Search Results (906)

Background: This work aimed to enhance the solubility of Tadalafil (TDL), a BCS Class II drug, by preparing Type IV lipid-based formulations. Methods: Type IV formulations were prepared using surfactants and/or hydrophilic co-surfactants, resulting in oil-free systems. Results: Based on the solubility test, Transcutol^® HP exhibited the highest solubility for TDL (48.33 ± 0.004 mg/mL) and was selected as the co-surfactant. Among surfactants, Kolliphor^® PS80 (42.74 ± 2.29 mg/mL), Kolliphor^® EL (41.87 ± 2.50 mg/mL), Kollisolv^® PEG 400 (40.70 ± 0.30 mg/mL), and Kolliphor^® HS15 (31.40 ± 3.63 mg/mL) demonstrated high solubilization capacity. These were used to prepare formulations without the addition of an oil phase. The developed formulations resulted in a system with a nano-droplet size (<50 nm) and PDI values < 0.3, which was clear, transparent, and resistant to pH dilutions. The optimum Type IV lipid formulations were further characterized and demonstrated good thermodynamic stability under temperature and pH changes. The optimized formulation was adsorbed onto different carriers and transformed into solid TDL-loaded formulations. The in vitro dissolution rate of the drug from the solidified lipid formulations was studied in various dissolution media. It was observed that the solid formulations prepared with Neusilin US2^® (2:1) exhibited a significantly higher dissolution of over 95% within 5 min compared to the marketed product. The in vitro lipolysis studies demonstrated that F2 formulation maintained TDL in a supersaturated state throughout digestion, with limited enzymatic degradation of the excipients. Cytotoxicity evaluation using the MTT assay in Caco-2 cells confirmed the biocompatibility of both drug-free and TDL-loaded formulations, with IC₅₀ values of 19.55 µg/mL and 17.55 µg/mL, respectively. Conclusions: The overall results suggested that the developed solid Type IV lipid formulations can improve the dissolution rate of TDL, which would potentially lead to an improvement in its oral bioavailability and, consequently, a reduction in the treatment dose as a safe delivery system. Full article

Bioactive peptides (BAPs) from dairy products have garnered increasing attention as natural agents with health-promoting properties, including antihypertensive, antioxidant, antimicrobial, immunomodulatory, opioid, and antidiabetic activities. This systematic review synthesizes research published between 2014 and 2024, retrieved from Scopus and PubMed, and selected according to PRISMA guidelines. A total of 192 studies met the inclusion criteria, collectively reporting over 3200 distinct peptides, with antihypertensive sequences, predominantly angiotensin-converting enzyme (ACE) inhibitors, constituting the largest category (n = 1237). β-casein was the principal precursor across bioactivities, followed by αs1-casein, β-lactoglobulin, and α-lactalbumin. Peptides were primarily produced via enzymatic hydrolysis, microbial fermentation, and gastrointestinal digestion, with peptide profiles influenced by the type of milk, microbial strains, and processing conditions. While cow’s milk remained the dominant source, investigations into goat, sheep, camel, buffalo, and donkey milk revealed species-specific biopeptides. Recent advances in proteomics have enhanced peptide identification and bioactivity prediction, enabling the discovery of novel sequences. These findings underscore the significant potential of dairy-derived BAPs as functional food components and nutraceutical ingredients, while highlighting the need for further in vivo validation, bioavailability studies, and broader exploration of underrepresented milk sources. Full article

Pacific pygmy octopus Paroctopus digueti is a promising model for cephalopod research and aquaculture; its feeding and nutritional biology remain poorly understood. The anterior salivary glands (ASG), posterior salivary glands (PSG), and digestive gland (DG) are central to these processes, but molecular comparisons are lacking. To address this gap, we performed a transcriptomic study to explore the enzymatic repertoire and functional specialization of these tissues. Total RNA was extracted from ASG, PSG, and DG of three pre-adult individuals collected in La Paz Bay, Mexico. RNA-Seq libraries were sequenced, and a non-redundant multi-tissue transcriptome was assembled. The ASG displayed high expression of neuropeptides, playing a role in neuroendocrine regulation. The PSG showed elevated protease expression, supporting its function in extracellular digestion, alongside toxins that reinforce its role as a venom gland. The DG was enriched in proteins linked to biomolecule catabolism and antimicrobial peptides, alluding to metabolic specialization and immune defense. These results were validated by qPCR, and target genes were also amplified in Octopus maya and O. hubbsorum, showing some similarities in expression patterns. Overall, our findings suggest strong glandular specialization in P. digueti, providing insights into cephalopod digestive physiology and supporting its value as a model species. Full article

