Search Results (969)

Background/Objectives: The barrier properties of the human small intestine play a crucial role in regulating digestion, nutrient absorption and drug metabolism. Current in vitro organotypic models consist only of an epithelium, which does not take into account the possible role of stromal cells such as fibroblasts or the extracellular matrix (ECM) which could contribute to epithelial barrier properties. Therefore, the aim of this study was to determine whether these stromal cells or ECM were beneficial or detrimental to barrier function when incorporated into an organotypic human small intestine model. Methods: Intestinal epithelial cell lines or primary cell organoids derived from the epithelial stem cells of the small intestine were cultivated either on a porous Transwell membrane (epithelial model) or on a primary small intestinal stromal cell-populated collagen-fibrin hydrogel (full thickness model). Results: Both models expressed villin (enterocytes), lysozyme (Paneth cells), Ki67 (proliferative cells) and zonula occludens-1 (tight junctions). The polarized epithelial barriers of the full thickness models demonstrated a significant decrease in transepithelial electrical resistance (TEER) with values comparable to that found in the native small intestine in contrast to the higher TEER values observed in the epithelial models. This correlated to an increase in secreted zonulin, a regulator of intestine permeability, in the full thickness models. The decreased TEER values were due to both the stromal cells and the choice of the hydrogel versus the Transwell membrane. Moreover, erythropoietin and epithelial growth factor secretion, which have roles in regulating barrier integrity, directly correlated with the changes in TEER and permeability. Conclusions: This study emphasizes the importance of different cell types being incorporated into small intestine models and, also, the influence of the scaffold or matrix used. Full article

In this study, new monolithic poly(9-anthracenylmethyl methacrylate-co-trimethylolpropane trimethacrylate (TRIM) columns, referred as ANM monoliths were prepared, for the first time, and were used for the separation media for biomolecules and proteomics analysis by nano-liquid chromatography (nano-LC). Monolithic columns were prepared by in situ polymerization of 9-anthracenylmethyl methacrylate (ANM) and trimethylolpropane trimethacrylate (TRIM) in a fused silica capillary column of 100 µm ID. Polymerization solution was optimized in relation to monomer and porogenic solvent. Scanning electron microscopy (SEM) and chromatographic analyses were performed for the characterization studies of ANM monoliths. The ANM monolith produced more than 46.220 plates/m, and the chromatographic evaluation of the optimized ANM monolith was carried out using homologous alkylbenzenes (ABs) and polyaromatic hydrocarbons (PAHs), allowing both strong hydrophobic and π-π interactions. Run-to-run and column-to-column reproducibility values were found as <2.91% and 2.9–3.2%, respectively. The final monolith was used for biomolecule separation, including both three dipeptides, including Alanine-Tyrosine (Ala-Tyr), Glycine-Phenylalanine (Gly-Phe), and L-carnosine and five standard proteins, including ribonuclease A (RNase A), α-chymotrypsinogen (α-chym), lysozyme (Lys), cytochrome C (Cyt C), and myoglobin (Mb) in order to evaluate its potential. Both peptides and proteins were baseline separated using the developed ANM monolith in nano-LC. The ANM monolith was then applied to the protein and peptide profiling of MCF-7 cell line, which allowed a high-resolution analysis of peptides, providing a high peak capacity. Full article

This study explored the effects of mango seed (MS) powder supplementation on the growth, immune response, gene expression, and intestinal morphology of Nile tilapia (Oreochromis niloticus) over an 8-week period. A total of 300 Nile tilapia fingerlings (average weight of 15.29 ± 0.05 g) were divided into five treatment groups and fed either a basal diet or one of four experimental diets containing MS powder at concentrations of 10 (MS10), 20 (MS20), 40 (MS40), and 80 (MS80) g kg⁻¹. The results demonstrated that Nile tilapia fed MS-supplemented diets experienced significant improvements (p < 0.05) in weight gain (WG), specific growth rate (SGR), and survival rate (SR) compared to the control group (0 g kg⁻¹ MS). The MS-treated groups also showed a significant increase (p < 0.05) in the height and branching of intestinal villi along the entire length of the intestine, as well as a significantly higher villus-to-crypt depth ratio (V/C), indicating enhanced intestinal health and functionality. Moreover, although MS supplementation did not increase peroxidase activity, it did lead to a significant increase (p < 0.05) in the activity of skin mucus and serum lysozyme, along with upregulated gene expression of immune-related (IL-1, IL-8, and LBP) and antioxidant genes (GST-α, GPX, and GSR). Polynomial regression analysis identified an optimal MS dosage of 36.43–45 g kg⁻¹ for effectively improving growth, immunity, and immuno-oxidant gene expression in Nile tilapia. These results emphasize mango seed (MS) as a promising natural supplement for improving the diet of Nile tilapia and, potentially, other freshwater fish widely used in aquaculture. Full article

