Search Results (328)

This study identified the optimal enzymatic treatment for improving the foaming characteristics of oat globulin, and alkaline protease was found to be the most effective enzyme. The impact of alkaline protease on the foaming properties and structural changes in oat globulin was explored. The results show that the foaming capacity of oat globulin hydrolysates is negatively correlated with surface hydrophobicity and positively correlated with the degree of hydrolysis. The results of circular dichroism (CD) and size-exclusion chromatography (SEC) indicate that hydrolysis generated smaller, disordered peptides. Under equilibrium conditions at a 2% concentration, a reduction of 1.62 mN/m in surface tension and an increase of 3.82 μm in foam film thickness were observed. These peptides reduce surface tension between air and water, forming larger, thicker, and more stable foams. Compared to untreated oat globulin, the foaming capacity of hydrolyzed ones increased by 87.17%. Under comparable conditions, these findings demonstrate that limited hydrolyzed oat globulin exhibits potential as an effective plant-based foaming agent up to a degree of hydrolysis of 15.06%. Full article

Titanium is widely recognized as an interesting material for electrodes due to its excellent corrosion resistance, mechanical strength, and biocompatibility. However, further functionalization is often necessary to impart advanced interfacial properties, such as selective ion transport or stimuli responsiveness. In this context, the integration of smart polymers, such as poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA)—noted for its dual pH- and thermo-responsive behavior—has emerged as a promising approach to tailor surface properties for next-generation devices. This work compares two covalent immobilization strategies for PDMAEMA on titanium: the “graft-to” method, involving the attachment of pre-synthesized polymer chains, and the “graft-from” method, based on surface-initiated polymerization. The resulting materials were characterized with size exclusion chromatography (SEC) for molecular weight, Fourier-transform infrared spectroscopy (FTIR) for chemical structure, scanning electron microscopy (SEM) for surface morphology, and contact angle measurements for wettability. Electrochemical impedance spectroscopy and polarization studies were used to assess electrochemical performance. Both strategies yielded uniform and stable coatings, with the mode of grafting influencing both surface morphology and functional stability. These findings provide valuable insights into the development of adaptive, stimuli-responsive titanium-based interfaces in advanced electrochemical systems. Full article

Background/Objectives: Feline panleukopenia, caused by FPV, is a highly contagious disease in cats. Current vaccines face challenges including complex production, high cost, and safety risks. Developing safer, more efficient alternatives is crucial. This study aimed to produce FPV virus-like particles (VLPs) using a recombinant baculovirus system expressing the VP2 gene and evaluate their immunogenicity and protective efficacy in cats. Methods: Sf9 insect cells were infected with recombinant baculovirus to express VP2 protein. The VP2 protein was purified using ultrafiltration and size-exclusion chromatography (SEC). Dynamic light scattering (DLS) and transmission electron microscopy (TEM) confirmed the assembly of VLPs. Twenty healthy cats were randomly divided into four groups; three groups received different doses (5 μg, 15 μg, and 45 μg) of FPV VLP vaccine, while the fourth group served as the control group immunized with PBS. Blood samples were collected on day 21 to measure hemagglutination inhibition (HI) and virus-neutralizing (VN) antibody responses. Cats in the 15 μg dose group were challenged with virulent FPV strain 708 on day 21, and clinical signs and white blood cell counts were monitored for 10 days. Results: Immunized cats exhibited significantly higher HI and VN antibody titers compared to controls. After challenge, vaccinated cats showed no clinical signs of disease, and their white blood cell counts remained stable. In contrast, control cats developed severe symptoms and experienced significant leukopenia. Conclusions: The FPV VLP vaccine generated in this study are highly immunogenic and provide effective protection against virulent FPV challenge, demonstrating their potential as a safer vaccine candidate for feline panleukopenia. Full article

