Strong Inhibition Research Articles

Clinical Assessment of the Drug-Drug Interaction Potential of Omaveloxolone in Healthy Adult Participants.

Omaveloxolone is approved in the United States and the European Union for the treatment of patients with Friedreich ataxia aged ≥16 years. It is mainly metabolized by cytochrome P450 (CYP) 3A4 in vitro. Two drug-drug interaction studies (NCT04008186 and NCT05909644) were performed to evaluate (1) the effect of drug-metabolizing enzymes (DMEs) and drug transporter (DT) modulators on the pharmacokinetics of omaveloxolone and (2) the effect of omaveloxolone on the pharmacokinetics of DME and DT substrates. Additionally, the safety of coadministering these drugs with omaveloxolone was assessed. Coadministration of the strong CYP3A4 inhibitor itraconazole significantly increased omaveloxolone maximum plasma concentration (Cmax) and area under the plasma concentration-time curve from time 0 extrapolated to infinity (AUC0-∞) by approximately 3- and 4-fold, respectively. Conversely, coadministration with the moderate CYP3A4 inducer efavirenz decreased Cmax and AUC0-∞ of omaveloxolone by 38.0% and 48.5%, respectively. Omaveloxolone exposure was also increased following coadministration with verapamil, a moderate CYP3A4 and P-glycoprotein (P-gp) inhibitor, but it was unaffected by the strong CYP2C8 inhibitor gemfibrozil. Coadministration of omaveloxolone reduced systemic exposure of the substrates of CYP3A4, CYP2C8, breast cancer resistance protein, and organic anion transporting polypeptide 1B1 but had no effect on those of P-gp and organic cation transporter 1. Omaveloxolone was well tolerated when administered alone and in combination with the DME and DT modulators or substrates. These findings support concomitant medication precautions and dosing recommendations for omaveloxolone when coadministered with a moderate or strong CYP3A4 inhibitor or inducer, as well as the substrates of certain CYP450 enzymesor transporters.

Lactic Acid Bacteria from Kiwi: Antifungal and Biofilm-Inhibitory Activities Against Candida albicans

Urogenital infections impact millions of individuals globally each year, with vulvovaginal candidiasis (VVC) being one of the most prevalent conditions affecting women. Candida albicans is the primary pathogen responsible for VVC. The utilization of probiotics as an alternative therapeutic approach to antibiotics in managing such infections has gained increasing attention. This study aimed to evaluate the potential of THY-F51, a lactic acid bacterium isolated from kiwi, as a probiotic to support vaginal health through its antifungal, anti-biofilm, and anti-inflammatory properties against C. albicans. The identification of THY-F51 was confirmed through 16S rRNA gene sequencing. A series of evaluations were performed to determine its antifungal efficacy against C. albicans, biofilm-inhibitory activity, antioxidant properties, and effects on inflammatory cytokines. Cytotoxicity assays and assessments of bacterial survival under vaginal pH conditions (pH 3.8–4.5) were also conducted. The results demonstrated that THY-F51, identified as Leuconostoc citreum, exhibited potent antifungal activity against C. albicans, with an MIC of 1.25 mg/mL and an MFC of 2.5 mg/mL. Furthermore, THY-F51 displayed a strong inhibition of C. albicans biofilm formation, as well as notable antioxidant activity in the supernatant. Additionally, THY-F51 demonstrated high survival rates under vaginal pH conditions, an absence of cytotoxic effects, and a significant reduction in C. albicans adhesion to HeLa cells. Moreover, THY-F51 effectively suppressed C. albicans-induced inflammatory cytokines, including TNF-α, IL-1β, IL-6, and IL-8. These findings suggest that THY-F51, isolated from kiwi, holds substantial promise as a safe and effective probiotic for reducing vaginal inflammation and promoting vaginal health.

Population Pharmacokinetics of Cobicistat and its Effect on the Pharmacokinetics of the Anticancer Drug Olaparib.

