Search Results (1,114)

Background/Objectives: Army Liposome Formulation with QS21 (ALFQ) is a vaccine adjuvant formulation consisting of liposomes that contain saturated zwitterionic and anionic phospholipids, 55 mol% cholesterol, and small molar amounts of monophosphoryl lipid A (MPLA) and QS21 saponin as adjuvants. A unique aspect of ALFQ is that after addition of QS21 to nanoliposomes (<100 nm), the liposomes self-assemble through fusion to form giant (≥1000 nm) unilamellar vesicles (GUVs). The purpose of this study was to introduce and investigate new intermediate structures in the fusion process that we term tethered incomplete microspheres (TIMs), which were discovered by us incidentally as structures that were visible by phase contrast microscopy. Methods: Differential centrifugation; phase contrast microscopy; confocal microscopy of vesicles or TIMs which contain fluorescent chromophores linked to phospholipids or cholesterol; ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis of lipid components of liposomes and TIMs; and dynamic light scattering were all used for the characterization of TIMS. Results and Conclusions: (A) Sizes of TIMs range from overall aggregated structural sizes of ~1 µm to mega sizes of ≥200 µm. (B) Stable TIM structures occur when a fusion process is stopped by depletion of a fusogenic lipid during an evolving fusing of a lipid bilayer membrane. (C) TIMs consist of long-term stable (>2 years), but also metastable, tightly aggregated tear-drop or spherical incomplete GUVs tethered to visible masses of underlying vesicles that are not individually visible. (D) The TIMs and GUVs all contain phospholipid and cholesterol (when present) as bulk lipids. (E) Lyophilized liposomes lacking QS21 saponin, but which still contain MPLA (ALF55lyo), also self-assemble to form GUVs and TIMs. (F) Cholesterol is a required component in nanoliposomes for generation of GUVs and TIMs by addition of QS21. (G) Cholesterol is not required for production of GUVs and TIMs in ALFlyo, but cholesterol greatly reduces and narrows the polydisperse vesicle distribution. Full article

Adequate bleeding control is crucial in surgical procedures. Surgifoam and Hemospon are absorbable hemostatic sponges made from lyophilized porcine gelatin, commonly used in oral surgery. This clinical study aimed to evaluate bleeding control, soft tissue healing, and postoperative pain in dental sockets after mandibular third molar extractions filled with Surgifoam and Hemospon. Twenty-five volunteers requiring extractions of both left and right lower third molars participated in this randomized, double-blind, split-mouth study. After extraction, each socket was randomly filled with Hemospon (test group) or Surgifoam (control group). Postoperative pain was assessed using the Visual Analog Scale (VAS) on days 1, 2, 3, and 7. Bleeding at 30 and 60 min (Souto and Mühlemann scales) and soft tissue healing at 7 and 14 days (Brancaccio scale) were evaluated using Fisher’s exact test. Bleeding scores at 30 min predominantly showed no bleeding (score 0) in 80% of participants using Surgifoam, compared to 60% in the Hemospon group. No significant differences in bleeding were observed between groups (p > 0.05), and both showed a similar reduction over time. Soft tissue healing was revealed at 14 days complete healing (score 0) in 90% of participants in both groups. No significant differences between Hemospon^® and Surgifoam^® were observed (p > 0.05). Postoperative pain evaluation showed highly variable scores on the first day (median: 2; range: 1–6) for both Surgifoam^® and Hemospon^®. By the seventh day, pain levels significantly reduced (median: 0), with no significant differences observed between the groups at any time point (p > 0.05). In conclusion, the results of this study suggest that Hemospon and Surgifoam are equally effective in bleeding control, healing, and pain control after third molar surgery. This research aims to guide surgeons on the clinical aspects of using these specific hemostatic sponges in post-extraction procedures for posterior molars and seeks to direct future clinical studies involving these materials. Full article