Background: Intracellular bacteria frequently result in chronic and recurrent infections. MRSA is one of the most prevalent facultative intracellular bacteria in clinical infections. The drug resistance of MRSA and the difficulty of most antibiotics in entering cells result in a suboptimal clinical efficacy of antibiotics in the treatment of intracellular MRSA. Bacteriophages represent a promising alternative therapy in the context of the current antimicrobial resistance crisis. Nevertheless, the low efficiency of phage entry into cells and their rapid inactivation remain challenges in the treatment of intracellular MRSA using phages. The utilization of functionalized carriers for the delivery of phages into cells and their protection represents a feasible strategy. Methods: In this study, a new MRSA bacteriophage (vB_SauS_PHM) was isolated from hospital sewage, exhibiting the characteristics of short incubation period, large lytic amount, and good environmental tolerance. Subsequently, vB_SauS_PHM was encapsulated by TAT peptide-functionalized liposomes through microfluidic technology and size-exclusion chromatography (SEC), forming a phage delivery system, designated TAT-Lip@PHM. Results: The encapsulation rate of the phage by TAT-Lip@PHM was 20.3%, and the cell entry efficiency was ≥90% after 8 h. The 24 h eradication rate of 300 μg/mL TAT-Lip@PHM against intracellular MRSA was 94.05% (superior to the 21.24% and 44.90% of vB_SauS_PHM and Lip@PHM, respectively), while the mammalian cell activity was >85% after 24 h incubation. Conclusions: The TAT-Lip@PHM effectively delivered the phage into the cell and showed an excellent killing effect on intracellular MRSA with low cytotoxicity. This work provides a technical reference for the application of phages in the treatment of intracellular bacterial infection. Full article

Background/Objectives: Long-acting injections (LAIs) are innovative drug delivery systems that improve patient compliance by maintaining therapeutic drug levels over extended periods. Micro- and nanosuspensions are commonly used in LAIs to enhance bioavailability, but their thermodynamic instability poses challenges, including particle aggregation and growth. This study aimed to evaluate the impact of two helping processes—vehicle thermal treatment and high-shear homogenization—on the stability and manufacturing efficiency of aripiprazole monohydrate (AM) suspensions. Methods: AM suspensions containing sodium carboxymethyl cellulose (CMCNa), mannitol and disodium phosphate in water for injections (WFIs) were prepared using a combination of thermal treatment of the vehicle solution, high-shear homogenization and bead milling. Four manufacturing variants were tested to assess the influence of these processes on particle size distribution (PSD), viscosity and stability during a 3-month accelerated stability study. Molecular weight changes in CMCNa from thermal treatment were analyzed using size exclusion chromatography with multiangle scattering (SEC-MALS), and PSD was measured using laser diffraction. Results: Thermal treatment of the vehicle solution had minimal impact on CMCNa molecular weight, preserving its functionality. High-shear homogenization during bead milling significantly reduced particle aggregation, resulting in improved PSD and reduced viscosity. Synergistic effects of the two helping processes used in one manufacturing process were observed, which led to superior stability and minimal changes in PSD and viscosity during storage. Batches without the helping processes exhibited increased particle size and viscosity over time, indicating reduced suspension stability. Conclusions: Incorporating vehicle thermal treatment and high-shear homogenization during bead milling enhances the stability and manufacturing efficiency of AM suspensions. These findings underscore the importance of optimizing laboratory-scale processes to ensure the quality and safety of pharmaceutical suspensions. Full article

Background: Exosomes are nanoscale extracellular vesicles actively secreted by cells that play critical roles in disease biomarker discovery, drug delivery, and direct therapeutic applications. However, in vivo pharmacokinetic (PK) studies of exosomes remain limited, hindering their clinical translation. Due to their complex and heterogeneous composition, most existing PK methods rely on chemical or genetic labeling, which may alter their native behavior and complicate accurate analysis. Methods: To address this challenge, we developed a label-free liquid chromatography–tandem mass spectrometry (LC-MS/MS) method to investigate the PK of naive exosome-based therapeutic modalities. Exosomes were isolated from rat plasma using size exclusion chromatography (SEC) and quantified using multiple reaction monitoring (MRM) targeting specific exosomal peptides as surrogate analytes. Following intravenous administration of exosomes via the tail vein, plasma concentrations were determined by peptide peak areas, and PK parameters were calculated using a non-compartmental model. Results: The method was rigorously validated for specificity, linearity, sensitivity, and reproducibility. Its feasibility was further confirmed by successfully characterizing the PK profile of HEK 293F-derived exosomes in rats. Conclusions: This analytical strategy enables direct, label-free quantification of exosomes in plasma and provides a robust platform for advancing exosome-based drug development and translational research. Full article

