Search Results (316)

Pinealectomy leads to melatonin deficiency, which is known to disrupt circadian clock regulation and may increase vulnerability of the hippocampus to oxidative stress and neuroinflammatory processes. The objective of this study was to examine the gene expression levels of circadian locomotor output cycles kaput (CLOCK), brain and muscle ARNT-like 1 (BMAL1), period circadian regulator 1 (PER1), cryptochrome circadian regulator 1 (CRY1), brain-derived neurotrophic factor (BDNF), and interleukin-6 (IL-6) in the hippocampus to elucidate the impact of pinealectomy-induced circadian dysregulation on these gene expressions and to assess its association with hippocampal alterations. A total of 30 Wistar rats were randomly divided into three groups: Control, Sham, and Pinealectomy (PNX) (n = 10 per group). Gene expression levels were analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). Immunohistochemical analysis was performed to assess caspase-3 and glial fibrillary acidic protein (GFAP) immunoreactivity. In addition, oxidative stress parameters, including malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH), as well as the inflammatory marker tumor necrosis factor-alpha (TNF-α), were measured. The pinealectomy group showed a significant downregulation of BMAL1, BDNF, CLOCK, CRY1, and PER1 gene expression levels, with decreases ranging from approximately 60% to 83% compared with the sham and control groups, whereas IL-6 expression was significantly increased by approximately 185% (p < 0.05). Immunohistochemical analysis demonstrated significantly increased caspase-3 and GFAP immunoreactivity in the PNX group. Furthermore, pinealectomy resulted in a significant increase in MDA and TNF-α levels, accompanied by marked decreases in SOD, CAT, and GSH levels (p < 0.05). In conclusion, pinealectomy is associated with significant disruption of hippocampal circadian clock gene expression, accompanied by oxidative stress, neuroinflammation, and histopathological alterations. These findings highlight the critical role of circadian regulation in maintaining hippocampal cellular integrity. Full article

Phyllotreta striolata is a global pest of cruciferous vegetables, and controlling its soil-dwelling larvae is challenging. The lack of standardized larval bioassay methods hinders the screening of effective biocontrol agents. In this study, we established a stable and standardized laboratory-efficacy trial system for P. striolata larvae. Indoor rearing techniques were optimized for Brassica juncea var. foliosa and Brassica juncea var. megarrhiza were identified as the optimal host plants, with ideal oviposition conditions at 26–28 °C using black flannel substrate, and soil-cultured Brassica rapa var. pekinensis as the host plant. Based on these findings, a larval bioactivity assay was established using B. juncea var. megarrhiza slices on water-agar. This system maintained a natural larval mortality rate below 5% within 48 h, meeting the bioassay requirements. The reliability of the system was validated by evaluating the activity of the engineered Bacillus thuringiensis (Bt) strain G033A against larvae, where the LC₅₀ value decreased from 23.013 mg/mL to 7.295 mg/mL with an extended treatment time (12–48 h). Using this standardized method, novel Cry proteins with high activity against P. striolata larvae were screened. Cry8Ca and Cry8Ga proteins exhibited LC₅₀ values of 2.243 mg/mL and 1.649 mg/mL, respectively. Full article

Background/Objectives: Growing evidence suggests that disruption of circadian rhythms contributes to the pathogenesis of inflammation and inflammatory bowel disease; however, clinical data linking circadian gene variants to ulcerative colitis remain limited. In this study, we aimed to investigate associations between key circadian rhythm gene polymorphisms and clinical and treatment-related characteristics in ulcerative colitis. Methods: A total of 107 patients with ulcerative colitis and 80 healthy controls were included in this single-center cross-sectional study. The BMAL1 rs7950226, CLOCK rs1801260, and CRY1 rs2287161 polymorphisms were analyzed using the polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) method. Genotype and allele frequency distributions were compared between patients and controls, and associations with clinical characteristics were evaluated within the ulcerative colitis cohort. Results: Genotype distributions of BMAL1 rs7950226 and CLOCK rs1801260 were similar between patients with ulcerative colitis and healthy controls; however, the G allele of BMAL1 was more frequent in patients (p = 0.028). Within the ulcerative colitis cohort, CLOCK rs1801260 genotypes were significantly associated with inflammatory and treatment-related characteristics, with the CC genotype linked to higher C-reactive protein levels (p = 0.021) and the TT genotype associated with increased azathioprine use (p = 0.006). Conclusions: These findings suggest a potential association between circadian rhythm gene variants and clinical features of ulcerative colitis, particularly in relation to inflammatory activity and treatment requirements, and provide preliminary clinical insight that warrants further investigation in larger and longitudinal studies. Full article