Pharmacokinetic (PK) boosting is the intentional use of strong inhibitors of metabolic enzymes or transporters to boost the systemic exposure of a therapeutic drug. PK boosting is expanding to therapeutic areas outside human immunodeficiency virus (HIV) therapy. Data on the PK of the booster cobicistat and its effect on CYP3A-substrates outside of HIV therapy are lacking. This study aimed to describe the PK of once- and twice-daily cobicistat regimens in healthy volunteers and patients with rheumatoid arthritis, cancer, or HIV infection and to investigate the interplay between cobicistat and the anticancer drug olaparib. Cobicistat levels from 683 samples from 66 subjects in four clinical trials were included in the analysis. For olaparib, 261 samples from 12 subjects from one trial were included. Population PK analysis was performed by nonlinear mixed-effects modelling. Both cobicistat and olaparib PK were adequately described by a well-stirred liver model with one central compartment and Erlang type absorption. Cobicistat PK was similar across patient populations and dosing regimens. Cobicistat increased olaparib prehepatic bioavailability 1.65-fold (RSE 6%) and decreased intrinsic clearance 0.34-fold (RSE 6.5%). A correlation between olaparib PK and cobicistat exposure could not be identified. The interindividual variability in olaparib clearance was lower with cobicistat than without cobicistat. The developed pharmacokinetic models adequately described cobicistat and olaparib plasma concentrations. PK boosting with cobicistat at 150 mg twice daily led to an increase in olaparib bioavailability and decrease in clearance. This effect was not correlated with cobicistat exposure, which may reflect saturation of the boosting effect of cobicistat at this dose. NCT02565888 (30-09-2015), NCT00825929 (19-01-2009), Netherlands Trial Register NL7766 (18-12-2018), NCT05078671 (22-09-2021).

Toxicokinetics and analytical toxicology of the phenmetrazine-derived new psychoactive substance 3,4-methylenedioxyphenmetrazine studied by means of in vitro systems.

Compounds derived from known drugs are usually brought on the new psychoactive substance (NPS) market without any previous toxicological risk assessment. The European Union Drugs Agency issued an EU early notification for 3,4-methylenedioxyphenmetrazine (MDPM) in 2024. It is structurally related to the stimulants amphetamine, 3,4-methylenedioxymethamphetamine (MDMA), and phenmetrazine and expected to have similar effects. So far, no scientific reports are available describing its toxicokinetic and analytical profile. This study aimed to provide such data to allow a thorough risk assessment and to ease its analytical detectability in forensic and clinical toxicology and doping control. Data reported include the in vitro plasma protein binding, the in vitro half-life and in vitro metabolism of MDPM by human liver microsomes and S9 fraction (pHLS9) and by HepaRG cells. A monooxygenase mapping and the in vitro cytochrome P450 inhibition of MDPM was elucidated. Results showed that HepaRG cells and pHLS9 formed the same MDPM metabolites via demethylenation and O-methylation and that MDPM has a low plasma protein binding and is a low-turnover drug. Monooxygenase mapping revealed that the demethylenation was exclusively CYP2D6-mediated. MDPM showed strong inhibition of CYP2D6 and moderate inhibition of CYP1A2 and CYP3A4. Polymorphisms or the simultaneous intake of substances that are also CYP2D6 substrates can have a considerable impact on the toxicity of MDPM. Based on in vitro data, the demethylenyl-methyl metabolite of MDPM and the parent compound are recommended as analytical urine screening targets.

DyphAI dynamic pharmacophore modeling with AI: a tool for efficient screening of new acetylcholinesterase inhibitors

Therapeutic strategies for Alzheimer’s disease (AD) often involve inhibiting acetylcholinesterase (AChE), underscoring the need for novel inhibitors with high selectivity and minimal side effects. A detailed analysis of the protein-ligand pharmacophore dynamics can facilitate this. In this study, we developed and employed dyphAI, an innovative approach integrating machine learning models, ligand-based pharmacophore models, and complex-based pharmacophore models into a pharmacophore model ensemble. This ensemble captures key protein-ligand interactions, including π-cation interactions with Trp-86 and several π-π interactions with residues Tyr-341, Tyr-337, Tyr-124, and Tyr-72. The protocol identified 18 novel molecules from the ZINC database with binding energy values ranging from −62 to −115 kJ/mol, suggesting their strong potential as AChE inhibitors. To further validate the predictions, nine molecules were acquired and tested for their inhibitory activity against human AChE. Experimental results revealed that molecules, 4 (P-1894047), with its complex multi-ring structure and numerous hydrogen bond acceptors, and 7 (P-2652815), characterized by a flexible, polar framework with ten hydrogen bond donors and acceptors, exhibited IC₅₀ values lower than or equal to that of the control (galantamine), indicating potent inhibitory activity. Similarly, molecules 5 (P-1205609), 6 (P-1206762), 8 (P-2026435), and 9 (P-533735) also demonstrated strong inhibition. In contrast, molecule 3 (P-617769798) showed a higher IC50 value, and molecules 1 (P-14421887) and 2 (P-25746649) yielded inconsistent results, likely due to solubility issues in the experimental setup. These findings underscore the value of integrating computational predictions with experimental validation, enhancing the reliability of virtual screening in the discovery of potent enzyme inhibitors.

