Metabolic adaptation enables cancer cells to persist under fluctuating nutrient conditions and is a major driver of tumor progression, including colorectal cancer (CRC). The contribution of diet, such as red and processed meat, in tumor progression remains poorly understood. Here, we investigated how repeated exposure to hemin (HEM) and kynurenine (KYN), alone and in combination, shapes CRC cell metabolism under nutrient deprivation. Using respirometry and transcriptomics in 3D spheroids, we found that HEM and KYN induced distinct, context-dependent metabolic adaptations, differentially affecting glycolytic and oxidative energy pathways depending on nutrient availability. Co-exposure to HEM + KYN maintained energy production during glucose deprivation through enhanced lipid storage, revealing a mechanism by which red meat–derived metabolites reinforce metabolic plasticity. These findings demonstrate that red meat–derived metabolites modulate key bioenergetic pathways in a complementary manner and that their combined presence amplifies cancer cell adaptability, highlighting a potential mechanism linking diet to CRC progression.

Curcumin exhibits broad biological activity, including anticancer effects, but its therapeutic potential is limited due to poor stability, rapid metabolism, and non-specific activity, prompting the development of structurally modified analogues with improved drug-like properties. However, how such modifications influence intracellular metabolism and subcellular behaviour remains insufficiently understood. In this study, the intracellular fate of the reference molecule curcumin and three promising curcumin-based benzothiazepane analogues with enhanced selective anticancer activity was investigated in a cancerous (HCT-116) and non-cancerous (IPEC-J2) intestinal cell line. An untargeted LC-QTOF-MS approach was applied to characterise degradation and metabolic products in cell lysates and extracellular medium, complemented by fluorescence-based imaging to explore their subcellular distribution. Interestingly, all compounds were detected intracellularly in both cell lines, but displayed distinct structure- and cell-dependent metabolic patterns. Sulphation was predominant in HCT-116 cells, whereas methylation was more abundant in IPEC-J2 cells. In addition, phase II modifications – methylation and sulphation – occurred preferentially at phenolic moieties. Fluorescence imaging revealed mainly cytosolic, nuclear-excluded distribution for all compounds, with limited evidence for selective organelle accumulation. These findings demonstrate that curcumin-based benzothiazepane analogues undergo distinct cell-dependent metabolic processing, highlighting intracellular metabolism as a potential contributor to their previously reported selective anticancer activity.

Curcumin exhibits broad biological activity, including anticancer effects, but its therapeutic potential is limited because of poor stability, rapid metabolism, and nonspecific activity, prompting the development of structurally modified analogs with improved drug-like properties. However, how such modifications influence intracellular metabolism and subcellular behavior remains insufficiently understood. In this study, the intracellular fate of the reference molecule curcumin and 3 promising curcumin-based benzothiazepine analogs with enhanced selective anticancer activity was investigated in cancerous (HCT-116) and noncancerous (IPEC-J2) intestinal cell lines. An untargeted liquid chromatography with quadrupole time-of-flight mass spectrometry approach was applied to characterize degradation and metabolic products in cell lysates and extracellular medium, complemented by fluorescence-based imaging to explore their subcellular distribution. Interestingly, all compounds were detected intracellularly in both cell lines but displayed distinct structure- and cell-dependent metabolic patterns. Sulfation was predominant in HCT-116 cells, whereas methylation was more abundant in IPEC-J2 cells. In addition, phase II modifications-methylation and sulfation-occurred preferentially at phenolic moieties. Fluorescence imaging revealed mainly cytosolic, nuclear-excluded distribution for all compounds, with limited evidence for selective organelle accumulation. These findings demonstrate that curcumin-based benzothiazepine analogs undergo distinct cell-dependent metabolic processing, highlighting intracellular metabolism as a potential contributor to their previously reported selective anticancer activity. SIGNIFICANCE STATEMENT: By combining liquid chromatography with quadrupole time-of-flight mass spectrometry and subcellular fluorescence microscopy, this study provides insight into the intracellular fate of curcumin and 3 analogs, which differed both between compounds and between cancerous HCT-116 and noncancerous IPEC-J2 cells. These findings highlight the importance of structural modifications and cell-dependent metabolic processing when evaluating bioactive compounds because such mechanisms may substantially influence biological activity and selectivity.

