Search Results (224)

Species of the Ferula genus are known for their traditional medicinal applications against diverse illnesses. Our previous study was the first to suggest the cholinesterase inhibitory activity of Ferula persica L. However, the neuroprotective efficacy of therapeutic molecules is often limited by their ability to cross the blood–brain barrier (BBB) and reach the brain. In the present study, the BBB permeability of the main molecules present in the aerial parts and roots of F. persica L. extracted under optimum conditions was assessed using two well-established methods: the parallel artificial membrane permeability assay (PAMPA) and the HBMEC cell culture in vitro model. The results demonstrated a high permeability of several neuroprotective compounds, such as apigenin, diosmetin, and α-cyperone. Additionally, the neuroprotective potential of F. persica extracts was evaluated using SH-SY5Y neuron-like cells exposed to different insults, including oxidative stress (H₂O₂), excitotoxicity (L-glutamate), and Aβ1-42 peptide toxicity. However, none of the obtained extracts provided significant protection. This study highlights the importance of in vitro cell culture models for a better understanding of BBB permeability mechanisms and reports the tentative identification of newly formed sulfated metabolites derived from the metabolism of ferulic acid, apigenin, and diosmetin by HBMEC cells. Full article

This study aimed to identify a probiotic bacterium with antagonistic activity against the foodborne pathogen Staphylococcus aureus (S. aureus) and investigate the mechanism of its antibacterial components. Growth kinetics were analyzed to assess bacterial proliferation. Acid and bile salt tolerance are vital indicators for evaluating probiotic survival in the gastrointestinal tract. The results indicated that Companilactobacillus farciminis (C. farciminis) YLR-1 not only had high tolerance to salt conditions (0.03%, 0.3%, and 0.5%) but also has a high survival rate at pH 3–4. The bacteriocin-like inhibitory substance (BLIS) isolated from C. farciminis YLR-1 was dialyzed using a membrane with a molecular weight cut-off (MWCO) of 500 Da, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The results indicate that the BLIS produced by C. farciminis YLR-1 is a small-molecule peptide. BLIS displayed pH tolerance within acidic and neutral environments (4–8) and exhibited thermostability. When treated with proteinase K, the antibacterial action of BLIS was found to be inactivated. Membrane disruption mechanisms were examined using fluorescence imaging and scanning electron microscopy (SEM). SEM and fluorescence imaging revealed that BLIS-induced membrane damage in S. aureus ATCC 25923 causes cytoplasmic leakage and cell death. Full article

The increasing popularity of “superseeds” such as flax, sesame, amaranth and quinoa as functional foods raises the need for robust analytical methods for authentication purposes. In this work, a standardized workflow for the extraction, characterization and identification of unique peptides that may be used as markers to distinguish superseed species was investigated. Ammonium bicarbonate/urea (Ambi/urea) extraction, sodium dodecyl sulfate (SDS) buffer and trichloroacetic acid (TCA) precipitation were initially implemented and, based on the level and composition of the extracted proteins, the SDS buffer protocol was selected. Electrophoresis analysis revealed consistent protein profiles between biological replicates from each of the eleven seed species, confirming the reproducibility of the SDS buffer protocol. Targeted mass spectrometry successfully identified species-specific peptide markers for six of eleven superseeds investigated, including peptides from conlinins in flaxseed (WVQQAK), 11S globulins in sesame (LVYIER), oleosin in quinoa (DVGQTIESK), agglutin-like lectins in amaranth (CAGVSVIR), as well as cupin-like proteins in poppy seeds (INIVNSQK) and edestins in hemp seeds (FLQLSAER). Moreover, proteome cross-analysis allowed us to disqualify the isomeric peptide LTALEPTNR from 11S globulins present in amaranth and quinoa. However, no reliable markers were identified for chia, canihua, basil, black cumin, and psyllium seeds under current conditions. While this targeted proteomics approach shows promise for superseed authentication, comprehensive method validation and alternative strategies for marker-deficient species are required before routine implementation. Full article