In this study, we explore the design of novel random poly(lauryl methacrylate-co-tert-butyl methacrylate-co-methacrylic acid), P(LMA-co-tBMA-co-MAA) copolymers via the RAFT copolymerization of LMA and tBMA followed by the selective hydrolysis of tBMA segments. For the molecular characterization of the novel copolymer, a series of physicochemical techniques were implemented, including size exclusion chromatography (SEC), proton nuclear magnetic resonance (¹H-NMR) and attenuated total reflectance–Fourier transform infrared (ATR–FTIR) spectroscopy. Our experimental results confirmed the successful synthesis of the targeted copolymers. The compositions were in accordance with the targeted differing fraction of hydrophobic tBMA/LMA elements, and hydrolysis resulted in at least 64% conversion to hydrophilic MAA units. The copolymers, bearing both an amphiphilic character and polyelectrolyte properties while being composed of randomly distributed monomeric segments of biocompatible materials, were subsequently investigated in terms of their self-assembly behavior in aqueous solutions. Dynamic light scattering and fluorescence spectroscopy experiments demonstrated the formation of self-assembled nanoaggregates (average hydrodynamic radii, R_h < 100 nm) that formed spontaneously, having low critical aggregation concentration (CAC) values (below 3.5 × 10⁻⁶ g/mL), and highlighted the feasibility of using these copolymer systems as nanocarriers for biomedical applications. Full article

This work is devoted to the changes in polysaccharides in thermally treated wood after its accelerated aging with the aim of its optimal utilization after its original use has ended. Spruce wood samples were treated by the Thermowood process at temperatures of 160 °C, 180 °C, and 210 °C and subjected to accelerated aging in wet mode. The influence of treatment temperature and accelerated aging was monitored by wet chemistry, high-performance liquid chromatography (HPLC), X-ray diffraction (XRD), size exclusion chromatography (SEC), and Fourier-transform infrared spectroscopy (FTIR). During thermal treatment, hemicelluloses are mainly degraded. At the temperature of 210 °C, aromatic compounds formed as degradation products of lignin and hemicelluloses bind to cellulose fibers and increase cellulose yield. Preferential decomposition of the amorphous portion of cellulose leads to an increase in its crystallinity, while higher temperatures cause degradation of the crystal lattice. The degree of polymerization in both cellulose and hemicelluloses decreases due to the cleavage of glycosidic bonds. Accelerated aging does not significantly affect the changes in polysaccharides. The results obtained can be used in the processing of cellulose and hemicelluloses from thermally modified wood at the end of its life cycle in various industrial fields. Full article

Applying physical modification methods to raise the resistant starch content is a feasible strategy for developing foods with a low glycemic index (GI) and regulating postprandial hyperglycemia. Here, broken rice starch (C) was modified via ultrasound and quercetin complexation (US-Q). The structure, physicochemical properties, and in vitro digestibility of the US-Q product were subsequently determined. Scanning electron microscopy (SEM) images showed that the modification changed the starch granules’ morphology, forming a more compact and stable structure. Fourier transform infrared (FTIR) spectroscopy images revealed the interaction between the starch and quercetin. An X-ray diffraction (XRD) analysis demonstrated that the crystallinity of the US-Q was lower than that of the C, indicating that the combined modification with ultrasound and quercetin disrupted the long-range ordered structure of the starch and facilitated the formation of a short-range ordered structure from amylose. Size exclusion chromatography (SEC) images showed that both the molecular weight (from 72,080.96 kDa to 85,141.95 kDa) and amylose content (from 15.94% to 26.76%) increased significantly, while the branching degree and average degree of polymerization of amylopectin decreased, suggesting that the ultrasonic treatment processing method had a significant impact on the formation of the quercetin–starch complexes. In terms of in vitro digestion, the resistant starch content of the US-Q was significantly increased from 6.57% to 20.23%, whereas the hydrolysis rate was decreased from 92.6% to 78.35%, indicating that the presence of quercetin reduced the digestibility of the starch complexes by inhibiting the starch-hydrolyzing enzyme activity. Overall, this study improves the understanding of ultrasound and quercetin dual treatment of broken rice starch, providing a theoretical basis for the development of low-GI starch foods for industrial applications. Full article

