Search Results (823)

The effects of static magnetic field (SMF) treatment on the solid-state culture of Sanghuangporus vaninii (SV) were investigated to enhance metabolite production and bioactivity. SMF parameters including intensity, exposure duration, and temperature were optimized, and treatment at 4 mT for 2 h per day produced the most pronounced effects, increasing total flavonoid (TFC), polyphenol (TPC), and triterpenoid (TTC) contents by 61–438% compared with the control. Ultrasonic extraction and semi-preparative chromatography enabled the isolation of three key compounds: D-(+)-trehalose (1), 5,7-dihydroxy-3,4′-dimethoxyflavone (2), and pinolenic acid (3), all of which were elevated following SMF treatment. Importantly, SMF exposure was associated with enhanced inhibitory activities against enzymes relevant to chronic metabolic disorders. The overall inhibitory activities against α-amylase, α-glucosidase, pancreatic lipase, and xanthine oxidase increased by 6–28% compared with the control, reaching a maximum inhibition of 97.60 ± 0.17%. Preliminary in vitro screening at 100 μg/mL showed that compounds 1 and 2 inhibited both α-amylase and α-glucosidase, whereas compound 3 selectively inhibited pancreatic lipase. Subsequent IC₅₀ analysis confirmed that compound 2 under SMF treatment exhibited inhibitory activity comparable to acarbose against α-amylase (45.62 μg/mL vs. 52.18 μg/mL) and α-glucosidase (38.74 μg/mL vs. 35.42 μg/mL). In addition, compound 3 showed moderate inhibition of pancreatic lipase with an IC₅₀ value of 42.15 μg/mL. These findings suggest that SMF treatment may enhance metabolite production and in vitro enzyme inhibitory activity in S. vaninii. However, these results are limited to in vitro assays, and further studies including cellular and in vivo validation, toxicity assessment, and pharmacokinetic evaluation, are required before any therapeutic or industrial applications can be considered. Full article

The industrial application of starter cultures requires stable physiological and genetic performance. In this study, Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus were continuously subcultured. Physiological stability was assessed through colony morphology, fermentation activity, and growth profiling. Genetic stability was evaluated through comparative genomics of carbohydrate metabolism networks and single-nucleotide polymorphism (SNP) analysis. The results showed that after 2000 generations, the cellular morphology of the strains remained intact. Additionally, the strains exhibited enhanced growth performance and fermentation capability. The Gompertz model revealed that adapted S. thermophilus A37 and L. bulgaricus B29 exhibited shortened lag phases, increased maximum specific growth rates, and high stationary-phase cell densities. Phenotypic microarray and comparative genomics revealed that S. thermophilus mainly used mono- and disaccharides, with impaired ribose metabolism due to the absence of the rbsk gene in the pentose phosphate pathway. In contrast, L. bulgaricus metabolized diverse oligosaccharides, sugar alcohols, and plant-derived substrates. Additionally, it effectively catabolized ribose through the phosphoketolase pathway and possessed a trehalose degradation cluster. All strains exhibited genomic stability, with SNPs revealing fewer than 21 variations per isolate. This study provides an important theoretical foundation for evaluating the stability of fermentation starter cultures. Full article

Probiotics hold considerable promise for treating and preventing inflammatory disease; however, their application is often limited by unclear anti-inflammatory mechanisms and reduced viability following lyophilization. In this study, I thoroughly evaluated the anti-inflammatory potential of Lactiplantibacillus plantarum FS4722 (L. plantarum FS4722) and substantially enhanced strain viability through optimization of the lyoprotectant formulation. Functional assays demonstrated that the fermented supernatant, heat-inactivated bacterial suspension, and cell lysate derived from L. plantarum FS4722 effectively suppressed transcription and expression of inflammatory cytokines in LPS-stimulated RAW 264.7 macrophages. The fermented supernatant exhibited the strongest inhibitory effects, surpassing the reference probiotic Lacticaseibacillus rhamnosus GG (LGG). Mechanistic investigations revealed that anti-inflammatory activity is primarily mediated via inhibition of the MAPK and NF-κB signaling pathways. Furthermore, using component screening combined with response surface methodology, the lyoprotectant formulation (10.00% trehalose, 1.00% sodium carboxymethyl cellulose, and 5.00% skim milk) was optimized, resulting in a lyophilization survival rate of 82.32% while maintaining cellular integrity; in this accelerated stability assessment, the strain retained 78.89% of its activity after 28 days of storage at 4 °C. Collectively, this study provides a robust and efficient approach for probiotic formulation while systematically elucidating the underlying anti-inflammatory mechanisms, thereby offering practical guidance for the development and clinical application of high-performance probiotic products. Full article

