Food Chemistry

Todays canola quality rapeseed press cake (RPC) is a protein-rich co-product with potential as human food, but its application is limited due to antinutritional compounds and bitter taste. It remains, however, unknown how introduction of raw RPC to a food matrix affects sensory perception and which metabolites drive the sensation. Here, raw RPC from whole or dehulled seeds was introduced into snack bars at 0%, 7%, 14%, and 21%, and sensory responses were correlated to selected known RPC-derived bitter compounds. A trained panel evaluated 13 RPC-characteristic sensory attributes, and the bitter compounds sinapic acid, kaempferol 3-O-(2'''-O-sinapoyl-{beta}-sophoroside) (KSS), KSS-hexose, selected bitter glucosinolates, and goitrin were quantified using targeted LC-MS/MS. Most dose-dependent sensory responses increased up to 14% RPC and then plateaued, whereas astringent mouthfeel increased almost linearly across the full dose range. Dehulling intensified several odor- and flavor-related attributes but did not increase bitterness or protein content in the final product. Principal component analysis linked bitterness and astringency positively with KSS, KSS-hexose, and goitrin. Dose-over-threshold analysis further showed that goitrin, but not progoitrin, reached concentrations relevant for bitterness perception. Together, the results demonstrate that raw RPC contributes distinct dose-dependent sensory attributes and that metabolite transformations in the food matrix shape final sensory profiles. These findings provide a basis for developing RPC-containing foods and for breeding rapeseed lines with improved sensory characteristics. HIGHLIGHTSO_LIThis study presents the first sensory panel assessment of rapeseed press cake (RPC)-containing in food products (snack bars) made from whole and dehulled seeds. C_LIO_LI13 RPC-characteristic sensory attributes are identified. C_LIO_LISensory profiles of the tasted snack bars differed significantly, influenced by the dosage of RPC and by the dehulling treatment. Bitterness and astringency are positively correlated with the RPC dosage. C_LIO_LIGoitrin, kaempferol 3-O-(2'''-O-sinapoyl-{beta}-sophoroside) (KSS) and sinapic acid are RPC-derived bitter compounds that correlate with bitter taste of RPC-containing snack bars. C_LIO_LIApproximately 90% of glucosinolates introduced with the RPC are not detected in the snack bars, and goitrin levels in snack bars accounts for only [~]10% of introduced progoitrin. C_LIO_LIGoitrin is - for the first time - reported to contribute to the perceived bitterness of an RPC-containing food product. C_LI

Milk microRNAs are believed to play gene regulatory functions in the consumers cells. Milk from different species is enriched in microRNAs predicted to influence immunity, metabolism, and intestinal homeostasis. For milk microRNAs to regulate gene expression in the consumer, they must survive digestion and be present at sufficient levels to influence intestinal cells and potentially beyond-intestinal cells. Milk microRNAs are proposed to be protected from degradation through their association with milk extracellular vesicles (EVs), which might also deliver them to cells. Studies on milk microRNA oral transfer and tissue bioavailability are limited by interspecies sequence homology, making it difficult to distinguish endogenous from exogenous microRNAs. Here, we used a transgenic (TG) cow model expressing four unique microRNA sequences (AmiRs) in its milk to study their association with milk EVs, their resistance to in vitro digestion, and AmiR uptake and regulatory activity in vitro. We confirmed the presence of the four milk EV populations in raw wild-type (WT) and TG cow milk, similar to those previously reported in commercial (pasteurized) cow milk, and confirmed their association with AmiRs and classical milk microRNAs. AmiRs showed differential resistance to simulated adult digestion. In vitro uptake studies showed a modest gene regulatory effect of AmiRs in Caco-2 cells incubated with TG milk EVs. The intent of using this model was to perform in vitro analysis which could lay the groundwork for later in vivo bioavailability studies, taking advantage of the uniqueness of the AmiRs sequences and bypassing the limitation of microRNA sequence homology. HighlightsO_LINew milk EV populations identified previously in commercial bovine milk were also identified in raw milk, indicating that they are not merely the result of processing C_LIO_LIThe routinely discarded EVs (12K and 35K) seem to be preferentially enriched with microRNAs in raw cow milk as was previously shown for commercial cow milk C_LIO_LIAmiRs in transgenic milk resist differentially to simulated digestion C_LI

