Search Results (127)

Background: The emergence of antifungal resistance and the limited number of clinically available antifungal drug classes necessitate the development of new agents with improved efficacy and safety. We investigated how linker architecture influences the antifungal activity and lead properties of hybrid triazole-thiazolidine derivatives. Methods: A focused library of triazole-thiazolidine hybrids incorporating alkylamine, amide, cyclic amine, 2-hydroxypropyl, and thiazepane linkers was synthesized and characterized. Antifungal activity was evaluated against reference strains and clinical isolates of Candida spp., Aspergillus fumigatus, dermatophytes, and Cryptococcus neoformans. Structure–activity relationships were analyzed by molecular docking. Selected compounds were further assessed by SCRAPPY profiling, fluorescence microscopy, mammalian-cell cytotoxicity assays, acute oral toxicity studies, and evaluation of microsomal stability and interactions with human CYP450 isoforms. Results: Linker architecture strongly influenced antifungal potency. Amide- and cyclic amine-containing hybrids were generally the most active, whereas simple alkylamide derivatives showed narrower activity profiles. Compound 28 emerged as the most promising lead, exhibiting sub-microgram MIC values against several Candida isolates, particularly C. parapsilosis, and retaining measurable activity against an azole-resistant C. albicans strain. Docking generated putative CYP51-binding models, while SCRAPPY profiling and fluorescence microscopy revealed an azole-like cellular response consistent with perturbation of sterol-associated homeostasis. Compound 28 was tolerated at 300 mg/kg in an acute oral study but showed concentration- and time-dependent cytotoxicity and rapid CYP3A4-mediated microsomal metabolism. Conclusions: Systematic variation of linker architecture identified compound 28 as a promising exploratory antifungal lead. Further optimization should focus on improving metabolic stability and cytotoxicity, together with direct target validation, pharmacokinetic characterization, and in vivo efficacy studies. Full article

Prabchompoothaweep (PCT) is an anti-allergic remedy on the Thailand National List of Essential Medicines, traditionally used to relieve common colds and allergic reactions in Thai traditional medicine (TTM). However, the anti-allergic activity of PCT and its plant ingredients had not been characterized. We evaluated the acute and chronic oral toxicity of PCT ethanolic extract in rats and assessed its in vitro and in vivo anti-allergic activities. Activity in vitro was measured as inhibition of antigen-induced β-hexosaminidase release in RBL-2H3 cells and in vivo using an ovalbumin (OVA)-induced allergic rhinitis mouse model. The extract produced no mortality or signs of toxicity in either study. Several plant ingredients and the PCT extract inhibited β-hexosaminidase release more effectively than chlorpheniramine. In OVA-induced mice, PCT (75, 150, and 300 mg/kg) did not significantly lower OVA-specific serum IgE or IgG2a (OVA-specific IgG1 was likewise not reduced); however, it reduced inflammatory cell infiltration, goblet cell hyperplasia and mast cell numbers in the nasal mucosa and downregulated the T helper 2 (Th2) cytokines IL-5 and IL-13. These findings support the traditional anti-allergic use of the PCT remedy. Full article