CRISPR-Cas12a enables rapid and specific detection of PCR/LAMP (loop-mediated isothermal amplification) reaction products; however, this approach often requires open-tube manipulation, rendering it prone to cross-contamination. Here, we developed a novel one-pot reaction system that eliminated carryover contamination and facilitated endpoint detection using a CRISPR/Cas12a-based system. We leveraged the dependence of the CRISPR-Cas12a cleavage system on the protospacer-adjacent motif (PAM) to design PCR/LAMP primers that incorporated the PAM site (TTT) into amplified DNA. Pre-incubation of Cas12a with crRNA1 and crRNA2 using PCR/LAMP resulted in efficient cleavage of cross-contaminating DNA, while the target gene remained intact due to the lack of PAM sites. Furthermore, a Cas12a-detection complex (comprising Cas12a, crRNA3, trehalose, and the ssDNA probe) pre-stored on the lid was introduced to mix with the PCR/LAMP amplicons, which triggered the non-specific cleavage of fluorescent probes for direct visual detection under a blue LED instrument. This method effectively degraded up to 10⁶ copies of carryover contaminants within one hour, demonstrating the potential of one-pot detection methods in complex samples. Full article

Chinese yam (Dioscorea opposita Thunb.) is a nutrient-rich tuber with recognized health benefits, yet its application in beverage products remains limited due to processing and formulation challenges. In this study, a sequential fermentation strategy was adopted, using Saccharomyces bayanus followed by Lactobacillus brevis to enhance microbial viability and metabolic activity in Chinese yam juice. Samples were collected as an unfermented control (CY), yeast-fermented juice (SP), and sequentially fermented juice (LB). Microbial analysis showed that sequential fermentation supported high LAB viability, reaching 8.92 log CFU/mL in LB, accompanied by a progressive decrease in pH from 5.67 (CY) to 4.27 (LB). Untargeted LC-MS/MS metabolomics identified 1442 metabolites and revealed distinct shifts in the metabolic composition of CY, SP, and LB, indicating stage-dependent modifications of metabolic pathways. Targeted analyses confirmed substantial depletion of sucrose and maltose during fermentation, while trehalose accumulated from undetectable levels in CY to 5.23 mg/g in SP and 7.49 mg/g in LB. Organic acid profiling demonstrated marked increases in lactic and succinic acids, consistent with microbial carbohydrate metabolism. Total phenolic and flavonoid contents increased by 58% and 30%, respectively, while antioxidant capacity (DPPH, ABTS, and FRAP) improved by up to 120% after sequential fermentation. The final fermented beverage (LB) contained a low ethanol concentration of 0.8% (v/v). Sensory evaluation indicated that sequential fermentation improved the overall flavor, aroma, and acceptability of the Chinese yam juice. These findings demonstrate that sequential fermentation with S. bayanus and L. brevis effectively enhances the bioactive composition and antioxidant potential of Chinese yam juice, supporting its development as a functional fermented beverage. Full article

Sclerodermus guani Xiao et Wu, 1983 plays a significant role in the biological control of agricultural and forestry pests. To investigate whether different sugar types significantly affect the longevity, fecundity, and nutrient reserves of female S. guani adults, this study provided 1 mol/L solutions of sucrose, fructose, glucose, mannose, or trehalose under laboratory conditions, with a distilled water group serving as the control. The longevity and nutrient content of parasitoids were measured after varying feeding durations, while fecundity was assessed in preliminary experiments. The results demonstrated that prolonged sugar feeding significantly extended parasitoid longevity, with fructose, glucose, and sucrose exhibiting the most pronounced effects and no significant differences among them. Nutrient analysis revealed that sugar consumption significantly increased total carbohydrate content, slowed lipid depletion, and promoted protein accumulation. Sucrose, fructose, and glucose outperformed other sugars and the control in these aspects. Fecundity assays indicated that glucose and trehalose significantly shortened the pre-oviposition period and enhanced egg production. In conclusion, nutritional supplementation markedly improves the longevity and reproductive performance of S. guani, with 1 mol/L glucose identified as the optimal dietary source. Full article