Pharmacokinetic Interactions of Paxlovid Involving CYP3A Enzymes and P-gp Transporter: An Overview of Clinical Data.

The US FDA has approved paxlovid, a combination of nirmatrelvir and ritonavir, as the first oral treatment for the management of mild-to-moderate COVID-19 patients. The purpose of this review article is to explore the clinical data that is currently available regarding the drug-drug interactions (DDIs) of paxlovid with various medications. Keywords, such as drug interactions, paxlovid, ritonavir, nirmatrelvir, pharmacokinetic interactions, CYP3A, and P-glycoprotein, were used to search online databases, including LitCOVID, Scopus, Embase, EBSCO host, Google Scholar, ScienceDirect, Cochrane Library, and reference lists. Paxlovid interacted with a variety of medications due to strong inhibition of CYP3A4 and P-gp transporter protein by ritonavir and the dual function of nirmatrelvir as a substrate and inhibitor of CYP3A enzymes and P-gp transporter protein. Numerous case reports and other studies determined that the risk of toxicities of several drugs, including anticoagulants (warfarin, rivaroxaban), calcium channel blockers (nifedipine, manidipine, verapamil), statins (atorvastatin), immunosuppressants (tacrolimus), antiarrhythmics (amiodarone), antipsychotics (clozapine, quetiapine), and ranolazine have been enhanced by the concomitant administration of paxlovid. Adverse effects of paxlovid from DDIs can range from less-than-ideal therapeutic responses to potentially fatal toxicities. Effective management requires close observation, adjustments to dosage, and assessment of substitute treatments. Collaboration between pharmacists and other medical professionals is necessary to guarantee effective and safe treatment outcomes of paxlovid therapy.

Protective and Anti-Inflammatory Effect of Novel Formulation Based on High and Low Molecular Weight Hyaluronic Acid and Salvia haenkei

Salvia haenkei (SH-Haenkenium®), a native plant of Bolivia, is known as strong inhibitor of senescence and recently exploited in wound healing and for its potential anti-inflammatory properties. Hyaluronan at high and low molecular weight (HCC), explored in diverse cell models, and recently used in clinical practice, showed beneficial effects in dermo aesthetic and regenerative injective treatments. In this research work a novel formulation based on HCC coupled SH was tested for its potentiality in counteracting dermal injury. In vitro wound healing has been used to demonstrate HCC + SH capacity to improve keratinocytes migration respects the sole HCC, supported also by positive modulation of remodeling and integrity biomarkers. In addition, an in vitro dehydration test showed its ability to defend the skin from dryness. Moreover, an in vitro inflammation model (with lipopolysaccharides derived from E. coli) was used to assess molecular fingerprint of the pathological model and compare the cell response after treatments. Inflammatory biomarkers (e.g., KRT6, TLR-4 and NF-κB) and specific cytokines (e.g., IL-6, IL-22, IL-23) proved the effect of HCC + SH, in reducing inflammatory mediators. A more complex model, 3D-FT skin, was used to better resemble an in vivo condition, and confirmed the efficacy of novel formulations to counteract inflammation. All results trigger the interest in the novel formulation based on SH extract and hyaluronan complexes for its potential efficacy as natural anti-inflammatory agent for damaged skin, for its healing and regenerative properties.