The growing global demand for clean energy and sustainability has increased interest in lignocellulosic biomass as a viable alternative to conventional fossil fuels. Among the various biomass resources, sugarcane bagasse, an abundant agro-industrial by-product, has emerged as a promising feedstock to produce renewable fuels and value-added chemicals. Its high carbohydrate content offers significant potential for bioconversion. However, its complex and recalcitrant lignocellulosic matrix presents significant challenges that necessitate advanced pretreatment techniques to improve enzymatic digestibility and fermentation efficiency. This review consolidates recent developments in the valorization of sugarcane bagasse focusing on innovative pretreatment and fermentation strategies for sustainable bioethanol production. It emphasizes the synergistic benefits of integrating various pretreatment and fermentation methods to improve bioethanol yields, reduce processing costs and enhance overall process sustainability. This review further explores recent technological advancements, the impact of fermentation inhibitor, and emerging strategies to overcome these challenges through microbial strains and innovative fermentation methods. Additionally, it highlights the multi-faceted advantages of bagasse valorization, including waste minimization, renewable energy production and the promotion of sustainable agricultural practices. By evaluating the current state of research and outlining future perspectives, this paper serves as a comprehensive guide to advancing the valorization of sugarcane bagasse in the transition towards a low-carbon economy. The novelty of this review lies in its holistic integration of technological, economic, and policy perspectives, uniquely addressing the scalability of integrated pretreatment and fermentation processes for sugarcane bagasse, and outlining practical pathways for their translation from laboratory to sustainable industrial biorefineries within the circular bioeconomy framework. Full article

Guava fruit is widely consumed in tropical countries and beyond. The phenolic fraction of guava pulp and processing waste (a single fraction containing seeds, skins, and pulp residues) have been reported to carry in vitro biological activities, acting on biomarkers of metabolic diseases such as type 2 diabetes and obesity (enzymatic inhibition of α-glucosidase and pancreatic lipase), atherosclerosis (mitigation of LDL-cholesterol oxidation), and mutagenesis (suppression of DNA strand scission). However, such bioactivities may be compromised by the exposure of guava phenolics to the harsh conditions found along the human gastrointestinal (GI) tract. To overcome this limitation, guava phenolic extracts were microencapsulated with maltodextrin through freeze-drying. The effect of crude and microencapsulated extracts on biomarkers of metabolic diseases was compared before and after in vitro simulated GI digestion. Moreover, guava waste extracts were tested for their ability to interfere with the intracellular redox status of Caco-2 and HeLa cells incubated with free radicals. Microencapsulation considerably improved the bioaccessibility of guava phenolics across digestion stages, which reflected on the enhancement of most bioactivities measured, with the exception of pancreatic lipase inhibition (both pulp and waste extracts) and LDL oxidative protection (pulp extract). Meanwhile, microencapsulation accentuated intracellular antioxidant activity in Caco-2 cells induced by guava waste extract whereas a prooxidant effect in HeLa cells was intensified. This highlights the selectivity of the same extract toward different cell lines. Overall, microencapsulation was demonstrated as a promising tool for protecting and even enhancing the nutraceutical power of guava phenolics, reinforcing their relevance in the development of functional foods and nutraceutical products. Full article

This review indicates that microalgae may serve as a sustainable supply of bioactive compounds and lipids over the long run. It also discusses the significance of lactic acid bacteria (LAB) and Saccharomyces cerevisiae in biotransformation processes. Microalgae contribute to food security and environmental sustainability due to their rapid growth and diverse applications, including food, feed, and biofuels. Fermentation with LAB and S. cerevisiae enhances the nutritional and functional properties of microalgal biomass, rendering it more digestible, bioactive, and palatable. This review discusses the metabolic characteristics of LAB and S. cerevisiae, their ability to modify microalgal components through enzymatic action, and the resultant products, including enhanced fatty acid profiles and bioactive compounds. Furthermore, the biotransformation of pigments during LAB fermentation is examined, revealing significant alterations in the hue and bioactivity of the pigments, hence enhancing the appeal of microalgal products. Future perspectives emphasize the necessity for further investigation to identify optimal fermentation conditions and to explore the synergistic interactions between LAB and S. cerevisiae in the production of novel beneficial components from microalgae using both microbes. Full article