AS1411 is a G-quadruplex (G4) aptamer that binds tightly to nucleolin (NCL) on the cell surface and has shown strong anticancer effects. However, this aptamer is highly polymorphic, presenting different types of G4s, which may hinder its preclinical application. Several modifications have been made to decrease the polymorphism of this aptamer. In this work, we designed six AS1411 derivatives by substituting guanine with thymine in the central linker and modifying the number of thymines either in the linker itself and/or at both ends of the sequence. The G4 formation, stability, and NCL binding were evaluated by several biophysical techniques and computational and cell studies. Overall, a decrease in polymorphism of G4-forming sequences compared to AS1411 is observed by size exclusion chromatography (SEC) and circular dichroism (CD) spectroscopy in the presence of potassium salt. The melting experiments reveal a higher ability of the derivatives without thymine at both sequence ends to form a G4, consistent with the G4H score predictions. Additionally, it is possible to conclude that deletions of T in the central core increase the ability to form G4. Moreover, the AS1411 derivatives bind NCL with high affinity (K_D values in the 10⁻⁹ M range), particularly the sequences with only thymine modifications in the central linker. In silico studies reveal structural insights and demonstrate that AS1411 derivatives interact with NCL, establishing multiple interactions with the different domains, thereby further supporting the experimental findings. By using a lung cancer cell line with high cell surface NCL expression, we evaluate the internalization and uptake of AS1411 derivatives, identifying the derivative-lacking thymines in the central core as the ones with the highest internalization and cellular uptake. Full article

Small extracellular vesicles (sEVs) play a crucial role in intercellular communication and have demonstrated significant relevance in reproductive biotechnology, particularly in in vitro maturation (IVM) and bovine embryo production. This study evaluates the effects of bovine follicular fluid-derived extracellular vesicles (ffsEVs) isolated using two methods: ultracentrifugation (UC) and size-exclusion chromatography (SEC) on oocyte maturation and preimplantational embryonic development. Significant differences in the size of ffsEVs obtained by both isolation methods were noted, with UC-derived ffsEVs (UC ffsEVs) being smaller than those isolated by SEC (SEC ffsEVs). UC ffsEVs were more effective in upregulating critical oocyte quality genes, such as HSF1 and CPT1B. However, no significant differences were observed in embryonic developmental rates. Furthermore, the expression of genes associated with preimplantational embryonic quality revealed that only the SEC ffsEVs group exhibited a significant increase in IFNT1 and SOX2 levels, indicating an enhancement in embryonic quality. Notably, blastocysts derived from SEC ffsEVs also showed a higher total cell count compared to those from UC ffsEVs. No differences were found in other critical genes like GLUT1 and CDX2. These results suggest that the use of SEC ffsEVs could improve the in vitro embryo production process, highlighting the importance of the isolation method in determining the functional efficacy of ffsEVs according to research objectives. Full article

Observational studies have shown that human digestive function declines naturally with age. Oral enzyme supplementation is a candidate strategy to enhance macronutrient digestion in older adults. The objective of this study was to test the effects of a mixture of six microbial enzyme preparations (ENZ) on nutrient bioaccessibility from a mixed meal in an in vitro model of digestive senescence. The mixed meal included chicken meat, peas, and potatoes. The INFOGEST 2.0 static simulation of oro-gastric digestion was used to model human digestive physiology along with a consensus protocol to model aging. Analytical testing of gastric digesta included measurements of free amino nitrogen (FAN), amino acid (AA), fatty acid (FA), glycerol, maltose, and glucose concentrations. Peptide distribution profiles were evaluated by size exclusion chromatography (SEC) and gel electrophoresis. After simulating digestion of the mixed meal, all nutrient bioaccessibility outcomes compared to pepsin-only controls, except glycerol, were further enhanced by ENZ in the aging condition compared to the standard condition (FAN: 77.1 vs. 39.3%; essential AA: 100.4 vs. 57.6%; total FA: 12.8- vs. 8.0-fold; maltose: 142.1 vs. 0.7%). SEC confirmed ENZ’s proteolytic capacity to generate more lower molecular weight peptides and free AAs in standard and aging conditions compared to pepsin alone. Gel electrophoresis confirmed proteolytic enhancement with ENZ. These data showcase ENZ’s hydrolytic activity toward macronutrients and suggest ENZ’s capacity to compensate for reduced pepsin activity in an aging-adapted oro-gastric digestion simulation. Full article

