Search Results (55,871)

Flavor esters are valuable compounds widely used in the food, beverage, and cosmetics industries for their aroma and flavor-enhancing properties. Traditional methods of obtaining these compounds, such as extraction from natural sources or chemical synthesis, present challenges related to cost and toxicity, respectively. Enzymatic synthesis, particularly using immobilized lipases, offers a sustainable and efficient alternative. This study investigates the application of CRL immobilized on Diaion HP-20 for geranyl butyrate synthesis via esterification of geraniol and butanoic acid using Candida rugosa lipase (CRL) immobilized on Diaion HP-20 (CRL-DHP-20). The immobilization process resulted in a protein loading of 29.6 ± 2.2 mg/g support from an initial 40 mg/g, and the immobilized biocatalyst exhibited a hydrolytic activity of 124.0 ± 2.5 U/g using olive oil emulsion. Reaction conditions were optimized through a central composite design, evaluating the influence of biocatalyst concentration, temperature, and agitation on ester conversion. The optimal conditions (13.4% CRL-DHP-20, 48.2 °C, and 220.1 rpm) led to 85.4% conversion in 360 min. Additionally, CRL-DHP-20 retained 84% of its initial activity after six reaction cycles, indicating good operational stability. These findings highlight the potential of CRL-DHP-20 as an effective and reusable biocatalyst for green synthesis of flavor esters. Full article

1-Deoxy-D-xylulose-5-phosphate synthase (DXS) serves as the initial rate-limiting enzyme in the methylerythritol phosphate (MEP) pathway, governing the biosynthesis of precursors for photosynthetic pigments and terpenoids. In this study, the LaDXS2-2 gene was cloned and functionally characterized in lavender (Lavandula angustifolia). The full-length coding sequence (CDS) of LaDXS2-2 spans 2178 base pairs, encoding a protein of 725 amino acids. Phylogenetic analysis revealed that LaDXS2-2 is most closely related to the DXS from Salvia miltiorrhiza. Expression profiling demonstrated that LaDXS2-2 was highly expressed in flower buds, and its transcript levels were significantly upregulated (p < 0.05) in response to ethephon, high light intensity, and low temperature, while exhibiting tissue-specific responses to gibberellin application. Subcellular localization assays confirmed LaDXS2-2 is targeted to the chloroplast. Heterologous overexpression of LaDXS2-2 in tobacco resulted in a marked increase in photosynthetic pigment content, enhanced the actual photochemical efficiency of photosystem II [Y(II)], and reduced non-photochemical quenching (NPQ). Integrated transcriptomic and metabolomic analyses further revealed that LaDXS2-2 overexpression activated the diterpenoid biosynthesis pathway and upregulated amino acid metabolism as well as the TCA cycle, while competitively suppressing phenylpropanoid and flavonoid biosynthesis pathways. These findings indicate that LaDXS2-2 not only enhances photosynthetic efficiency by promoting the synthesis of photosynthetic pigments but also suggests a potential role in influencing primary carbon and nitrogen metabolism, as inferred from transcriptomic and metabolomic data. This functionality may ultimately influence plant growth and metabolic homeostasis. Overall, this study provides a theoretical foundation for the synergistic improvement of photosynthetic efficiency and secondary metabolism in crops. Full article