Pulses and pseudocereals are sustainable protein sources of bioactive peptides (BAPs) with potential antioxidant, antihypertensive, antidiabetic, antimicrobial, and immunomodulatory activities. BAPs are typically liberated during gastrointestinal digestion or through bio-based processes, among which enzymatic hydrolysis and microbial fermentation represent the most widely applied strategies. Enzymatic hydrolysis provides controlled and reproducible release of short peptide motifs; recent advances such as ultrasound- or high-pressure–assisted hydrolysis enhance yield and bioactivity. Fermentation exploits microbial proteolytic activity to generate complex peptide mixtures, while improving sensory quality, reducing antinutritional compounds, and responding to consumer demand for natural and “clean-label” products. In silico tools increasingly complement these approaches by accelerating peptide discovery, predicting interactions with molecular targets, and guiding process design. This review provides an updated overview of bio-based methods to produce BAPs from pulses and pseudocereals, emphasizing the comparative advantages of enzymatic and fermentation technologies and their integration with computational tools. Moreover, it examines regulatory frameworks in the European Union, the United States, Japan, and China, while discussing current challenges for industrial scale-up and application in functional foods and nutraceuticals. These combined strategies offer a promising pathway to unlock the health and sustainability potential of plant proteins. Full article

Phenol red is a widely used, low-cost, label-free colorimetric pH indicator that bridges traditional colorimetric assays with modern quantitative imaging and cell-based screening platforms. Its protonation-dependent absorbance shift (430–560 nm) allows for the real-time monitoring of extracellular acidification, which indirectly reflects cellular metabolism, growth, and respiration. Although phenol red lacks the molecular specificity of genetically encoded or fluorogenic biosensors, it remains useful in systems where pH changes are effective proxies for physiological processes. Existing tissue digestion protocols often overlook key parameters, especially pH control and enzyme cofactor use. This study presents a straightforward, spectrophotometric method to monitor and adjust the pH of low-volume (1 mL) buffered enzymatic dissociation media using phenol red and a plate reader. We titrated dissociation solutions to physiological pH (~7.4) using spectrophotometric pH measurements validated against conventional glass pH probe readings, confirming method reliability. Accurate pH assessment is critical for isolating viable primary cells for downstream applications such as tissue engineering, single-cell omics, and neurophysiological assays. We highlight that papain-based dissociation media supplemented with L-cysteine can be acidic (pH 6.6) if unadjusted, compromising cell viability. This accessible approach enhances reproducibility by promoting pH documentation concerning dissociation conditions that contribute to advancing consistency in biomedical, cellular, neuronal, and tissue engineering research. Full article

This research evaluated the influence of lactic acid bacteria and cellulase, individually or in combination, on the quality of mixed amaranth and sweet potato vine silages. The experiment included four groups: control group with no additives addition (CG), added cellulase group (AS1), added lactic acid bacteria group (AS2) and combined supplementation group (AS3), with five replicates per group. The ensiling period lasted for 60 days. Parameters of silage, including chemical components, fermentation profile, aerobic stability, and in vitro nutrient digestibility, were determined. The results revealed that the quality of amaranth and sweet potato vine mixed silage was improved to a certain degree after addition of two additives individually. Combining these additives observably increased (p < 0.05) the lactic acid and crude protein contents and decreased the pH, ratio of ammonia nitrogen to total nitrogen and neutral detergent fiber content of mixed silage. Compared with the CG and AS1 groups, the number of lactic acid bacteria in the AS3 group increased significantly (p < 0.05), while aerobic bacteria and mold counts showed the opposite tendency. Also, the in vitro dry matter, crude protein and neutral detergent fiber digestibility of the AS3 group were higher (p < 0.05) than those of the CG group. Combined inoculation observably reduced (p < 0.05) the ammonia nitrogen concentration and increased (p < 0.05) the propionic and butyric acid concentrations of mixed silage under in vitro incubation. In summary, the inoculation of lactic acid bacteria and cellulase can enhance the fermentation profile and nutritional values of mixed silage made from amaranth and sweet potato vine, and the best improvement effects are obtained by the combined utilization of the two additives. Full article