This study investigates the antidiabetic activity, gastrointestinal stability, and permeability of a modified yeast-derived peptide, VLSTSFPPW (VW9), using in vitro gastrointestinal digestion and Caco-2 cell models. LC-MS/MS analysis revealed that VW9 with DPP-IV inhibitory activity, was extensively cleaved during digestion into smaller fragments like PPW, FPPW, FPP, ST, and SFPP. Among these, PPW displayed the highest activity in vitro (IC50: 22.60 ± 0.18 μM) and in situ (IC50: 474 ± 10.2 μM). Kinetic and molecular docking studies indicated that PPW may inhibit the DPP-IV enzyme in a competitive way mainly due to hydrogen-bond interactions between Trp residue at C-terminus of PPW and the active site residues of DPP-IV. PPW could not penetrate epithelial cells, but a modified derivative was found on both apical and basolateral sides without DPP-IV inhibitory activity. These findings highlight the challenges of stability, activity, and permeability of VW9 and its derived peptides for potential therapeutic applications.

Type 2 diabetes mellitus (T2DM) has become a serious and growing public health concern. Marine-derived bioactive peptides (MDBPs), characterized by structural diversity and a high content of hydrophobic amino acids, offer a valuable reservoir for identifying novel antidiabetic compounds. This work aims to explore the T2DM therapeutic potential of MDBPs. It provides an overview preparation of MDBPs, and investigates their interactions with key therapeutic targets in T2DM, and examines their structure-activity relationships. Additionally, safety considerations of MDBPs and strategies for improving bioavailability of MDBPs are addressed. MDBPs contribute to T2DM management by modulating key metabolic enzymes through hydrogen bonding and hydrophobic interactions, thereby influencing glucose metabolism and insulin signaling. The therapeutic potential of MDBPs is influenced by intrinsic factors such as peptide sequence, molecular weight, and hydrophobicity, which determine stability, bioactivity, and target specificity. Advanced delivery systems, including microencapsulation and nanocarriers, have been employed to enhance their bioavailability and controlled release. However, rigorous safety evaluations and clinical studies remain essential to facilitate their successful translation into viable therapeutic options for T2DM.

The combination of heat and enzymatic treatment on ovalbumin (OVA) has been shown to lead to a mixture of amyloid-like fibrils (ALFs) and peptides. Due to their gelling behavior, these mixtures are able to stabilize oil-inwater (O/W) emulsions. As peptides present alongside the OVA ALFs can impact interfacial behavior, it is necessary to remove them. Here, the ability to separate peptides from OVA ALFs by ultracentrifugation and dialysis was investigated. Size exclusion-high performance liquid chromatography results showed that dialysis produced pure OVA ALFs, while ultracentrifugation resulted in both a fibril- and a peptide-enriched fraction. Drop shape tensiometry confirmed that dialysis removed peptides, as a delayed decrease in interfacial tension indicated slower adsorption kinetics of larger structures. This was further supported by an increase in dilatational elasticity compared to samples containing peptides. Emulsions [10.0 % (O/W)] with only fibrils (obtained by dialysis) contained oil droplets that were noticeably larger than those in emulsions containing peptides. However, these emulsions exhibited high creaming and coalescence stability. In contrast, emulsions with peptideenriched OVA dispersions contained smaller oil droplets but were prone to coalescence due to the lack of a thick viscoelastic layer or a highly viscous continuous phase. The obtained results suggest that peptides, when present, preferentially adsorb at the interface, favoring small emulsion droplets, while the long OVA ALFs form a gel-like network in the continuous phase. This distinct interfacial behavior of peptides and ALFs, in the presence or absence of each other, may be useful when considering their use in food products.