Background: Bacillus clausii SKB/BCL21 (MCC 0569) is a novel strain that shows promise as a probiotic for both human and animal healthcare. Objectives: The objective is to evaluate the safety profile of B. clausii SKB/BCL21 through genomic and toxicity assessments in Wistar rats, as well as to assess its efficacy as a probiotic in broiler chickens challenged with Clostridium perfringens. Methods: The identification of genus and species was performed using 16S rRNA and whole genome sequencing (WGS). A genomic analysis was conducted through bioinformatic screening of the B. clausii SKB/BCL21 genome to identify virulence factors, genes encoding toxins, mobile genetic elements, and antibiotic resistance genes. In vitro biosafety assays were conducted to evaluate mucin degradation, gelatinase, hemolytic activity, and DNase activity. The in vivo safety evaluation was performed by acute and subacute oral toxicity studies as per the OECD 423 guidelines. In efficacy testing broilers challenged with C. perfringens were administered with low dose (1 × 10⁸ cfu/kg of feed) and high dose (1 × 10⁹ cfu/kg of feed) of B. clausii SKB/BCL21. Performance metrics, such as average weight gain, feed conversion ratio (FCR), and mortality rates, were evaluated in comparison to a positive control group that received Virginiamycin 50% (15 ppm). Results: The isolate SKB/BCL21 was identified as Bacillus clausii based on 16S rRNA and whole genome sequencing (WGS). The bioinformatic analysis of the B. clausii SKB/BCL21 genome reveals that it lacks genes associated with toxins, mobile genetic elements, and virulence factors. However, it does contain intrinsic and non-transferable antibiotic resistance genes within its chromosomal DNA. In the acute toxicity study, an oral dose of 2000 mg/kg (400 billion cfu/kg) body weight was found to be nontoxic. The No Observed Adverse Effect Level (NOAEL) for B. clausii SKB/BCL21 was found to be 1000 mg/kg (200 billion cfu) body weight/day by oral route in the subacute toxicity study. The findings of in vivo toxicity studies indicate that there were no treatment-related changes in any of the endpoints assessed. The effects of low (1 × 10⁸ cfu/kg of feed) and high (1 × 10⁹ cfu/kg of feed) doses of B. clausii SKB/BCL21 on the growth performance metrics, including average weight gain, feed conversion ratio, and mortality rates in broiler chickens infected with C. perfringens, showed results similar to those of the positive control (Virginiamycin 50%, 15 ppm). Conclusions: Based on these preliminary studies, B. clausii SKB/BCL21 can serve as a potential alternative to antibiotic growth promotors in broiler production. These results suggest that the B. clausii SKB/BCL21 is safe and could be a potential probiotic for animal feed supplements. Full article

Curcumin, a natural polyphenol with well-established antioxidant and anti-inflammatory properties, was evaluated for its protective effects against glyphosate (GLY)-induced mammary toxicity. Pregnant mice received GLY (50 or 250 mg/kg/day) with or without curcumin (150 mg/kg/day) by oral gavage; GLY dose-dependently disrupted alveolar architecture, caused marked inflammatory infiltration, and downregulated tight-junction proteins. In parallel, goat mammary epithelial cells (GMECs) were treated with GLY (1 or 7 mM) and curcumin (1 µM) to dissect acute cellular stress pathways. These complementary models address distinct aspects of GLY toxicity: the in vivo system reflects defined oral exposure, whereas the in vitro setting employs high concentrations to probe mechanistic events. Transcriptomic profiling combined with functional assays demonstrated that GLY triggered oxidative stress, endoplasmic reticulum stress, and intracellular Ca²⁺ overload, activated the TNFα/MAPK axis, suppressed STAT3 phosphorylation, and ultimately promoted apoptosis. Curcumin co-treatment alleviated these alterations at both structural and molecular levels. These findings indicate that curcumin can combat GLY-induced mammary injury by restoring cellular homeostasis and inhibiting apoptotic and inflammatory signaling, thereby protecting breast health. Full article

Background: Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by oxidative stress, cholinergic dysfunction, and amyloid-β (Aβ) aggregation. Tri-Sannibat-Phol (TSB), a classical Thai polyherbal formulation comprising Piper retrofractum fruit, Ocimum tenuiflorum root, and Piper nigrum root, has been traditionally used for its medicinal properties, yet its anti-AD potential has never been scientifically evaluated. Methods: The therapeutic potential of TSB was investigated through in vitro bioassays including antioxidant, acetylcholinesterase (AChE) inhibitory, and anti-Aβ aggregation assays, alongside neuroprotective evaluation in H₂O₂-induced SH-SY5Y neuroblastoma cells. Acute oral toxicity was assessed in male ICR mice in accordance with OECD Guideline 420. Cognitive-enhancing effects were evaluated using the modified Y-maze, Novel Object Recognition, and Morris Water Maze tests in a scopolamine-induced amnesic mouse model. LC-MS/MS analysis was performed for phytochemical characterization and chemical standardization of the formulation. Results: TSB demonstrated significant antioxidant activity, AChE inhibitory activity, and anti-Aβ aggregation effects, with P. nigrum and O. tenuiflorum identified as the primary contributing components. Neuroprotective effects were confirmed in H₂O₂-induced SH-SY5Y cells, where TSB significantly improved cell viability across concentrations of 1–100 µg/mL. Acute oral toxicity assessment revealed an LD₅₀ exceeding 2000 mg/kg, indicating a favorable safety profile. In vivo behavioral studies demonstrated that TSB at medium-to-high doses significantly reversed scopolamine-induced cognitive deficits across all three behavioral tests. LC-MS/MS analysis identified thirteen piperidine alkaloids, with piperine as the dominant constituent at 17.61 ± 0.80% w/w, proposed as the primary bioactive driver and chemical marker for future quality standardization. Conclusions: These findings suggest that TSB exerts multi-targeted anti-AD effects through complementary mechanisms, supporting its potential as a traditional medicine-based therapeutic candidate for further preclinical and clinical investigation. Full article

