Anthriscus sylvestris (wild chervil) leaf extracts represent a promising natural resource for cosmeceutical biotechnology applications. This study systematically evaluated polarity-based extracts, including hot water, 40
The biopharmaceutical industry has relied heavily on Chinese hamster ovary (CHO) cell lines to produce therapies using primarily fed-batch processes. Exploration of intensification strategies based on perfusion processes to increase volumetric productivity to reduce manufacturing footprint is an area of active investigation. Continuous perfusion processes provide significant increases in volumetric productivity. Operation of cell culture processes in a continuous mode presents unique automation challenges, to drive towards high yields while assuring cell health to enable extended run durations. Typical operating strategies for continuous bioprocesses include the use of a bleed stream to remove cells to maintain a target viable cell density known to provide stable cell health while maintaining a perfusion rate sufficient to supply required substrates. Here we propose a multi-strategy nonlinear model predictive control (NMPC) to allow switching between objectives ranging from stable operation to economic optimization. Economic optimization is primarily achieved through allowing the controller to operate at elevated cell densities and is enabled through use of the cell culture model that tracks accumulation of biomaterials and toxins and their influence on cell growth and death dynamics, allowing accurate prediction of maximum stable viable cell density. It is shown that, compared to a standard reference recipe, economic optimization results in over 68
Sponge‑derived siliceous spicules have emerged as “liquid microneedling” systems that enhance transdermal delivery by forming dense microchannels in the stratum corneum while remaining embedded within the viable epidermis. Unlike solid microneedle arrays, which are removed immediately and whose microchannels rapidly close, individual spicules persist for days, extending the window for permeation of hydrophilic macromolecules that otherwise poorly cross intact skin. Beyond passive barrier disruption, rigid biogenic silica structures modify local tissue mechanics and elicit a self‑limited foreign body response, raising the possibility that they engage platforms with proposed mechanotransductive and immunomodulatory roles relevant to dermal remodeling. This review contextualizes spicule-based liquid microneedling relative to established physical enhancement techniques and summarizes evidence supporting its role as a physical penetration enhancer, while presenting mechanotransduction and immunomodulation as proposed but still incompletely validated mechanisms. We integrate current evidence on mechanotransduction and extracellular matrix homeostasis, with emphasis on YAP/TAZ‑dependent signaling in dermal fibroblasts, and outline how spicule‑induced micro‑injury and foreign body signaling may influence macrophage polarization and local immune microenvironments. Clinical data from a phase 2b randomized, vehicle‑controlled trial of a once‑weekly Spongilla lacustris–based formulation in moderate‑to‑severe acne provide proof‑of‑concept for therapeutic use of sponge‑derived spicules in inflammatory dermatoses. Finally, we highlight emerging functionalized spicule platforms and key biosafety and regulatory considerations that will shape their translation into dermatologic practice.
Molecular dynamics (MD) simulation is a powerful tool for generating protein conformational ensembles, yet the resulting high-dimensional data present significant challenges for traditional atomic-level structural analysis. To address this, we present a complementary and scalable method: the joint-based descriptor (JBD), a macroscopic geometric framework that encodes protein conformation through a minimal set of dihedral angles measured at the joints of secondary structures. Human vitamin K reductase (hVKOR), an essential three-transmembrane (TM) anticoagulant membrane enzyme, served as a model system. We analyzed approximately 2,250 structural models generated from a 200-ns MD simulation. Using three specific joint dihedral angles (Ω1, λ1, and Ω2), the JBD mapped the dynamic topology of the hVKOR ensemble. The analysis revealed tightly restricted helical arrangements and identified a discrete and characteristic λ1 range (–80 to –50) as a characteristic topological signature for hVKOR. We further computed a Jscore metric to quantify conformational distances between hVKOR and other 3TM protein families, enabling hierarchical clustering of closest relatives while separating distant topologies. Finally, we implemented a conformational mimicry bias analysis based on directional Jtotal differences to resolve non-reciprocal structural relationships across the 3TM proteome, revealing asymmetric conformational mimicry in which specific families exhibit a directional bias to adopt the hVKOR signature. Our results establish Jscore quantification as a scalable approach for model curation and proteome-wide topology mapping, bridging static AI-predicted structures and dynamic conformational landscapes of proteins.