Discovery of a Novel Multitarget Analgesic Through an In Vivo-Guided Approach

Background: Pain is a complex condition influenced by peripheral, central, immune, and psychological factors. Multitarget approaches offer a more effective and safer alternative to single-target analgesics by enhancing efficacy, reducing side effects, and minimizing tolerance. This study aimed to identify a novel multitarget analgesic with improved pharmacological properties. Methods: An in vivo-guided screening approach was used to discover a new analgesic compound. Compound 29, derived from a novel scaffold inspired by opiranserin and vilazodone pharmacophores, was identified through analog screening in the formalin test. Its efficacy was further evaluated in the spinal nerve ligation (SNL) model of neuropathic pain. Mechanistic studies explored its interaction with neurotransmitter transporters and receptors, while pharmacokinetic and safety assessments were conducted to determine its stability, brain penetration, and potential toxicity. Results: Compound 29 demonstrated high potency in the formalin test, with an ED50 of 0.78 mg/kg in the second phase and a concentration-dependent effect in the first phase. In the SNL model, it produced dose-dependent analgesic effects, increasing withdrawal thresholds by 24% and 45% maximum possible effect (MPE) at 50 and 100 mg/kg, respectively. Mechanistic studies revealed strong triple uptake inhibition, particularly at dopamine (DAT) and serotonin (SERT) transporters, alongside high-affinity 5-HT2A receptor antagonism. Pharmacokinetic analysis indicated enhanced stability and blood–brain barrier permeability. In vitro studies confirmed its nontoxicity to HT-22 cells but revealed potential hERG inhibition and strong CYP3A4 inhibition. Conclusions: Compound 29 is a promising multitarget analgesic with potent efficacy and favorable pharmacokinetics. Ongoing optimization efforts aim to mitigate side effects and enhance its therapeutic profile for clinical application.

Translation elongation defects activate the Caenorhabditis elegans ZIP-2 bZIP transcription factor-mediated toxin defense.

The Caenorhabditis elegans bZIP transcription factor ZIP-2 is activated by toxins or mutations that inhibit translational elongation. The zip-2 DNA-binding protein is encoded in a downstream main open reading frame (mORF), but under normal translation elongation conditions only an upstream overlapping oORF -1 frameshifted from mORF is translated. Mutations or toxins that slow translational elongation, but not inhibitors of translational initiation or termination, activate ZIP-2. An mORF initiation codon mutation does not disrupt the normal zip-2 response to translational elongation defects, suggesting that zip-2 activation does not depend on this ATG. An mORF early termination mutant can be activated by strong translation elongation inhibition, suggesting that translation initiated upstream on oORF +1 frameshifts when elongation is inhibited to the mORF reading frame downstream of the stop codon to activate a fused oORF/mORF ZIP-2 transcription factor. The protein and DNA sequences of zip-2 oORF and mORF are conserved across the Caenorhabditis, suggesting selection for particular codons sensitive to translational elongation defects. Mutations that disrupt the oORF initiation codon constitutively activate zip-2, but not if the mORF initiation codon is also mutant, showing that zip-2 oORF competes with mORF for translational initiation. oORF initiation codon mutation-activated zip-2 slows C. elegans growth, and this slow growth is suppressed by a zip-2 null mutation. A zip-2 null mutant also strongly suppresses the growth arrest caused by translational elongation inhibitors. Thus, ZIP-2 is both a sensor of translational elongation attack, and a defense regulatory output via its activation of response genes.

Mechanistic insights into alkaloid-based inhibition of squalene epoxidase: A combined in silico and experimental approach for targeting cholesterol biosynthesis.

Squalene epoxidase (SQLE) is a key enzyme in the cholesterol biosynthesis pathway and an attractive therapeutic target for hypercholesterolemia and antifungal treatment. In this study, we investigated the inhibitory potential of six alkaloids-berberine, evodiamine, harmine, reserpine, matrine, and sanguinarine-against SQLE using a combined in silico and in vitro approach. Molecular docking revealed strong binding affinities ranging from -8.1 to -11.0 kcal/mol, with evodiamine demonstrating the highest affinity, followed by sanguinarine and berberine. 200 ns MD simulations confirmed stable interactions for all alkaloid-enzyme complexes, characterized by low RMSD values, robust hydrogen bonding, and favorable free energy landscapes, as supported by MM/PBSA analysis. Experimental validation through in vitro inhibition assays revealed that evodiamine (IC₅₀ = 2.87 ± 0.08 μM) exhibited potent inhibition comparable to the standard inhibitor TNSCPA (IC₅₀ = 2.65 ± 0.18 μM), while berberine (IC₅₀ = 3.57 ± 0.18 μM) and reserpine (IC₅₀ = 4.91 ± 0.34 μM) showed strong and moderate inhibition, respectively. Harmine, matrine, and sanguinarine were less effective. Enzyme kinetics studies demonstrated that berberine and reserpine act as noncompetitive inhibitors, binding to allosteric sites, whereas evodiamine exhibited competitive inhibition at the active site. ADMET analysis highlighted favorable pharmacokinetic properties for berberine, evodiamine, and sanguinarine, while reserpine and matrine exhibited limited bioavailability due to solubility and size constraints. The unique inhibitory mechanisms observed were consistent with the structural and physicochemical properties of the compounds. These findings establish berberine, evodiamine, and reserpine as promising SQLE inhibitors, with evodiamine emerging as a lead candidate.