Exogenous protein degradation dynamics during transgenic maize straw degradation in soil and the mechanisms underlying soil microbial community construction remain unclear. Applying null-model analysis to determine these mechanisms is important for assessing transgenic crop straw return-to-field-related impacts on dynamic soil quality and microbial ecological function changes. A laboratory leaf degradation burial simulation was conducted to establish an exogenous protein Cry1A.401 soil degradation model and clarify its behaviors. Coupled Illumina MiSeq 16S rDNA sequencing–soil physicochemical factor analysis was used to evaluate soil microbial community characteristic and diversity changes during leaf degradation and explore soil microbial community construction mechanisms and driving factors. The results revealed that exogenous protein Cry1A.401 released from transgenic insect-resistant maize leaves exhibited consistent degradation characteristics, decreasing rapidly at the initial stage but slowly at the middle/late stages. The diversity levels within/between soil microbial community groups did not significantly differ. Coexistence was the dominant interaction type among soil microbial communities. Community assembly occurred stochastically and was limited primarily by diffusion. Insights into the putative mechanistic links among Bacillus thuringiensis (Bt) proteins, soil properties, and microorganisms are provided. Our understanding of the ecological impacts of exogenous Bt proteins released into soil via leaves on soil ecosystems was enhanced. Full article

Monochamus alternatus larvae, as concealed trunk-boring pests, evade conventional insecticide contact due to their cryptic feeding niche. To overcome this limitation, previous studies have engineered strains of the naturally entomopathogenic bacterium Yersinia entomophaga. The lethality of these strains against M. alternatus was enhanced by incorporating extracellular secretion systems and enriching insecticidal proteins within the larval midgut. However, plasmid loss occurs during serial subculturing. Here, we established an engineered strain that expresses the red fluorescent protein gene mCherry to explore the applicability of bacterial conjugation transfer to Yersinia. We then constructed a chromosomally integrated strain (CSLH88-pCHSW) that incorporates extracellular secretion systems. The results of stability assays demonstrated 100% retention of the mCherry and Cry3Aa-T-HasA genes over 78 generations. SDS-PAGE and Western blot analyses confirmed the extracellular secretion of the Cry3Aa-T protein in the CSLH88-pCHSW strain. Bioassays revealed that the CSLH88-pCHSW strain was significantly more virulent against M. alternatus larvae than both the wild-type strain (CSLH88) and the plasmid-transformed strain (CSLH88-pCHKW), and exhibited markedly faster insecticidal kinetics. Our study reveals the application of bacterial conjugation transfer technology for constructing biocontrol strains. This genomically stabilized Yersinia strain eliminates the risks of failure associated with plasmid loss in the field, enabling the sustainable control of M. alternatus. Full article

Insecticidal proteins derived from Bacillus thuringiensis (Bt) have been effectively employed in controlling lepidopteran pests, notably in transgenic crops targeting Spodoptera species. However, concerns have arisen regarding the long-term efficacy due to the emergence of tolerant and resistant insect populations. Prior research suggested that repeated exposures to Bt, which contains a mixture of spores and crystals, may contribute to the development of tolerance; however, the specific effects of sequential exposure to purified Cry1 and Vip3Aa proteins remain unclear. This study aimed to assess whether prior exposure of Spodoptera exigua neonate larvae to sublethal concentrations of Cry1Ab, Cry1Ca or Vip3Aa proteins would heighten their tolerance upon subsequent exposure, and whether such effects would extend to their offspring. Pre-exposure to Cry1Ab or Vip3Aa did not affect larval responses to the toxin. For Cry1Ca, a slight increase was observed under one treatment condition, but the effect was not considered biologically relevant. Transgenerational analysis revealed no enhancement of tolerance; rather, there was a negative impact on the offspring’s response in some cases. These findings indicate that although previous studies have documented that sublethal contact with bacterial preparations may significantly affect insect tolerance, exposure to sublethal doses of purified Cry1 and Vip3Aa proteins is unlikely to lead to the development of tolerance in S. exigua. Full article

