Search Results (373)

Background: Brucellosis remains a significant zoonotic infection affecting approximately 500,000 people worldwide annually, with no licensed human vaccine available. Recombinant influenza virus vectors expressing Brucella spp. antigens represent a promising vaccine platform. However, transitioning from laboratory constructs to clinical candidates requires validation of scalable purification methods compliant with Good Manufacturing Practice (GMP) standards. This study aimed to develop and optimize a pilot-scale purification protocol for these vectors. Methods: Recombinant influenza A viruses (H5N1) expressing Brucella spp. antigens (Omp16, Omp19, L7/L12, Cu-Zn SOD) were propagated in MDCK cell culture. The optimized purification process included: (1) clarification; (2) ultrafiltration/diafiltration (100 kDa MWCO); (3) two-step chromatography (anion-exchange Q-Sepharose^® Fast Flow and multimodal Capto™ Core 700); and (4) sterile filtration. Process validation was performed across three independent pilot-scale batches (20 L each). Results: The purification process demonstrated high reproducibility for all constructs. Final preparations met established quality criteria: infectious titer ≥ 5.2 log₁₀ TCID₅₀/mL, hemagglutination activity 7.33 ± 0.58–8.33 ± 0.58 log₂, total protein content 157–305 μg/mL, residual host cell DNA < 10 ng/dose, and bacterial endotoxin levels ≤ 0.15 IU/mL. The overall recovery of infectious virus was 20–24%, an optimal value for multi-stage bioprocessing. Preservation of the target genetic insert was confirmed in all final preparations by PCR and sequencing. Conclusions: The developed integrated purification protocol yields vectors with high purification efficiency, preserving biological activity and meeting regulatory quality requirements for residual host cell DNA and endotoxins. The technological platform demonstrated versatility, robustness (inter-batch coefficient of variation for yield did not exceed 10–12%), and scalability, establishing a foundation for preclinical and clinical studies of candidate brucellosis vaccines. Full article

Proteases are among the most commercially important industrial enzymes, yet their large-scale production is often limited by complex and costly downstream processing. In this study, an integrated bioprocess was developed for the production, in situ recovery, and purification of an extracellular protease produced by Aspergillus sp. (SIS 22/UCP 1287) under submerged fermentation. Enzyme extraction was coupled directly to fermentation using a polyethylene glycol (PEG)–phosphate aqueous two-phase system (ATPS), aiming to enhance recovery while preserving enzymatic activity. The effects of PEG molecular weight, polymer and phosphate concentrations, and pH on enzyme partitioning were systematically investigated through a full factorial experimental design. Low-molecular-weight PEG and near-neutral pH conditions significantly favored enzyme migration to the PEG-rich phase. Under optimized conditions (15% PEG 3500, 20% phosphate, pH 7.0), the ATPS achieved a partition coefficient of 65.55, enzyme recovery of 209%, and a purification factor of 1.64. Subsequent purification by DEAE–Sephadex ion-exchange chromatography yielded a tenfold increase in specific activity, with optimal elution at 0.5 M NaCl. SDS–PAGE analysis confirmed the homogeneity of the purified protease, revealing a single band at approximately 59 kDa. Overall, the proposed integrated ATPS–chromatography strategy represents a robust, scalable, and environmentally friendly platform that significantly simplifies downstream processing while maintaining high enzyme activity, highlighting its potential for industrial and biotechnological applications. Full article

Interest in plant proteins and functional foods in the consumer diet is growing rapidly. One way to ensure food security is to diversify protein sources through the development of new plant varieties. This is of particular importance in the current era of climate change, when many historically cultivated varieties may be at risk. Due to their high protein content and favorable amino acid profile, soybeans have a wide range of nutritional and technological applications. The objective of this study was to asses a novel European black soybean variety that was obtained through phenotypic selection and stabilized across successive generations. The present study undertook a thorough evaluation of the chemical composition of the black soybean breeding line PLBPB1/24 and the commercial yellow soybean variety “Abelina”. The study, which employed analytical methods such as GC-FID, ICP-OES, ion-exchange chromatography, and spectrophotometric assays, revealed that PLBPB1/24 had a higher fat content, twice the amount of free amino acids (0.564 vs. 0.279 g/100 g), and an increased iron content (by 8%), while having a lower content of undesirable trypsin inhibitors (by 26%) compared to “Abelina”. These results confirm the potential of this new variety as a valuable source of nutrients, particularly in functional foods and plant-based diets. Full article

