A series of novel semisynthetic indole-fused taraxerone derivatives (2a-2f) were synthesised from taraxerone isolated from Rhizophora stylosa Griff. and characterised by 1H NMR,13C NMR, and HRMS analyses. Incorporation of the pharmacologically important indole moiety was aimed at enhancing protein-ligand interactions and antibacterial activity. The synthesised compounds, 2a-2f, were evaluated against Gram-positive bacteria (Lysinibacillus sp. and Bacillus circulans) and Gram-negative bacteria (Pseudomonas putida and Salmonella paratyphi). All derivatives exhibited notable antibacterial activity, with minimum inhibitory concentration (MIC) values ranging from 3.12 to 25 µg mL-1. Among them, compound 2e showed the highest potency, with an MIC of 3.12 µg mL-1, approximately two-fold more active than streptomycin sulphate (MIC = 6.25 µg mL-1). Molecular docking studies against the bacterial target protein (PDB ID: 5JZX) revealed favourable binding affinities for compounds 2d-2f. These results identify indole-fused taraxerone derivatives as promising leads for antibacterial drug development.

Phytochemical investigation of the n-hexane extract of Cynanchum acutum afforded one known lupeol fatty acid ester (1), reported here for the first time from C. acutum, and two previously undescribed lupane/cycloartane-type triterpenoid derivatives, including a lupane-type unsaturated fatty acid ester (2) and a C7 alkyl ether cycloartane derivative (3), together with three known triterpenoids (4-6) and one known sterol (7). The structures of the isolated compounds were established by comprehensive analyses of 1D and 2D NMR, EI-MS, and HRESIMS data. Antimicrobial assays using the agar well diffusion and broth microdilution methods revealed marked antimicrobial activities against selected bacterial and fungal strains. Notably, the acetylated lupane derivative 4 exhibited potent antibacterial activity against Bacillus cereus (MIC = 3.2 ± 0.2 μM; MBC = 6.4 ± 0.4 μM), exceeding that of the reference antibiotic amoxicillin. Additionally, the previously undescribed unsaturated fatty acid ester 2 showed strong antifungal activity against Aspergillus niger (MIC = 1.9 ± 0.1 μM). DPPH radical-scavenging evaluation revealed that the norditerpene ketone 6 possessed the highest antioxidant activity (IC₅₀ = 151.7 ± 3.5 μM), supporting the importance of a free C-3 hydroxyl group in radical-scavenging activity. Cytotoxic evaluation against MCF-7 and WI-38 cell lines showed moderate antiproliferative effects, with compound 2 displaying the highest activity against MCF-7 cells (IC₅₀ = 76.8 ± 4.7 μM). Overall, this study expands the chemical diversity of C. acutum and provides further evidence for the biological activities associated with structurally diverse lupane- and cycloartane-type triterpenoids.

Ribosome profiling is a powerful technique to discover translation events, and to reveal translational dynamics at gene and codon level. The procedure includes laborious ribosome isolation steps using density gradient ultracentrifugation. However, ultracentrifuges are not available in every laboratory, and ultracentrifugation is not suitable for high-throughput experimentation. PEG precipitation has been used to purify ribosomes and in this study we have tested whether PEG precipitation can be used as an alternative method to isolate ribosomes for ribosome profiling. We have benchmarked PEG precipitation against spin column-based size-exclusion and standard sucrose density ultracentrifugation methods for ribosome isolation. The methods were tested using the bacterial model system Bacillus subtilis treated with mupirocin, which stalls ribosomes at isoleucine codons, providing a testable reference for ribosome profile comparison. We found that the use of PEG precipitation, size-exclusion spin columns, and a combination of both, yielded ribosome profiles that were comparable to the standard procedure. With these methods, ultracentrifugation can be omitted, facilitating ribosome profiling and enabling high-throughput analyses.