Background: Heparan sulfate (HS) is widely implicated as a receptor for Chlamydia cell attachment and infectivity. However, the enzymatic modification of HS modified by the 3-O sulfotransferase-3 (3-OST-3) enzyme in chlamydial cell entry remains unknown. Methodology: To rule out the possibility that host cell 3-O sulfated heparan sulfate (3-OS HS) plays a significant role in C. muridarum entry, a Chinese hamster ovary (CHO-K1) cell model lacking endogenous 3-OST-3 was used. In addition, we further tested the efficacy of the phage-display-derived cationic peptides recognizing heparan sulfate (G1 peptide) and the moieties of 3-O sulfated heparan sulfate (G2 peptide) against C. muridarum entry using human cervical adenocarcinoma (HeLa 229) and human vaginal epithelial (VK2/E6E7) cell lines. Furthermore, molecular dynamics simulations were conducted to investigate the interactions of the Chlamydia lipid bilayer membrane with the G1 and G2 peptides, focusing on their binding modes and affinities. Results: The converse effect of 3-OST-3 expression in the CHO-K1 cells had no enhancing effect on C. muridarum entry. The G2 peptide significantly (>80%) affected the cell infectivity of the elementary bodies (EBs) at all the tested concentrations, as evident from the reduced fluorescent staining in the number of inclusion bodies. The observed neutralization effect of G2 peptide on C. muridarum entry suggests the possibility of sulfated-like domains being present on the EBs. In addition, data generated from our in silico computational structural modeling indicated that the G2 peptide ligand had significant affinity towards the C. muridarum lipid bilayer. Conclusions: Taken together, our findings show that the pretreatment of C. muridarum with 3-O sulfated heparan sulfate recognizing G2 peptide significantly prevents the entry of EBs into host cells. Full article

Diabetic foot osteomyelitis (DFO) is a severe complication of diabetes mellitus and a leading cause of non-traumatic lower extremity amputation. Treatment remains clinically challenging with high recurrence rates despite standard antibiotic therapy and surgical debridement. This narrative review synthesizes current evidence on novel operative and nonoperative therapies for DFO, focusing on emerging biomaterials, local antibiotic delivery systems, innovative surgical techniques, and adjunctive topical agents. Studies examining bioabsorbable and nonabsorbable antibiotic carriers, such as calcium sulfate beads, collagen sponges, and bioactive glass, demonstrate promising infection resolution rates and a potential to reduce the surgical burden, though most are limited by small cohorts and observational designs. Similarly, alternative surgical approaches (i.e., cancelloplasty, conservative bone excision, and tibial cortex distraction) have shown early success in limb preservation. Nonoperative strategies, including adjunct antimicrobials, antimicrobial peptides, and topical oxygen, offer additional options, particularly for patients unfit for surgery. While initial outcomes are encouraging, the supporting evidence is heterogeneous and primarily limited to case series and small, noncomparative trials. Overall, these novel therapies show potential as adjuncts to established DFO management, but further prospective research is indicated to define their long-term efficacy, safety, and role in clinical practice. Full article

Tofu quality is critically influenced by coagulants, though their impact on protein digestibility remains underexplored. This study aimed to investigate the effects of calcium sulfate (CaSO₄), magnesium chloride (MgCl₂), and their combination (CaSO₄ + MgCl₂) on the physicochemical properties and protein digestibility of tofu. Water-holding capacity, cooking loss, texture, protein composition, and protein digestibility were analyzed. The results showed that the CaSO₄ + MgCl₂ combination yielded a water-holding capacity of 99.16%, significantly higher than CaSO₄ tofu (93.73%) and MgCl₂ tofu (96.82%), while reducing cooking loss to 2.03% and yielding the highest hardness (897.27 g) and gumminess (765.72). Electrophoresis revealed distinct protein retention patterns, with MgCl₂ (0.6% w/v) forming denser gels that minimized protein leakage into soy whey. During in vitro digestion, MgCl₂-coagulated tofu exhibited superior soluble protein release (5.33 mg/mL after gastric digestion) and higher intestinal peptide (5.89 mg/mL) and total amino acid (123.06 μmol/mL) levels, indicating enhanced digestibility. Conversely, the CaSO₄ + MgCl₂ combination showed delayed proteolysis in electrophoresis analysis. These findings demonstrate that coagulant selection directly modulates tofu’s texture, water retention, and protein bioavailability, with MgCl₂ favoring digestibility and the hybrid coagulant optimizing physical properties. This provides strategic insights for developing nutritionally enhanced tofu products. Full article