Pyriproxyfen, a juvenile hormone analog insecticide, poses severe risks to non-target silkworms (Bombyx mori), as evidenced by disrupted metamorphosis—a process strictly dependent on chitin synthesis and its precursor trehalose. However, the specific molecular interference of pyriproxyfen in these metabolic pathways remains unclear. This study investigated the transcriptional response of silkworm midguts to pyriproxyfen using RNA-Seq and validated spatiotemporal gene expression via qRT-PCR. By integrating transcriptomic data with long-term spatiotemporal profiling, we revealed novel tissue-specific expression dynamics. RNA-Seq identified 2059 differentially expressed genes, primarily enriched in metabolic pathways. Spatiotemporal analysis revealed that most chitin- and trehalose-related genes generally exhibited a biphasic “suppression–compensation” trend (initial downregulation followed by upregulation). Notably, tissue-specific responses were evident, with ChsA being continuously suppressed in the middle silk gland, which may be associated with impaired sericin secretion, while showing abnormal upregulation in the posterior silk gland. Additionally, trehalose metabolism genes (Treh and Tret) paralleled the fluctuation of chitin genes, indicating systemic metabolic reprogramming. These results suggest that the toxicity of pyriproxyfen is associated with a decoupling of trehalose metabolism from chitin synthesis and the induction of tissue-specific metabolic disorders. The tissue-specific, long-term spatiotemporal profiling of chitin and trehalose genes presented in this study fills a critical knowledge gap. This study characterizes the transcriptional profile associated with pyriproxyfen toxicity and provides a robust molecular reference for assessing its environmental risks to beneficial insects. Full article

Vitis amurensis is a cold-hardy wild grape species and represents valuable germplasm for breeding cold-tolerant grapevines. In this study, we identified a highly expressed gene (VaTPS9) in one-year-old shoots of V. amurensis ‘Shuangfeng’ during overwintering, but its biological function remained unclear. Temporal and spatial expression analyses revealed distinct expression patterns of VaTPS9 among different tissues from June to November, with the highest transcript abundance detected in one-year-old shoots in November. Gene cloning and sequence alignment showed that VaTPS9 encoded a type II trehalose-6-phosphate synthase (TPS) and was designated as VaTPS9. Functional analyses demonstrated that overexpression of VaTPS9 enhanced cold tolerance in yeast, Arabidopsis thaliana, and V. amurensis callus tissues. Conversely, virus-induced gene silencing (VIGS) of VaTPS9 in grapevine plantlets markedly increased cold sensitivity under low-temperature stress. These reciprocal gain- and loss-of-function phenotypes indicate that VaTPS9 positively regulates cold tolerance, likely by modulating trehalose metabolism and associated physiological responses, including reactive oxygen species (ROS) homeostasis. Collectively, our findings provide new insights into the molecular basis of cold adaptation in wild grape species and highlight VaTPS9 as a promising candidate gene for improving cold tolerance in cultivated grapevine. Full article

Tuberculosis, caused by Mycobacterium tuberculosis, remains a global health challenge, particularly due to multidrug-resistant strains. Photodynamic therapy using porphyrin-based photosensitizers offers a promising alternative by targeting the trehalose-rich cell wall of the bacillus. Motivated by prior experimental observations that shorter linkers improve efficacy, this study probes the molecular basis of linker-length-dependent activity in trehalose–porphyrin glycoconjugates. Here, we show that shorter linker lengths are consistent with improved activity in vitro and, in an Ag85B docking model, constrain conformational flexibility, reduce solvent exposure, and promote tighter packing consistent with stronger predicted interactions. Using computational docking, we analyzed binding scores, RMSD variability, steric clashes, and protein–ligand interactions for conjugates docked into Ag85B, a key enzyme in cell wall synthesis. Shorter linkers (0–2 carbons) were found to exhibit superior binding scores, lower RMSD variability, and stronger interactions with residues such as ARG 43, including unique π–cation interactions. In contrast, longer linkers displayed increased flexibility, reduced binding specificity, and greater solvent exposure. These findings, which support our experimental observations, suggest a molecular basis for linker-dependent efficacy and provide a framework for designing next-generation porphyrin-based therapeutics for tuberculosis treatment. Full article