Malondialdehyde (MDA) is a reactive carbonyl compound produced through lipid peroxidation during feed storage, which poses a significant threat to fish health. This study aimed to evaluate the effects of dietary MDA on the growth rate, gastrointestinal health, and muscle quality of striped catfish (Pangasianodon hypophthalmus). A basal diet (M0) containing 34% crude protein and 10.5% crude lipid was formulated. Each group was sprayed with malondialdehyde solution (0, 5, 10, 20, 40, and 80 mg/kg, on dietary crude lipid basis; 0, 0.53, 1.07, 2.13, 4.26, and 8.52 mg/kg, on dietary basis) before feeding, respectively. Each diet was randomly assigned to triplicates of 30 striped catfish (initial weight 31.38 g) per net cage. After 8 weeks, dietary inclusion of MDA regardless of level significantly depressed the growth rate and feed utilization. The extent of structural damage to the gastrointestinal tract increased progressively with increasing dietary MDA levels. The extent of damage to the intestinal biological barrier (intestinal microbial structure), chemical barrier (trypsin, lipase, amylase, and maltase activity), physical barrier (zonula occludent-2, occludin, claudin 7α, and claudin 12 relative expression), and immune barrier (contents of complement 4, complement 3, immunoglobulin M, and lysozyme activity) was dose-related to dietary MDA. Moreover, a linear decline in the activities of intestinal antioxidant enzymes (catalas, superoxide dismutase, et al.) and anti-inflammatory factor (transforming growth factor beta1, interleukin 10) relative expression was noted alongside an increase in dietary MDA content. In contrast, the relative expression levels of intestinal inflammatory factor (interleukin 8, transcription factor p65, tumor necrosis factor alpha) relative expression displayed an opposing trend. Additionally, dietary MDA exerted a linear influence on muscle color and texture characteristics. In conclusion, high doses of MDA (5–80 mg/kg) reduced the growth performance of striped catfish, attributed to linear damage to the gastrointestinal tract, a linear decrease in antioxidant function, and the occurrence of an inflammatory response. High doses of MDA (>40 mg/kg) were observed to significantly increase dorsal muscle b-value and induce muscle yellowing. Full article

This study describes the applicability of the fluorescence polarization assay (FPA) based on the use of FITC-labeled oligosaccharide tracers of defined structure for the measurement of active lysozyme in hen egg white. Depending on the oligosaccharide chain length of the tracer, this method detects both the formation of the enzyme-to-tracer complex (because of lectin-like, i.e., carbohydrate-binding action of lysozyme) and tracer splitting (because of chitinase activity of lysozyme). Evaluation of the fluorescence polarization dynamics enables simultaneous measurement of the chitinase and lectin activities of lysozyme, which is crucial for its detection in complex biological systems. Hen egg white lysozyme (HEWL), unlike human lysozyme (HL), formed a stable complex with the chitotriose tracer that underwent no further transformations. This fact allows for easy measurement of the carbohydrate-binding activity of the HEWL. The results of the lysozyme activity measurement for hen egg samples obtained through the FPA correlated with the results obtained using the traditional turbidimetry method. The FPA does not have the drawbacks of turbidimetry, which are associated with the need to use bacterial cells that cannot be precisely standardized. Additionally, FPA offers advantages such as rapid analysis, the use of compact equipment, and standardized reagents. Therefore, the new express technique for measuring the lysozyme activity is applicable for evaluating the complex biomaterial, including for the purposes of food product quality control. Full article

We investigated the effect of feed deprivation for 45 days on the growth, immunity, and health of 0⁺marron (Cherax cainii) initially fed for 110 days on various protein sources including fishmeal (FM), poultry by-product meal (PBM), black soldier fly meal (BSFM), soybean meal (SBM), lupin meal (LM), and tuna hydrolysate. The marron were weighed and sacrificed immediately after feeding stopped (day 0) and at days 15, 30, and 45 after the feed deprivation trial commenced. Total haemolymph count, differential haemocyte count, lysozyme activity, protease activity, total bacterial count in the digestive tract, and organosomatic indices were analysed. Initially feeding marron any protein sources did not influence the percentage of weight gain and specific growth rates of marron. All marron showed more than 83% survival; however, marron fed soybean meal showed significantly lower survival than others. Dietary sources of protein altered organosomatic indices of starved marron during various starvation periods and resulted in a significant decrease in total haemocyte counts, lysozyme activity, protease activity, and bacterial count in the digestive tract of marron. Starved marron initially fed PBM and BSFM showed higher tolerance to starvation, followed by marron initially fed FM and SBM, while marron initially fed TH and LM showed the highest susceptibility to starvation. Full article