Alzheimer’s disease (AD) is characterized by β-amyloid plaques, neurofibrillary tangles, neuroinflammation, and oxidative stress—pathological features that pose significant challenges for the development of therapeutic interventions. Given these challenges, this review comprehensively evaluates the neuroprotective mechanisms of bioactive compounds derived from red algae, including polysaccharides and phycobiliproteins, which are considered a promising source of natural therapeutics for AD. Red algal constituents exhibit neuroprotective activities through multiple mechanisms. Sulfated polysaccharides (e.g., carrageenan, porphyran) suppress NF-κB-mediated neuroinflammation, modulate mitochondrial function, and enhance brain-derived neurotrophic factor (BDNF) expression. Phycobiliproteins (phycoerythrin, phycocyanin) and peptides derived from their degradation scavenge reactive oxygen species (ROS) and activate antioxidant pathways (e.g., Nrf2/HO-1), thus mitigating oxidative damage. Carotenoids (lutein, zeaxanthin) improve cognitive function through the inhibition of acetylcholinesterase and pro-inflammatory cytokines (TNF-α, IL-1β), while phenolic compounds (bromophenols, diphlorethol) provide protection by targeting multiple pathways involved in dopaminergic system modulation and Nrf2 pathway activation. Emerging extraction technologies—including microwave- and enzyme-assisted methods—have been shown to optimize the yield and maintain the bioactivity of these compounds. However, the precise identification of molecular targets and the standardization of extraction techniques remain critical research priorities. Overall, red algae-derived compounds hold significant potential for multi-mechanism AD interventions, providing novel insights for the development of therapeutic strategies with low toxicity. Full article

Zn is a trace element necessary for the functioning of about 300 enzymes. It plays a biochemical, structural, and regulatory role. It participates in the immune response, proper functioning of the endocrine system, and regulation of gene expression. Its deficiencies are most often caused by the mismatch between dietary intake and the body’s needs. Bioavailability of zinc depends on interactions with other food components. Phytates negatively affect this element’s absorption, whereas proteins, peptides, and amino acids increase its bioavailability. It has been proven that organic forms of zinc are better absorbed than inorganic compounds, like zinc oxide and sulfate. Amino acid combinations with zinc can use amino acid transporters in the absorption process. Estimation of Zn bioavailability and bioaccessibility are based on in vivo and in vitro studies, each having their advantages and disadvantages. The current review aims to gather and summarize recent research on the dietary role of Zn, especially data on bioavailability from food substances promoting/inhibiting absorption, and the latest methods for determining the level of bioavailability of this nutrient. Full article

Novel peptide-zinc chelates (DSPs-Zn) with a zinc content of 186.94 mg/g were synthesized from deer tendon peptides at pH 6, 60 °C, 60 min, and peptide-zinc mass ratio of 1:3. Ultraviolet-visible absorption spectroscopy (UV) and Fourier transform infrared spectroscopy (FTIR) demonstrated that the chelation sites of the deer tendon polypeptides (DSPs) with zinc ions were located at the carboxyl oxygen and amino nitrogen atoms of the peptides. Amino acid analysis showed that aspartic acid, glutamic acid, lysine, and arginine play important roles in the chelation process. In vitro simulated gastrointestinal digestion studies showed that DSPs-zinc exhibited higher stability than zinc sulfate and zinc gluconate in the pH range 2–8 and in a simulated gastrointestinal digestion environment. The above experimental results suggest that DSPs-Zn has the potential to be used as a novel zinc nutritional supplement. Full article