Yttrium-90 (⁹⁰Y) is a pivotal pure beta-emitting radionuclide extensively employed in the targeted therapy of malignant tumors, such as hepatocellular carcinoma and lymphoma. The ⁹⁰Sr-⁹⁰Y generator system represents the most effective method for producing no-carrier-added (NCA) ⁹⁰Y to meet escalating clinical demands. However, safe clinical application necessitates the stringent separation of its parent isotope, ⁹⁰Sr, which poses significant radiotoxicological risks due to its long half-life and bone-seeking behavior. This review comprehensively summarizes recent advances in solid-phase adsorbent materials developed for the high-efficiency separation of Y³⁺ and Sr²⁺. We systematically analyze the design strategies, molecular recognition mechanisms, and performance evaluation metrics of various functional systems. Key materials discussed include extraction chromatography (EXC) resins based on organophosphorus extractants, diglycolamide (DGA) derivatives, and crown ethers, as well as inorganic ion exchangers such as antimony-based materials, manganese oxides, and zeolite-like molecular sieves. Special attention is given to composite modification strategies, including silica-based and polymer-matrix composites, and metal doping techniques aimed at enhancing radiation resistance, acid stability, and Sr-Y separation factors (SF). Finally, we provide an outlook on the future development of next-generation ⁹⁰Sr-⁹⁰Y generator materials, highlighting the imperative of transitioning from idealized simulated environments to robust, field-ready applications. Full article

Glycosaminoglycans (GAGs) are the carbohydrate portion of proteoglycans (PGS), a family of complex biomacromolecules ubiquitously found in the extracellular matrix and on cell surfaces that play critical roles in a plethora of physiological and pathological processes. In the present work, chondroitin sulfate (CS) and dermatan sulfate (DS) were extracted and purified from the head (GCB) and skin (GDB) of blue runner fish (Caranx crysos) to explore their structural features and biological properties. GCB and GDB were purified by ion-exchange chromatography with yields of 0.82% and 0.61%, respectively. Chemical and structural analysis showed that GCB and GDD demonstrated quite similar sulfation degrees (4.45% and 4.24%, respectively). The molecular weight values obtained for GCB and GDB as estimated by high-performance size exclusion chromatography coupled with a triple detector array (HP-SEC-TDA) were 48.9 and 28.54 KDa, respectively. Structural features were elucidated using FT-IR and 2D NMR spectroscopy. GCB was mainly identified as chondroitin sulfate, containing 82% GlcA and minor proportions of IdoA and IdoA2S (scoring 18% dermatan-like structures). In contrast, GDB was predominantly dermatan sulfate, with a higher unsulfated IdoA content (54%) and a lower GlcA percentage (17%). In vitro anticoagulant activity, evaluated using APTT and PT assays, demonstrated that both GAGs exhibit significant anticoagulant potential. In addition, both fractions exhibited no antiplatelet activity, suggesting that the isolated glycosaminoglycans selectively target the coagulation cascade without affecting platelet aggregation. Furthermore, hemolytic assays confirmed that neither GCB nor GDB showed any hemolytic activity at the tested concentrations. Cytotoxicity assessment in HEK293 and HUVEK cell lines further confirmed the absence of detectable toxicity even at high concentration. Overall, these marine-derived GAGs present promising therapeutic potential as a source of anticoagulant drugs. Full article

Broiler meat is an important source of protein with a balanced essential amino acid profile; however, the environmental stress associated with a high stocking density can reduce its nutritional value. Adaptogen supplementation represents a promising strategy to mitigate this negative impact. This study aimed to evaluate the amino acid composition of the breast and thigh muscles of Smena-9 broiler chickens under a high stocking density and dietary supplementation with an adaptogens complex (dihydroquercetin + vitamins E and C). The experiment was conducted on 120 broilers allocated to four groups: recommended stocking density (negative control), high stocking density (positive control), high stocking density with adaptogens from day 21, and high stocking density with adaptogens from day 1 of the experiment. The stocking density changed as the bird grew. On day 21, the bird stocking density in the S(-)CON group was 38 heads per 1 m², in the other groups it was increased by 10% and amounted to 42 heads per 1 m². Each week, the stocking density changed and amounted to 32 and 35 heads per 1 m² in 28–35 days, 21 and 23 heads per 1 m² in 35–42 days, and 17 and 19 heads per 1 m² in 43 days and until the end of fattening, respectively. The amino acid profile was determined by ion-exchange chromatography with post-column derivatization using ninhydrin. Age was the primary factor influencing the amino acid composition, followed by muscle type and stocking density. Sex-related effects were less pronounced and were observed at 24 days of age. Breast meat exhibited higher amino acid content than thigh meat (82.4–89.9 vs. 65,8–77.8 g/100 g dry matter for total amino acids, respectively). Under a high stocking density, the most notable changes were observed in the thigh muscle at early ages. By day 52, the birds receiving the adaptogen complex from day 1 showed an increase in the sum of essential amino acids in the breast meat (+2.4%, p < 0.05), with the most pronounced improvements observed for methionine (+5.9%, p < 0.001) and lysine (+3.9%, p < 0.05) compared with the recommended-density control group (S(-)CON). Additionally, early adaptogen administration preserved the levels of flavor-related amino acids between days 34 and 52, whereas a decline was observed in other groups, indicating maintained sensory quality with age. Thus, the early administration of the adaptogen complex appears to mitigate for high stocking density negative effects and contributes to preserving the nutritional value of broiler meat, although the underlying mechanisms remain hypothetical and require further. Full article