Enrichment of antioxidant peptides by interfacial modification of oat polypeptides induced by zinc ions.

The pursuit of methods to enhance the purity of food-sourced bioactive peptides continues to pose significant challenges. This study introduces an innovative approach to enrich antioxidant peptides by using zinc ion coordination to augment the foaming capabilities of oat peptides. The resulting antioxidant peptide fraction (AF) accounted for 18% of the oat globulin hydrolysates, with a significant increase (22-47%) in scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH), OH, and O2- radicals. Proteomics identified 479 peptide segments within AF, and the HipHop analysis further identified 340 antioxidant peptides. Notably, the larger peptides (7-23 amino acids) were the primary contributors to the antioxidant activity, featuring key pharmacophores, i.e., charge centers, hydrophobic centers, and hydrogen bond acceptors. The AF and its key monomers (DDTKTWPEDL, YSTDPANPTKSA, NKREQQSGNNIF, and QVGQSPQYQEG) exhibited potent inhibitory effects on tyrosinase (IC50, 18.60-46.20μg/mL) and provided strong inhibition against lipid oxidation, indicating great potential for applications in health supplements and food preservation.

P-1239. Systemic Exposure of Olorofim in Children with Invasive Fungal Infections

Abstract Background Olorofim (OLO) is a novel antifungal active vs. Aspergillus (including azole-resistant strains), resistant moulds (e.g., Lomentospora prolificans [LoPro]), and dimorphic moulds. Treatment with OLO in children with invasive fungal infection (IFI) is permitted under the Managed Access Program (MAP), where monitoring of trough concentrations is required due to sparsity of data in this population. Methods To be eligible for the MAP, children were aged 6 to 17 years, had an IFI infection that was shown to be or expected to be susceptible to OLO and had limited or no treatment options. Trough plasma samples (collected after an overnight fast) were sent to the central laboratory for analysis of OLO using a validated mass spectrometry assay. As a minimum, samples were taken after at least 7 days dosing and following any dose changes. The paediatric dose was informed by a population pharmacokinetic (PK) model built using PK data from 52 adult patients with IFI. This model was used to predict systemic exposure in children aged 6 to 17 years based on PK parameters adjusted by weight. Paediatric doses were chosen based on a goal of Cmin ≥ 0.1µg/mL in over 90% of population, whilst Cmax and AUC were to be kept as low as possible. A further dose adjustment was required if OLO was given with strong cytochrome P450 3A4 inhibitors or inducers. Results To date, 22 children have received OLO, aged 6 to 17 years (mean = 11.0 y), with body weights of 20 to 80 kg (mean = 37.4 kg). IFIs treated include: IA (7), LoPro (4), Scedosporium spp. (4), coccidiomycosis (2), and other rare fungi (5). Trough level data are available for 19 subjects; the predicted maintenance dose resulted in trough levels ≥ 0.1 µg/mL in 15 subjects (79%). For the remaining 4 subjects (21%) OLO dose was increased. After dose adjustment (if required), observed trough plasma concentrations exceeded the threshold needed for efficacy (i.e., > 0.1 µg/mL) on all sampling occasions. Conclusion With the relatively small population and sparse PK sampling data available, the predicted body-weight dependent doses for children aged 6 to 17 y are considered reasonable. The dose predictions will be updated as the PopPK model is refined and trough levels will continue to be monitored pending conduct of a formal paediatric study. Disclosures Karen Cornelissen, PhD, F2G: Stocks/Bonds (Private Company) John H. Rex, MD, F2G: Employee Daniela Zinzi, MD. Infectious Diseases Specialist, VP Clinical R&D at F2G: Stocks/Bonds (Private Company) Roger Bruggemann, Pharm, PhD, F2G: Advisor/Consultant|F2G: Grant/Research Support|Gilead: Advisor/Consultant|Gilead: Grant/Research Support|Mundipharma: Advisor/Consultant|Mundipharma: Grant/Research Support|Pfizer: Advisor/Consultant|Pfizer: Grant/Research Support James Wright, n/a, F2G: Advisor/Consultant

Analysis of Protein Inhibitors of Trypsin in Quinoa, Amaranth and Lupine Seeds. Selection and Deep Structure–Function Characterization of the Amaranthus caudatus Species