Developing edible, cell-interactive, animal-free scaffolds and replicating the structure and composition of natural meat, including its fat content, remain major challenges in the development of cultured meat. This review explores recent advances in scaffold materials, focusing on plant-based proteins and polysaccharides supporting muscle cell adhesion, proliferation, and differentiation. Techniques such as micropatterning, directional freezing, and extrusion, along with functionalization using adhesion peptides, are reviewed to enhance cell alignment and interaction. The review also highlights the development of microcarriers and the challenges of bioreactor-based cell culture using plant-derived materials. Additionally, the integration of structured plant-based lipids is examined as a viable strategy to mimic animal fat, thereby enhancing texture, flavor, and nutritional value. Despite encouraging progress, data on long-term culture and larger-scale applications are often missing. Further research is needed to ensure scalability and validate results with primary livestock cells. Moreover, the impact of processing conditions on the cell-interactive properties of plant-derived proteins remains poorly understood. Additionally, specific components responsible for influencing cell adhesion and growth are often not identified. Future work should assess the organoleptic properties, digestibility, and regulatory considerations of the cultured hybrid product. Ultimately, plant-based scaffolds and fats offer a promising path toward the production of cultured meat products that do not require animal-derived scaffolding materials or fats, thereby reducing the reliance on animal inputs beyond the initial cell source.

β-carotene is a carotenoid with provitamin A activity whose digestive stability and bioaccessibility prior to intestinal absorption are important to fully exploit its health benefits. Microencapsulation protects carotenoids, but there is a lack of information on the extent to which its characteristics and interactions with complex food matrices could impact the carotenoid micellization during digestion. We evaluated the effect of milk fat content on the in vitro bioaccessibility of β-carotene from microparticles containing carotenoids from mango peel. The microparticles tested contained solvent-extracted carotenoids and supercritical fluid-extracted carotenoids, and were separately co-digested with whole, semi-skimmed and skimmed milks. Bioaccessibility was assessed using an in vitro digestion method adapted to carotenoids. β-carotene recoveries after in vitro digestions ranged from 79.6 to 102.2 %, with the highest values corresponding mainly to microparticles with the lowest initial β-carotene concentration. β-carotene bioaccessibilities ranged from 8.8 to 75.5 %, the highest values being obtained mainly when the microparticles were co-digested with whole milk, especially when those containing supercritical fluid-extracted carotenoids were used. The bioaccessibility-enhancing effect of the milk fat was explained by the higher concentration of free fatty acids in the micellar phase, while the better results in the microparticles containing supercritical-fluid-extract was attributed to the lower initial concentration of β-carotene. In conclusion, increasing the milk fat content increased the bioaccessibility of encapsulated β-carotene from mango peel, further determining that, a lower initial concentration of β-carotene in the microparticles resulted in higher bioaccessibility.

Scaffold development for muscle cell growth in cultured meat production requires understanding the mechanical properties of the extracellular matrix (ECM) in bovine muscle tissue. However, previous studies have focused on non-bovine samples and have altered the native ECM structure by processing it into hydrogels. This study characterizes the native ECM of bovine muscle tissues, providing reference data for designing ECM alternatives while preserving its structural and mechanical features. Protocols were optimized for thin samples (similar to 1.5 mm), corresponding to the cultured meat products currently under development. ECM was isolated from fresh bovine sirloin and tenderloin using 0.5 % SDS for decellularization. An extensive sample quality validation was performed, including cryo-SEM to visualize internal structures, Picogreen assays to quantify residual dsDNA and confirm effective cell removal, and proteomic and glycosaminoglycan analyses to verify retention of essential ECM components. Subsequent mechanical analyses included amplitude sweeps (G', G", linear visco-elastic region, cross-over point) and texture analyses (total extension, maximal load). Results confirmed efficient decellularization, preserving the ECM's structural and compositional integrity. The storage modulus (G') was 15.2 kPa for sirloin and 12.3 kPa for tenderloin, decreasing significantly to 0.3 kPa (p <0.01) and 1.0 kPa (p <0.001) for their decellularized counterparts. Texture analyses revealed no significant differences, suggesting ECM primarily determines these properties. Maximal load ranged from 1.4 N to 3.1 N, while total extension varied between 9.2 mm and 11.6 mm. These findings provide reference data for scaffolds replicating natural ECM properties and establish target mechanical properties for cultured muscle fiber constructs.