1.BackgroundManihot esculenta (Cassava) is a vital staple in Sub-Saharan Africa, yet its high levels of cyanogenic glycosides and anti-nutrients pose health risks. While boiling is common, its holistic impact on the nutritional biochemistry and antioxidant profile of the "Farmers Pride" (IBA 961632) variety remains under-characterized. This study evaluated the sequential impact of food processing -boiling and multi-stage fermentation -on cassavas toxicological and bioactive profiles. MethodsFresh tubers were boiled for 10 minutes and fermented for 24, 48, and 72 hours. Proximate composition, vitamins, and anti-nutritional factors (cyanide, oxalate, phytate) were quantified. Linamarase activity and total phenolic and flavonoid contents were measured to assess enzymatic detoxification and phytotherapeutic potential. ResultsBoiling concentrated carbohydrates but created a "nutrient void," leaching 93% of Vitamin C. However, fermentation acted as a biochemical refinery; by 72 hours, total cyanide plummeted from 98.15 to 0.54 mg/100g, meeting WHO safety standards. Concurrently, fermentation triggered a resurgence in bioactives, significantly increasing phenolic and flavonoid levels. ConclusionBoiling alone is insufficient for detoxification. Sequential fermentation beyond 48 hours is essential to "rescue" antioxidant potential and ensure safety. The 72-hour fermented tuber represents an optimized bioactive food vehicle for managing oxidative stress-related pathologies like prostatic hyperplasia

In vegetarian diets, phytate is known to disrupt the adsorption of minerals. Fortifying foods with phytase, a therapeutic enzyme known to mitigate phytate, might increase the uptake of important nutrients. Phytase is susceptible to environmental stress such as heat and acidic conditions encountered during food processing. Therefore, we developed and optimized a core-shell microparticle composed of a phytase-chitosan core and a shell consisting of cross-linked alginate-{kappa}-carrageenan. Ethanol was used to precipitate the microparticles, and the ethanol concentration was optimized along with the chitosan and phytase ratio and the alginate-carrageenan concentration, to form stable core-shell microparticles. The optimized core-shell microparticles have a loading capacity of 32.7% with a high encapsulation efficiency of 80.3% and uniform micro-size with a diameter of 3.2 {micro}m and a poly-dispersity index of 0.178. Loaded phytase retained 62.7% enzymatic activity after heat treatment and digestion conditions. These results indicate that core-shell microparticles are suitable for retaining enzyme activity within the food matrix under typical food processing conditions. HighlightsO_LIDevelopment of size-controlled core-shell microparticles to protect phytase C_LIO_LIPhytase-chitosan microparticles are surrounded by an alginate-{kappa}-carrageenan shell C_LIO_LIOptimization achieved 32.7% loading capacity with a uniform size of 3.2 {micro}m C_LIO_LICore-shell microparticles retained 62.7% enzyme activity after heat and digestion C_LIO_LIPhytase powder (2 mg) is required for a single maize meal C_LI

The objective of this research was to characterize the proteome of the Apis mellifera royal pupae to evaluate its potential as a nutraceutical and functional food. Six pupal instars (E1-E6) were analyzed using liquid chromatography, mass spectrometry, and bioinformatics techniques to determine their properties and biological functions. The results showed 15 proteins across the different instars. In E1, the Isoform X2 of the Caf1 protein and the vitellogenin precursor were found, both critical in genetic regulation and nutrient transport. E2 revealed three proteins linked to energy and genetic processes. Proteins identified in E3 were associated with sugar metabolism and cellular structure. E4 presented proteins related to cellular stress and oxidative processes. In E5, three proteins were identified, associated with molecular transport and energy metabolism. Results for instar E6 were inconclusive since the complexity of peptide identification. From a nutraceutical and functional perspective, the identified proteins show significant potential due to their antioxidant activities, metabolic control, and cellular regulation. Noteworthy proteins include aldose reductase for its role in diabetes management, glutamate dehydrogenase for its importance in amino acid metabolism, vitellogenin as a nutrient source and immune system stimulant, and heat shock protein 60 A, with therapeutic potential in cardiovascular diseases.