The importance of protein-based materials in agricultural pest control has received increasing attention in recent years. Herein, Plutella xylostella brush-border membrane vesicles (BBMVs) were used as a target to screen for human domain antibodies with insecticidal activity. Three rounds of panning of the phage display library yielded the domain antibody C4D, which competed with the Cry1Ac toxin to bind to P. xylostella BBMVs. Against P. xylostella larvae, the recombinant soluble C4D protein showed an LC₅₀ of 1.57 μg/cm² (95% fiducial limits: 0.83–2.54). Using pull-down assays and liquid chromatography–tandem mass spectrometry, we identified the C4D binding partner in P. xylostella midgut BBMVs to be a cadherin-like protein. Bio-Layer Interferometry assay revealed that the dissociation constant between soluble C4D and P. xylostella cadherin-like protein was 2.99 × 10⁻⁶ M. Thus, the present study explored strategies to generate insecticidal antibodies, and the human domain antibody C4D identified and characterized in this study can serve as a framework for generating novel insecticidal agents. Full article

Background: Deprivation of sleep (DS) might affect mood and cognitive abilities, including psychomotor functions (PF). Molecular mechanisms underlying these effects remain unclear, though studies suggest that the circadian rhythm plays a role. Methods: Seventy participants underwent polysomnography (PSG) and DS. PF was evaluated using Bimanual Eye–Hand Coordination Test (BEHCT). Mood, PF, and clock gene expression (Circadian Locomotor Output Cycles Kaput (CLOCK), Brain and Muscle ARNT-Like 1 (BMAL1), Period Circadian Regulator 1 (PER1), Cryptochrome Circadian Regulator 1 (CRY1), Nuclear Receptor Subfamily 1 Group D Member 1 (NR1D1), and Neuronal PAS Domain Protein 2 (NPAS2)) were analyzed post-PSG and post-DS. Mood changes after DS classified participants as responders (RE) or non-responders (NR). Results: In NRs, but not REs, the BEHCT error count positively correlated with the expression of BMAL1, CRY1, PER1, NR1D1 (R = 0.60, p = 0.002; R = 0.49, p = 0.018; R = 0.57, p = 0.023; and R = 0.53, p = 0.011, respectively), with PER1 explaining its variability in 57.8% (b = 0.174, R² = 0.578, F = 20.144, and p < 0.001). Conclusions: Obtained results suggest that altered clock gene expression may contribute to individual differences in mood and PF following DS. Full article

To explore the function of the period gene (Ec-per) in Exopalaemon carinicauda, we cloned the gene of 4611 bp with a 5′UTR of 201 bp, a 3′UTR of 813 bp, and an ORF of 3597 bp encoding 1198 amino acids. The predicted protein includes two PAS and one PERIOD domain. qPCR analysis revealed that Ec-per was expressed across all tissues tested at different developmental stages and during both embryonic and larval stages. Moreover, Ec-per oscillated rhythmically under different conditions of light-to-dark (L:D) ratios, including continuous darkness (0 L:24 D), where changes in the photoperiod influenced amplitude and phase shifts. The knockdown of Ec-per mRNA significantly reduced the expression of the circadian-related genes timeless (tim) and cryptochrome 1 (cry1) (p < 0.05). This suggests that Ec-per is an endogenous clock gene that may participate in molecular feedback loops and synergistically regulate the circadian rhythms through interacting with tim and cry1. RNA interference of Ec-per also markedly downregulated ecdysone receptor mRNA (p < 0.05), suggesting a positive role in the ovarian development of E. carinicauda. In situ hybridization further demonstrated that Ec-per is involved in oocyte proliferation and the accumulation of exogenous nutrients. This study provides new insights for promoting ovarian development and artificial breeding in crustaceans through optimized light-cycle management. Full article