While wheat is a good source of iron (Fe) and zinc (Zn), their bioaccessibility is limited due to chelation with phytic acid. Sprouting wheat at 26 °C reduced the phytate content by 25-40 % and increased bioaccessibility (determined with an in vitro digestion assay) 1.5-2.7 times (Fe) and 1.6-2.3 times (Zn), depending on the sprouting time (48-120 h). Subsequent application of in vitro digests to Caco-2 cells showed that sprouting for 120 h at 26 °C did not enhance wheat Fe bioavailability, while it increased Zn bioavailability 1.6-fold. When the in vitro digests were exposed to diffusive gradients in thin film (DGT) with Chelex resin, which binds free mineral ions, sprouting was found to have caused release of non-labile Fe complexes and some labile Zn complexes. The latter resulted in higher levels of bioavailable forms of Zn in wheat.

Despite extensive investigation into the anti-cancer activity of the natural polyphenol curcumin, its therapeutic application is restricted by its inherent physicochemical properties. Synthetic curcumin analogues, however, offer a promising strategy to improve the drug-like potential of curcumin. In this study, we evaluated three curcumin-based benzothiazepane analogues for their ability to selectively target colon cancer cells. Their cytotoxicity was assessed on intestinal cancerous HCT-116 and non-cancerous IPEC-J2 cells using cell viability assays and microscopic imaging. Two analogues, AT007 and AT096, demonstrated enhanced anti-cancer selectivity compared to curcumin. Interestingly, this effect correlated with the aggregation of these compounds in cell medium, which was influenced by compound concentration and medium composition (particularly the presence of albumin). Confocal microscopy confirmed the presence of particles up to 12 µm inside both cell lines, yet downstream metabolic and transcriptomic responses revealed distinct coping mechanisms that may underlie the higher survival of IPEC-J2 cells. Rather than direct molecular interactions typical of soluble compounds, the observed selectivity appears to result from indirect, particle-driven physical effects, potentially involving (intracellular) membrane disruption. Our findings suggest that aggregation behaviour can be a key determinant in improving the potency and selectivity of bioactive compounds, opening new opportunities for the design and screening of more selective anti-cancer therapeutics.

Iminosugars have a carbohydrate-like backbone in which the ring oxygen is replaced by nitrogen. They are naturally found in foods such as rice, buckwheat, mulberries, and fermented vegetables, and are reported to exert anti-hyperlipidemic and anti-hyperglycemic effects due to the inhibition of cellular glycosidases. This mechanism suggests their potential role in cancer treatment and prevention. In this study, two natural iminosugars, D-fagomine (FGM) and 1-deoxynojirimycin (DNJ), and their synthetic derivatives were screened for potential anticancer properties using Caco-2 and HCT-116 cells as models for the early and late stages of colon cancer, respectively. Iminosugars were found to decrease cell viability, with effects varying based on the type of iminosugar, cell type, growth condition (glucose concentration), exposure time (1 vs. 13 days), and tissue architecture (monolayer vs. spheroid). The combined use of innovative techniques, such as IncuCyte® live cell imaging and Seahorse real-time cellular metabolic analysis, and microscopic observation after staining enabled us to detect changes in substrate utilization for energy metabolism, including increased glycolysis and alterations in lipid and glycogen stores. The evidence that iminosugars, both natural and synthetic, influence cellular bioenergetics paves the way for their potential use in various applications, including cancer treatment.

Large-scale production of cultured meat requires muscle cell culture in bioreactors, where microcarriers (MCs) support cell attachment, growth, and differentiation. However, most MCs are composed of inedible materials, requiring a cell detachment step, and/or contain animal-derived components, which are undesirable for cultured meat production. Therefore, we developed animal-free edible microcarriers based on soy protein isolate (SPI) that support muscle cell growth. SPI MCs supported cell attachment and growth similar to commercial collagen-coated dextran MCs, as bovine myoblasts expanded 24-fold over 8 days in a bioreactor. Moreover, myoblasts could differentiate into myotubes on the SPI-MCs. Importantly, SPI supported cell attachment in serum-free medium, as opposed to methacrylated gelatin (GelMA). Proteomics analysis revealed that, during SPI processing, cell adhesion peptides become available on the biomaterial, which also partially leach into the cell culture medium and replace serum components. To conclude, our study demonstrates the feasibility of growing and differentiating bovine muscle cells on edible, fully plant-based MCs, providing a scalable system for the production of cultured meat.