Plant-based foods are complex systems, where a multitude of bioactive molecules, such as (poly)phenols and carotenoids are the outcome of endless interactions defining food chemical composition. Significant progress has been made to develop reliable food composition databases that can be used to assess the intake of dietary plant bioactives. However, many lesser-known phytochemicals, like glucosinolates and monoterpenoids are often excluded, also due to the fragmented information available in the literature. Therefore, we present PhytoFooD, a comprehensive phytochemical food database that collects qualitative and quantitative information on 1,067 bioactive compounds in 1,410 plant-based foods. We evaluated the intake of main plant bioactives in European diets and demonstrated the role of concentration variability within foods in intake assessments. This database represents a promising tool for dietary intake assessors and researchers in nutrition, paving the way for a comprehensive and accurate knowledge of our diet and the interconnected health effects of plant bioactives.

Managing post-weaning diarrhoea (PWD) in piglets is difficult due to limits on antibiotics and zinc. Chitosan is emerging as a potential feed additive. We analysed a chito-oligosaccharide hydrochloride (COS-HCl), a low molecular weight (LMW) chitosan, and a medium molecular weight (MMW) chitosan, and assessed their effects on growth, faecal consistency, microbiota, and potential interference with enterotoxigenic Escherichia coli (ETEC). The three chitosans were characterised using {superscript 1}H-NMR, SEC-RI-MS, and SEC-RI-MALLS. COS-HCl had an Mw of 0.824 kDa; LMW and MMW showed Mw ranges of 14.4 kDa (0.3-30 kDa) and 116 kDa (15-600 kDa). Degrees of acetylation were 9.5%, 6.5%, and 15%. Two 42-day field studies evaluated average daily gain (ADG), faecal consistency, and microbiota. In the first trial, COS-HCl at 0.025-0.1% did not significantly affect ADG (-33 to - 12 g/d). In the second, LMW and MMW at 0.01% did not significantly change ADG (-7 and +3 g/d). Faecal consistency, ETEC shedding, and microbiota composition were similar to controls. An enzymatic HPLC-MS method enabled quantification of MMW chitosan in premix. Our results highlight the importance of advanced chitosan characterisation for precision nutrition and suggest that a threshold dosemay be needed to benefit growth and gut health in PWD management. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/714014v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@19c9e23org.highwire.dtl.DTLVardef@152461aorg.highwire.dtl.DTLVardef@7886e0org.highwire.dtl.DTLVardef@df0d9b_HPS_FORMAT_FIGEXP M_FIG C_FIG

To evaluate the international interoperability of food composition databases, we assessed the compatibility of seven national food composition tables with USDA FoodData Central (FDC) using the LLM-based matching method reported previously (Nakagawa and Yamamoto, 2026). Databases from four English-speaking countries (Canada, United Kingdom, Australia, and New Zealand), South Korea, and Japan were compared with 8,158 USDA FDC entries (SR Legacy and Foundation Foods, excluding Survey/FNDDS). Match rates varied by country (62.0-89.7%) and food category. After excluding six USDA categories unsuitable for cross-national comparison, 45.2% of the remaining 6,290 entries were not matched by any country. Canada showed the highest concordance, reflecting shared North American food supply. Japan and South Korea showed similar low coverage for vegetables and spices. These findings suggest that while USDA FDC represents a practical foundation for a globally comprehensive food composition database given its breadth, systematic incorporation of country-specific foods and classification schemes will be necessary to achieve true international interoperability.

Steroidal alkaloids may be responsible for some of the health benefits of a tomato rich diet, but little is known about their metabolic fate after consumption. The objective of this study was to elucidate the pharmacokinetic parameters of plasma steroidal alkaloids and to define their bioavailability and metabolism following a single tomato containing meal. Healthy subjects (n = 11, 6M/5F) consumed 505 g of tomato juice following a two-week tomato washout and blood plasma were collected post-prandially at 11 time points over 12-hours. Plasma steroidal alkaloids were analyzed using UHPLC-MS. The fractional absorption of steroidal alkaloids was 11.8 {+/-} 7% and over 99% of the absorbed dose were present as metabolized products. The maximum concentration of total plasma steroidal alkaloids in subjects was 406.5 {+/-} 377.0 nmol/L occurring at 6 hours after consumption, with an AUC0-12hr of 2529.0 {+/-} 1644.8 nmol*h/L. Liver S9 enzymatic synthesis of steroidal alkaloid metabolites including trihydroxy-tomatidine and sulfonated dihydroxy-tomatidine improved confidence in compound identification. This study reports the first pharmacokinetic data for tomato steroidal alkaloids, demonstrating moderate absorption and extensive metabolism after tomato juice consumption. These data provide context for future studies investigating the potential role that these compounds may play in human health.