Anaerobic germination tolerance (AGT) is a critical adaptive trait for rice establishment in flood-prone environments and direct-seeded systems. Here, we identified and validated the quantitative trait locus qAG2.1 for AGT and introgressed it into the elite lowland rice variety CR Dhan 801 through marker-assisted backcross breeding. The introgressed lines exhibited significantly improved germination under anaerobic conditions, demonstrating the effectiveness of qAG2.1 in a high-yielding genetic background. While CR Dhan 801 showed a low anaerobic germination percentage (17.6%), the donor ARC10424 exhibited 82.6%, and the best-performing introgressed line (22009-3) achieved 49.2%. Importantly, the improved lines maintained agronomic performance comparable to CR Dhan 801 under non-stress conditions, indicating minimal yield penalty. To gain mechanistic insight, the qAG2.1 interval was dissected in silico to prioritise candidate genes putatively associated with AGT. This analysis highlighted genes linked to ethylene biosynthesis and signalling (e.g., OsACO3, OsERF109), abscisic acid biosynthesis (OsNCED1), gibberellin homeostasis (OsGA2ox9), trehalose metabolism (OsTPS5, OsTPP1), detoxification of anaerobic by-products (OsALDH2A), and water transport (OsPIP1;3). Collectively, these results validate qAG2.1 as a further deployable locus for improving anaerobic germination in elite rice backgrounds and provide a set of putative candidate genes for future functional characterisation. Full article

Crystal dumpling wrapper production is hampered by rapid surface dehydration, severe freeze-cracking propensity, and storage-induced retrogradation. Modulation of blended starch properties through functional additives was investigated. This study systematically evaluated the impact of hydroxypropyl distarch phosphate (HPDSP), trehalose (TRE), guar gum (GG), and composite phosphates (CP) on physicochemical and structural properties of wheat–potato starch composite gel. Concurrently, the effects of additives on the cracking rate of crystal dumplings and texture of wrappers were investigated. Analysis revealed that apparent viscosity was increased by all additives except CP. Different additives significantly improved the freeze–thaw stability of the composite gel during the first three cycles. GG maintained enhanced freeze–thaw stability throughout the entire freeze–thaw cycle (dehydration shrinkage rate: 2.69–40.55%). Multivariate analytical techniques (SEM, FTIR, XRD, DSC) collectively indicated that the additives effectively inhibited starch retrogradation, whilst HPDSP showed the strongest retrogradation inhibition. CP enhanced water-retention capacity and produced a softer blended gel (hardness at 21 days was 100.56 gf). Furthermore, additives significantly reduced the freezing cracking rate of crystal dumplings and improved the textural properties of dumpling wrappers. Full article

Background: Thermostability remains a key bottleneck for equitable access to mRNA-LNPs vaccines, mainly due to cold-chain requirements. Objectives and methods: Here, we optimized freeze-drying formulations by screening excipients (sugars, sugar alcohols, and proteins) and buffers to preserve mRNA-LNPs as solid formulations under ambient and refrigerated conditions. Physicochemical properties (size, polydispersity index [PDI], and encapsulation efficiency [EE]) and functional integrity, assessed by fluorescence-based in vitro transfection assays, were evaluated during long-term storage of up to six months. Results: Preliminary screening identified 20% sucrose and trehalose with Tris or histidine buffers as optimal for preserving physicochemical properties during freeze-drying, including high encapsulation efficiency (>90%), particle size (~200 nm), and low polydispersity (PDI < 0.2). Mannitol, gelatin, and PBS-based buffers showed adverse effects. At 4 °C, formulations F1–F3 maintained physicochemical stability and functional transfection activity for up to four months. In contrast, 20 °C storage caused progressive destabilization, with increased size, PDI, and encapsulation loss (>60% by six months). Among all formulations, 20% sucrose with 5 mM Tris (F1) showed the most robust preservation of physicochemical integrity and in vitro transfection efficiency under refrigerated and ambient conditions. Conclusions: Sugars outperformed sugar alcohols and gelatin as cryoprotectants. All formulations were stable, including functionally active at 4 °C for up to four months, while a sucrose/Tris formulation retained acceptable stability at 20 °C. Overall, the results demonstrate the feasibility of storing mRNA drug products as solid formulations at non-freezing temperatures. Full article