The biosynthesis of sex pheromones in lepidopteran pheromone glands is tightly regulated by pheromone biosynthesis-activating neuropeptide (PBAN) signaling; yet the contribution of non-coding RNA-mediated post-transcriptional regulation remains largely unclear. This study aimed to characterize temporal transcriptomic changes, candidate non-coding RNA-mediated regulatory associations, and temporal molecular dynamics underlying transcriptional remodeling after PBAN treatment in Ostrinia furnacalis. First, we performed comprehensive whole-transcriptome sequencing (WTS) on 18 biologically independent samples collected at six time points (0, 20, 40, 60, 90, and 120 min) after PBAN injection. Then, we systematically identified and quantified the dynamic expression patterns of differentially expressed (DE) mRNAs, miRNAs, lncRNAs, and circRNAs in response to PBAN stimulation. By integratively analyzing these multidimensional omics datasets and inferring sequence-based interaction relationships, we inferred a dynamic candidate competing endogenous RNA (ceRNA) like regulatory network. The candidate ceRNA network anchored four core node genes: the PBAN receptor (PBANR), the rate-limiting enzyme acetyl-CoA carboxylase (ACC), and the terminal biosynthetic enzymes desaturase (DES) and fatty acyl-CoA reductase (FAR). The qRT-PCR results further support the temporal expression pattern of key genes during the PBAN response, suggesting that this network can provide a valuable resource for further functional studies. Full article

High-Mobility Group Box (HMGB) proteins belong to the family of high-mobility proteins characterized by two DNA-binding domains and an unstructured, negatively charged C-terminal domain that modulates DNA–protein and protein–protein interactions. These proteins participate in multiple cellular processes, including DNA replication, transcription, recombination, and repair. The functional activity of HMGB proteins is associated with various physiological and pathological conditions, including malignant tumors and cardiovascular diseases, highlighting the need for strict regulation of their levels and activity to maintain cellular homeostasis. Such regulation can occur at multiple levels, including proteolytic degradation. In recent years, a number of E3 ubiquitin ligases that promote the degradation of HMGB proteins, as well as deubiquitinases (DUBs) that stabilize them by removing ubiquitin tags, have been identified. This review summarizes these enzymes and their proposed roles in controlling the functions of the HMGB family proteins, both through direct interaction with these proteins and via mediator proteins. Full article

Endogenous gene tagging using CRISPR has changed the understanding of the role played by different proteins due to the ability to track and study proteins in their natural state. With CRISPR-based gene tagging, it is possible to insert fluorescent, luminescent, epitope, affinity, and proximity labels into the target protein at its endogenous genomic location without affecting its physiological expression and dynamics. Here, we discuss the DNA-repair mechanisms employed in endogenous gene tagging, including homology-dependent repair, NHEJ-based integration, and alternative approaches that can be used with challenging cell types. Key aspects of efficient CRISPR tagging experiments are also described. Additionally, we review recent advances in the increasing array of protein tag technologies, including fluorescent proteins, split-reporter technologies, NanoLuc/HiBiT, peptide epitopes, and proximity biotinylation enzymes. Lastly, we review the scalability of endogenous tagging approaches using multiplex editing, atlas-scale proteome tagging, iPSC-based disease modeling, and drug discovery platforms for assessing target engagement, protein degradation, phenotype screening, and mechanism of action of compounds. Although difficult in primary and pluripotent cells, new methods based on avoiding double-strand breaks, such as prime editing, PASTE, and CRISPR associated transposases, will drive the future expansion of endogenous tagging approaches. Such developments firmly set up CRISPR gene tagging as a fundamental technology in quantitative cell biology and translational pharmacology. Full article

Demand for greener and safer chemistries has driven the innovation of bioinspired and enzyme-mimicking catalysts for selective and efficient oxidation and hydrogenation under mild conditions. Natural catalysts, including peroxidases, oxidases, hydrogenases, oxygenases and dehydrogenases, boast remarkable activity, specificity, stability, selectivity, low energy requirements and atom economy. Disadvantages of enzymes, such as poor thermal stability, a narrow operational range, low recovery yield and the expense of purification, are motivating the discovery and design of enzyme substitutes. Several artificial platforms have appeared recently: nanozymes, artificial metalloenzymes, biomimetic metal Complexes, MOFs, atomic catalysts, bioinorganic hybrid systems, among others. These systems aim to replicate key structural and mechanistic features of enzymes while providing greater operational stability, recyclability, and scalability. Recent work has demonstrated the benefit of enzyme mimics in increasing eco-sustainability in reactions such as alcohol oxidation, selective alkane oxidation, waste degradation, catalytic photooxygen activation and biomass waste conversion. Similarly, biomimetic hydrogenation catalysts have shown outstanding activity in asymmetrically hydrogenating chemicals, reducing CO₂ into chemicals, hydrogenation by hydrogen transfer and creating hydrogen through water. Through control of active sites, second coordination sites, defects and electrons/protons in the system, significant gains have been seen in reaction selectivity and frequency of turning over substrate into product. Nanozymes, biohybrid catalysis and artificial catalysts guided by deep learning are further broadening the applications of biomimetic catalysis in oxidation and hydrogenation. The article review aims to provide a summary of the most current progress with bioinspired and enzyme-mimicking catalysts, focusing on catalytic mechanisms, how to design such catalysts, how green chemistry benefits from their development and where further application is likely in the coming years. Full article