Phosphorus is a major building block for life and is used as an essential fertilizer in modern agriculture. With the recent depletion of phosphate rocks, the main source of phosphorus, cyanobacteria have attracted attention as valuable alternatives for phosphate production. They exhibit “luxury phosphorus uptake” and accumulate excess phosphate as polyphosphate within their cells. This study aimed to enable the model cyanobacterium Synechocystis sp. to produce polyphosphate without experiencing phosphate deficiency. Mutant strains PP and PS were constructed using psbA2 and psbA2 small promoters, respectively, to overexpress the polyphosphate kinase proteins that elongate the polyphosphate chain. Upon the addition of excess phosphate without phosphate deficiency, the fresh cell weight of the mutant strains PP and PS increased by 233
The proliferation of multidrug-resistant Klebsiella pneumoniae strains underscores the urgent need to decipher the transcriptional mechanisms governing their pathogenicity and resistance. This study characterizes the genome-wide regulatory architecture of K. pneumoniae MGH 78578 by mapping the binding landscapes of the housekeeping sigma factor RpoD and the general stress response sigma factor RpoS using high-resolution chromatin immunoprecipitation with exonuclease treatment (ChIP-exo). Contrasting with prior low-resolution studies, RpoD was identified as the dominant transcriptional orchestrator, encompassing a larger sigmulon than RpoS across both mid-exponential and stationary phases. While RpoS exhibited a high number of binding sites, it displayed relaxed promoter specificity characterized by the absence of a conserved −35 motif, reflecting a poised and plastic regulatory potential. A distinct evolutionary signature was observed on plasmids, where sigma factor binding density matched or exceeded that of the chromosome, but a smaller proportion of these binding events occurred at regulatory positions, indicating that these horizontally acquired sequences are not yet fully adapted to the host’s sigma factor recognition patterns. In the context of pathogenicity, distinct binding patterns were observed: RpoD was predominantly associated with core antimicrobial resistance determinants, including plasmid-encoded β-lactamases, whereas RpoS was preferentially linked to surface-remodeling virulence factors. These findings provide a high-resolution reconstruction of the K. pneumoniae transcriptional network, revealing how evolutionary history and sigma factor specificity coordinate the complex interplay between multidrug resistance and virulence.
Intracellular lipid droplets (LDs) in oleaginous yeasts store excessive carbon, which is important for biofuel and dietary lipid production. Studying the LDs’ morphologies in living cells is of importance for harvesting and screening promising isolates along with their culture conditions. In this study, a single-cell–based deep learning segmentation model was developed to detect and analyze individual cells and LDs from optical microscopy images of the oleaginous yeast strain KACC 46,215. The model achieved high detection accuracy, with AP50 values of 90.27 for cell identification and 89.94 for LD segmentation. Using this model, the average size of LDs and the LD-to-cell size ratio were quantitatively extracted and found to increase in correlation with actual lipid content determined by gas chromatography after cultivation. The developed segmentation model enables automated calculation of LD morphological metrics from optical images and provides a non-invasive strategy to predict the timing of lipid accumulation and determine optimal cell harvesting points during cultivation.
The increasing environmental problem of conventional plastics has driven interest in biodegradable alternatives and their sustainable monomer production. 1,4-butanediol (1,4-BDO) is one of the building blocks of polybutylene adipate terephthalate, a biodegradable polymer. In this study, L-glutamate-based whole-cell conversion system for 1,4-BDO production was developed using an engineered Escherichia coli strain. A synthetic metabolic pathway was composed of gadB, gabT, yqhD, and car3/sfp genes. The maximum titer of 1,4-BDO production was 48 mM from 100 mM L-glutamate as a substrate. Polyphosphate kinase 2 (PPK2) was introduced into the system together with polyphosphate (polyP) as a phosphate donor. Under the PPK2-expressing system, the 1,4-BDO titer increased up to 55 mM depending on polyP availability. Further analysis of metal ions identified Mg2+ as the most effective cofactor, and under Mg2+-optimized conditions, a 1,4-BDO titer of approximately 71 mM was obtained from 100 mM L-glutamate. These results suggest that PPK2 introduction combined with Mg2+ optimization is an effective strategy for enhancing 1,4-BDO production in an E. coli whole-cell conversion system.