Protease inhibitors are biomolecules with growing biotechnological and biomedical relevance, including those derived from plants. This study investigated strong trypsin inhibitors in quinoa, amaranth, and lupine seeds, plant grains traditionally used in Andean South America. Amaranth seeds displayed the highest trypsin inhibitory activity, despite having the lowest content of aqueous soluble and thermostable protein material. This activity, directly identified by enzymatic assay, HPLC, intensity-fading mass spectrometry (IF-MS), and MS/MS, was attributed to a single protein of 7889.1 Da, identified as identical in Amaranthus caudatus and A. hybridus, with a Ki of 1.2 nM for the canonical bovine trypsin. This form of the inhibitor, which is highly homogeneous and scalable, was selected, purified, and structurally–functionally characterized due to the high nutritional quality of amaranth seeds as well as its promising agriculture–biotech–biomed applicability. The protein was crystallized in complex with bovine trypsin, and its 3D crystal structure resolved at 2.85 Å, revealing a substrate-like transition state interaction. This verified its classification within the potato I inhibitor family. It also evidenced that the single disulfide bond of the inhibitor constrains its binding loop, which is a key feature. Cell culture assays showed that the inhibitor did not affect the growth of distinct plant microbial pathogen models, including diverse bacteria, fungi, and parasite models, such as Mycoplasma genitalium and Plasmodium falciparum. These findings disfavour the notion that the inhibitor plays an antimicrobial role, favouring its potential as an agricultural insect deterrent and prompting a redirection of its functional research.

Abstract A013: Shikonin, a strong inhibitor of Phosphoinositide 3-Kinase (PI3K) and Mitogen-Activated Protein (MAP) Kinase pathways, induces cell death in UV-B irradiated B16F10 melanoma cells

Abstract Ultraviolet (UV) radiation-induced skin damage plays a pivotal role in the pathogenesis of melanoma, a highly aggressive and fatal form of skin cancer. Shikonin, naturally occurring nathoquinone , is well-known for its inhibitory effects on phosphoinositide 3-kinase (PI3K) and mitogen-activated protein (MAP) kinase signaling pathways. It has previously been demonstrated to exert effects against various non-melanoma skin cancers. In this study, we investigated the effects of Shikonin treatment on UVB-irradiated B16F10 melanoma cells, revealing a dose-dependent reduction in cell viability and an enhancement of apoptosis. Invitro findings indicate that the pro-apoptotic effects of shikonin in UVB-exposed B16F10 cells are mediated by several mechanisms, including the generation of reactive oxygen species (ROS), modulation of the antioxidant defense response, and mitochondrial membrane potential (ΔΨM) depolarization. Further supporting the apoptotic role of Shikonin, we observed DNA fragmentation, caspase activation, poly(ADP-ribose) polymerase (PARP) cleavage, and an increase in sub-G2 cell populations. Shikonin treatment also significantly attenuated MEK-ERK signaling, influenced the PI3K/Akt pathway, and potentially inhibited UVB-induced nuclear translocation of NF-κB. Additionally, co-treatment with UVB and Shikonin led to a decrease in the Bcl-2/Bax ratio and upregulated the expression of pro-apoptotic proteins Bax and caspases. Collectively, these data suggest that Shikonin may act as a potential therapeutic agent for melanoma when combined with UVB exposure, providing a novel avenue for future treatment strategies. Citation Format: Aalim Maqsood Bhat, Sheikh Tasduq Abdullah. Shikonin, a strong inhibitor of Phosphoinositide 3-Kinase (PI3K) and Mitogen-Activated Protein (MAP) Kinase pathways, induces cell death in UV-B irradiated B16F10 melanoma cells. [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Targeted Therapies in Combination with Radiotherapy; 2025 Jan 26-29; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(2_Suppl):Abstract nr A013.

Actual Use of Budesonide Enteric-Coated Capsules for Crohn’s Disease in Japan: Analysis of Health Insurance Big Data

Using a large health insurance database in Japan, we examined the real-world usage of budesonide enteric-coated capsules (BUD) in treating Crohn's disease. We analyzed data from the Japan Medical Data Center claims database for Crohn's disease patients prescribed BUD from April 2016 to March 2021, focusing on prescription status, adverse events (AEs), monitoring tests, and concomitant medications over 2 years following BUD initiation. Patients were categorized into two groups based on BUD usage duration: ≤1 year and >1 year. Of the 7364 registered patients, 1049 (14.2%) were prescribed BUD. Among the 562 followed for 2 years, 505 (89.9%) used BUD for ≤1 year and 57 (10.1%) for >1 year. Over 70% of the patients used at least one biologic, and more than 20% used at least two. The proportions of new thiopurine initiation were 22 and 9% in the ≤1-year and >1-year groups, respectively (p = 0.0181). We did not identify any obvious increase in AEs from long-term BUD use within the confines of our study design. However, regardless of prescription duration, over half of the patients lacked hepatitis B virus screening, glycated hemoglobin measurement, adrenal function quantification, or bone densitometry. Usage of strong CYP3A4 inhibitors was more frequent among patients in the BUD >1-year group. This study revealed that numerous Japanese patients received long-term BUD prescriptions. Although no apparent increase in AEs from long-term BUD was detected, we identified inadequate monitoring of AEs and drug interactions, as well as insufficient use of steroid-sparing agents.