Pineapple biomass represents an abundant renewable source of carbon and a promising feedstock with considerable potential for the production of sustainable fuels. In the present study, the influence of liquid hot water (LHW) pretreatment on the pineapple mother plant was investigated at different controlled severities, then characterized by Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). Results show that LHW pretreatment causes structural changes, leading to lignin and hemicellulose depolymerization up to a severity factor of 2.36–3.55, whereas at severity factors in the range of 4.13–5.90, cellulose, hemicellulose, and lignin appear to repolymerize. This pretreatment resulted in a higher hydrolysis efficiency (94.92 ± 0.04%) at 50 °C for 72 h. Compared with the untreated sample, the hydrolysis rate under these conditions increased by a factor of 2.16. SEM imaging revealed significant disruption of the PMP microstructure following LHW treatment, while XRD data confirmed an increase in the crystallinity index. FTIR analysis further indicated modifications in functional group profiles, supporting the structural and compositional changes induced by pretreatment. Overall, this study demonstrates the effectiveness of LHW pretreatment in enhancing the enzymatic digestibility and modifying the physicochemical properties of PMP biomass, providing a foundation for its valorization into high value bioproducts. Full article

Background/Objectives: Obesity and dysglycemia are increasingly associated with intestinal barrier dysfunction and alterations in gut microbiota. Intestinal hyperpermeability is emerging as a therapeutic target in metabolic disorders, but human data integrating barrier biomarkers, epithelial morphology, and microbial composition remain scarce. Methods: Forty-six adults (82.6% female; 38.3 ± 7.8 years) were stratified into lean normoglycemic controls (CON), individuals with obesity and normoglycemia (NOB), and those with obesity and dysglycemia (DOB). Biochemical/inflammatory biomarkers, such as lipopolysaccharide (LPS) and LPS-binding protein (LBP), were measured. Duodenal biopsies were obtained by upper digestive videoendoscopy. Histomorphometry, expression of junctional and cytoskeletal proteins, and enzymatic activity of the duodenal epithelium were used as markers of intestinal permeability. Fecal microbiota composition (FMC) was analyzed by amplifying the V4 region of the 16S rRNA gene, which was sequenced using next-generation sequencing technology. Results: Duodenal histomorphometry did not differ across groups. Intestinal alkaline phosphatase (IAP) was significantly lower in DOB compared to CON. LPS correlated positively with fat mass, and LBP with the waist-to-hip ratio. The villus-to-crypt ratio correlated negatively with BMI, while IAP correlated inversely with fasting glucose and HbA1c. β-actin expression was inversely associated with BMI, glucose, insulin, and HOMA-IR. Microbiota diversity indices were similar between groups, although specific taxa, particularly within the Clostridiales order, were reduced in dysglycemia. Conclusions: Reduced IAP activity and consistent correlations between barrier biomarkers and metabolic parameters highlight intestinal barrier dysfunction as a relevant feature of obesity and dysglycemia. Subtle microbiota alterations further support a link between gut ecology and metabolic control. These findings underscore the intestinal barrier as a promising therapeutic target in metabolic disorders. Full article

A growing scientific interest in bioactive compounds from sea cucumbers is contributing to a broader recognition even in regions where their consumption is not common. This study evaluated the biological potential of a Holothuria forskali extract obtained through different extraction methods, including water extraction, ethanol–water extraction, and enzymatic hydrolysis. The hydrolysate (H), rich in low-molecular-weight peptides, yielded the highest antioxidant (30.6 ± 0.6 mg VitC Eq/g sample for ABTS and 10.7 ± 0.1 mg GAEs/g sample for Folin-reactive substances) and ACE-inhibitory (82.6%) activities. Based on these results, the hydrolysate was selected for encapsulation in two nanostructured delivery systems for comparative purposes: chitosan nanoparticles (NPs) and rapeseed lecithin liposomes (LPs). Both nanostructures were characterized in terms of size, ζ-potential, and polydispersity and subjected to simulated in vitro gastrointestinal digestion (GIDv) to assess their stability and mucoadhesive properties. After digestion, antioxidant activity increased in both systems, particularly in liposomes. Although encapsulation initially reduced ACE-inhibitory activity, gastrointestinal digestion restored or enhanced it, especially in liposomal formulations (≈37% inhibition). The mucoadhesive potential of the nanostructures after DGIv, focusing on their interactions with mucin, was assessed. Liposomal digests significantly increased viscosity in the presence of mucin, while chitosan nanoparticles decreased it, suggesting the formation of soluble complexes with reduced hydrodynamic volume. Electrostatic and hydrogen bonding interactions between chitosan and mucin were particularly evident in the NPH formulation. The rheological synergism parameter (Δη) revealed more negative values for NPs and NPHs, indicating stronger mucoadhesive interactions compared to controls and suggesting their suitability for mucosal delivery. These findings support the use of H. forskali hydrolysates as a source of functional bioactive compounds and highlight the potential of chitosan-based nanocarriers for enhancing their stability, bioaccessibility, and mucoadhesive properties in functional food or nutraceutical applications. Full article

