.54). These results indicated that black seed coat color was dominant and consistent with the control by a single dominant locus, designated VrScL115, in the LZL115 × LZL156 genetic background. Bulked segregant analysis sequencing (BSA-seq) initially mapped VrScL115 to an approximately 2.90 Mb region on chromosome 4. Using newly developed KASP markers and recombinant screening in expanded F₂ populations, the locus was further delimited to a 121.79 kb interval between markers LS_K3333 and LS_K3379. Of the 11 annotated genes within this interval, LOC106758748 was the only gene containing high-confidence coding-sequence variants between the parents. This gene encodes a putative R2R3-MYB transcription factor homologous to MYB90. Comparative sequence analysis identified several allelic variants potentially associated with black seed coat color, and protein structure prediction indicated local structural differences between the parental proteins. LOC106758748 showed consistently higher expression in the developing seed coats of LZL115 than in those of LZL156 at 10, 15, and 20 days after pollination, with expression peaking at 15 days. Haplotype analysis of 246 mung bean accessions showed that the LZL115-associated allele combination at LS_K3352, LS_K3365, LS_K3367, and LS_K3370 was present in 21 of 27 black-seeded accessions (77.8%) and absent from all 219 non-black accessions, corresponding to a specificity of 100% and a false-negative rate of 22.2%. These findings support LOC106758748 as the leading candidate gene for VrScL115; however, direct in vivo functional validation is still required to confirm its causal role in black seed coat formation. The four-marker combination may be useful for identifying germplasm carrying the LZL115-associated allele, although further validation in independent germplasm populations is required. Full article

Auxin response factors (ARFs) are key transcriptional regulators of the auxin signaling pathway and play important roles in plant organogenesis and regeneration. However, the functions of ARF family genes in lily scale-derived bulblet regeneration remain largely unclear. In this study, 24 LdARF genes were identified from the genome of Lilium davidii var. unicolor. Phylogenetic analysis revealed that LdARF proteins showed evolutionary conservation with ARF homologs from other monocot species. Genome-wide identification, phylogenetic analysis, and expression profiling revealed functional divergence among LdARF genes during scale-derived bulblet regeneration. Among them, LdARF17 exhibited a distinct regeneration-associated expression pattern, characterized by rapid induction after scale excision and sustained high expression during subsequent bulblet initiation and formation. Subcellular localization analysis demonstrated that LdARF17 is localized in the nucleus. Transient overexpression of LdARF17 significantly promoted bulblet regeneration and was associated with increased expression of auxin-responsive and regeneration-related genes, including IAA14, LBD16, and LBD29. These findings suggest that LdARF17 acts as a positive regulator of lily scale regeneration and may influence auxin-responsive transcriptional processes associated with early cell proliferation, providing new insights into the molecular mechanisms underlying vegetative regeneration in lilies. Full article

Background: Delirium management remains inconsistently implemented in intensive care units (ICUs), and existing studies provide limited insight into how assessment capability, organisational opportunity, and emotional motivation interact in Chinese tertiary care settings. Aim: To explore ICU nurses’ perceived barriers and facilitators to delirium assessment and management using the Capability, Opportunity and Motivation model of Behaviour (COM-B). Methods: A qualitative descriptive exploratory study was conducted with 14 registered ICU nurses from two tertiary hospitals in Chongqing, China, between December 2025 and February 2026. Purposive sampling captured variation in clinical experience, educational background, and ICU type. The first author conducted face-to-face interviews in Mandarin Chinese and completed the initial coding. COM-B informed both the interview guide and the deductive organisation of the analysis, while six themes and 33 lowest-level factors were developed within the three domains. During revision, the fixed analytic structure was audited against all 14 Mandarin transcripts, and the second author independently applied the fixed codebook to five complete transcripts selected for maximum variation; disagreements were resolved by returning to the original Mandarin context and predefined coding boundaries. Results: Eighteen barriers and fifteen facilitators were identified across six themes within the COM-B domains. Capability was strengthened by clinical experience, senior nurses’ intuitive pattern recognition, and conversational assessment, but constrained by difficulty recognising hypoactive delirium, uncertainty when assessing older adults, inconsistent use of CAM-ICU, and insufficient targeted training. Opportunity was enhanced by staffing, environmental control, peer support, family engagement, and physician collaboration, but restricted by fragmented visibility, sensory overstimulation, workload, restrictive visiting, and delayed medical responses. Motivation reflected the tension between professional duty and accountability for safety on one hand and emotional burden, verbal abuse, and violence on the other. Cross-domain accounts showed that opportunity constraints could amplify anxiety and safety-first motivation, narrowing feasible behaviour toward containment, whereas social support could sustain relational care. Conclusions: Delirium management was shaped by interacting behavioural, organisational, and emotional conditions. Strengthening assessment capability alone is unlikely to produce consistent practice without staffing, workflow, environmental, and interprofessional support. Mapping the findings to Behaviour Change Wheel intervention functions highlighted training, environmental restructuring, enablement, and modelling as priority candidate functions for prospective evaluation; education alone was unlikely to be sufficient. Reporting Method: This study was reported in accordance with the Consolidated Criteria for Reporting Qualitative Research checklist. Patient or Public Contribution: No patient or public contribution was involved in the design, conduct or reporting of this study. Participants were registered intensive care unit nurses. Full article

