This study evaluated the effects of essential oils, such as limonene (LIM) and a cinnamaldehyde–carvacrol blend (CCB), on the fermentation, chemical composition, in vitro digestibility, and aerobic stability of corn silage stored for 150 and 200 days. Treatments included a control, CCB (100 and 200 mg/kg DM), and LIM (100 and 200 mg/kg DM). Essential oils improved fermentation by increasing lactic acid (up to 7.46% of DM) and reducing proteolysis (NH3-N: 0.46–0.59% of total N). Limonene, particularly at 100 mg/kg DM, enhanced the lactic:acetic acid ratio (up to 3.07), better preserved non-fiber carbohydrates (≈32.7%), and increased in vitro DM digestibility (up to 81.5%) compared to the control (≈76.0%). The CCB treatment raised acetic acid concentrations (up to 3.04% of DM). Extending storage to 200 days reduced DM recovery (≈84.0%) versus 150 days (92.5%). Treated silages showed greater aerobic stability, with a lower pH increase after 72 h of air exposure, most notably in the LIM 200 treatment (pH 4.10) compared to the control (pH 5.40). Essential oils, particularly limonene, effectively improve the fermentative quality, nutritional value, and aerobic stability of corn silage under prolonged storage.

There is a lack of comprehension of the structure related properties of specific matrixes important for efficient packaging application. Therefore, this study examines structural, surface and barrier properties of bioactive films composed of two marine issued biopolymers: iota-carrageenan (Car) and sodium alginate (Alg), enriched with natural antioxidants and cross-linking agents (CaCl 2 ). The active films were formulated with chlorogenic acid (CA), D -limonene (DL) and sea fennel (SF) ( Crithmum maritimum ), known for their bioactive potential (against oxygen radicals and microbes). In addition, CaCl₂ was used as a crosslinker to improve the polymer networking, and even more important, moisture resistance. Results showed that the combination of Car and Alg formed interpenetrating polymer networks, improving the water vapor and oxygen barrier. FTIR analysis identified interactions between biopolymer matrices and bioactive molecules. UV-Visible spectroscopy showed improved UV barrier in CA-enriched films. Contact angle measurements and surface free energy analysis confirmed a dominant polar character, particularly in alginate-based blends, with no clear effects of the active molecules. Furthermore, Car-Alg films showed the lowest water vapour permeability, highlighting the role of polymer interactions in reducing permeability despite increased surface hydrophilicity. Crosslinking with Ca²⁺ resulted in higher film cohesion, reducing oxygen permeability. • Interactions between carrageenan and alginate or calcium chloride induce structural changes. • Incorporation of bioactive (antioxidant or essential oil) impacts the network organization. • Carrageenan and alginate complex networks reduce moisture transfers. • CaCl 2 as crosslinker reduces oxygen transfers. • UV barrier properties enhanced by the antioxidant incorporation.

Terpenes such as Limonene (LIMO) and β-Caryophyllene (BC) have demonstrated pain-modulating properties, potentially through interactions with the Transient receptor potential vanilloid 1 (TRPV1) receptor. This study examines the antinociceptive effects of four terpenes derived from Cannabis sativa: Limonene, β-Caryophyllene, α-Humulene (HUM), and α-Myrcene (MYR) using Caenorhabditis elegans (C. elegans). The primary objective was to characterize terpene-induced modulation of nocifensive responses to noxious heat, and to elucidate their influence on molecular pathways via specific receptor targets. Thermotaxis assays quantified the antinociceptive activity of increasing terpene concentrations in wild-type nematodes. To assess receptor-specific mechanisms, assays were performed in mutant strains lacking functional OCR-2 and OSM-9 (TRPV-like nociceptors) and NPR-19 and NPR-32 (encoding cannabinoid-like receptors). Proteomic profiling coupled with bioinformatics analysis identified terpene-induced alterations in signaling pathways and biological processes. All four terpenes exhibited significant antinociceptive activity in wild-type C. elegans, with impaired effects observed in vanilloid receptor mutants, implicating TRPV-like channels in their mechanism of action. Proteomic and pathway analyses revealed terpene-specific molecular signatures, highlighting differential modulation of neuronal and stress-responsive signaling cascades. By elucidating the molecular mechanisms underlying terpene-induced nociceptive modulation, this work strengthens the growing body of evidence supporting the therapeutic promise of terpenes in pain management outside the effect referred to as the "entourage effect".