Background and Objectives: Synthetic cannabinoids (SCs) are new psychoactive substances associated with acute intoxications. Experimental data obtained under controlled and comparable conditions remain limited for this category of compounds. This descriptive, hypothesis-generating screening study aimed to characterize the acute toxicity profile of three SCs, JWH-007, AM-694, and MAB-CHMINACA. Materials and Methods: Acute toxicity was evaluated in female Swiss Albino mice, in accordance with the OECD 423 guideline, following oral and intraperitoneal administration. Animals were monitored for 14 days for behavioral and clinical signs of toxicity. At the end, histopathological examination was performed to describe organ-level changes. Serum concentrations of the tested compounds were quantified by LC-ESI-MS/MS 24 h after intraperitoneal administration. Results: The three compounds were associated with distinct behavioral, clinical, and histopathological observations. JWH-007 was associated with transient behavioral depression and histopathological changes in peripheral organs. AM-694 was associated with histopathological changes in systemic organs and limited behavioral manifestations. MAB-CHMINACA was associated with acute behavioral toxicity and central nervous system lesions, including neuronal vacuolization, necrosis, oedema, and inflammatory changes. Conclusions: These preliminary findings describe compound-specific in vivo toxicity patterns and may inform the design of future confirmatory studies on SCs’ toxicity. The observed behavioral and histopathological changes should be interpreted as hypothesis-generating and require statistical validation before conclusions can be drawn regarding comparative toxicity, structural class effects, or predictive value for risk stratification. Full article

Prospero autumnale L. is a Mediterranean medicinal plant traditionally employed for inflammatory and neurological disorders. Nonetheless, its safety profile, toxicity, and application for treating inflammation and pain are yet to be comprehensively established. This investigation aimed to assess the bioactivity and toxicity of extracts derived from its aerial (AgP) and underground (UgP) parts. The phytochemical constituents of various P. autumnale extracts were analyzed using LC-MS/MS, and their phenolic content was quantified. The biological activities were evaluated through in vitro assays—including antioxidant, anti-inflammatory, acetylcholinesterase-inhibitory, and photoprotection assessments—and in vivo experiments, including evaluations of acute oral toxicity, anti-inflammatory, and analgesic effects. UgP extracts demonstrated significant antioxidant activity, with the methanolic extract exhibiting the highest reducing and superoxide scavenging capacities. Dichloromethane and ethyl acetate extracts performed exceptionally well in ABTS and DPPH assays. The aqueous extract from AgP exhibited noteworthy anti-inflammatory and analgesic effects, surpassing diclofenac in vitro and demonstrating efficacy in vivo. It also showed considerable acetylcholinesterase inhibition, while the ethyl acetate extract displayed high photoprotective potential. The acute toxicity was moderate (LD₅₀: 300–400 mg/kg), indicating dose-dependent risks. LC-MS/MS analysis revealed diverse phenolics potentially contributing to both therapeutic and adverse effects. This research enhances the medicinal prospects of P. autumnale, provides new perspectives on plant utilization, and suggests its potential as a natural anti-inflammatory agent. However, due to moderate toxicity and dose-dependent effects, cautious application is advised. These findings underscore the importance of toxicological evaluation alongside bioactivity screening in ethnopharmacology to ensure safety. Full article