The genus Tamarix L. includes several species widely used in traditional medicine for their therapeutic properties. This study aims to evaluate the bioactive potential of Tamarix gallica extracts from Western Algeria using an integrated in vitro and in silico approach. GC–MS analysis with BSTFA derivatization was performed to characterize the chemical profile of the methanolic fraction. In addition, total phenolic, flavonoid, and tannin contents were determined in methanolic extracts of leaves and stems. The biological activities were assessed using antioxidant (DPPH, ABTS, β-carotene, FRAP, O-phenanthroline, and cupric reducing assays), antimicrobial, antidiabetic, and anti-Alzheimer in vitro assays. Molecular docking was conducted to evaluate the inhibitory potential of selected flavonoids against α-amylase, acetylcholinesterase, and butyrylcholinesterase. Results revealed a rich metabolite profile dominated by long-chain aliphatic alcohols (including hentriacontan-12-ol), phytosterols (β-sitosterol), fatty acids, phenolic derivatives, and sugar alcohols. The extracts exhibited strong antioxidant activity (IC₅₀ = 1.34 ± 0.43 and 12.32 ± 0.36 μg·mL⁻¹), significant antimicrobial effects against the tested pathogens, and notable antidiabetic and anticholinesterase activities (IC₅₀ = 78.65 ± 1.43 and 98.37 ± 1.07 μg·mL⁻¹). Molecular docking analysis supported these findings, showing strong binding affinities of quercetin and rhamnetin toward the target enzymes. Overall, T. gallica exhibits promising multifunctional bioactivities with potential pharmaceutical relevance. Full article

Chilling injury is a major problem limiting the postharvest storage and marketability of mango fruit at low temperature. The present study investigated the individual and combined effects of sodium nitroprusside (SNP), L-arginine (Arg) and salicylic acid (SA) on chilling tolerance, regulation of oxidative stress and the postharvest quality of ‘Keitt’ mango fruit stored at 5 ± 1 °C for 28 days followed by 4 days of shelf life at 23 °C. Fruits were pre-treated with 1 mM SNP, 1 mM Arg, 2 mM SA or their binary combinations before storage. The chilling injury, membrane damage, lipid peroxidation, protein oxidation and fruit softening were greatly enhanced by cold storage in untreated fruits. In contrast, all the treatments significantly ameliorated these deteriorative changes, and the combined treatments were superiorly effective. Among these, SNP + Arg was the most effective treatment, which reduced the chilling injury index from 4.05 in control fruits to 1.00 after shelf life, completely inhibiting the incidence of decay and reducing electrolyte leakage and malondialdehyde accumulation by 47.4 and 48.2%, respectively. The same treatment also maintained higher firmness, titratable acidity, visual appearance and ascorbic acid content than untreated fruits. The enhanced chilling tolerance was accompanied by increased antioxidant defense, as SNP + Arg significantly stimulated the activities of superoxide dismutase, catalase and peroxidase, but suppressed the activity of pectin methylesterase. Multivariate analyses, such as PCA, clustered heatmap and integrated stress index, demonstrated a strong negative relationship between oxidative stress markers and antioxidant metabolism. The results showed that combined SNP and Arg treatments enhanced chilling tolerance through increasing antioxidant capacity, preserving membrane integrity, and retarding ripening-related metabolism, which provides an effective way to maintain the postharvest quality of cold-stored mango fruit. Full article