This study investigates the integration of Bayesian networks (BN) and structural equation models (SEM) to explore genomic relationships among nine traits related to productivity, defense, and climate-adaptability in an interior lodgepole pine breeding program. Data from 392 open-pollinated trees, genotyped with 25,099 SNP markers, were analyzed. The traditional multitrait model (MTM) served as a benchmark for comparing SEM in estimating genetic (co)variance components, genetic correlations, breeding value (BV) predictions, and predictive ability, using both pedigree- (ABLUP) and genomic-based (GBLUP) individual-tree mixed models. The Hill-Climbing algorithm identified 12 significant causal structures (λ) among traits. Strong positive causal effects included tree height (HT) on wood density (WD) (λHT→WD = 0.413) and on stable carbon isotope ratio (C13) (λHT→C13 = 0.565), and limonene (LIMO) on carbon assimilation rate (CAR) (λLIMO→CAR = 0.368). The most influential causal relationship was HT → C13, followed by resistance to western gall rust (WGR) → CAR, CAR → LIMO, and WGR → C13. SEM incorporated these relationships, capturing both direct and indirect effects. Compared with MTM, SEM yielded lower residual variances, higher additive variances, and higher heritability estimates for all traits. The λ values from SEM correlated strongly with genetic correlations (0.932), with similarly high correlations between models (0.929), though SEM produced lower posterior mean correlations. BV correlations between models were high (ABLUP > 0.82, GBLUP > 0.84), but some reranking occurred among the top 39 trees (ABLUP > 0.71, GBLUP > 0.42). ABLUP and GBLUP-SEM models outperformed MTM in predictive ability, with mean gains of 6.62 and 6.03%, mainly for conditioned traits. BN-SEM enhances understanding of trait networks, improving genomic evaluations and breeding strategies in forest trees.

Abstract Traditional gel electrolytes based on weak interactions possess insufficient self‐healing property and also tend to rigidify during usage, which largely compromises the electrochemical performances of flexible supercapacitors (FSCs). Herein, a multifunctional ionic gel electrolyte, formed by the polymerization of limonene (LIM) and lipoic acid (LA) is proposed, displaying a collaborative self‐healing mechanism based on dual dynamic bonds, i.e., hydrogen bonds and disulfide bonds, and excellent persistence when combined with ionic liquids (ILs). In situ characterizations verify the exceptional self‐healing capabilities arising from dynamic disulfide and H‐bonds, which maintain structural integrity of the device upon deformation and even mechanical defects. ILs with electrostatic interactions and low saturation vapor pressure further strengthen the internal forces of the gel, meanwhile providing high ionic conductivity and durability upon varied temperature. Subsequently, the assembled FSCs with N‐doped carbon electrodes and P(LA‐LIM)‐IL electrolyte exhibit an impressive energy density of 22 µWh cm −2 and cyclability without attenuation of capacitance after 10 000 cycles. It also maintains 85% of capacitance after 100 cycles of mechanical deformation and a remarkable recovery of capacitance (92%) after 20 cutting/healing cycles. This multifunctional ionic gel electrolyte may broaden the way to design flexible SCs with superior self‐healing and resistant properties toward long lifespan.