M8OI is a cytotoxic methylimidazolium ionic liquid solvent through its binding to the ubiquinone binding site on complex I of the mitochondrial electron transport chain. Given the overlap in terms of toxic mechanism of action with the pesticide rotenone, the potential neurotoxic effects of M8OI were examined. In vitro, cytotoxicity and mitochondrial function were assessed in SH-SY5Y cells by measuring MTT reduction and oxygen consumption/extracellular acidification using a Seahorse analyser. SH-SY5Y cells were sensitised to M8OI toxicity by replacing medium glucose with galactose. Glucose protected the cells from M8OI toxicity, whereas galactose showed no clear dose–response protection. M8OI induced a dose-dependent reduction in oxygen consumption rate with a compensatory increase in extracellular acidification rate, consistent with inhibition of mitochondrial oxidative phosphorylation and a shift toward glycolysis. In vivo, rats were orally exposed via drinking water for 20 weeks and assessed using behavioural tests. In addition, the concentrations of M8OI and its metabolites were quantified by LC–MS in rat brain and other tissues. In rats, M8OI concentrations were ~30-fold higher in kidney than brain, and brain levels were at least 100-fold lower than the concentrations that affected SH-SY5Y cell viability in vitro. However, based on open field tests, M8OI exposure suppressed motor activity without any anxious behaviours. The cytotoxicity of M8OI in SH-SY5Y neuroblastoma cells was associated with metabolic mitochondrial dysfunction. However, the neurobehavioural changes observed in orally exposed rats occurred at significantly lower brain concentrations than would be predicted to lead to neural cell death. Nevertheless, direct comparisons between acute in vitro exposures and chronic in vivo outcomes should be interpreted cautiously. Full article

Aspergillus cristatus is a key microorganism involved in the fermentation of Fu brick tea, but systematic strain-level safety data remain limited despite its long history of use in dark tea production. In this study, the toxicological safety of A. cristatus CCNH008 was assessed through acute oral toxicity assay, a 90-day repeated oral toxicity study with a recovery period and genotoxicity assays. In acute oral toxicity tests, CCNH008 caused no mortality, no adverse clinical signs, and no treatment-related effects on bodyweight or pathology in mice or rats, with an LD₅₀ greater than 10 g/kg bodyweight. In the 90-day repeated oral toxicity study in rats, the oral administration of CCNH008 at doses up to 1.67 g/kg bodyweight/day produced no treatment-related changes in clinical signs, bodyweight, food consumption, hematological or biochemical parameters, urinalysis, organ weights, or histopathology, including during a 28-day recovery period. Genotoxicity was evaluated using a bacterial reverse mutation assay, an in vitro mammalian chromosome aberration test, and an in vivo mammalian erythrocyte micronucleus test; no mutagenic or clastogenic effects were observed. Collectively, these findings demonstrate that CCNH008 shows no evidence of acute, subchronic oral toxicity, or genotoxicity, providing important strain-level safety evidence to support its potential application in food-related products. Full article

The growing global protein demand and environmental concerns from conventional animal agriculture have driven the exploration of sustainable alternative protein sources. Single-cell proteins (SCPs) from microbial fermentation offer a promising solution. This study comprehensively evaluated the nutritional value and safety profile of SCP produced from Ralstonia eutropha H16 through integrated in vitro and in vivo assessments. Nutritional analyses revealed a high crude protein content of 71.87 ± 5.05 g/100 g dry weight, with total amino acids of 53.67 ± 1.05 g/100 g. The essential amino acid content was 24.38 ± 0.51 g/100 g, accounting for 45% of the total amino acids. An essential amino acid index (EAAI) of 1.46 ± 0.04 and an amino acid score (AAS) of 0.83 ± 0.06 confirmed its classification as a high-quality protein source according to FAO/WHO standards. In vivo rat feeding trials demonstrated an adjusted protein efficiency ratio (PER) of 1.81, exceeding common plant proteins such as wheat (0.8–1.1). True digestibility (TD) reached 85.73%, with a biological value (BV) of 49.37%, net protein utilization (NPU) of 42.33%, and protein digestibility-corrected amino acid score (PDCAAS) of 0.71. Comprehensive safety assessments included chemical contaminant screening, acute oral toxicity studies in rats and mice, in vitro chromosome aberration tests, and erythrocyte micronucleus tests. Heavy metals and aflatoxin B₁ levels were below regulatory limits. Acute oral toxicity studies established LD₅₀ values exceeding 10,000 mg/kg body weight in both rodent species, classifying this protein source as practically non-toxic. The 28-day sub-acute toxicity study showed no significant adverse effects at low doses (6.25% protein replacement). Both genotoxicity assays (mammalian cell chromosome aberration assay and mammalian erythrocyte micronucleus test) returned negative results. These findings establish R. eutropha H16-derived SCP as a safe, nutritious, and sustainable protein source with considerable potential for feed and food applications, contributing to global food security and environmental sustainability. Full article