Plant-derived bioactive compounds are recognised for their antioxidant potential and benefit for human diseases, including age-related diseases caused by oxidative stress. However, their antioxidant composition and safety profiles remain insufficiently understood. This study integrates phytochemical profiling, antioxidant evaluation, and in vivo toxicological assessment of Warrigal spinach (Tetragonia tetragonioides) and Kensington Pride mango (Mangifera indica). Spinach exhibited greater antioxidant capacity, and higher total phenolic and flavonoid content than mango: TPC (14.2 {+/-} 0.6 mg GAE/g vs 1.30 {+/-} 0.07 mg GAE/g) and TFC (9.61 {+/-} 0.39 mg QE/g vs 0.08 {+/-} 0.0 mg QE/g). LC-ESI-QTOF-MS/MS identified 187 metabolites dominated by flavonoids (53.5%) and phenolic acids (16%), with spinach showing greater chemical diversity. Quantitative analysis revealed higher levels of hydroxycinnamic acids and flavonoid glycosides in spinach, whereas mango contained distinct metabolites, including mangiferin and pyrogallol. Zebrafish embryo / larval assays demonstrated high safety margins, with LC50 values of 478.8 mg/L (spinach) and >480 mg/L (mango). At 480 mg/L spinach displayed developmental abnormalities and malformations. These findings demonstrate that antioxidant capacity is linked to phenolic composition, but does not predict toxicity. Thus, integrated phytochemical and safety evaluation for extracts with complex compound mixtures are critical to identify botanicals suitable for future drug development. HighlightsO_LIWarrigal spinach exhibited >10-fold higher phenolic content and antioxidant capacity than Kensington Pride mango. C_LIO_LILC-ESI-QTOF-MS/MS identified 187 metabolites, with flavonoids as the dominant phytochemical class. C_LIO_LIZebrafish assays confirmed high safety margins, demonstrating no direct correlation between antioxidant capacity and toxicity. C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=114 SRC="FIGDIR/small/720960v1_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@c885dborg.highwire.dtl.DTLVardef@cbed43org.highwire.dtl.DTLVardef@460182org.highwire.dtl.DTLVardef@d1ec5_HPS_FORMAT_FIGEXP M_FIG Graphical abstract C_FIG

Reported avocado dietary fiber (DF) content and composition are inconsistently reported, particularly during ripening. Thus, this study aimed to characterize the amount and type of DF in Hass avocados and evaluate DF changes during ripening. Unripe (day 0), ripe (day 5), and overripe (day 12) Hass avocados were freeze-dried and defatted. DF was analyzed using non-starch polysaccharide (NSP) enzymatic-chemical methods. Per 100g of as-is avocado, unripe contained 3.96g total DF, ripe 3.68g, and overripe 3.26g. In ripe avocados, DF comprised 43% soluble (SDF) and 57% insoluble dietary fiber (IDF). SDF consisted primarily of rhamnogalacturonan-1 and arabinan pectins, while IDF was predominantly cellulose (32%), hemicelluloses (23%), and lignin (2%). Total DF decreased with ripening, with pectin undergoing solubilization and depolymerization, while cellulose and hemicelluloses remained stable. These findings are important as dietary fibers differentially influence intestinal microbial fermentation and health benefits.

Phytohormones are key players in the regulation of plant development and metabolism. The different phytohormone classes comprise numerous chemically very diverse compounds, which are often present at very low concentrations. The chemical properties of phytohormones range from acidic to basic and from polar to non-polar. Furthermore, concentration varies strongly among different phytohormones, between plant species, tissues and developmental stages. Challenges often arise when only small amounts of plant material are available and when plant species are investigated in which the phytohormone profile has not yet been characterized. To establish a method for comprehensive phytohormone analysis we addressed these challenges by choosing and optimizing a suitable extraction method followed by optimized HPLC separation. We compared the most widely-used mass spectrometric detection methods, multiple reaction monitoring (MRM) on a triple quad instrument with high-resolution mass spectrometry (HRMS) on a Q-TOF instrument, and discuss the advantages of both methods and their limitations. O_LIWe compared various methods described in literature for the extraction of six phytohormone classes by liquid-liquid extraction and solid phase extraction purification and describe our optimizations to the selected method. C_LIO_LIWe optimized HPLC separation for 50 different phytohormones. C_LIO_LIWe evaluated the application of MRM and HRMS detection strategies. C_LI