The effect of sodium alginate on the denaturation and aggregation behavior of bovine serum albumin and hen egg-white lysozyme was studied. Large amounts of polysaccharide increase the thermal stability of albumin due to the weak binding interactions. At the same time, sodium alginate can reduce the quantity of amyloid fibrils formed by albumin under denaturing conditions, which is a consequence of the stabilization of the native protein form by glycan binding. In the case of lysozyme, the polysaccharide has no influence on the thermal stability of the protein in 2 M guanidinium hydrochloride. However, the inhibition of fibril formation with an increase in the lag time was observed, which is explained by the binding of sodium alginate to lysozyme fibrils, but not to the protein monomer. The molecular nature of the binding interactions between alginate and the studied proteins was elucidated using molecular docking and known experimental structures of glycan–protein complexes. Full article

Synchronously recovering phosphorus as vivianite and humic acids (HAs) from waste activated sludge (WAS) is of great significance for the carbon neutralization of wastewater. In this study, flocculation, enzyme degradation (lysozyme/protease/amylase/cellulase in a 1:1:1:1 ratio), and pH adjustment were used to reclaim vivianite and HAs. After FeCl₃ coagulation–precipitation and enzymatic hydrolysis of the sludge for 11 h, the supernatant was enriched with Fe²⁺ and PO₄³⁻, with the molar ratio of Fe²⁺:PO₄³⁻ of 2.21. To improve the purity of the vivianite, the crude protein was separated at pI 6.0. The purity of the crystals reached a peak of 97.44 ± 0.04% at pH 7.5. HAs extracted from the residuals had a high affinity for metal adsorption, and the adsorption process was both endothermic and efficient. Overall, this study demonstrates the feasibility and effectiveness of the joint reclaiming of vivianite and HAs, providing new insights into multiple resource recovery from WAS. Full article

To investigate the dietary effects of chlorogenic acid (CGA) on the growth performance, lipid metabolism, antioxidant activity, and non-specific immunity of Asian swamp eel (Monopterus albus) during the domestication stage, a 28-day feeding experiment was conducted to supplement with CGA at levels of 0 (Cont.), 250 (CGA 0.50%), 500 (CGA 1.00%), and 750 (CGA 1.50%) mg/kg·bw by feeding with yellow mealworm (Tenebrio molitor). Compared with the control group, the addition of 250–750 mg/kg of CGA significantly increased the weight-gain rate (WG) of M. albus, and the CGA 1.0% group displayed the highest value. The content of hemoglobin and high-density lipoprotein in all CGA groups was markedly elevated (p < 0.05), while the triglyceride, glucose, low-density lipoprotein, and glycosylated serum protein levels were lowered (p < 0.05). Among the antioxidant enzymes, the glutathione peroxidase and catalase activity was significantly higher in all experimental groups than that of the control group, whereas the malondialdehyde activity was significantly reduced (p < 0.05). For a non-specific immune enzyme system, the lysozyme and alkaline phosphatase activity in all treatments and the superoxide dismutase and acid phosphatase activity in the CGA 0.5% group was markedly increased (p < 0.05). In conclusion, supplementation with CGA can enhance the growth performance and improve the lipid metabolism, antioxidant capacity, and non-specific immunity of M. albus during the domestication stage, and the optimal CGA supplementation for T. molitor as biocarrier bait is 500 mg/kg, corresponding to 405 mg/kg. Full article

Taking into account the trends in the field of green chemistry and the desire to use natural materials in biomedical applications, (bio)polyelectrolyte complexes ((bio)PECs) based on a mixture of chitosan and gelatin seem to be relevant systems. Using the approach of self-assembly from the dispersion of the coacervate phase of a (bio)PEC at different ratios of ionized functional groups of chitosan and gelatin (z), hydrogels with increased resistance to mechanical deformations and resorption in liquid media were obtained in this work in comparison to a hydrogel from gelatin. It was found that at z ≥ 1 a four-fold increase in the elastic modulus of the hydrogel occurred in comparison to a hydrogel based on gelatin. It was shown that hydrogels at z ≈ 1 had an increased sorption capacity and water sorption rate, as well as increased resistance to the in vitro model environment of phosphate-buffered saline (PBS) solution containing lysozyme at 37 °C. It was also shown that in PBS and simulated gastric fluid (SGF) solutions, the effect of the polyelectrolyte swelling of the hydrogels was significantly suppressed; however, at z ≥ 1, the (bio)PEC hydrogels had increased stability compared to the samples at z < 1 and based on gelatin. Full article