Pollen allergy represents a growing public health concern, yet the role of microplastic pollution in modulating allergen behavior remains largely unresolved. In this study, we investigated interactions between polyethylene terephthalate (PET) microplastics (0.2–12 µm; predominantly 0.4–1 µm) and cedar pollen proteins, with emphasis on the major allergen Cry j 1. Surface charge characterization using the pH drift method revealed two apparent points of zero charge in the acidic (pH 3.0–3.8) and near-neutral (~7.5) regions, indicating surface chemical heterogeneity. Protein adsorption experiments conducted at physiological pH (7.4) showed concentration-dependent and saturable removal of proteins from solution with increasing PET mass and a 3.10-fold preferential enrichment of aromatic-rich protein fractions. Spectroscopic analyses revealed adsorption-induced but non-denaturing structural perturbations, including increased exposure of aromatic residues and partial β-sheet destabilization. Complementary all-atom molecular dynamics simulations showed rapid and stable Cry j 1 adsorption onto PET, anisotropic surface accommodation, modest increases in solvent accessibility, and subtle secondary structure rearrangements without global unfolding. Together, these findings indicate that PET microplastics can selectively bind and structurally modulate pollen allergens in ways that may influence allergen persistence and epitope presentation, with potential implications for IgE-mediated sensitization in polluted environments. Full article

Bacillus thuringiensis (Bt) is one of the most extensively used microbial insecticides, attributed to the action of insecticidal crystal proteins (ICPs), primarily Cry toxins, which mediate damage to the insect midgut epithelium. Recent evidence suggests that Bt toxicity is also strongly influenced by its physiological state and interactions with the host gut environment. Biofilm formation represents an important adaptive strategy that enhances bacterial stress tolerance and may modulate insecticidal performance, although the underlying mechanisms remain unclear. However, it is still unclear how Bt in the biofilm state alters host responses at the structural and transcriptomic levels. Using the tea plantation pest Ectropis grisescens as a model, we systematically evaluated the insecticidal efficacy of biofilm-state Bt formulations and their synergistic effects with a biofilm inducer system composed of Tween-80, tea saponin, matrine, and tea polyphenols. Bioassays showed that the biofilm-state Bt supplemented with composite inducers achieved the highest corrected mortality and reduced the LC₅₀ against neonate larvae by 2.88-fold compared with conventional planktonic Bt. Histopathological, biochemical, and transcriptomic analyses further revealed that biofilm-state Bt caused more severe midgut damage and induced extensive remodeling of detoxification- and stress-response-related pathways. These findings highlight Bt physiological state as a critical determinant of formulation efficacy and provide a novel framework for Bt optimization through microbial physiological regulation. Full article

Field-evolved resistance of Helicoverpa zea to crops expressing Cry insecticidal proteins from Bacillus thuringiensis (Bt) is widespread across the United States. To comparatively evaluate physiological factors associated with Bt susceptibility, we analyzed two laboratory strains (Benzon and SIMRU) and one field colony obtained from a commercial corn field near Pickens, Arkansas. Biochemical assays of larval midgut extracts showed that Pickens exhibited significantly altered activities of chymotrypsin-like proteases, aminopeptidase N (APN), and alkaline phosphatase (ALP) compared with the SIMRU or Benzon colonies, with differences varying by larval instar. In contrast, trypsin-like protease activities did not differ significantly among the three colonies. Gene expression analyses of ten serine protease genes and seven candidate Cry receptor genes (including cadherin, ATP-binding cassette family C2, ALP, and four APN genes) revealed significant transcriptional differences in the Pickens relative to the lab colonies. Collectively, these results suggest that chymotrypsin-like proteases may play an important role in the activation of Cry toxins in H. zea. Altered chymotrypsin and APN activities, together with differential gene expressions in the Pickens population, likely contribute to reduced Bt susceptibility. The biochemical and molecular differences provide insight into potential physiological factors underlying reduced Bt susceptibility and may inform future Bt resistance monitoring and management strategies. Full article