Introduction: Herbs and their products are a source for drug discovery, and most of all synthetic drugs originate from them. The present study was designed to evaluate the in vitro and invivo efficacy, as well as the potential mechanisms, of Rhanterium epapposum essential oil (REE), β-myrcene (MC), camphene (CP), and limonene (LN) alone and in combination with pyrimethamine (PYM) against Toxoplasma gondii. Materials and Methods: In vitro, the effectiveness of REE and its components on tachyzoites, the infectivity rate, caspase-3 activity, and nitric oxide (NO) and expression levels of inducible NO synthase and gamma interferon (IFN-γ) genes were evaluated. In the invivo assays, infected mice received REE, MC, CP, and LN either alone or in combination with PYM for 2 weeks. Subsequently, the number and diameter of tissue cysts, oxidant/antioxidant enzyme levels, inflammatory cytokines, and bradyzoite surface antigen 1 (BAG1) gene expression were examined. Results: The best 50% inhibitory concentration values were reported for MC + LN, PYM + MC, and MC + CP, with values of 12.1, 15.4, and 16.6 µg/mL, respectively. REE and its primary compounds significantly increased (p < 0.001) NO release (6.6-14.1 nM), caspase-3 activity, and the expression levels of inducible NO synthase and IFN-γ genes in cells. In addition, significantly decreased (p < 0.001) the number (0.0-115.3 cysts) and size (0.0-83.1 nm) of tissue cysts, oxidative stress markers, decreased the expression levels of IL-1β and TNF-α, and BAG1 (0.48-5.10 fold change), while, significantly increased (p < 0.001) glutathione peroxidase (1.65-5.12 U/mg) and superoxide dismutase (31.9-74.1 U/mg) levels in the infected mice. Conclusions: We demonstrated the high in vitro and invivo efficacy of REE and its principal compounds against chronic toxoplasmosis. The highest efficacy was observed following combination therapy of T. gondii-infected mice with PYM and these compounds, with a significant difference noted compared with PYM alone. However, further trials must validate these findings and clarify the underlying mechanisms.

Cellulose-based aerogels modulate fragrance adsorption and controlled release by carbonization/in-situ aromatization.

Cutaneous cancer is a constantly progressive health challenge as life-threatening in men and women around the world. This study addressed repurposed ciclopirox olamine (COA) through the utilization of different essential oils limonene (LIM), eugenol (EU), and olive oil (OLO) as natural lipids possessing innate anticancer potential. HSPiP software provided preliminary screening of oils and solvents based on the theoretical solubility of COA. Placebo microemulsions (ME1-ME3) were prepared after optimizing Smix at varied ratios (surfactant to co-surfactant ratio) and evaluated for size, zeta potential, and polydispersity index (PDI). EU, LIM, and OLO (as microemulsions) were used to assess anticancer potential against A431, A375, and B16-F10 cell lines. Moreover, HSF (human fibroblast cells) was used to assess the safety of these excipients at the same concentrations. It was required to carry out concentration-dependent cytotoxic potential of EU and LIM against A431 and B16-F10 as compared to COA. Finally, an apoptosis study was conducted to understand the mechanistic perspective of EU and LIM against A431 and B16-F10. Results showed an excellent correlation (r2 = 0.97) of the predicted theoretical solubility of COA in the explored excipients to the experimental solubility data. UV scanning of the COA-solubilized oils negated any chemical interactions. ME1-ME3 showed tween-80 (10.9–21.6%) dependent size reduction (510–339 nm) and PDI values (0.28–0.31). In the cytotoxicity study, EU, LIM, and OLO elicited concentration-dependent cellular killing. LIM was found to be relatively sensitive to A431 and B16-F10 compared to EU. Notably, LIM resulted in dramatically aggressive induced early and late apoptosis in both cell lines at the explored IC50. Thus, the topical nanoformulations of COA containing anticancer essential oils can offer a promising approach for treating cutaneous melanoma.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-18141-4.

This study investigates a multifunctional hydrogel system integrating carboxymethyl cellulose (CMC) in a 3D-printed limonene (LIM) scaffold coated with poly(3,4-ethylenedioxythiophene): polystyrene sulfonate (PEDOT:PSS). The system allows to enhance wound healing, prevent infections, and monitor the healing progress. CMC is crosslinked with citric acid (CA) to form the hydrogel matrix (CMC-CA), while the 3D-printed limonene (LIM) scaffold is embedded within the hydrogel to provide mechanical support. PEDOT:PSS and curcumin-loaded PEDOT (PEDOT:CUR) nanoparticles are integrated into the hydrogel-membrane system for electrochemical detection of bacterial infection and controlled delivery of the antibacterial drug. The CMC-CA hydrogel exhibits excellent mechanical properties, suitable for conforming to irregular wound surfaces. In addition to provide additional mechanical support, the LIM scaffold is used as a pillar for the incorporation of PEDOT The integration of PEDOT:PSS and PEDOT:CUR enable not only real-time monitoring of bacterial growth but also the electrostimulated release of curcumin, which demonstrates antibacterial activity against Escherichia coli and Staphylococcus aureus. Electrostimulation of the CMC-CA/LIM/PEDOT system promotes cell proliferation, supporting accelerated wound healing. In conclusion, the CMC-CA/LIM/PEDOT system combines mechanical support, infection monitoring, and enhanced healing through controlled drug delivery and electrical stimulation, addressing critical challenges in wound management.