Induction of amyloid-like morphology in food proteins offers high potential to induce new techno-functional properties in food products (e.g. use as emulsifier, thickener or gelling agent in e.g. bakery and confectionery products). However, the health impact of amyloid-like fibril (ALF) consumption remains widely understudied and merits additional research. The aim of this study was to (partially) elucidate the general health impact of food-borne ALF consumption, using egg white ovalbumin as a case study. Based on in vitro cell culture models it was demonstrated that ovalbumin ALFs (i) do not induce direct cytotoxic effects on intestinal (Caco-2, IPEC-J2) and neuronal (SH-SY5Y) cell lines, but (ii) are able to induce a Toll-like-receptor-mediated innate immune response, similar to endogenous amyloids, in activated THP-1 cells. Furthermore, the consecutive in vitro digestion and absorption (enterocyte and M-cell) experiments demonstrated that ovalbumin ALFs (i) do not completely lose their ALF morphology upon in vitro gastrointestinal digestion, and that (ii) the ALF core sequences, located at the center of the ALF structure, are transported across Caco-2 based cell models, suggesting aggregate transport. In vivo, intestinal translocation of ingested ALFs would imply potential cross-seeding of endogenous, disease-related precursor proteins. The ability of ovalbumin ALFs to induce aggregation of a disease-related precursor protein, αSyn, was evaluated in a precursor overexpressing cell model. Here, it was illustrated that only homologous (αSyn) - but not heterologous (ovalbumin) - seeding resulted in intracellular aggregation bodies of (phosphorylated) αSyn. The lack of cross-seeding supports the assumption that ovalbumin ALF consumption is not a risk factor for the development of α-synucleinopathies like Parkinson's disease.

The liver's complex microenvironment and spatially zonated functions present major challenges for in vitro modeling, particularly in drug development and disease research. While oxygen and nutrient gradients have been used to create zonation, the potential of the surrounding biomaterial, i.e. the extracellular matrix (ECM), remains relatively underexplored. Recently, native ECM components and/or binding motifs, such as decellularized ECM (dECM) or arginine-glycine-aspartate (RGD) peptides, are increasingly integrated to improve in vitro hepatocyte functionality. However, the biological underpinning of ECM-cell interactions and resulting hepatocyte behavior are often poorly understood, which hampers to exploit the full potential of biomaterial-based strategies for relevant liver tissue modeling. Within this context, the spatial ECM characteristics within the Space of Disse are of critical importance. In this review, we therefore first outline how ECM-receptor interactions influence hepatocyte function and then review how biomaterial composition and mechanics steer zone-specific cell functionality. We propose six practical design principles to guide biomaterial engineering for future applications, including mechanical tuning, affinity-guided selection, zone-specific ligand selection, enhanced ligand diversity, binding site accessibility, and native molecular environment preservation. These strategies will not necessarily increase model complexity, but may support intentional biology-driven biomaterial design beyond conventional scaffold macro-engineering, and enhance the physiological relevance of 3D liver models without sacrificing scalability, simplicity, or reproducibility. STATEMENT OF SIGNIFICANCE: There is an urgent demand for in vitro liver models that accurately reflect human biology for drug testing, disease modeling, and regenerative medicine. Existing systems rarely capture the liver's complexity, while animal models face ethical and translational constraints. A crucial but largely neglected feature is liver zonation, the region-specific variation in hepatocyte function. While oxygen and nutrient gradients are known to influence zonation, the role of extracellular matrix (ECM) cues remains underexplored. This review integrates evidence from hepatology, ECM-receptor biology, and biomaterials to propose an evidence-based, zonation-aware design framework. By translating biological principles into actionable material guidelines, it offers researchers the tools to develop next-generation liver models that better predict human outcomes and accelerate progress across the whole field.