Bt Cry toxins remain the cornerstone of transgenic crop protection against Lepidopteran pests, yet field-evolved resistance, particularly in invasive species such as Spodoptera frugiperda and Helicoverpa armigera, can threaten their long-term efficacy. This review presents a comprehensive and unified mechanistic framework that synthesizes current understanding of Bt Cry toxin modes of action and the complex, multilayered regulatory mechanisms of field-evolved resistance. Beyond the classical pore-formation model, emerging evidence highlights signal transduction cascades, immune evasion via suppression of Toll/IMD pathways, and tripartite toxin–host–microbiota interactions that can dynamically modulate protoxin activation and receptor accessibility. Resistance arises from target-site alterations (e.g., ABCC2/ABCC3, Cadherin mutations), altered midgut protease profiles, enhanced immune regeneration, and microbiota-mediated detoxification, orchestrated by transcription factor networks (GATA, FoxA, FTZ-F1), constitutive MAPK hyperactivation (especially MAP4K4-driven cascades), along with preliminary emerging findings on non-coding RNA involvement. Countermeasures now integrate synergistic Cry/Vip pyramiding, CRISPR/Cas9-validated receptor knockouts revealing functional redundancy, Domain III chimerization (e.g., Cry1A.105), phage-assisted continuous evolution (PACE), and the emerging application of AlphaFold3 for structure-guided rational redesign of resistance-breaking variants. Future sustainability hinges on system-level integration of single-cell transcriptomics, midgut-specific CRISPR screens, microbiome engineering, and AI-accelerated protein design to preempt resistance trajectories and secure Bt biotechnology within integrated resistance and pest management frameworks. Full article

Stem mustard (Brassica juncea var. tumida Tsen et Lee) is an important economic vegetable in China. Premature bolting induced by temperature fluctuations has become a major cultivation constraint. Photoreceptors (PHRs) serve as critical photosensor proteins that interpret light signals and regulate physiological responses in plants. In this study, five core PHR families, namely F-box-containing flavin binding proteins (ZTL/FKF1/LKP2), phytochrome (PHY), cryptochrome (CRY), phototropin (PHOT) and UV RESISTANCE LOCUS 8 (UVR8) were identified in Brassica species. RNA-seq analysis revealed their expression patterns during organogenesis in B. juncea. Seven candidate PHRs were validated by qRT-PCR in B. juncea early-bolting (‘YA-1’) and late-bolting (‘ZT-1’) cultivars. Agrobacterium-mediated BjuFKF1_1 overexpression (OE) lines resulted in significantly earlier flowering under field conditions. Histochemical GUS staining indicated that BjuFKF1_1 was expressed in seedlings, leaves, flower buds and siliques. Transcript analysis revealed that the expression level of BjuFKF1_1 was up-regulated in all tissues at both the vegetative and reproductive stages, whereas the expression of BjuFKF1_1 interacting protein-encoding genes were down-regulated in flowers. Under blue light, genes encoding interacting proteins (BjuCOL5, BjuSKP1, BjuCOL3, BjuAP2, BjuAP2-1 and BjuLKP2) were up-regulated in flower buds, whereas BjuCOL and BjuPP2C52 were down-regulated in flowers. Developmental stage analysis revealed the up-regulation of five (BjuAP2, BjuCOL3, BjuCOL5, BjuAP2-1 and BjuLKP2) and four (BjuCOL, BjuCOL5, BjuAP2 and BjuLKP2) interaction protein-encoding genes during the reproductive stage under white and blue light, respectively. These findings elucidate the role of BjuFKF1_1 in flowering regulation and provide molecular targets for B. juncea bolting-resistant variety breeding. Full article

Chronotype, an individual’s preferred timing of sleep and activity within a 24 h cycle, significantly influences metabolic health, muscle function, and body composition. This review explores the interplay between circadian rhythms, hormonal fluctuations, and behavioral patterns—such as nutrition timing, physical activity and sleep quality—and their impact on muscle mass, strength, and quality. Evening chronotypes (ETs) are consistently associated with poorer sleep, irregular eating habits, reduced physical activity, and increased risk of obesity, sarcopenia and metabolic disorders compared to morning types (MTs). At the molecular level, disruptions in circadian clock gene expression (e.g., BMAL1, PER2, CRY1) affect protein synthesis, insulin sensitivity, and energy metabolism, contributing to muscle degradation and impaired recovery. The review highlights critical components—targeting chrono-nutrition, sleep quality, and exercise timing—to align lifestyle behaviors with circadian biology, thereby preserving muscle health and improving overall metabolic outcomes. Full article