The incorporation of graphene into cellulose matrices has emerged as a promising strategy for enhancing the structural and functional properties of composite materials. This comprehensive review provides a critical analysis of recent advances in optimizing graphene content in cellulose matrices and its impact on composite performance. Various optimization techniques, including response surface methodology, particle swarm optimization, and artificial neural networks, have been employed to identify optimal graphene concentrations and processing conditions. Quantitative analyses demonstrate significant improvements in mechanical properties, with notable increases in tensile strength and Young’s modulus reported for graphene/microfibrillated cellulose composites. Substantial enhancements in thermal stability have been observed in lysozyme-modified graphene nanoplatelet–cellulose composites. Electrical conductivity has been achieved at low graphene loading levels. Additionally, barrier properties, biocompatibility, and functionality for applications such as energy storage and environmental remediation have been substantially improved. This review explores case studies encompassing the optimization of thermal conductivity, viscosity, durability behaviors, pollutant removal, and various other properties. Despite promising results, challenges remain, including uniform dispersion, scalability, cost-effectiveness, and long-term stability. Strategies such as surface functionalization, solvent selection, and protective coatings are discussed. Future research directions, including novel processing techniques like 3D printing and electrospinning, as well as the incorporation of additional functional materials, are outlined. This review synthesizes current knowledge, identifies emerging trends, and provides a roadmap for future research in the rapidly evolving field of graphene–cellulose composites. Full article

Nile tilapia (Oreochromis niloticus) and European sea bass (Dicentrarchus labrax) are economically significant species in Mediterranean countries, serving essential roles in the aquaculture industry due to high market demand and nutritional value. They experience substantial losses from bacterial pathogens Vibrio anguillarum and Streptococcus iniae, particularly at the onset of the summer season. The immune mechanisms involved in fish infections by V. anguillarum and S. iniae remain poorly understood. This study investigated their impact through experiments with control and V. anguillarum- and S. iniae-infected groups for each species. Blood samples were collected at 1, 3, and 7 days post bacterial injection to assess biochemical and immunological parameters, including enzyme activities (AST and ALT), oxidative markers (SOD, GPX, CAT, and MDA), and leukocyte counts. Further analyses included phagocyte activity, lysozyme activity, IgM levels, and complement C3 and C4 levels. Muscle tissues were sampled at 1, 3, and 7 days post injection to assess mRNA expression levels of 18 immune-relevant genes. The focus was on cytokines and immune-related genes, including pro-inflammatory cytokines (TNF-α, TNF-β, IL-2, IL-6, IL-8, IL-12, and IFN-γ), major histocompatibility complex components (MHC-IIα and MHC-IIβ), cytokine receptors (CXCL-10 and CD4-L2), antimicrobial peptides (Pleurocidin and β-defensin), immune regulatory peptides (Thymosin β12, Leap 2, and Lysozyme g), and Galectins (Galectin-8 and Galectin-9). β-actin was used as the housekeeping gene for normalization. Significant species-specific responses were observed in N. Tilapia and E. Sea Bass when infected with V. anguillarum and S. iniae, highlighting differences in biochemical, immune, and gene expression profiles. Notably, in N. Tilapia, AST levels significantly increased by day 7 during S. iniae infection, reaching 45.00 ± 3.00 (p < 0.05), indicating late-stage acute stress or tissue damage. Conversely, E. Sea Bass exhibited a significant rise in ALT levels by day 7 in the S. iniae group, peaking at 33.5 ± 3.20 (p < 0.05), suggesting liver distress or a systemic inflammatory response. On the immunological front, N. Tilapia showed significant increases in respiratory burst activity on day 1 for both pathogens, with values of 0.28 ± 0.03 for V. anguillarum and 0.25 ± 0.02 for S. iniae (p < 0.05), indicating robust initial immune activation. Finally, the gene expression analysis revealed a pronounced peak of TNF-α in E. Sea Bass by day 7 post V. anguillarum infection with a fold change of 6.120, suggesting a strong species-specific pro-inflammatory response strategy. Understanding these responses provides critical insights for enhancing disease management and productivity in aquaculture operations. Full article