Volatile organic compounds (VOCs) released from cancer cells are being explored as non-invasive readouts for cancer-associated states. However, translating complex VOC profiles into practical and biologically interpretable detection platforms remains challenging. Here, we developed a Caenorhabditis elegans-based whole-organism microfluidic biosensing platform for lung cancer cell-derived VOC detection. The polydimethylsiloxane chip operates on standard nematode growth medium (NGM) plates, preserving crawling-based chemotaxis while providing a standardized geometry for paired comparison of a matched reference medium or vehicle and the corresponding conditioned-medium or VOC test sample. Using this platform, wild-type C. elegans generated a quantifiable chemotactic response that distinguished SK-MES-1 lung cancer cell-conditioned medium from MRC-5 non-malignant lung fibroblast-conditioned medium and DMEM-only controls, with attraction observed over a dilution range of 10⁻³ to 10⁻⁷. Gas chromatography-mass spectrometry profiling revealed candidate VOC cues enriched in SK-MES-1-conditioned medium, which were putatively assigned to ethylbenzene, p-xylene, cyclohexanol, o-xylene, and decamethylcyclopentasiloxane. Purified-compound assays revealed compound-specific behavioral responses, with ethylbenzene and o-xylene producing attraction-like responses and p-xylene, cyclohexanol, and decamethylcyclopentasiloxane producing avoidance-associated responses. odr-3(n2150) mutants showed loss or reversal of the wild-type attraction response, supporting the involvement of ODR-3-dependent olfactory signaling. Early-life olfactory imprinting further amplified adult chemotactic responses. These results support the feasibility of a C. elegans-based whole-organism biosensing framework that integrates microfluidic standardization, candidate VOC profiling, genetic pathway analysis, and olfactory plasticity, providing a foundation for future lung cancer-associated VOC detection using patient-derived samples.
Polymers are crucial in promoting bone tissue engineering, presenting flexible solutions for bone repair and regeneration. Composite biomaterials combine two or more biomaterials to obtain specific properties tailored to patient-specific needs rather than using them separately. Due to their biocompatibility and biodegradability, they are excellent choices for producing scaffolds that replicate the body’s natural extracellular matrix. Scaffolds must possess non-hazardous characteristics like biodegradability and biocompatibility for the human body, along with essential mechanical behavior to provision body weight or fulfil other functions depending on the tissue type. Currently, the prevalence of bone disorders and defects due to tumour, injuries, microbial infections, and degenerative or inflammatory conditions is increasing. Thus, the advancement of bone repair and replacement has progressed alongside the enhancement of orthopedic technologies and high-quality biomaterials. However, advancements, major challenges in bone tissue engineering remain, such as the generation of unfavourable degradation by-products, limited mechanical strength, and inadequate cellular interactions. This review focuses on key aspects of scaffold design, including biodegradability, mechanical performance, fabrication approaches, and the choice of biomaterials. Further evaluates scaffold properties in relation to biological functions, structural demands, material composition, and both conventional and advanced manufacturing methods. Additionally, current limitations and future outlooks are discussed. Moreover, the paper highlights that the continued development of bio-scaffolds could open new avenues in tissue engineering, leading to more advanced biomimetic bone substitutes with strong potential for clinical translation.
Microbial fuel cell (MFC) offers a promising approach to improve wastewater quality and generate bioenergy from dark fermented effluents. In this study, the use of dark-fermented palm oil mill effluent as an electron donor for bioelectricity generation was investigated using a double-chambered MFC. The MFCs were operated at room temperature (29 ± 2℃), anode electrolytes adjusted to pH 7, and a chemical catholyte as the oxidizing agent. The maximum power and current densities of 63.31 ± 8.07 mW/m2 and 155.16 ± 12.88 mA/m2, respectively, were generated from the MFCs inoculated with sludge, which was 5.9 times higher than control without inoculum. Microbial community analysis revealed the enrichment of fermentative and electrogenic representative taxa from the phyla Bacillota, Bacteroidota and Pseudomonadota on the anode electrodes. Optimizations of the running conditions were carried out, suggesting the optimum parameters of 0.5 kΩ external resistance, anolyte initial pH 9, and 75
Echinatin (Ecn), a potent bioactive compound extracted from the roots and rhizomes of licorice, demonstrates anti-oxidant, anti-cancer, cardioprotective, anti-neurotoxic, and hepatoprotective properties. This study aims to investigate the antithrombotic potential of Ecn and clarify its mechanisms of action. Its anticoagulant effects were assessed via activated partial thromboplastin time, prothrombin time, thrombin and factor Xa (FXa) activities, as well as fibrin formation and platelet aggregation. Additionally, the effect of plasminogen activator inhibitor type 1 and tissue-type plasminogen activator was evaluated in tumor necrosis factor (TNF)-α–stimulated human endothelial cells. Our experiments revealed that Ecn exhibited antithrombotic activity comparable to that of rivaroxaban, a well-established direct FXa inhibitor, particularly in suppressing FXa activity and platelet aggregation induced by adenosine diphosphate (ADP) and U46619, a synthetic thromboxane A2 analog. Ecn also reduced the surface expression of P-selectin, inhibited phosphorylation of myristoylated alanine-rich C kinase substrate, and decreased PAC-1 activation following ADP or U46619 stimulation. Furthermore, Ecn enhanced nitric oxide production while preventing excess endothelin-1 release in HUVECs exposed to these agonists. In vivo experiments in mouse models of arterial and pulmonary thrombosis demonstrated that Ecn is a potent anticoagulant and antithrombotic agent. These findings indicate that Ecn could serve as a promising lead compound for developing innovative anti-FXa and antiplatelet therapeutics.