Certain sacoglossan sea slugs, often known as “solar-powered sea slugs”, are a group of marine gastropods that have the unique ability to photosynthesize by stealing functional chloroplasts from algae. The sacoglossan Elysia papillosa can maintain functional chloroplasts for up to two weeks after feeding. The microbiome of these slugs may play a crucial role in their metabolism, immunity, development, but more importantly their photosynthesis. Shotgun metagenomic sequencing was conducted on four samples of E. papillosa in order to characterize their microbiome. Sequences were classified and relative abundance was quantified with Centrifuger and functional data was examined using SqueezeMeta. Bacteria were analyzed by taxonomic groups and hypothesized function to the sea slug was determined with literature analysis. All samples were dominated by phyla Actinomycetota, Bacillota, Patescibacteriota, and Pseudomonadota. The presence of the phyla Bacteroidota and Bacillota was notable in all samples, which contain species known to produce enzymes that break down polysaccharides. It is possible that these bacteria could assist in degradation of the polysaccharide xylan found in the cell walls of Penicillus, the algal food source of E. papillosa. One species that was found in all samples was Cutibacterium acnes which has been shown to be an important component of the gut microbiota in other marine invertebrates and may provide the host with vitamin B12 and other beneficial nutrients. Many of these bacteria may be opportunistic rather than commensal. As a result, more research is required to describe the interactions between the slug and its microbiome, but this preliminary report provides a valuable starting point for identifying the microbiome make-up to further understanding of these relationships. Full article

Mounting acaricide resistance in Tetranychus mexicanus (McGregor) (Acari: Tetranychidae), among the most damaging phytophagous mites in tropical and subtropical crops, has intensified the search for botanical alternatives. An oil-in-water nanoemulsion of Thymus vulgaris essential oil (TVEO-NE) was developed and evaluated for lethal and sublethal effects on adult females of T. mexicanus. TVEO, composed mainly of thymol (45%) and p-cymene (37%), was formulated by low-energy emulsification yielding stable dispersions (~200 nm; PDI < 0.25; zeta potential of −22.2 mV). At 30.0 mg a.i./mL, TVEO-NE caused 68.3% corrected mortality at 72 h and suppressed fecundity by ~44–52%; vehicle controls exerted only moderate effects, identifying the essential oil as the primary bioactive driver. Morphological examination revealed collapse of female idiosomata and disruption of excretory pellet architecture, corroborating the bioassay data. Molecular docking against a cathepsin L homology model revealed that thymol and p-cymene interact exclusively via hydrophobic contacts and display substantially lower ChemPLP fitness scores than the reference cysteine protease inhibitor E64, indicating weak predicted binding affinity and arguing against enzyme inhibition as the primary mechanism. Taken together, bioassay, morphological, and docking are consistent with supporting membrane partitioning as a plausible primary mode of action, positioning TVEO-based nanoemulsions as promising botanical tools for T. mexicanus management. Full article

Bonellia viridis, an echiuran polychaete that inhabits infralittoral rocky habitats around the Atlantic, Mediterranean, and Southeastern Pacific coastlines, exhibits environmentally mediated sexual dimorphism: planktonic larvae develop into dwarf males after exposure to bonellin, a green pigment produced by adult females. Bonellin is a chlorin with a structure consistent with derivation from uroporphyrinogen III, the last universal precursor of all known tetrapyrroles, yet its biosynthesis remains unknown. Here, the de novo genome assembly for a single adult female specimen of B. viridis isolated from Okinawa has been generated (via Illumina sequencing) and found to comprise 429.95 Mb across 95,859 contigs, with an N50 of 6505 bp, recovering 83.3% of near-universal metazoan BUSCO orthologs. Homologs of all canonical enzymes of the heme biosynthetic pathway (termed hem genes) were identified across the genome. The genomic resources establish a foundation for research into the biochemical basis of pigment production, chemically mediated sex determination, and the distinct biology of B. viridis. Full article