Tachykinins (TKs), a conserved family of neuropeptides, play critical roles in regulating multiple physiological processes such as feeding and energy metabolism in insects. This study identified the TK gene (ApTK) from the Chinese oak silkworm, Antheraea pernyi, an economically important insect species. Bioinformatic analysis showed that ApTK possesses four FX₁GX₂R motifs (X₁ and X₂ represent variable amino acid residues), comprising FMGVR, FYGVR, FIGVR, and FFGMR, in the C-terminus and shares a close phylogenetic relationship with TKs from Bombyx mori and Manduca sexta. Tissue-specific expression profiling demonstrated that ApTK was mainly expressed in the brain and midgut. Starvation–refeeding experiments showed that the expression of ApTK was significantly upregulated during food deprivation and returned to baseline after refeeding, evincing its involvement in hunger signaling. RNA interference (RNAi)-mediated knockdown of ApTK led to a significant increase in larval body weight and increased levels of triglyceride, glycogen, and trehalose, indicating enhanced energy storage. Collectively, these results demonstrate that ApTK acts as a key regulator in restraining feeding and modulating energy homeostasis in A. pernyi. Our findings provide insights into the neuroendocrine mechanisms underlying feeding behavior and energy metabolism in A. pernyi. Full article

Cold tolerance of natural enemy insects is a critical determinant of their overwintering survival and efficacy in biological control. The green lacewing (Chrysoperla nipponensis) is an important natural enemy insect that overwinters as adults in nature; however, its high overwintering mortality severely limits its effective application in spring. To investigate the molecular mechanisms underlying low-temperature adaptation, this study focuses on the temperature-sensitive Transient Receptor Potential (TRP) channels and their roles in the cold tolerance of C. nipponensis. The TRPA subfamily gene, Pyrexia-1, was identified and found to be significantly downregulated upon cold exposure. A functional analysis indicates RNAi-mediated knockdown of Pyrexia-1 significantly lowered both the supercooling point and the freezing point of C. nipponensis adults, enhancing their survival rate at −10 °C. These results indicate Pyrexia-1 as a negative regulator of cold tolerance. Further mechanistic investigation revealed that inhibition of Pyrexia-1 function specifically down regulates the expression of trehalase (TRE1) genes, resulting in a marked accumulation of the cryoprotectant trehalose in adults. This metabolic adjustment was accompanied by the upregulation of heat shock protein Hsp70. Overall, these findings establish Pyrexia-1 as a critical molecular switch linking temperature-sensing signals to the metabolic pathways governing freeze resistance, thereby orchestrating the systemic cold adaptation in C. nipponensis. This discovery provides novel insights into the molecular basis of insect low-temperature adaptation and suggests a potential strategy for enhancing the overwintering capacity of natural enemy insects by targeting this regulatory node. Full article

Mesenchymal stem cells (MSCs) are candidates for the treatment of palatal wounds in combination with biomaterials. In this study, we developed a method for the production of a ready-to-use mucoadhesive hydrogel containing MSCs for palatal wounds and evaluated its healing effects. Dental follicle MSCs (DFMSCs) were isolated from the dental follicle tissue of a healthy twenty-year-old donor. DFMSCs were suspended in a cell-preserving solution containing platelet-rich plasma, trehalose, and DMSO, and loaded into a catechol–chitosan hydrogel solution at a ratio of 1:400 (v/v) with 5 × 10⁵ or 6 × 10⁶ cells per hydrogel to create a novel lyophilization method for cell integration into the biomaterial. Hydrogels were fabricated as scaffolds with a diameter of 5 mm and a depth of 4 mm. After lyophilization of the hydrogels with cells, a viability test was performed after the production of hydrogels on the seventh day and the fifth month. Palatal wounds were created as full-thickness wounds in rats using a 5 mm diameter punch. The hydrogels were applied to the palatal wounds of rats, and histochemical and immunohistochemical analyses were performed. The results showed that, after rehydration of the hydrogels, DFMSCs had over 80% viability and were homogeneously distributed in the hydrogels. After the application of DFMSC-loaded hydrogels, palatal wounds healed within 7 days, and inflammatory cell infiltration, fibroblastic proliferation, and ulceration were significantly reduced, while epithelial regeneration was significantly increased compared to the control group. The viability ratio of DFMSCs was 83.7% on the seventh day and 71.3% in the fifth month. Hydrogels loaded with DFMSCs represent a promising, ready-to-use biomaterial approach for supporting palatal wound healing. Full article