Background: Understanding how aroma compounds enhance monosodium glutamate (MSG) umami perception remains a critical challenge in flavor science. Methods: The umami-enhancing effects of meaty flavorings were investigated using nasal clip sensory evaluation (orthonasal blockage). Active aroma compounds were subsequently identified using gas chromatography-mass spectrometry (GC-MS). The three-dimensional structure of the umami receptor T1R1/T1R3 was constructed by homology modeling. The interaction mechanism was deciphered using molecular dynamics (MD) simulations. Results: Seafood essence S demonstrated the most potent umami enhancement. Five key compounds significantly intensified the MSG umami intensity: methional, dimethyl sulfide (DMS), D-limonene (DLE), 2,3-dimethylpyrazine, and dimethyl trisulfide. Notably, this enhancement persisted even under nasal clip conditions, revealing a novel mechanism independent of cross-modal interactions. Sulfur-containing compounds consistently demonstrated umami-enhancing effects across the evaluation conditions. MD simulations showed that aroma compounds induced allosteric remodeling of T1R1/T1R3, strengthening MSG-receptor hydrogen bonding (1.8–2.6-fold increase), reducing receptor flexibility, and stabilizing the ternary complex. Binding affinity was highest for DMS, followed by DLE and methional. Conclusion: This study provides the first receptor-level evidence that aroma compounds directly modulate MSG-taste receptor interactions through allosteric regulation, offering a novel theoretical framework for odor–taste interactions with significant implications for umami enhancer design and flavor research.

Objective D-Limonene (D-LMN) is a potential anti-oral cancer agent. The incorporation of D-LMN into a nanoemulsion enhances its potential efficacy against oral cancer cells by improving its aqueous compatibility. This study evaluated the impact of surfactant type and concentration on their properties through experimental and molecular dynamics studies. Methods D-LMN nanoemulsions were produced using different polyoxyethylene nonionic surfactants. The nanoemulsions were compared on droplet size, size distribution, zeta potential, droplet morphology, stability, interaction among molecules, anticancer activity, and cell death mechanism. Molecular dynamics simulations were also conducted to elucidate the dynamics of the components. Results Cremophor RH40 and Tween 60 yielded a nanoemulsion with a small droplet size (<100 nm). Cremophor RH40 also produced the nanoemulsion with the narrowest size distribution. Two-dimensional nuclear Overhauser effect spectroscopy–nuclear magnetic resonance and molecular dynamic simulation studies revealed that Cremophor RH40 effectively produced nanoemulsions with small droplets and high stability. The nanoemulsions formulated with Tween 60 exhibited more potent inhibitory effects against oral cancer cells than those with Cremophor RH40. However, surfactant type did not influence the mechanism underlying the apoptotic. Conclusions This study demonstrated that surfactants with an intermediate hydrophilic–lipophilic balance of approximately 15, particularly those with large polar heads and multiple hydrocarbon chains, were suitable for the production of anti-oral cancer D-LMN nanoemulsions by phase-inversion temperature technique.

The aim of this study was to investigate the effect of Ostwald ripening inhibitors on D-limonene (D-LMN) nanoemulsions and to elucidate their impact on oral cancer cells. Various inhibitors, including olive oil, soybean oil, and perilla oil, were incorporated into D-LMN nanoemulsions at different ratios (25:75-75:25, D-LMN to inhibitor). The resulting nanoemulsions were evaluated for droplet size, size distribution, zeta potential, stability, droplet morphology, cytotoxicity, antimetastatic and anti-invasive activities, apoptosis induction, and cell cycle arrest. Results showed that the 75:25 D-LMN to inhibitor ratio produced the smallest droplet size and exhibited great stability, particularly with perilla oil. Notably, D-LMN nanoemulsions displayed strong anti-oral cancer effects by reducing cell viability, metastasis, and invasion. Apoptosis was induced, as evidenced by nuclear fragmentation, Annexin V binding, and altered expression of BAX, BCL-XL, Cytochrome c, and Caspase-9. Additionally, the nanoemulsions caused cell cycle arrest via downregulation of Cyclin D1, CDK2, CDK4, and CDK6. These findings highlight the potential of D-LMN nanoemulsions as a promising alternative therapeutic strategy for oral cancer treatment.