Background: Cognitive impairment is a common complication after ischemic stroke and affects patients’ quality of life. Elevated glial fibrillary acidic protein (GFAP) and low vitamin D levels may contribute to neuroinflammation and impaired neuroplasticity, but their association with post-stroke cognitive impairment remains unclear. This study aimed to determine whether high serum GFAP and low vitamin D levels are risk factors for cognitive impairment in ischemic stroke patients. Methods: A prospective cohort study was conducted in patients with acute ischemic stroke. Serum GFAP and vitamin D levels were measured on the third day after stroke onset using an enzyme-linked immunosorbent assay (ELISA). Cognitive function was assessed two weeks after stroke onset using the Indonesian version of the Montreal Cognitive Assessment (MoCA-Ina). Data were analyzed using the chi-square test and multivariate logistic regression. Results: Seventy-six subjects were included in this study, of which 55 (72.4%) developed cognitive impairment. High serum GFAP (≥1.885 ng/mL) (RR = 1.755; 95% CI: 1.252–2.459; p = 0.001) and low vitamin D levels (<16.185 ng/mL) (RR = 1.773; 95% CI: 1.234–2.547; p = 0.001) were both associated with cognitive impairment. Multivariate analysis showed that high GFAP (AOR = 10.039; 95% CI: 2.484–40.569; p = 0.001) and low vitamin D levels (AOR = 6.640; 95% CI: 1.798–24.518; p = 0.005) were independent risk factors. Conclusions: Elevated serum GFAP and low vitamin D levels were independently associated with cognitive impairment after ischemic stroke and may serve as potential biomarkers for early risk stratification. Full article

Vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic (VEXAS) syndrome is an adult-onset inflammatory disorder caused by acquired mutations in UBA1, the gene encoding the primary ubiquitin-activating enzyme. The recognition of VEXAS has transformed the current understanding of autoinflammatory disease by demonstrating that somatic alterations arising within hematopoietic stem cells can precipitate severe, multisystem inflammation in later life. While pathogenic UBA1 variants are essential to disease pathogenesis, many affected individuals also harbor additional somatic mutations associated with clonal hematopoiesis, most commonly involving DNMT3A and TET2. These concurrent mutations may contribute to clonal architecture; however, their independent impact on inflammatory phenotype and hematologic manifestations remains incompletely defined. Emerging evidence suggests that co-occurring clonal hematopoiesis mutations may be independently associated with poorer overall survival, though their causal role remains unestablished. This review examines the evolving genetic framework of VEXAS syndrome, emphasizing UBA1 as the obligate driver mutation while reviewing current evidence regarding non-Met41 UBA1 variants and co-occurring somatic mutations. Full article