Accurate, simultaneous, and efficient quantification of chemically diverse phytohormone species is a critical task towards understanding the complex system of phytohormone signaling pathways. Quantification of phytohormones with the commonly used technique liquid chromatography coupled to tandem mass spectrometry is susceptible to the influence of non-phytohormone components present in the sample, a phenomenon referred to as matrix effect. To reduce matrix effect, some phytohormone quantification methods include additional steps of cleanup of crude extracts. However, to what extent additional purification steps provide increased accuracy compared to simpler, less laborious methods is seldomly evaluated. We evaluated three previously described phytohormone extraction methods, two of which include solid-phase extraction and one that does not, in their ability to minimize matrix effect and generate accurate estimates of phytohormone species spanning six classifications, from fruit and leaf tissue of Solanum lycopersicum cv. Micro-Tom (tomato). Our results show that, while the methods that included solid phase extraction occasionally outperformed each other regarding matrix effect and/or recovery efficiency for broad range of phytohormones, they rarely outperformed the simpler single-phase extraction method. Short AbstractAccurate, simultaneous quantification of chemically diverse phytohormones by LC-MS/MS is frequently confounded by matrix effects, leading to the incorporation of additional purification steps. We systematically compared three published extraction protocols with or without solid-phase extraction in tomato tissues across six hormone classes. Solid-phase methods occasionally improved matrix suppression or recovery, but did not consistently outperform the single-phase approach, questioning the added value of extra cleanup steps, particularly when high-throughput is desired, as in the case of systems biology interrogations.

This work explores the dietary intake of plant bioactives in the European adult population. The information available in the scientific literature is quite fragmented, with only partial knowledge of dietary bioactive intake and their health effects, and without harmonised figures across populations and phytochemical families. In this context, we comprehensively evaluated the intake of (poly)phenols, terpenoids, N-containing compounds, and miscellaneous phytochemicals in the European adult population, using public data from 26 countries reporting on 38,944 individuals. Further research was conducted to investigate the contributions of classes, subclasses, and individual compounds, as well as their relationships. Main food sources of each class and subclass of phytochemicals were also identified. Finally, variability in phytochemical intake across European countries was evaluated. This work significantly advances the current knowledge of plant bioactive intake and sets the stage for future research in nutrition and health fields.

The production of petroleum-based plastics used for packaging has led to significant environmental challenges in both aquatic and terrestrial ecosystems. Consequently, there is a growing need to explore viable alternatives to the usage of these conventional plastics. This study investigates the utilization of cellulose powder for producing of biodegradable plastics as a more sustainable substitute for petroleum-based materials. Bioplastic films were formulated with varying glycerol contents ranging from 0.5ml - 2.0ml. The glycerol served as a plasticizer to improve the mechanical properties of the films, which were subsequently subjected to biodegradability and tensile strength tests. Biodegradability was evaluated through soil burial tests, which revealed that higher glycerol concentrations accelerated rate of weight loss, with the 2.0 ml formulation exhibiting the fastest degradation rate. Tensile strength increased with glycerol content up to 1.5 ml, where a maximum strength of 7.23 N/mm2 was recorded, but declined at 2.0 ml. The findings indicate that a glycerol concentration of 1.5 ml yields the most optimal bioplastic formulation for short-term packaging applications.