Background: Glioblastoma is a primary aggressive brain tumor, with an average survival rate of ~14.6 months. The low therapeutic benefit of current treatments is in part due to glioblastoma-initiating cells hijacking microglia/macrophages to support tumor growth, prompting the development of multitargeted therapeutic approaches. One such therapeutic target is transcription factor Nrf2. High Nrf2 activity is associated with high-grade tumors, and high Nrf2 levels in microglia/macrophages lead to polarization toward an immunosuppressive profile, supporting glioblastoma progression and therapy resistance. Interestingly, however, sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables and a potent Nrf2 activator, has anti-carcinogenic effects in multiple animal models. Methods: We utilized the zebrafish glioblastoma initiation model of human AKT1 overexpression in neuronal progenitors to capture the intermediate progenitor-cell-like state of glioblastoma-initiating/pre-neoplastic cells and evaluated the therapeutic potential of SFN and VVD130037, an Nrf2 inhibitor currently in clinical trials. Results: Initial findings suggest that SFN, individually and in combination with VVD130037, had the potential to decrease the levels of AKT1. Surprisingly, VVD130037 tended to increase AKT1. The pAKT1 levels also increased in Nrf2-deficient human cells. Moreover, failure to activate Nrf2 in neural progenitors in response to SFN, while a preliminary observation that warrants further investigation, suggests that the decrease in AKT1 was not potentially mediated by Nrf2 activation. The combined effect of SFN and VVD130037 on AKT1 was particularly strong in irf8-/- mutant larvae, which lack a microglia/macrophage population, indicating the existence of a non-tumorigenic cell population(s) sensitive to changes in Nrf2 activity. Conclusions: AKT1 inhibition in glioblastoma-initiating cells by the phytochemical SFN, combined with Nrf2 inhibition in the surrounding cells, may impede glioma progression. Full article

Background: Previous Australian child developmental assessment service (CDAS) cohorts have identified a small proportion of children experiencing extreme waiting times (≥730 days) for standardised developmental assessments. However, factors associated with these extreme waits remain poorly understood. Aim: To examine factors associated with a priori defined extreme waiting times (≥730 days) by comparing affected children with the remaining CDAS cohort, while validating administrative data and exploring caregiver experiences. Methods: An embedded mixed-methods service evaluation was undertaken. Quantitative data were extracted from the CDAS database and electronic medical records for children assessed between October 2022 and September 2024. Children with validated extreme waiting times were compared with the remaining CDAS cohort. Records initially identified as waiting ≥730 days underwent manual validation of referral and assessment dates. Caregivers of children with validated extreme waits were purposively recruited for semi-structured interviews. Fifteen caregivers participated; twelve families did not respond. Interview transcripts underwent thematic analysis. Results: Of 67 children initially identified as waiting ≥730 days, 20 (29.9%) were excluded following manual validation because of administrative recording inaccuracies, leaving 47 children with confirmed extreme waiting times. Of these, 46 completed CDAS assessment and had available validated waiting-time data, with a median wait of 859 days (IQR 775–932), compared with 254 days (IQR 111–355) in the comparison cohort of 2039 other children. Children in the extreme-wait cohort were more often from culturally and linguistically diverse backgrounds (76.6% vs. 53.5%; p = 0.002), required interpreters more frequently (23.4% vs. 9.0%; p = 0.003), and were more likely to be referred by paediatricians (89.4% vs. 74.1%; p = 0.017). Over one-third (n = 17) were prioritised for assessment within six months at triage. Review assessments accounted for 25/47 (53.2%) of the extreme-wait cohort. Caregivers described acceptance of lengthy public-sector waits, communication gaps, uncertainty regarding service pathways, reliance on interim supports, financial barriers to private assessment, and positive experiences once assessments were completed. Conclusions: Children experiencing extreme waits had distinct characteristics. Extreme waiting reflected genuine service delays, planned clinical review pathways, and administrative recording inaccuracies. Administrative data should be validated before waiting-time metrics are used for service evaluation. Improved communication, service navigation, equitable access to interim supports, and targeted service redesign may improve caregiver experiences and reduce extreme waits in publicly funded developmental assessment services. Full article