Cancer chemotherapy is continuously challenged by serious complications like pronounced side effects and multidrug resistance (MDR). Natural products, such as curcumin, offer promising alternatives due to their diverse biological applications and low toxicity. However, curcumin's clinical utility is limited by poor bioavailability, rapid metabolism, and non-specific (PAINS) activity. Building on previous findings, this study explored the structural modification of curcumin-inspired benzothiazepane derivatives in an attempt to enhance their therapeutic potential through modifications of the two peripheral (hetero)aromatic rings and the benzothiazepane scaffold. In this way, eight new 2-(hetero)aryl-4-(4-(hetero)aryl-2-oxobut-3-en-1-ylidene)benzothiazepanes and two 4-thiobutan-2-one "double Michael addition" derivatives were synthesized and tested for cytotoxicity against a panel of eight cancer cell lines. The screening results indicated that bis-(4-hydroxyphenyl) analogs bearing a chlorinated benzothiazepane ring exhibited the highest potency and broad-spectrum activity at the low micromolar range. Bis-substitutions with 3-pyridinyl and 2-furyl groups showed less potent but more specific activity profiles, potentially reducing PAINS effects. 2-Aminothiophenol-derived double Michael addition products demonstrated increased broad-spectrum activity, highlighting the importance of the free aniline amino group for targeted effects. This study underscores the potential of benzothiazepane derivatives as viable cancer cell cytotoxic agents and provides useful insights for future optimization and evaluation.

Curcumin is a natural product displaying a broad range of biological activities, including anticancer properties. It is, however, poorly absorbed by the human body and, as a so-called pan-assay interference compound, it exhibits non-specific activity leading to false positive results in biological assays. Nonetheless, different structural modifications of the curcumin scaffold have previously shown to lead to an improved biological and specificity profile without losing antiproliferative activity. In that respect, recent research in our group culminated in unprecedented benzothiazepane-based hit molecules with promising biological and drug-like properties. In the present hit expansion study, 14 new 2-aryl-4-(4-aryl-2-oxobut-3-en-1-ylidene)benzothiazepanes were successfully synthesized through the implementation of various aromatic ring modifications and subsequently tested for cancer cell cytotoxicity using eight different cancer cell lines, revealing useful structure-activity relationship insights for this new class of compounds.

Foam mat drying was used for the dehydration of a phenolic-rich rambutan peel extract. Foam products were evaluated for their physicochemical stability during storage. Experiments were carried out to determine the effect of foaming (10% of egg albumin/soy protein/pea protein) and stabilizing agents (0.5% of methylcellulose); storage conditions i.e. type of packages (polyethylene, glass), packaging techniques (modified atmosphere packaging, vacuum) under light or dark at 30 degrees C on phenolic compositions, antioxidant activity, water activity (aw), and color of foam products during 5 months storage. Foam products produced from egg albumin showed the highest total phenolic content (TPC) (8.73 +/- 0.17 mg GAE/g) and antioxidant activity (7.47-9.70 mg TE/g). During storage, TPC, antioxidant activity, and the amount of individual phenolic compounds (geraniin, corilagin, rutin, ellagic acid, and quercetin) mostly decreased independent of the presence of light. Aw in all samples increased from 0.25 to 0.38 - 0.45 (except for the samples in glass packaging); whereas the color of the samples was insignificantly changed for 5 months. Rambutan peel phenolic-rich extract encapsulated within egg albumin can be stored at least 5 months with minimal quality changes and might be used as a food preservative ingredient.

1,5-Benzothiazepines represent key scaffolds in medicinal chemistry because of their applicability in a wide variety of pharmaceuticals. Many cardiovascular, antidepressant, and antipsychotic drugs on the market accommodate this privileged structure, but the biological effects of 1,5-benzothiazepines are much more diverse, including enzyme inhibitory, antimicrobial, central nervous system depressant, anti-inflammatory, and anticancer activity. The introduction of a heterocyclic substituent onto a chemical structure can be beneficial in terms of bioactivity and drug-like properties. In this review, the biological activities of 1,5-benzothiazepines with and without heterocyclic substituents are discussed and compared. In some cases, the addition of a heterocycle proved to be beneficial for enhancing activity, especially the introduction of a 2-thienyl group, and also the position of heteroatoms in the substituent can have an impact on the overall properties of the resulting compounds.