Freshwater algae represent an underexplored source of naturally occurring bioactive metabolites with potential applications in pharmaceutical and biomedical research. This study investigated the phytochemical composition, antioxidant capacity, and preliminary cytotoxic potential of ethanolic and n-hexane extracts of freshwater algal species collected at Jilani Park, Lahore, Pakistan. Algal species were identified morphologically by Dr. Ghazal Yasmeen (Institute of Botany, Punjab University, Lahore). Extracts were analyzed using gas chromatography-mass spectrometry (GC-MS) and qualitative phytochemical screening. Antioxidant activity was evaluated using DPPH radical scavenging, hydrogen peroxide scavenging, and reducing power assays. Cytotoxic potential was assessed using MTT and cell adhesion assays on HeLa and SF767 cell lines as preliminary indicators of bioactivity. GC-MS analysis identified 25 compounds, including sterols, fatty acid esters, terpenoids, phenolic compounds, and volatile metabolites. Phytochemical screening confirmed the presence of flavonoids, phenolics, tannins, and terpenoids in the extracts. Among the tested extracts, the n-hexane fraction demonstrated comparatively higher antioxidant activity across multiple assays. Ethanolic extracts showed moderate reductions in HeLa cell viability, whereas limited effects were observed in SF767 cells. These findings suggest that freshwater algae are promising natural reservoirs of antioxidant metabolites with potential relevance for future isolation and characterization of bioactive compounds for biomedical applications. Further purification and mechanistic studies are required to identify specific active constituents.

This study revealed Quantitative trait loci (QTL) controlling oil sorption capacity in kenaf which could assist in improving genotypes through marker assisted selection (MAS) for effective oil spill clean-up. Two accessions with extreme sorption capacity were selected and crossed to generate F1 which were selfed for linkage and QTL analyses. The oil sorption capacity of the F2 progeny was determined phenotypically and scored as a morphological marker to allow mapping of the markers associated with oil sorption gene(s). The genomic DNA extraction was carried out using a modified CTAB (cyltrimethyl-ammonium bromide) procedure. Diversity Array Technology Sequence (DArTSeq) platform was used to sequence 96 DNA samples prior to linkage and QTL analyses. Genotypic data were analysed using the JoinMap 4.1 Software to construct a genetic map of F2 segregating populations whereas MapQTL1 was used to detect QTL. The mapping population which included the two parents [NHC5(1) and NHC12(2)] together with 72 F2 progeny were genotyped with selected polymorphic SNP markers. Mapping of markers was performed using the regression mapping algorithm. 18 linkage groups were generated from the linkage analysis with Linkage group 1 (LG1) having the longest map length of 138.93 centimorgan (cM) whereas LG 16 had the shortest map length of 83.05 cM. This genetic map has a total map length of 1888.40 cM giving an average length of 104.91 cM per group and an average of one marker for every 1.39 cM. A total of 3 significant and 8 putative QTL were detected by MQM mapping and interval mapping method.

Catechinopyranocyanidins (Cpcs) which consist of diastereomers A and B are pigments derived from adzuki beans and are compounds in which the catechin and cyanidin skeletons are condensed to a pyrano ring. While catechins and anthocyanidins possess high antioxidant capacity, the physiological functions of Cpcs remains unclear. In this study, the antioxidant capacity of Cpcs was evaluated by in vitro antioxidant assays and by assessing their cytoprotective activity against oxidative stress in normal human dermal fibroblasts (NHDFs). Antioxidant capacity based on the hydrogen atom transfer (HAT) mechanism, as assessed by the ORAC assay revealed that Cpcs exhibit 14.1 mol TE/mol (Trolox equivalent antioxidant capacity: TEAC). Meanwhile, capacity based on the single electron transfer (SET) mechanism, as assessed by the DPPH, ABTS and CUPRAC assays revealed, they exhibit 2.1-3.6 mol TE/mol. Since TEAC value of Cpcs demonstrated by the HAT based mechanism higher than its SET based oxidative capacity suggesting that the antioxidant capacity of Cpcs is driven by the HAT mechanism. In cell culture experiments, Cpcs ameliorate cell toxicity in rotenone-induced injury model, suggesting to cytoprotective activity against mitochondrial dysfunction-dependent apoptosis. These results reveal novel physiological functions of Cpcs which may serve as a design guideline for elucidating in vivo dynamics based on antioxidant mechanisms.