A growing body of evidence suggests the potent anti-inflammatory properties of the newly discovered arm of the renin–angiotensin–aldosterone system, ACE2/Ang-(1–7)/MasR, in various disease conditions. Our group was the first to report the anti-inflammatory properties of the Ang-(1–7) polypeptide in the murine dextran sulfate sodium (DSS) colitis model. Both its short half-life and high degradation rate limit the clinical use of Ang-(1–7). One way to compensate for these limitations is through the use of the non-peptide MasR agonist AVE0991. Herein, we aimed to study the anti-inflammatory effects of AVE0991 using the DSS model and the possible synergistic effects with other clinically available medications. Colitis severity was determined using both prophylactic and treatment approaches by gross anatomical and histological assessments and daily weight changes. The colonic expression level/activity of various pro-inflammatory and adhesion molecules was determined by western blotting, immunofluorescence, and proteomic profiling. We showed that AVE0991 treatment significantly reduced colitis severity more effectively with the prophylactic than the treatment approach. An additive anti-inflammatory effect was observed in the combination regimen with AVE0991 plus azathioprine, which was mediated through an increased colonic expression level of mucins and focal adhesion kinase, decreased colonic activity of p38 MAPK and Akt, and decreased colonic expression level of various pro-inflammatory mediators. In conclusion, these data suggest a promising potential for the non-peptide MasR agonist AVE0991 in the treatment of inflammatory bowel disease. Full article

Mucopolysaccharidosis type IVA (MPS IVA, Morquio A syndrome) is a rare inherited disorder characterized by skeletal dysplasia due to deficient N-acetylgalactosamine-6-sulfate sulfatase activity, resulting in glycosaminoglycan (GAG) accumulation. Identifying accurate biomarkers reflecting clinical severity and therapeutic response remains challenging. This study evaluated potential surrogate biomarkers, including N-terminal pro-C-type natriuretic peptide (NT-proCNP), collagen types I and II, mono-sulfated keratan sulfate (KS), di-sulfated KS, and chondroitin-6-sulfate (C6S), in blood and urine samples from 60 patients ranging from 1 to 62 years of age. NT-proCNP levels were significantly elevated in patients of all ages and negatively correlated with growth impairment, especially after 8 years of age. Collagen type I levels significantly increased in adult patients, whereas collagen type II showed age-dependent elevations. Urinary KS, in mono- and di-sulfated forms, demonstrated moderate negative correlations with growth impairment. Moreover, NT-proCNP, mono- and di-sulfated KS in plasma, and urinary di-sulfated KS were not affected by enzyme replacement therapy in patients younger than 12 years, unlike urinary mono-sulfated KS. In conclusion, NT-proCNP has emerged as a promising independent biomarker reflecting the severity of skeletal dysplasia and possibly the near-future growth rate. These findings highlight the potential role of NT-proCNP in clinical assessment and monitoring therapeutic efficacy, addressing current unmet needs in MPS IVA management. Full article

The functional exploration of marine-derived proteins is at the forefront of nutritional research. The Argentine squid protein (ASP) was extracted from Argentine squid carcasses and was hydrolyzed using neutral protease, with the degree of hydrolysis serving as the response variable. Using single-factor experiments and response surface methodology, we identified optimal conditions for preparing Argentine squid protein peptides (ASPP). The hydrolysis degree reached 41.32% ± 0.27 under the conditions of 7% enzyme preparation addition, 2.4 h enzyme digestion time, and 6% substrate concentration. The ASPP was subsequently chelated with zinc sulfate to produce Zn-ASPP, whose structural and functional properties—including particle size, FTIR, DSC, viscosity, SEM, solubility, emulsibility, foamability, and antioxidant capacity—were systematically characterized. The results indicate that Zn-ASPP forms stable nanoparticles with strong antioxidant activity. The strongest antioxidant capacity reached 73.79% at a solution pH of 8, making it particularly valuable for food industry applications. This work may provide a theoretical basis and practical guidance for the development of zinc-fortified marine protein supplements with enhanced antioxidant properties. Full article