Many Greek islands host geothermal springs whose waters can be desalinated to produce drinking water. Some of these waters contain meaningful concentrations of the valuable Rb⁺ and Cs⁺ ions, which, when extracted from the desalination brine as RbCl and CsCl salts, can yield revenues exceeding the freshwater production costs. We demonstrate the use of reverse osmosis (RO) to produce freshwater and apply theoretical simulations to assess a proven extraction method applied to the RO retentate of geothermal water from Samothrace, characterized by [Rb⁺] = 2.72, [Cs⁺] = 0.55, [K⁺] = 514, [Na⁺] = 3759 (all in mg/L) and pH 6. The extraction method, developed by the authors, relies on ion exchange using a PES-coated Zn-hexacyanoferrate sorbent with high affinity for monovalent cations (no affinity for multi-valent cations), followed by a unique ion-chromatography separation. We show that the production cost remains <25% of the salts’ market price, with ROI of ~4.5 years. Full article

The scientific literature lacks sufficient data on the transport of various toxic pollutants by polymer particles. Investigating how the structure of microplastic particles formed during the degradation of polymeric materials affects pollutant sorption processes will improve our ability to predict environmental behavior. General-purpose polystyrene, expanded polystyrene, ABS plastic (acrylonitrile–butadiene–styrene) and crosslinked polystyrene are produced on an industrial scale. Copolymers of styrene with divinylbenzene are used on a large scale as sorbents for gel permeation chromatography (Styragel brand sorbents), in the production of catalysts on a polymer substrate or ion-exchange resins. In this study, non-spherical, crosslinked polystyrene microparticles with varying polystyrene chain packing densities were used as model microplastic particles representative of crosslinked polystyrene. It was shown that the adsorption of a hazardous chemical rhodamine B was influenced by both the packing density of the polystyrene chains and the presence of ionic functional groups, i.e., the “degree of aging” of the microplastic particles. The sorption capacities of these model microparticles were compared with those of natural origin (silicon dioxide, quartz powder, and microcrystalline cellulose). A viability assay using HEK293 and HeLa cell lines exposed to leachates from both pristine and rhodamine B-loaded microparticles revealed that all unmodified microparticles, regardless of their nature, exhibited no cytotoxicity at concentrations up to 1000 μg/mL. In contrast, microparticles with adsorbed rhodamine B significantly reduced cell viability to 20–40% at concentrations of 100 μg/mL. Full article

Aerogels derived from microbial exopolysaccharides are useful in the food, pharmaceutical, and environmental sectors, but their application in anticancer therapy is constrained by inadequate characterization, especially regarding effects on normal cells. This study used ethanol precipitation and trichloroacetic acid deproteinization to extract crude exopolysaccharide from the fermentation broth of Bacillus amyloliquefaciens SQ-2. The pure fraction, EPS-3791, was obtained using Sephadex G-100 gel filtration chromatography and DEAE cellulose ion exchange. The weight–average molecular weight of EPS-3791 was 64.4 kDa. Monosaccharide analysis indicated fructan as the dominant component, which was consistent with the results of methylation analysis and NMR spectroscopy, confirming that EPS-3791 is a fructan mainly linked by →1)–Fruf–(2→bonds. UV scanning indicated high purity. FTIR analysis revealed functional groups including hydroxyl, carbonyl, and C–O–C groups. EPS-3791 exhibited a porous three-dimensional network morphology by SEM, with a decomposition temperature of 191.61 °C by TGA. Additionally, aerogels were prepared by freeze drying. EPS-3791 aerogels demonstrated minimal cytotoxicity to normal L929 cells while inhibiting the growth of human lung cancer A549, breast cancer MCF–7, and cervical cancer HeLa cells in a dose-dependent manner. Scratch wound healing experiments revealed that EPS-3791 aerogels hindered HeLa cell migration while promoting L929 wound closure. These findings identify EPS-3791 as a fructan-type exopolysaccharide aerogel with specific anticancer properties. Full article