Flax bast biomass is a phloem-derived lignocellulosic substrate whose enzymatic conversion to reducing sugars is limited by a compact non-cellulosic matrix and restricted interfacial accessibility. A sequential plasma-freeze pretreatment (PF) was developed to reduce this recalcitrance by combining plasma-induced surface modification with a subsequent aqueous freezing followed by freeze-drying sequence. Under optimized plasma conditions (215 W, 52 sccm O2, and 4 min), the mass loss was 8.690%. At 72 h, PF achieved a cumulative reducing-sugar release of 174.83 ± 5.90 mg glucose equivalents/g initial dry fiber, 95.8% higher than that of raw flax bast biomass (RF) and 40.2% higher than that of the low-temperature plasma-only control (LTP); its mean value also exceeded those of ultrasonic and supercritical CO2 pretreatments. Comparisons with LTP and the aqueous freezing followed by freeze-drying control (FFD) indicated that plasma mainly increased the exposure of oxygen-containing functional groups and generated fine-scale etching, whereas the subsequent sequence promoted fiber-bundle separation and pore enlargement. PF increased the relative cellulose proportion from 62.53% to 71.08%, while decreasing pectin, lignin, and wax contents to 6.94%, 3.41%, and 2.93%, respectively. The surface O/C ratio increased from 0.2687 to 0.4700, and PF produced the highest specific surface area and total pore volume of 1.433 m2/g and 0.003394 cm3/g, respectively. The cellulose I crystalline structure was retained. PF showed higher Bacillus subtilis OD600 than RF and a larger late-stage rebound in liquid-phase reducing-sugar concentration during Aspergillus niger cultivation. PF offers a low-chemical-input pretreatment route for enhancing enzymatic reducing-sugar release from flax bast biomass.

Chronic inflammation underlies several common diseases although it is inadequately managed by conventional anti-inflammatory therapies that often suppress immune responses and have long-term adverse effects. Innate immune memory encompasses trained immunity and tolerance, which provide a comprehensive framework for persistent maladaptive innate responses that drive the pathogenesis of neurodegenerative disorders, oncological, cardiometabolic, and autoimmune diseases. This Invited Mini-Review elucidates the molecular mechanisms of innate immune memory including epigenetic reprogramming of chromatin configurations, metabolic rewiring towards glycolytic or oxidative metabolic pathways and the signaling of pattern recognition receptors (PRRs) and the NLRP3 inflammasome that reinforce either enhanced or diminished innate responsiveness. We subsequently discuss pharmacological strategies for selective modulation of innate immune memory. Epigenetic modulators including histone deacetylase (HDAC) and Bromodomain and Extra- Terminal Domain (BET) inhibitors may modify aberrant chromatin programs, while metabolic modulators like metformin and statins can influence their cellular bioenergetics and inflammatory response. Direct NLRP3 inflammasome inhibitors including MCC950 and INF-39 target maladaptive NLRP3 inflammasome signaling. MCC 950 was stopped at the preclinical stage due to hepatotoxicity. Early clinical studies of trained immunity include Bacillus Calmette-Guerin (BCG) vaccine and metabolic modulation. These studies provide evidence that innate immunity programming can be altered in humans, but clinical disease modifying efficacy has not yet been established. We examine translational obstacles and provide a strategy for precision immunopharmacology that incorporates molecular biomarkers, cell-type- and disease-specific targeting, and combinatorial methods to achieve selective immune reprogramming. The development of targeted modulators for innate immune memory will require robust biomarker frameworks. Such approaches may provide more effective and durable therapeutic advances for chronic inflammatory disorders while reducing global immunosuppression.

The present study aimed to develop a fermented cucumber juice beverage using Lactobacillus rhamnosus NCDC-347. The fermentation procedure was optimized using Response Surface Methodology (RSM). The independent variables used for optimization were inoculum size (2-5%), pH (5-9), and incubation temperature (30-45 °C), while the responses were Total Phenolic Content (TPC), antioxidant activity, total sugar content and microbial growth. The optimized fermentation conditions were 37.5 °C, pH 7.0, and an inoculum size of 3.5%. Under these conditions, L. rhamnosus NCDC-347 reached approximately 8.0 ± 0.1 Log CFU/mL after 48 h of fermentation. Under the optimized conditions, fermentation significantly reduced the pH from 7.00 ± 00 to 3.29 ± 0.13, and total soluble solids 3.3 ± 0.15 to 1.2 ± 0.05, along with a reduction in total sugar content from 17.94 ± 1.10 g/L to 4.40 ± 0.45 g/L, confirming effective microbial metabolism. A significant increase in TPC from 1.02 ± 0.06 g GAE/L, to 1.5 ± 0.06 g GAE/L was observed which was associated with enhanced antioxidant activity as determined by DPPH (80.79 ± 1.81%), and ABTS (93.43 ± 0.90%) radical scavenging assays. The fermented cucumber juice showed significant inhibitory activity against α-glucosidase (41.66 ± 1.52%), and α-amylase (57.33 ± 2.51%), indicating potential α-amylase and α-glucosidase inhibitory activity in vitro. Gas chromatography-mass spectrometry (GC-MS) was employed for the volatile profiling of fermented and unfermented cucumber juice, which revealed the production of several fermentation-derived compounds. Cell free supernatant of cucumber juice exhibited antimicrobial activity with highest and lowest inhibition against Bacillus subtilis MTCC 441 (16 mm), and pseudomonas aeruginosa ATCC 15692 (09 mm) respectively. The fermented cucumber juice showed cytotoxic activity against MCF-7 cells under the tested conditions. Sensory assessment using a 7-point hedonic scale showed higher mean scores for most evaluated attributes in the fermented juice than in the unfermented juice.