Leghemoglobin (LegHb) is a plant-derived heme-containing globin found in the root nodules of legumes like soybean that can be used as a food additive for red color and meaty flavor as a plant-based meat alternative. However, conventional extraction methods face challenges of low yield and high costs. To address this issue, precision fermentation with recombinant microorganisms has been applied for the sustainable large-scale production of plant leghemoglobins. This study attempted the production of plant legHbs using recombinant yeast strains, Saccharomyces cerevisiae and Komagatella phaffii. The plant legHb genes were identified from the genome of legumes such as soybean, chickpea, mung bean and overexpressed in yeast via extracellular secretion by the signal peptide and inducible promoters. Subsequently, hemin as a heme provider was added to the fermentation, resulting in increased levels of plant legHbs. In S. cerevisiae, gmaLegHb expression reached up to 398.1 mg/L, while in K. phaffii, gmaLegHb showed the highest production level, reaching up to 1652.7 mg/L. The secretory production of plant legHbs was further enhanced by replacing the signal peptide in the recombinant yeast. The secreted plant legHbs were purified by His-Tag from a culture supernatant or concentrated via precipitation using ammonium sulfate. These results suggest that the production of plant legHbs is significantly influenced by hemin and signal peptide. This study successfully demonstrates the production of the various plant legHbs other than soy legHb that can be used as natural colors and flavors for plant-based meat alternatives. Full article

Ulcerative colitis (UC), an idiopathic and recurrent ailment, substantially influences a patient’s health. Mung bean peptides (MBPs) are bioactive substances derived from mung bean protein that possess notable anti-inflammatory properties. However, their efficacy and underlying mechanisms in UC treatment remain unclear. In this study, the structural characteristics of MBPs were examined by determining various parameters, such as amino acid composition, molecular weight distribution, and peptide sequences, thereby structurally demonstrating their anti-inflammatory potential. The therapeutic effectiveness of MBPs in UC treatment was evaluated by assessing its influence on colon length, histological damage to colonic tissue, and disease activity index of mice suffering from colitis induced by dextran sulfate sodium (DSS). Additionally, the study explored the potential mechanism of action of MBPs in UC by analyzing the intestinal microbiota, inflammatory cytokines in serum, and tight junction (TJ) proteins in the colon tissue of mice. The results revealed that MBPs significantly increased colon length, reduced colonic tissue damage, and decreased the disease activity index in mice with UC. MBPs restored intestinal barrier function by upregulating the expression of ZO-1 and claudin-1 proteins within the colonic tissue of mice with DSS-induced colitis, thereby treating UC. MBPs exerted anti-inflammatory effects by downregulating the amplification of inflammatory cytokines in the serum, improving the gut microbiota structure in mice with colitis, and regulating immune-related signaling pathways. Therefore, there is an experimental basis for the potential use of MBPs as adjunctive therapy in UC. Full article

Composite hydrogels with improved mechanical and chemical properties can be formed by non-covalently decorating the nanofibrillar structures formed by the self-assembly of peptides with fucoidan. Nevertheless, the precise interactions, and the electrochemical and thermodynamic stability of these composite materials have not been determined. Here, we present a thermodynamic analysis of the interacting forces that drive the formation of a composite fucoidan/9-fluorenylmethoxycarbonyl-phenylalanine-arginine-glycine-aspartic acid-phenylalanine (Fmoc-FRGDF) hydrogel. The results showed that the co-assembly of fucoidan and Fmoc-FRGDF was spontaneous and exothermic. The melting point increased from 87.0 °C to 107.7 °C for Fmoc-FRGDF with 8 mg/mL of added fucoidan. A complex network of hydrogen bonds formed between the molecules of Fmoc-FRGDF, and electrostatic, hydrogen bond, and van der Waals interactions were the main interactions driving the co-assembly of fucoidan and Fmoc-FRGDF. Furthermore, the sulfate group of fucoidan formed a strong salt bridge with the arginine of Fmoc-FRGDF. This study provides useful biomedical engineering design parameters for the inclusion of other highly soluble biopolymers into these types of hydrogel vectors. Full article