Antimicrobial resistance (AMR) has reached a critical inflection point, with the World Health Organization projecting that drug-resistant infections could claim 10 million lives annually by 2050 if left unaddressed. Central to combating AMR is antimicrobial susceptibility testing (AST), which guides rational antibiotic prescribing, yet conventional phenotypic and genotypic methods remain fundamentally constrained by prolonged turnaround times (16–72 h), dependency on pure culture isolation, high infrastructure costs, and the inability to capture complex in vivo resistance dynamics. These limitations perpetuate empirical antibiotic prescribing, with 30–50
The production of petrochemical substitutes through the biological conversion of renewable biomass is gaining attention as an eco-friendly process. Surfactin is highly regarded as an effective biosurfactant for commercial uses. In this study, we developed a fed-batch fermentation process for lipopeptide production using turnip peel, an agricultural waste biomass rich in neutral sugars, as a co-substrate. Fermentation was initiated in a medium containing both turnip peel and glucose as co-substrates. Overflowing foam was collected into a foam tank, and the reduced volume was replenished with fresh media containing turnip peel. Compared with glucose-only fermentation, the co-substrate system increased total lipopeptide production and appeared to promote a relative increase in the proportion of surfactin among the produced lipopeptides. Moreover, the lipopeptides obtained from the co-substrate fermentation exhibited a lower critical micelle concentration and a higher emulsification index, suggesting enhanced surface activity. These findings demonstrate that turnip peel can serve as a cost-effective and sustainable resource for improving surfactin production while maintaining product distribution and enhancing functional performance.
Bioconversion process suggests a sustainable and green approach to producing high-value compounds from food waste. In this study, pineapple crown (PC) were utilized as a feedstock for lactic acid (LA) production through enzymatic hydrolysis and fermentation. Potassium hydroxide (KOH) pretreatment was performed to improve the enzymatic digestibility (ED) of PC, and the effect of KOH concentration on PC to glucose conversion (PtGC) was investigated. As a result, the 2
Fungal biosynthetic gene clusters (BGCs) are often co-expressed, creating competition for shared precursors and limiting desired metabolite accumulation. The marine-derived fungus Penicillium sp. KWF31 produces paraherquamide A (PHQA) alongside penicillic acid derivatives and pigments. Using a phenotype-guided strategy and comparative BGCs analysis, we disrupted the penicillic acid backbone gene g322 and pigment-associated polyketide synthase (PKS) genes to relieve pathway competition. While Δg322 abolished dihydropenicillic acid but intensified pigmentation, deleting the pigment PKS gene g4411 eliminated colony pigmentation yet; neither single mutant increased PHQA. In contrast, the double mutant Δg322Δg4411 removed both major byproduct branches and increased PHQA production by 42.3
Trans-2-nonenal is a volatile aldehyde associated with aging-related body odor generated by oxidative degradation of human skin lipids and is a key target compound in cosmetics, textile, and food quality control. Owing to its high volatility, quantitative analysis of trans-2-nonenal often requires additional pretreatment steps and careful control of experimental conditions. However, it still has limited reliability and demands extensive laboratory equipment for testing. To cover these issues, a solvent-assisted analytical strategy was proposed for stable analysis with small scale by introducing solvent filling the headspace with trans-2-nonenal like makeup gas of gas analysis. To find out applicable solvent, solvent screening was performed and showed cyclohexane was the suitable solvent giving the high gas chromatography signal of trans-2-nonenal at 20 °C for 15 min. It resulted in 56 ppm of theoretical limit of detection and high linearity (r2 = 0.99), and the property of solvent dipolarity/polarizability (π*) (r2 = 0.92) was found to influence co-evaporation of trans-2-nonenal with solvent analyzed by linear solvation energy relationship analysis. When the interaction of various carbohydrate based candidates with trans-2-nonenal was examined, acidic polysaccharide (A.P) from Rhizobium leguminosarum VF39 exhibited 63.0
The valorization of food processing residues for renewable bioethanol production contributes to sustainable development and resource recovery. In this study, aronia-derived biomass (ADB), a carbohydrate-rich by-product from aronia juice manufacturing, was utilized as a renewable feedstock through an integrated biorefinery approach. To enhance enzymatic digestibility (ED), alkali pretreatment conditions were optimized using response surface methodology by varying KOH concentration, temperature, and reaction time. The optimized conditions (3