Bacillus anthracis, a Gram-positive, spore-forming bacterium, poses significant public health risks due to its zoonotic potential and historical use in bioterrorism. Pathogenicity of B. anthracis is primarily attributed to its genetic components, including the essential plasmids pXO1 and pXO2. The pXO1 plasmid encodes toxic factors, while pXO2 facilitates immune evasion. The zoonotic potential, as well as potential bio-threat, necessitates investigation into novel therapeutic targets. Through metabolic network analysis, docking, and notably subtractive genomics, researchers have effectively pinpointed pathogen-specific target proteins essential for drug development. Subtractive genomics, in particular, stands out for its cost-effectiveness and ability to identify non-host homologous targets. In this study, we used a core proteome immunoinformatic approach to identify potential therapeutic targets in B. anthracis. A total of 3987 proteins were found to be conserved in B. anthracis and among them, 388 were essential, non-human homologous proteins. These proteins were then subjected to localization and pathway analysis which resulted in 79 unique pathway proteins. Through PPI and druggability analysis, we had identified 17 strain-independent therapeutic target proteins. All the identified proteins are non-homologous to human proteins and, therefore, are potential targets for drug development. These potential target proteins are involved primarily in cell division, peptidoglycan biosynthesis, and putative flagellar assembly, which are essential for bacterial survival and pathogenicity. These identified target proteins could be the foundation for the development of therapeutics against human anthrax.

The increasing prevalence of multidrug-resistant (MDR) bacterial pathogens highlights the need to explore microorganisms from under-investigated environments as potential sources of antibacterial metabolites. In the present study, marine Bacillus isolates were recovered from seawater and screened for antibacterial activity against three MDR Escherichia coli isolates. Twenty isolates were obtained, of which eight exhibited antibacterial activity and were subjected to secondary screening following solvent extraction. Among the tested extracts, the chloroform extract of isolate MBCP-8 showed the strongest inhibitory activity against all three MDR E. coli strains. Based on 16 S rRNA gene sequencing and phylogenetic analysis, MBCP-8 was identified as Bacillus subtilis strain SC2 and deposited in GenBank under accession number PP854588. The active chloroform extract was further fractionated by silica-gel column chromatography, and Fraction 3 exhibited the highest antibacterial activity. Broth microdilution testing was subsequently performed to determine the minimum inhibitory concentration (MIC) of Fraction 3. UPLC-UV-QTOF-MS profiling revealed two prominent molecular features tentatively annotated as discodermolide and 2-undecyl-4(1 H)-quinolinone N-oxide based on mass spectral/database matching. However, these annotations require confirmation using authentic standards and orthogonal structural techniques, including high-resolution MS/MS and 1D/2D NMR spectroscopy, before definitive compound identification. Overall, the findings demonstrate the antibacterial potential of seawater-derived B. subtilis SC2 and provide a basis for further purification, structural validation, mechanistic investigation, and evaluation of its metabolites against MDR pathogens.

High-carbohydrate-high-fat (HCHF) diets help reduce feed costs in intensive aquaculture, but they often induce glycolipid metabolic disorders and hepatopancreatic steatosis in farmed fish. Although probiotics have shown promise in ameliorating metabolic disorders, the mechanisms by which they interact with the gut microbiota-bile acid (BA) axis in fish remain poorly understood. In this study, common carp were fed an HCHF diet to establish a glycolipid disorder model, followed by dietary supplementation with Lactobacillus plantarum (LP) and Bacillus licheniformis (BL) for 8 weeks. Multiomics approaches, including 16S rRNA sequencing, targeted BA metabolomics, and untargeted intestinal metabolomics, were integrated with phenotypic, biochemical, histomorphological, and molecular analyses. HCHF feeding significantly promoted growth but was associated with systemic glycolipid dysfunction. Probiotic supplementation improved glycolipid profiles and antioxidant capacity, alleviated hepatopancreatic lipid accumulation and damage, and activated the hepatic farnesoid X receptor (FXR)/small heterodimer partner (SHP) signaling pathway. Both probiotics reshaped the gut microbiota by enriching the fish-specific beneficial bacterium Cetobacterium somerae and taxa with potential bile salt hydrolase activity, while also modulating key intestinal metabolic pathways. Notably, BL exerted a more pronounced regulatory effect on BA profiles. These findings suggest that LP and BL alleviate HCHF-induced glycolipid disorders in common carp, with the gut microbiota-BA axis serving as a potential key regulatory node, providing a theoretical basis for probiotic application in functional aquafeeds.