Mantis shrimp-inspired functionalized plant fibers to fabricate a soy protein adhesive with high strength and mildew resistance.

Gas sensor arrays designed for pattern recognition face persistent challenges in achieving high sensitivity and selectivity for multiple volatile organic compounds (VOCs), particularly under varying environmental conditions. To address these limitations, we developed multimodal intelligent MEMS gas sensors by precisely tailoring the nanocomposite ratio of NiO and ZnO components. These sensors demonstrate enhanced responses to ethylene glycol (EG) and limonene (LM) at different operating temperatures, demonstrating material-specific selectivity. Additionally, a multitask deep learning model is employed for real-time, quantitative detection of VOCs, accurately predicting their concentration and type. These results showcase the effectiveness of combining material optimization with advanced algorithms for real-world VOCs detection, advancing the field of odor analysis tools.

Background/Object: This study investigates the in vitro antiviral potential of D-limonene (DLM), monolaurin (ML), and cetylpyridinium chloride (CPC) in formulations targeting SARS-CoV-2 and influenza viruses. The aim was to develop oral and nasal formulations with optimized concentrations of these active ingredients to evaluate their efficacy, safety, and stability. Methods: Oral (formulation D) and nasal (formulation E) products were developed using specific concentrations of DLM (0.2-0.3% w/w), ML (0.1-0.2% w/w), and CPC (0.05-0.075% w/w). In vitro virucidal activity assays were conducted to assess the antiviral efficacy of the formulations against SARS-CoV-2 and influenza viruses. Stability testing was also performed under various storage conditions. Results: Formulation D (0.3% w/w DLM, 0.2% w/w ML, 0.05% w/w CPC, and 1.5% w/w Cremophor RH40) demonstrated a 3.875 ± 0.1021 log reduction and 99.99 ± 0.0032% efficacy against SARS-CoV-2 within 120 s. Formulation E (0.2% w/w DLM, 0.05% w/w CPC, and 0.75% w/w Cremophor RH40) showed a 2.9063 ± 0.1197 log reduction and 99.87 ± 0.0369% efficacy against SARS-CoV-2. Both formulations achieved >99.99% efficacy and log reductions exceeding 4.000 against various influenza strains. Stability testing confirmed optimal performance at 4 °C with no microbial contamination. Conclusions: The findings suggest that both formulations exhibit broad-spectrum antiviral activity against SARS-CoV-2 and influenza viruses in vitro. These results support their potential for further clinical evaluations and therapeutic applications, particularly in oral and nasal spray formulations.

This study aimed to evaluate the effect of limonene (LIM) on the bond strength of a universal adhesive system in self-etch mode to caries-affected dentin (CAD). Class I cavities were prepared in eighty sound human molars and divided into four groups (n = 20): Sound dentin (SD), SD + LIM (SD treated with LIM), CAD, and CAD + LIM (CAD treated with LIM). Following pH cycling, two layers of a universal adhesive system (Single Bond Universal, 3MESPE) were applied. After restoration (Filtek Z350, 3MESPE), samples were sectioned into 0.9 mm2 sticks and stored in distilled water at 37°C for 24 h and 6 months. Subsequently, sticks were subjected to microtensile bond strength testing. Data were analyzed using 2-way ANOVA and Bonferroni's test (p < 0.05). Fracture patterns were examined under optical and scanning electron microscopes. After 24 h, SD + LIM showed lower bond strength than CAD but remained higher than CAD + LIM. CAD + LIM exhibited the lowest bond strength, regardless of aging. In SD + LIM, widened dentinal tubules were observed, accompanied by resin tags. In CAD + LIM, disorganized and unprotected collagen fibrils were identified, with fewer resin tags. Limonene decreased the bond strength of the universal adhesive system in self-etch mode to caries-affected dentin. Limonene as a pretreatment in self-etch adhesive protocols shows potential for enhancing the durability of adhesive restorations on sound dentin. However, its use on caries-affected dentin could compromise adhesion, highlighting the need for careful consideration when applying it to compromised substrates.