A gastric–intestinal two-step enzymatic hydrolysis in vitro digestion simulation system was used to systematically investigate the digestion kinetics and amino acid release characteristics of five plant protein sources: soybean meal, rapeseed meal, corn DDGS, corn gluten meal, and corn germ meal. The results showed that in the gastric digestion phase (120 min), the protein hydrolysis degree of soybean meal was the highest (61.8%, p < 0.001), which was 4.4 times that of corn gluten meal (14.0%). In the intestinal digestion phase (240 min), the low-molecular-weight peptide release of corn gluten meal (31.2 mg/g) was significantly higher than that of corn DDGS (17.4 mg/g), showing a “weak in the stomach but strong in the intestine” characteristic. The “nutritional value equivalence” model constructed with soybean meal as the reference showed that the gastric digestion phase equivalence of rapeseed meal was only 32.2% (significantly lower than other materials), and the intestinal digestion phase equivalence of corn gluten meal was 62.9%. This study clarified the differences in digestion characteristics and key related indicators of different plant protein sources, providing quantitative references and scientific support for the food and feed industries to precisely select protein sources according to digestion phases and optimize the formula design. Full article

This study presents a novel approach to intensifying the anaerobic digestion of sewage sludge through enzymatic pretreatment using hydrolytic enzymes—cellulase and lysozyme. It aims to determine how enzymatic activation affects the efficiency of methane fermentation, defined as the degree of organic matter decomposition and yield and composition of biogas. An experiment was carried out under mesophilic conditions over 20 days, analyzing the physicochemical properties of sludge, biogas production, methane content, and sanitary parameters. The addition of cellulase and lysozyme significantly enhanced process efficiency, increasing both the rate of organic matter degradation and biogas yield. The highest biogas production values (0.73 L·g⁻¹ d.m. for cellulase and 0.72 L·g⁻¹ d.m. for lysozyme) were obtained at a 4% (w/w) enzyme concentration, with a corresponding increase in the degree of organic matter decomposition to 78.7% and 80.0%, respectively. The produced biogas contained 58–61% methane, exceeding the values observed in the control sample, which indicates a positive effect of enzymatic activation on methane selectivity. Enhanced biogas production was attributed to improved hydrolysis of complex organic compounds, resulting in greater substrate bioavailability for methanogenic microorganisms. Moreover, methane fermentation led to the complete elimination of E. coli from all supernatants, confirming the hygienization potential of the process. The results of this study indicate that enzymatic pretreatment may serve as a viable strategy to improve both the energy efficiency and hygienic safety of anaerobic digestion processes, with relevance for future optimization and full-scale wastewater treatment applications. Full article

The growing consumption of oilseed-pressed cakes (OSCs), a largely underutilized feedstock, plays a significant role in animal feed. The study evaluates the use of three OSCs—flax (FSC), pumpkin (PSC), and hemp (HSC)—as substrates for Bacillus subtilis ATCC 6051a (BS) in a solid-state fermentation (SSF) to enhance enzyme production and probiotic viability. The SSF process was assessed to evaluate the microbial growth, sporulation efficiency, enzymatic activity (protease, cellulase, xylanase, and phytase), and in vitro digestibility of fermented substrates. The results indicate that bacterial growth and sporulation varied significantly among substrates (p < 0.05). FSC presents the highest spore resistance (86.52%), followed by PSC (82.87%) and HSC (81.29%). Notably, protease was highest in HSC (184.67 U/g), while FSC supported maximum cellulase activity. HSC exhibited superior xylanase (1.86 ± 0.043 U/g DW, p < 0.05) and phytase production, while pH analysis indicated a shift toward alkalinity in PSC and HSC due to proteolytic activity. FSC maintained the most stable bacterial population during digestion, suggesting its potential as a probiotic carrier. These findings highlight that fermentation of OSCs with BS improved their nutritional value and can be used as a sustainable solution in feeding programs for piglets. Full article