Background/Objectives: Onion (Allium cepa) peems are an underutilized by-product rich in polyphenols. This study evaluated the physicochemical profile, and bioactive potential (antidiabetic, antimicrobial, antioxidant, and anticoagulant) of Moroccan red onion peels using integrated in vivo, in vitro, and in silico approaches. Methods: Moisture, pH, ash content, and mineral elements were determined, followed by phytochemical screening and three extractions: decoction E0, aqueous Soxhlet E1, and hydroethanolic Soxhlet E2 (70/30; ethanol/water, v/v). The measurement of polyphenols, flavonoids, and tannins was carried out using colorimetric methods, while the molecular profile was studied by high-performance liquid chromatography coupled to ultraviolet detection and electrospray ionization mass spectrometry (HPLC/UV-ESI-MS). Biological activities were determined using 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power, and total antioxidant capacity assays (in vitro antioxidant); microdilution (antimicrobial); prothrombin time and activated partial thromboplastin time (anticoagulant); and α-amylase/α-glucosidase enzymatic inhibition and oral glucose tolerance tests on normoglycemic rats. Also, acute toxicity was evaluated, and molecular interactions between these proteins and ligands (docking, molecular dynamics, and MM-PBSA) were analyzed. Results: Physicochemical analyses showed an acidic pH (3.06) and high ash content (15.21%), with the concentration of regulated elements remaining within FAO/WHO limits. The extractive content was between 6.90% E0 and 19.18% E2. The E1 extract had the maximum amount of total polyphenols (178.95 mg GAE/g); on the other hand, E2 was the richest in flavonoids by 121.43 mg QE/g. The HPLC/ESI-MS analysis of E0 revealed 20 compounds, among which flavonoids (84.93%) were predominant, with isorhamnetin (30.26%), followed by quercetin and its glycosylated forms. E1 showed the most potent antioxidant effects (IC₅₀ DPPH, 22.38 µg/mL, as that of ascorbic acid). The antibacterial activity of E0 was especially potent towards Enterobacter cloacae and Pseudomonas aeruginosa (MIC 75 µg/mL). A mild dose-dependent anticoagulant effect was seen. Antidiabetic activity was found to be outstanding: α-amylase (IC₅₀ 62.75 µg/mL) and α-glucosidase (IC₅₀ 8.49 µg/mL, stronger than acarbose) inhibitions were corroborated in vivo by a considerable decrease in the glycemic area under the curve. The molecular docking study in silico demonstrated strong molecular interactions, especially for quercetin 4′-O-glucoside with good binding energies. Conclusions: A. cepa peels from Morocco can be considered a safe plant matrix containing bioactive flavonoids with strong antioxidant and selective antimicrobial activities and promising antidiabetic effects, supported by molecular modeling. Full article