The GacS/GacA two-component system strongly regulates antimicrobial competition mechanisms of Pseudomonas fluorescens MFE01 strain.

Our model strain Pseudomonas fluorescens MFE01 uses an active type VI secretion system (T6SS) and volatile compounds (VCs) to outcompete other microorganisms in the natural environment. By investigating the cellular mechanism regulating the production of these weapons, we identified the two-component system GacS/GacA. Indeed, GacS cellular membrane sensor plays a crucial role in regulating T6SS activity and VC emission. Among the latter, 1-undecene and hydrogen cyanide are strong aerial inhibitors of the Legionella human pathogen and the Phytophtora infestans major plant pest, respectively. The aim is to improve the understanding of the regulation of these volatile molecule emission and the critical role of a global regulator in both plant and human health.

Development of Pectin-Based Films with Encapsulated Lemon Essential Oil for Active Food Packaging: Improved Antioxidant Activity and Biodegradation

This study evaluated the physicochemical, morphological, and functional properties of pectin-based films incorporated with lemon essential oil (EO) to assess their potential as biodegradable food packaging materials. The results showed that EO incorporation significantly influenced the film’s characteristics. The control film exhibited a smooth surface, while the EO-containing film had a rougher texture with globular structures and interconnected channels, likely representing dispersed EO droplets and matrix alterations. The mechanical analysis revealed increased elongation at break (20.05 ± 0.784%) for EO-incorporated films, indicating improved flexibility, while tensile strength and Young’s modulus decreased, suggesting reduced stiffness. Film thickness increased slightly with EO (0.097 ± 0.008 mm) compared to the control (0.089 ± 0.001 mm), though the difference was not statistically significant (p > 0.05). Moisture content decreased in EO-containing films (28.894%) compared to the control (35.236%), enhancing water resistance. Water solubility increased slightly (16.046 ± 0.003% vs. 15.315 ± 0.040%), while the swelling rate decreased significantly (0.189 ± 0.003 vs. 0.228 ± 0.040; p < 0.05), indicating greater structural stability in aqueous environments due to the hydrophobic nature of EO. Transparency tests showed that EO slightly increased film opacity (0.350 ± 0.02 vs. 0.290 ± 0.012), aligning with trends in UV-protective materials. The EO-incorporated films also exhibited moderate antibacterial activity against Staphylococcus aureus and Escherichia coli. Antifungal tests revealed strong inhibition of Botrytis cinerea (100%) and moderate inhibition of Alternaria alternata (50%) in EO-containing films. These results demonstrate that EO incorporation improves the functional properties of pectin films, enhancing their flexibility, antimicrobial activity, and environmental stability, making them promising candidates for sustainable food packaging applications. These novel active food packaging materials exhibit strong physical properties and significant potential in maintaining food quality and prolonging shelf life.

Alkaloid Profile, Anticholinesterase and Antioxidant Activities, and Sexual Propagation in Hieronymiella peruviana (Amaryllidaceae).