The marine benthic invertebrate Urechis unicinctus exhibits extraordinary tolerance to hypoxic environments, making its coelomic fluid a unique and promising biological source for discovering novel stress-adapting macromolecules. Polysaccharides derived from the coelomic fluid of U. unicinctus were systematically extracted, fractionated, and characterized to investigate their structural features and associated biological activities. Gradient ethanol precipitation (30–80%) combined with DEAE-52 ion exchange chromatography yielded twelve fractions with distinct physicochemical properties. Significant variations were observed in molecular weight (10³–10⁵ Da), sulfate content (3.77–24.26%), and monosaccharide composition. High-ethanol fractions, particularly U68P and U18P (extracted at 60 °C and 100 °C, respectively, and both precipitated with 80% ethanol), were enriched in low-molecular-weight, highly sulfated heteropolysaccharides composed of galactose, fucose, glucosamine, and ribose. These fractions exhibited superior antioxidant activities, including strong scavenging effects against DPPH, ABTS, and hydroxyl radicals. Moreover, they demonstrated pronounced neuroprotective effects in the oxygen–glucose deprivation/reoxygenation (OGD/R) model using SH-SY5Y cells, significantly improving cell viability. Structure–activity relationship analysis revealed that reduced molecular weight, increased sulfation degree, and more diverse monosaccharide composition (e.g., more diverse monosaccharide composition) synergistically contribute to improved bioactivity by facilitating cellular uptake and exposing functional groups. In contrast, high-molecular-weight homoglucan fractions showed relatively weak effects. Overall, this study identifies U. unicinctus coelomic fluid as a promising source of bioactive polysaccharides and provides a theoretical basis for the development of marine-derived anti-hypoxic and antioxidant agents. Full article

Blood collection tubes are widely used medical devices. Inaccurate citrate anticoagulant concentration can influence the results of coagulation tests. The producer’s expertise and responsibility are considered the quality safeguards. However, the tubes undergo changes during their lifecycle, partly due to storage conditions, and the end user or a third party has no comprehensive insight. A methodology is necessary to reveal the tube’s inherent characteristics. We provide insight into the anionic–cationic composition and pH of anticoagulant solutions in commercial tubes using high-performance ion exchange chromatography on a purified water model, making the anticoagulant volume accuracy assessment possible through a direct dye-dilution method. The results revealed differences between the tubes of two producers, Greiner BIO-ONE (A and A(nr)) and BD (C). Tube C has the most accurate anticoagulant amount. Both brands contain buffered citrate. The method of buffer preparation is not a source of interferant for the spectrometric method of the tubes’ quality evaluation. Acetate, formate, chloride, nitrite, sulfate, oxalate, bromide, and nitrate impurities were determined in anticoagulant solutions, all in tube A and some in the others. Tubes C exhibit the highest contamination with cations. Full article

Background/Objectives: Cervical cancer, predominantly driven by persistent infection with high-risk human papillomaviruses (HPVs), is one of the most common malignancies and an important cause of cancer-related mortality among women worldwide. Although existing licensed prophylactic HPV vaccines confer excellent protection, their global use remains suboptimal due to concentrated manufacturing capacity and high production costs. This study aimed to establish a cost-effective multivalent HPV virus-like particle (VLP) vaccine platform. Specifically, we used an enhanced baculovirus expression vector system to produce a 12-valent HPV VLP vaccine to improve antigen yield, thereby reducing manufacturing costs and ultimately improving affordability and availability in low- and middle-income countries. Methods: Optimized expression cassettes and an insect cell culture process were designed to enhance productivity across 12 HPV L1 genotypes. A scalable purification scheme integrating ion-exchange and adsorption chromatography was developed to produce high-purity VLPs with consistent structural integrity. Immunogenicity was assessed in a murine model. Elicited HPV type-specific IgG antibody responses were compared with those induced by the licensed 9-valent HPV vaccine. Results: The assembled 12-valent VLPs were comprehensively characterized using biophysical and immunochemical analyses, confirming structural stability and correct antigenicity. In vivo immunogenicity studies in mice showed strong and serotype-specific IgG responses, comparable or superior to those induced by the licensed 9-valent commercial vaccine. Conclusions: The enhanced baculovirus expression vector system is a versatile and economically sustainable platform for next-generation HPV vaccine production. This technology offers a promising approach to lowering vaccine manufacturing costs and improving global access, particularly in low- and middle-income regions heavily burdened by HPV-associated diseases. Full article