Adipose tissue is responsible for fat storage and is an important producer of extracellular vesicles (EVs). The biological content of exosomes, one kind of EV, provides information on aspects such as immunometabolic alterations. This study aimed to compare three plasma exosome isolation methods—using a commercial kit (CK), size exclusion chromatography (SEC), and differential centrifugation (DC)—and select the best one. Individuals categorized by normal and high body fat percentages were used. The DC and CK were proven to be the most advantageous out of the exosome isolation methods, so we suggest these methods for further protein and molecular analyses, respectively. Still, we emphasize the importance of selecting an appropriate methodology depending on the specific research objectives. At the same time, no statistical differences in exosome quality, morphology, total protein, or microRNA concentration were observed between individuals categorized by body fat percentage, so we suggest that the exosomal cargo varies in individuals with normal and high fat percentages. Full article

Amphiphilic statistical copolymers can be utilized for the formulation of nanocarriers for the drug delivery of insoluble substances. Oligoethylene glycol methylether methacrylate and methyl methacrylate are two biocompatible monomers that can be used for biological applications. In this work, the synthesis of linear poly(oligoethylene glycol methylether methacrylate-co-methyl methacrylate), P(OEGMA-co-MMA), and statistical copolymers via reversible addition fragmentation chain transfer (RAFT) polymerization is reported. P(OEGMA-co-MMA) copolymers with different comonomer compositions were synthesized and characterized by size exclusion chromatography (SEC), ¹H-NMR, and ATR-FTIR spectroscopy. Self-assembly studies were carried out by the dissolution of polymers in water and via the co-solvent protocol. For the characterization of the formed nanoaggregates, DLS, zeta potential, and fluorescence spectroscopy (FS) experiments were performed. Such measurements delineate the association of copolymers into aggregates with structural characteristics dependent on copolymer composition. In order to investigate the drug encapsulation properties of the formed nanoparticles, curcumin and quercetin were loaded into them. The co-solvent protocol was followed for the encapsulation of varying concentrations of the two drugs. Nanocarrier formulation properties were confirmed by DLS while UV–Vis and FS experiments revealed the encapsulation loading and the optical properties of the drug-loaded nanosystems in each case. The maximum encapsulation efficiency was found to be 54% for curcumin and 49% for quercetin. For all nanocarriers, preliminary qualitive biocompatibility studies were conducted by the addition of FBS medium in the copolymer aqueous solutions which resulted in no significant interactions between copolymer aggregates and serum proteins. Novel nanocarriers of curcumin and quercetin were fabricated as a first step for the utilization of these statistical copolymer nanosystems in nanomedicine. Full article

Size-exclusion chromatography (SEC) is presently a widely used and very informative technique for the characterization of macromolecules in solution. Beyond the first implementations of SEC—which required cumbersome column calibrations and were mainly intended for the determination of molecular weights—the modern SEC approach involving multiple detectors (md-SEC) is based on solution properties such as intrinsic viscosity and light scattering. Thus, md-SEC enables the direct and more efficient determination of molecular weights, as well as the determination of relationships between property and molecular weight, which can be quite useful in structural studies. Here, we first present a review of the fundamental aspects of the dilute-solution properties of macromolecules—particularly the differential refractive index, intrinsic viscosity, and scattering-related properties—on which the various detectors involved in md-SEC are based. Then, we developed SECtools, a suite of public-domain, open-source computer programs, which allow for the full analysis of md-SEC chromatograms. These analyses range from just the recorded raw signals (mV) of the detectors to a full determination of molecular weight averages and distributions. The use of these programs is illustrated through experimental studies using various samples. Full article