Hieronymiella peruviana, a recently described endemic species from southern Peru, belongs to the Amaryllidaceae family and is known for its diversity of alkaloids. Amaryllidoideae have been studied for their diverse biological activities, particularly for their properties in treating neurodegenerative diseases. This work examines the alkaloidal profile using GC-MS and UPLC-MS/MS of alkaloid-enriched extracts obtained from the leaves and bulbs of H. peruviana and their inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes. In addition, the phenolic and flavonoid content in the methanolic extract from bulbs was quantified and the antioxidant capacity was evaluated. Moreover, the seed germination was investigated under four temperature conditions (15, 20, 25, and 30 °C). Twenty-two alkaloids, most of them of the homolycorine- and galanthamine-type, including galanthamine, were identified in the alkaloid extracts by means of GC-MS and UPLC-MS/MS analysis. Lycorine and 8-O-Demethylhomolycorine were the most abundant alkaloids in the bulbs and leaves, respectively. The leaves and bulbs alkaloid extracts demonstrated strong AChE inhibition (IC50 = 5.20-8.60 µg/mL) and moderate BuChE inhibition (IC50 = 90.20-122.76 µg/mL). The bulbs' methanolic extract exhibited mild antioxidant activity, showing 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) values of 16.36% at 500 μg/mL and 58.31 mg Trolox equivalents (TE)/g, respectively. Seed germination was most effective at 20 °C and 25 °C. Finally, 60 days after germination, the survival rate of H. peruviana seedlings was 48.33%. These findings establish H. peruviana as a promising source of bioactive alkaloids with potential pharmacological and therapeutic applications, as well as providing critical insights for its propagation and conservation.

Interaction between nitrate and trichloroethene bioreduction in mixed anaerobic cultures.

Bioremediation of trichloroethene (TCE)-contaminated sites often leads to groundwater acidification, while nitrate-polluted sites tend to generate alkalization. TCE and nitrate often coexist at contaminated sites; however, the pH variation caused by nitrate self-alkalization and TCE self-acidification and how these processes affect nitrate reduction and reductive dichlorination, have not been studied. This study investigated the interaction between nitrate and TCE, two common groundwater co-contaminants, during bioreduction in serum bottles containing synthetic mineral salt media and microbial consortia. Our results showed that TCE concentrations up to 0.3 mM stimulated nitrate reduction, while the effect of nitrate on TCE reductive dechlorination was more complex. Nitrate primarily inhibited the reduction of TCE to dichloroethene (DCE) but enhanced the reduction of vinyl chloride (VC) to ethene. Mechanistic analysis suggested that this inhibition was due to the thermodynamic favorability of nitrate reduction over TCE reduction, while the promotion of VC reduction was linked to pH stabilization via self-alkalization. As the initial nitrate concentration increased from 0 to 3 mM, the relative abundance of putatively denitrifying genera, such as Petrimonas and Trichlorobacter, increased. However, the abundance of fermentative Clostridium sharply declined from 31.11 to 1.51%, indicating strong nitrate inhibition. Additionally, the relative abundance of Dehalococcoides, a genus capable of reducing TCE to ethene, slightly increased from 23.91 to 24.26% at nitrate concentrations up to 0.3 mM but decreased to 18.65% as the nitrate concentration increased to 3 mM, suggesting that Dehalococcoides exhibits a degree of tolerance to high nitrate concentrations under specific conditions. Overall, our findings highlight the potential for simultaneous reduction of TCE and nitrate, even at elevated concentrations, facilitated by self-regulating pH control in anaerobic mixed dechlorinating consortia. This study provides novel insights into bioremediation strategies for addressing co-contaminated sites.

Chemical Composition, Antimicrobial, and Enzyme Inhibitory Properties of Elaeagnus angustifolia L.

Research on natural antioxidants derived from plants has surged due to their potential health benefits. In the current study, the chemical composition, enzyme inhibitory activity, and antimicrobial effects of the Elaeagnus angustifolia L. plant, including leaves, flowers, and flower stalk (FS) extracts, were analyzed. A total of 28 compounds were identified across the different plant parts, with 24 in flowers (F), 9 in FSs, and 14 in leaves (L). The antimicrobial activity of the extracts was evaluated using the disk diffusion method against two Gram-positive (Staphylococcus aureus and Bacillus cereus) and two Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae). The minimum inhibitory concentration (MIC) was determined as 3.13-12.50mg/mL. The measured zone diameters were also in the range of 5-8mm. The results indicated that the methanol extracts exhibited significant antimicrobial properties. Additionally, the extracts demonstrated strong inhibition of cholinesterases (acetylcholinesterase [AChE]: 11.14±0.11-18.11±0.12µg/mL; butyrylcholinesterase [BChE]: 32.54±0.55-44.61±0.17µg/mL) and moderate inhibition against α-glycosidase and tyrosinase enzymes. Moreover, the molecular docking interaction of trans-ferulic, shikimic, and vanillic acids, the most abundant phenolic compounds in the extracts, with AChE, BChE, α-glycosidase, and tyrosinase, was assessed using AutoDock Vina software. E. angustifolia L. possesses a high phenolic content and exhibits significant antioxidant and enzyme inhibitory capabilities, which may contribute to its extensive utilization in food and traditional medicine.