Background: Oral squamous cell carcinoma (OSCC) is an aggressive cancer closely associated with smoking and alcohol consumption, with a higher incidence in men. Despite changes in treatment strategies, poor survival persists in most patients, highlighting the need for novel and improved therapeutic options. Naphthoquinone analogs are being investigated because of their active redox structure and broad pharmacological profile; they demonstrate cytotoxic antitumor activity, making them potential candidates for new drug agents. Objective: This study investigated new benzyl-naphthoquinone compounds as potential anticancer agents for various genotypes of oral squamous cell carcinoma (OSCC) and other cancer cells. Methods: This study reports the synthesis and evaluation of a series of eight benzyl-naphthoquinone compounds against oral squamous cell carcinoma. Results: Four compounds 1–4 showed the best cytotoxic profiles, with a selectivity index ≥ 3 for all OSCC cell lines tested. Compound 1 was the most selective compound in all OSCC models, showing a higher selectivity index than both carboplatin and shikonin. Furthermore, compound 1 induced DNA fragmentation, cell-cycle arrest, and caspase-3/7 activation, changes consistent with apoptosis, and time-lapse imaging corroborated the apoptotic phenotype. Hemolysis assays showed minimal toxicity in human erythrocytes, and acute in vivo evaluation in mice revealed no evident adverse effects under the conditions tested, indicating low acute toxicity, although more detailed histopathological and biochemical studies will be required to fully establish the safety profile. Molecular modeling suggested that compound 1 may interact with topoisomerase II, RSK2, and PKM2, which could contribute to the activation of apoptotic pathways, although these interactions remain predictive and require biochemical validation. Finally, in silico analysis of physicochemical and ADMET parameters indicated properties compatible with oral absorption and systemic exposure, together with predicted low toxicity; however, these results are model-based and should be confirmed experimentally. Conclusions: Based on these findings, compound 1 emerges as a promising lead candidate for the development of a novel chemotherapeutic agent against OSCC, with potential therapeutic efficacy against other cancer types. Full article

Background/Objectives: The use of the drug delivery system is notable for the systemic improvement of low orally bioavailable compounds, such as the bioactive phenolic acid, syringic acid. Innovative techniques are employed to enhance the performance of certain drug delivery systems. In connection with our previously reported journal with the use of MIL-100(Fe) as a drug carrier for syringic acid, this study utilized a mixed-linker synthesis of syringic acid and trimesic acid and characterized the properties in comparison with the unmodified MIL-100(Fe) through a solid solution approach. Methods: Modified MIL-100(Fe) was synthesized by substituting different molar concentrations of syringic acid for trimesic acid through de novo synthesis. Simple impregnation of syringic acid was carried out at 12, 24, 36, and 48 h and at 1:1 and 1:2 molar ratios of MIL-100(Fe) to syringic acid. Characterization was performed via PXRD, FTIR, BET, SEM, and DLS. In vivo studies included acute oral toxicity testing (OECD 425) and bioavailability assessment in Sprague Dawley rats. Results: The optimized amount of syringic acid to be substituted for trimesic acid is 0.10 mmol, as confirmed by the value of the PXRD. Optimized drug loading of 66.85 ± 0.004% was achieved using a 1:2 ratio of syringic acid to MIL-100(Fe)-10% over 36 h. Structural modifications were confirmed via FTIR, specifically through shifts at 1239.2 cm⁻¹, while TGA demonstrated thermal stability up to approximately 350 °C. Morphological analysis by SEM showed octahedral particles (210.70 ± 1.23 nm), and a decrease in BET surface area post-loading verified successful encapsulation. While in vitro release was media-dependent, toxicity studies at 2000 mg/kg showed no adverse effects; notably, SGOT and SGPT levels decreased, though BUN and creatinine levels rose. Compared to pure oral syringic acid, the SYA@MIL-100(Fe)-10% formulation demonstrated a 5.09-fold increase in relative bioavailability. Furthermore, it outperformed intraperitoneal administration of the drug by 1.65-fold. Conclusions: Modification of MIL-100(Fe) by incorporating syringic acid into the framework as a substituted organic linker indicates that SYA@MIL-100(Fe)-10% is a safe and effective delivery system for syringic acid, enhancing oral bioavailability. To the best of our knowledge, this is the first study to investigate the mixed-linker synthesis of MIL-100(Fe) by utilizing syringic acid as a structural co-ligand, rather than solely as an encapsulated guest. While MIL-100(Fe) has been extensively employed as a carrier for various therapeutics, this research uniquely integrates the active agent into the framework lattice itself to modulate porosity and loading capacity, subsequently evaluating its systemic performance in an in vivo model. Full article