Clinically available therapies often inadequately address severe chronic cutaneous pain due to short anesthetic duration, insufficient intensity, or side effects. This study introduces a pen device delivering tetrodotoxin (TTX), a potent neurotoxin targeting nerve voltage-gated sodium channels, as a safe and effective topical anesthetic to treat severe chronic cutaneous pain. Chemical permeation enhancers, such as sodium dodecyl sulfate (SDS) and limonene (LIM), are incorporated to enhance TTX skin permeability. The device ensures precise TTX dosing down to the nanogram level, essential to avoid TTX overdose. In rats, the pen device treatment produces TTX-dose-dependent anesthetic effectiveness. An administration of 900ng of TTX with SDS and LIM to the rat back skin produces a 393.25% increase (measurement limit) in the nociceptive skin pressure threshold, and the hypoalgesia lasts for 11.25h, outperforming bupivacaine (28µg), of which are 25.24% and under 1 h. Moreover, the pen device provides on-demand therapy for multiple treatments, consistently achieving prolonged anesthesia over ten sessions (1 treatment per day) without noted toxicity. Furthermore, a single topical administration of 16µg of TTX exhibits no TTX-related toxicity in rats. The TTX delivery pen paves the way for clinical trials, offering a promising solution for severe cutaneous pain.

Background and aims: Excessive oxidative stress and increased apoptosis are the main culprits to heart damage associated with ischemic heart disease. This study aimed to demonstrate the cardioprotective properties of limonene (LIM), a monoterpenoid compound, in isoproterenol-induced ischemia (ISO-I) in rats. Methods: Forty male Wistar rats were divided into five groups (n=8). Normal saline (1 mL/kg) and LIM at doses of 10 mg/kg, 20 mg/kg, and 40 mg/kg were administered for seven days before the induction of ischemia by ISO. Creatine kinase-MB (CK-MB), creatine phosphokinase (CPK), lactate dehydrogenase (LDH), malondialdehyde (MDA), total antioxidant capability (TAC), and relative expression of BCL2 and BAX genes underwent measurement. Results: CK-MB (3.73±0.560 versus 0.888±0.317), CPK (901±72.2 versus 397±33.5), LDH (1992±176 versus 821±94.1), and MDA levels in the serum (65.6±11.0 versus 28.3±4.75) and heart (104±8.70 versus 65.3±7.46) samples significantly increased following ischemia in the treatment groups compared to the control group (P&lt;0.001 for all markers). Ischemia decreased TAC in the heart (324±45.0 versus 588±64.5) and serum (202±23.0 versus 388±45.9) samples compared to the control counterparts (P&lt;0.001 for both samples). ISO-I is associated with a decrease in Bcl-2 (0.880±0.118 versus 1.74±0.178) and an increase in BAX (2.75±0.124 versus 1.08±0.0938) gene expression in comparison to the control group (P&lt;0.001 for both genes). LIM could reverse the aforementioned heart damage-related biomarkers in the serum and heart samples. Finally, LIM improved histopathological disarrangement following ischemia. Conclusion: The findings revealed that LIM, at least partially due to its antioxidant and anti-apoptotic potentials, exerted cardioprotection against ISO-induced heart ischemia in rats.

Terpenes are widely used in cosmetic formulations as chemical penetration enhancers (CPEs). However, the molecular mechanisms underlying the skin-penetration-enhancing effect have not been completely understood. In this work, molecular dynamics (MD) simulations were used to explore the influence of terpineol (TER), 1,8-cineole (CIN), and limonene (LIM) on the stratum corneum (SC) model. The results indicated that terpenes affected lipid membranes in a concentration-dependent manner and promoted skin permeation by disorganizing the cholesterol (CHOL) portion. The penetration of CPEs across the skin membrane also differed, with diffusion rates of limonene > 1,8-cineole > terpineol. The limonene molecules could penetrate the bilayer's center, forming a "triple layer" membrane structure. Furthermore, constrained simulations showed that the most favorable penetration pathway is via areas rich in CHOL. Terpineol could lower the energy barrier of the hydrophilic molecule (caffeic acid) across the cholesterol region. For the lipophilic molecule (osthole), limonene and 1,8-cineole could reduce the energy barrier across the cholesterol region. Each of the results provides novel insights into the mechanism of penetration of CPEs in the skin.

Antileishmanial and synergic effects of Rhanterium epapposum essential oil and its main compounds alone and combined with glucantime against Leishmania major infection