2-Amino-3-methylhexanoic acid (AMHA), as a naturally occurring α-amino acid, exhibits excellent bioactivity in inducing plant resistance to biotic and abiotic stresses, promoting plant growth and alleviating pesticide damage. However, its low yield in nature has hindered its industrialization. This study aims to address the current challenges in AMHA production, including synthetic difficulties, low yield, and poor production safety. We investigated the production capacity of Serratia marcescens NAU002-4 using high-performance liquid chromatography (HPLC), orthogonal experiments, and ion exchange chromatography (IEC), supplemented by precursor feeding assays. Optimization of the fermentation medium revealed that 10 g/L corn steep liquor as the nitrogen source and 10 g/L threonine as the precursor resulted in maximal yield. Further optimization of fermentation parameters—initial pH 7.2, inoculation volume 6%, stirring speed 180 rpm, and aeration rate 350 L/h—led to an AMHA titer of 504 mg/L. In parallel, field trials were conducted to evaluate the effects of pure, biosynthetic AMHA on tea plant growth and photosynthetic performance, using plant physiological data and chlorophyll fluorescence rise kinetics. The results demonstrated that biosynthetic AMHA significantly enhances tea plant growth and photosynthetic efficiency, providing a scientific basis for its development and application as a novel biostimulant. Full article

Acidic polysaccharides, valued for their outstanding bioactivity and physicochemical properties, represent a promising strategy for metabolic disease intervention. In this study, three acidic polysaccharide fractions (BRP-1, BRP-2, and BRP-3) were isolated from Brassica rapa L. using membrane filtration and ion-exchange chromatography. BRP-3, notable for its high galacturonic acid content (76.64%), was further purified to yield the homogeneous fraction BRP-3-1 (Mw = 22.3 kDa). Combining GC-MS, FTIR, and NMR analyses, we report for the first time the detailed structure of BRP-3-1—a heteropolysaccharide composed of rhamnose (1.687%), galacturonic acid (75.584%), galactose (14.452%), and arabinose (8.277%)—with a backbone composed with T-α-L-Araf-(1 → 5)-α-L- Araf -(1 → 4)-α-D-GalpA-(1 → 4)-α-D-2-O- GalpA Me-(1 → 4)-α-D-GalpA-(1 → 4)-α-D-GalpA-(1 → 3)-Galp-(1 → 4)-α-D-GalpA, and T-Rhap, T-Galp as well as T-GalpA for branched chain and terminals. In HepG2 insulin-resistant cells, BRP-3-1 demonstrated potent dual regulation of glucose and lipid metabolism—enhancing glucose consumption, lowering total cholesterol, and significantly reducing triglyceride levels in the high-dose group (800 μg/mL), outperforming BRP-2. This work systematically defines the structure of a highly bioactive acidic polysaccharide from B. rapa L. and confirms its metabolic regulatory effects, offering a strong scientific foundation for its application in functional foods and as an adjuvant therapeutic for metabolic disorders. Full article

Background: Polygonatum sibiricum (PS), a perennial herbaceous plant belonging to the Liliaceae family, is widely distributed in China and other East Asian countries. PS has been used as food and medicine for thousands of years, and its rhizomes are rich in Polygonatum sibiricum polysaccharides (PSP), which exhibit various bioactivities, yet their structural features and therapeutic mechanisms against ulcerative colitis (UC) remain unclear. Methods: A homogeneous polysaccharide, PSP-1b (57.45 kDa), was isolated from the rhizomes of PS via ion-exchange and gel filtration chromatography and structurally characterized using chromatographic and spectroscopic methods. In vivo, its effects were evaluated in a dextran sulfate sodium (DSS)-induced mouse model of UC, while in vitro mechanisms were explored using macrophages stimulated with lipopolysaccharide (LPS) and neutrophil extracellular traps (NETs). Results: PSP-1b was identified as a neutral polysaccharide with minimal branching. Its primary structural backbone was largely composed of →4)-β-D-Galp-(1→ residues. A portion of these backbone residues was substituted at the O-6 position by side chains primarily composed of β-D-Galp-(1→ units. In vivo, PSP-1b significantly alleviated DSS-induced colitis by reducing inflammatory cytokine secretion, suppressing colonic macrophage infiltration, and reversing neutrophil extracellular traps (NETs) deposition. In vitro, PSP-1b directly interacted with TLR4, inhibited the MAPK/NF-κB signaling pathway, and attenuated LPS- and NET-induced macrophage polarization and inflammation. Conclusions: PSP-1b as a promising candidate for functional foods or therapeutic agents targeting inflammatory bowel disease. Full article