The effect of gravity based on the vector sum of gravity and buoyancy forces working spontaneously for all species was introduced into capillary electrophoresis (CE) as another important force which cooperated with electrophoretic flow and electro-osmotic flow. Their portion was adjusted by simply tilting the whole of the capillary at an angle during CE running. The related formula was proposed and verified through a series of experiments. After investigating the related parameters, results showed that the gravity effect was significantly affected by additives in the buffer, the length, and the inner diameter of the capillary, and the size of the sample molecule. This made the different ions with opposite or significantly different mobilities to be observed at a CE run. It significantly improved separation efficiencies of some small molecules, chiral compounds, macromolecules and cells when the tilt angles of the capillary were adjusted to a special range predicted through the fitting curve. In addition, micrometer level microspheres and cells were firstly separated by the new CE strategy and the resolutions were more than 1.0. After ingeniously designing the gradient of the tilting angle with time, we were able to further enhance the separation efficiency of the targets. For example, the resolution of lysozyme and ribonuclease A could be increased from 3.691 to 7.342. These indicated the huge potential of the new CE strategy and its gradient mode in separation. Full article

This study evaluated bioactive compounds in blood orange (Citrus sinensis (L.) Osbeck) peel (BOP) as dietary additives. An 8-week feeding trial was conducted to investigate the effects of dietary supplementation on the growth performance, body composition, digestive enzyme activity, antioxidant capacity, and immune response of juvenile black rockfish. A total of 1260 juvenile rockfish (1.4 ± 0.01 g) were randomly distributed into seven treatment groups, each with 50 fish per circular tank. The groups were fed seven different diets containing graded levels of 0 (control, BOP0), 1 (BOP1), 2 (BOP2), 3 (BOP3), 5 (BOP5), 7 (BOP7), and 10 (BOP10) g kg⁻¹, respectively. The BOP10 diet significantly enhanced the final weight, weight gain, specific growth rate, protein efficiency ratio, and protein retention in fish. The BOP treatments notably affected the fishes’ whole-body crude protein and lipid contents. Plasma total cholesterol levels of fish fed the BOP0 and BOP1 diets were significantly higher than those fed the BOP7 and BOP10 diets. The activities of trypsin and lipase were significantly affected by dietary BOP levels. The antioxidant enzyme activity in the plasma of fish fed the BOP10 diet was significantly higher than those fed the BOP0 diet. The lysozyme activity and levels of immunoglobulin M and G in fish fed the BOP0 diet were significantly lower than those in fish fed the BOP10 diet. In conclusion, dietary supplementation of BOP at 10 g kg⁻¹ improved the growth performance and overall health of juvenile black rockfish. Full article

The development of effective and innovative vaccination strategies is urgently needed to better control the spread and transmission of the low-pathogenic avian influenza H9N2 subtype (LPAI-H9N2) in poultry. In addition, the enhancement of innate immunity by some of these innovative inactivated vaccines has not yet been investigated. Here, an experiment was conducted in commercial broiler chickens to compare the immune response to two different inactivated H9N2 vaccines. For this, Group 1 (G1) broilers were vaccinated with vaccine 1 [Nobilis^® H9N2-P (pathogen-associated molecular patterns—PAMP) technology], broilers in G2 were vaccinated with vaccine 2 [an inactivated whole H9N2 virus (IWV) autogenous oil emulsion vaccine], while birds in G3 were not vaccinated. The study lasted 34 days. Innate immune parameters (phagocytic activity, nitric oxide, and lysozyme), cytokine signaling (IL-1β, IL-6, IL-8), humoral immunity using the hemagglutination inhibition (HI) test, and the gene expressions of IFN-γ and TLR-21 were assessed. The results showed a significant increase in innate immunity and modulatory cytokines at 24–48 h after the vaccination of G1 broilers, with a continuous increase until the end of the experiment. In addition, a significant increase in geometric mean HI titers was observed in G1 at 11 days post-vaccination (dpv), and a significant (p ˂ 0.05) upregulation of IFN-γ and TLR-21 was observed in the same group, G1, at 31 dpv compared to G2 and G3. Nobilis^® H9N2-P may induce faster and stronger innate and active humoral immunity compared to another IWV, which may contribute to the protection of broilers against early H9N2 infections. However, challenge protection studies for several IWV vaccines, including PAMP-H9N2 against LPAI-H9N2, should be further evaluated in both specific pathogen-free (SPF) and commercial broilers. Full article