Background/Objectives: Tordylium trachycarpum Boiss. (Apiaceae) is traditionally used in Kurdish ethnomedicine for the management of gastrointestinal disorders; however, its pharmacological efficacy and safety profile remain insufficiently investigated. This study evaluated, for the first time, the gastroprotective activity and associated antioxidant, inflammatory, and apoptotic responses of the methanolic extract of T. trachycarpum using an ethanol-induced gastric ulcer model in Sprague–Dawley rats. Methods: Preliminary phytochemical screening revealed the presence of phenolics, flavonoids, terpenoids, tannins, coumarins, and glycosides. Acute oral toxicity testing demonstrated no signs of toxicity at doses up to 5 g/kg. Gastric ulceration was induced by absolute ethanol, and animals were pretreated with the extract (250 and 500 mg/kg) or omeprazole (20 mg/kg). Results: The extract significantly decreased the gastric lesion area from 258.50 ± 6.38 mm² in the ulcer control group to 143.70 ± 0.76 mm² and 115.50 ± 0.76 mm², corresponding to ulcer inhibition rates of 44.41% and 55.31%. Additionally, the extract increased mucus production, maintained mucosal structure, and raised stomach pH. Biochemical analysis showed a significant increase in antioxidant enzymes [superoxide dismutase (SOD) and catalase (CAT)] and a reduction in malondialdehyde (MDA) levels, indicating attenuation of oxidative stress. In addition, the extract modulated pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-10). Blood-based ELISA analysis demonstrated increased expression of heat shock protein 70 (HSP70) and reduced Bax levels, suggesting anti-apoptotic activity. Conclusions: These findings indicate that T. trachycarpum exerts significant gastroprotective activity through antioxidant, anti-inflammatory, and anti-apoptotic mechanisms, supporting its traditional use and highlighting its potential as a natural therapeutic candidate for the management of gastric ulcers. Full article

Background: Type 2 diabetes mellitus (T2DM) is a global health challenge characterized by chronic hyperglycemia and oxidative stress. Pithecellobium dulce root has long been recognized for its antidiabetic potential; however, its specific bioactive constituents and mechanisms of action remain poorly defined. This study aimed to evaluate the antidiabetic and antioxidant properties of extracts and isolated molecules from P. dulce root bark. Methods: The DCM/MeOH crude extract of P. dulce root bark was fractionated with n-hexane (PDEH) and ethyl acetate (PDAE), followed by chromatographic purification and spectroscopic characterization, yielding seventeen compounds (1–17). The antioxidant activity (DPPH, ABTS, FRAP) and antidiabetic potential of PDEH, PDAE, and 1–17 were assessed in vitro using yeast-derived enzymes and in silico (targeting human α-glucosidase [PDB: 2QLY] and human α-amylase [PDB: 4GQR]). The in vitro α-glucosidase experiments used saccharomyces cerevisiae enzyme, which varies from the human target. Therefore, these results should be taken as preliminary screening data that needs confirmation with human enzymes. Results: Compound 1 was identified as new, while 2 was isolated for the first time from a natural source. The cell-free chemical tests DPPH, ABTS, and FRAP measured antioxidant capability. These tests quantify radical-scavenging and electron-transfer capabilities in vitro and are preliminary chemical screening methods. They do not directly represent biological antioxidant activity in cells or organisms. PDEH demonstrated strong radical scavenging against DPPH (IC₅₀ = 15.30 μg/mL) and ABTS (IC₅₀ = 12.80 μg/mL), while pristriol (16) showed ferric reducing power (EC₅₀ = 4200 μM FeSO₄/g). Enzyme inhibition assays demonstrated activity against α-amylase (IC₅₀ 53.88–112.24 µg/mL; acarbose IC₅₀ = 91.20 µg/mL) and α-glucosidase (IC₅₀ 18.38–136.88 µg/mL; acarbose IC₅₀ = 11.31 µg/mL). Compounds 15, 1, and 2 showed superior activity compared to acarbose for α-amylase, with effect sizes (Cohen’s d) of 2.15, 0.94, and 0.82, respectively, and IC₅₀ values of 53.88, 88.15, and 92.62 µg/mL; for α-glucosidase, IC₅₀ values were 18.38, 39.25, and 36.40 µg/mL, respectively. Docking studies supported these findings, revealing binding energies of −9.08, −8.34, and −7.22 kcal/mol for compounds 1, 2, and 15 with α-amylase, and −10.35 and −9.79 kcal/mol for compounds 1 and 2 with α-glucosidase. ADME profiling further identified 1 and 2 as promising lead candidates for dual-enzyme inhibition. Conclusions: P. dulce root bark represents a potent source of bioactive molecules with both antioxidant and dual-enzyme-inhibitory properties. These findings validate its traditional use and highlight its potential in the development of multitarget therapies for T2DM management. Full article