Background/Objectives: Branched-chain fatty acids (BCFAs) exhibit a range of biological activities; however, their limited natural abundance and high cost have constrained in vivo research. Lanolin represents a promising source for enriching BCFAs. Nevertheless, the in vivo application, safety, and dose-effect relationship of BCFAs derived from lanolin (BCFAs-DFL) remain unassessed. Methods: In this study, the acute toxicity in C57BL/6J mice was first evaluated for 7 days by a single oral administration of 5000 mg/kg BW of BCFAs-DFL. Subsequently, 40 mice were divided into four groups (control group, low dose of 100 mg/kg BW, medium dose of 300 mg/kg BW, and high dose of 600 mg/kg BW) and were continuously administered by gavage for 28 days to study the effects of BCFAs-DFL on the growth, blood biochemistry, intestinal morphology, and intestinal flora of the mice. Results: In the acute toxicity test, BCFAs-DFL exhibited no lethality or abnormalities in mice, indicating its non-toxic nature. Throughout the 28-day trial, mice in the medium- and high-dose groups experienced a notable decrease in average daily feed intake (p < 0.05), yet their weight gain remained unaffected (p > 0.05). Hemoglobin and hematocrit levels declined in the high-dose group (p < 0.05). Conversely, serum aspartate aminotransferase and total bilirubin levels escalated in the medium- and high-dose groups, while triglycerides and urea nitrogen levels decreased (p < 0.05). The serum’s total antioxidant capacity and immunoglobulin levels (IgA, IgG) rose in proportion to the dosage (p < 0.05). BCFAs-DFL notably enhanced the villus height of the jejunum and ileum in mice (p < 0.05). Gut microbiota analysis indicated no significant impact on overall α and β diversity. Conclusions: The 28-day intervention revealed that BCFAs-DFL can modulate feeding behavior, TG, T-AOC, and immunoglobulin levels in mice. Additionally, it promotes the development of intestinal villi. Based on various indicators, a dosage of 100 mg/kg BW effectively induces beneficial metabolic regulation, such as the reduction of triglycerides, without causing a burden on liver metabolism. This dosage may represent a more suitable application for potential use. Full article

This study evaluated two formulations of L-carnitine, which were developed and impregnated in an oil-based self-emulsifying system (SEDDS), the first with tyrphostin AG17 and the second without the addition of tyrphostin AG17. The formulation with tyrphostin AG17 showed the presence of stable microvesicles up to 498 h after its preparation. To establish a robust safety profile in compliance with modern regulatory frameworks and the 3Rs principle (replacement, reduction, and refinement), a toxicological evaluation was conducted integrating an in silico quantitative structure–activity relationship (QSAR) analysis with confirmatory in vivo subchronic toxicity studies. The QSAR analysis, performed using the OECD QSAR Toolbox and strictly adhering to Organization for Economic Co-operation and Development (OECD) validation principles, predicted an acute oral LD50 of 91.5 mg/kg in rats, a value showing high concordance with the historical experimental data (87 mg/kg). Furthermore, computational modeling for repeated-dose toxicity yielded a no-observed-adverse-effect level (NOAEL) of 80.0 mg/kg bw/day, a no-observed-effect level (NOEL) of 60.4 mg/kg bw/day, and an ADI = 56 mg/day. These computational findings were substantiated by a 90-day subchronic toxicity study in male Wistar rats, where daily intragastric administration of tyrphostin AG17 at doses up to 1.75 mg/kg resulted in not statistically significant hematotoxic activity (p < 0.05), with a maximum cumulative dose over 90 days of 157.5 mg/kg. Collectively, these data indicate that tyrphostin AG17 combines high stabilizing efficacy with a manageable safety profile, supporting its proposed regulatory status as a functional food additive. Based on these results, it is concluded that tyrphostin AG17 shows promising characteristics for use as a stabilizer in food and other substances. Full article