{beta}-Carotene is a strong antioxidant and immunomodulator, but it breaks down in the presence of oxygen and isnt readily bioavailable. Nutrishield(R) {beta}-Carotene is a special starch-based microencapsulation system designed to protect {beta}-carotene from oxidation, make it easier to spread in the gastrointestinal environment, and improve its effectiveness. This study reported Nutrishield(R) as a formulation platform by using {beta}-carotene as a model compound. We compared how well it worked in vitro and in vivo with {beta}-carotene that wasnt encapsulated. We randomly put thirty-two male BALB/c mice into four groups (n=8): Control, LPS, LPS + {beta}-Carotene (10 mg/kg/day), and LPS + Nutrishield(R) (the same dose of {beta}-Carotene). After an intraperitoneal injection of LPS (1 mg/kg, twice weekly), the treatments were administered orally for 28 days. LPS administration significantly elevated TNF- (142 {+/-} 8 pg/mL), IL-6 (118 {+/-} 7 pg/mL), and serum 8-OHdG (5.4 {+/-} 0.3 ng/mL) in comparison to controls (TNF- = 62 {+/-} 5 pg/mL; IL-6 = 51 {+/-} 4 pg/mL; 8-OHdG = 2.1 {+/-} 0.2 ng/mL). Nutrishield(R) {beta}-Carotene supplementation significantly normalized cytokine profiles (TNF- = 69 {+/-} 4 pg/mL; IL-6 = 56 {+/-} 3 pg/mL; IL-10 increased from 38 {+/-} 2 to 76 {+/-} 3 pg/mL; p < 0.001 vs. LPS). Histological analysis demonstrated reduced hepatocellular vacuolation and renal tubular degeneration in the Nutrishield(R) {beta}-Carotene group relative to the {beta}-carotene and LPS groups. These findings demonstrate that Nutrishield(R) microencapsulation significantly enhances the functional antioxidant and anti-inflammatory efficacy of {beta}-carotene in a murine model of systemic inflammation. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=134 SRC="FIGDIR/small/711625v1_ufig1.gif" ALT="Figure 1"> View larger version (48K): org.highwire.dtl.DTLVardef@1da9828org.highwire.dtl.DTLVardef@3b8ef2org.highwire.dtl.DTLVardef@25e339org.highwire.dtl.DTLVardef@168e793_HPS_FORMAT_FIGEXP M_FIG C_FIG

Adipogenesis is the process of adipose-derived stem cells (ADSCs) responding to extracellular signals from the stem cell niche to differentiate into adipocytes (fat cells) and may be studied in vitro using a cocktail of chemicals that promote adipogenic differentiation to produce differentiated ADSCs (dADSCs). The global membrane N- and O-glycosylation changes of this process have been previously analysed and compared to native adipocytes as a benchmark for a true adipocyte profile, and revealed that bisecting GlcNAc type N-glycans are characteristic of adipogenesis. As stem cell differentiation has been widely reported to result in cellular protein changes, the same cells (ADSCs, dADSCs and mature adipocytes) were characterised for their membrane proteome here using label-free quantitative shotgun proteomics analysis. The membrane proteome displayed more differences in protein numbers between the cell types compared to the previously reported N-glycome which had shown high identical glycomes between stem cells and in vitro dADSCs, suggesting that the proteome is more dynamic during in vitro adipogenesis. Following the global shotgun proteomics analysis, a more targeted approach of carrying out proteomic analysis of de-N-glycosylated peptides of gel-separated proteins unearthed new glycoproteins not detected in the shotgun proteomic analysis. This approach identified the adipogenic marker, CD36, to be under-represented in the shotgun proteome analysis, but as the dominant (glyco)protein in the adipocyte membrane proteome that was also up-regulated at the mRNA transcript level in both the in vitro differentiated ADSCs (7.1-fold increase) and mature adipocytes (102.9-fold increase). A comparison of CD36 sequence coverage in the global shotgun analysis with the de-N-glycosylated CD36 revealed a 41% increase when N-glycans were removed prior to trypsin digestion, explaining its observed increased abundance and highlights the crucial need for de-N-glycosylation of proteins in proteomics experiments for increased identification of glycoproteins. The systems glycobiology approach by the integration of previously reported glycomics data and the proteomics and transcriptomics analyses in this work extended the investigation of membrane protein glycosylation changes in adipose-derived stem cell differentiation. The work provides a framework for future glycoproteomics-based investigations into the differentiation of stem cells into adipocytes, and will allow their related pathologies and potential therapeutic applications to be discovered. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=121 SRC="FIGDIR/small/722121v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@189a786org.highwire.dtl.DTLVardef@5563b8org.highwire.dtl.DTLVardef@5cb5borg.highwire.dtl.DTLVardef@69e11f_HPS_FORMAT_FIGEXP M_FIG C_FIG