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related death worldwide, and treatment remains challenging due to high recurrence rates, resistance to chemotherapy, and severe side effects. Dasatinib (Das) has shown therapeutic potential against HCC, but its clinical use is limited by poor bioavailability and short half-life (~3–4 h). Here, we developed a hyaluronic acid (HA)-coated sterosome for targeted and sustained delivery of Das to CD44-overexpressing HCC cells. Sterosomes composed of octadecylamine and cholesterol at a 5:5 (v/v) ratio were prepared via thin-film hydration and sonication, yielding stable particles (~90 nm) with high encapsulation efficiency (EE ~72%) for uncoated vesicles and ~58% after HA coating. HA-sterosomes (HA-St-Das) exhibited a uniform size (≈200 nm) and negative surface charge (–26 mV), with improved storage stability and resistance to lyophilization. In vitro release studies demonstrated pH-responsive Das release accelerated under acidic conditions (pH 6.0–5.0), mimicking tumor and lysosomal environments. In HepG2 cells, HA-St-Das exhibited enhanced cytotoxicity (IC50 ~7.0 μM) and prolonged intracellular retention compared to free Das and uncoated carriers. Fluorescence microscopy confirmed receptor-mediated uptake via CD44, leading to gradual and sustained intracellular delivery. Overall, the HA-St-Das system provides biocompatible, targeted, and controlled Das delivery, addressing key limitations of current liver cancer therapies and representing a promising nanomedicine platform for further development. Full article

Functional beverages are increasingly sought as components of a healthy diet, and goat milk offers a nutritious base with unique sensory attributes. This study aimed to develop a novel fermented goat milk beverage enriched with spent coffee grounds (SCG) extract, utilizing SCG’s high natural antioxidant content to improve nutritional and functional properties. SCG was extracted via aqueous solid–liquid extraction and lyophilized; its extract was incorporated into goat milk–fructose blends at 0%, 1%, and 2% (w/v). Analyses included physicochemical characterization (pH, acidity, fat, and protein), total phenolic content, and antioxidant capacity via DPPH assay, alongside consumer sensory evaluation for acceptance and purchase intent. Results demonstrated that higher SCG extract levels significantly increased pH, phenolic concentrations, and radical scavenging activity while reducing titratable acidity. The 2% SCG formulation achieved the highest overall, taste, and aftertaste acceptance and purchase intention. These findings suggest that SCG-enriched goat dairy beverages are feasible functional foods with enhanced antioxidant properties and consumer appeal, promoting valorization of coffee by-products. Full article

Anthrax remains a serious bioterrorism threat for which new and thermostable vaccines are needed. We previously demonstrated that immunization of rabbits with multiple-antigenic-peptide (MAP) vaccines elicit antibody (Ab) against the loop-neutralizing-determinant (LND), a cryptic linear neutralizing epitope in the 2β2-2β3 loop of protective antigen (PA) from Bacillus anthracis (B. anthracis), which mediates the complete protection of rabbits from inhalation spore challenge with B. anthracis Ames strain. Importantly, LND-specific Ab is not significantly elicited with PA-based vaccines. In the current study, we sought to identify a second unique neutralizing epitope which would also not overlap with the neutralizing specificities elicited by PA-based vaccines, and which could be combined with an LND vaccine as a prototype bivalent vaccine for anthrax. We evaluated linear peptide sequences in the α2-α3 helices of domain 3 of lethal factor (LF) in the form of virus-like particle (VLP) vaccines. Immunogenicity studies confirmed the presence of a 20-mer peptide sequence that is capable of eliciting protective levels of neutralizing Ab following two immunizations of rabbits using human-use adjuvants, and lyophilization of the VLPs did not diminish their immunogenicity. To our knowledge, this is the first demonstration that immunization with linear peptide sequences from LF can elicit protective levels of neutralizing Ab in vivo. Full article

Veterinary vaccines are essential tools for controlling infectious and zoonotic diseases, safeguarding animal welfare, and ensuring global food security. However, conventional vaccines are hindered by cold-chain dependence, thermal instability, and logistical challenges, particularly in low- and middle-income countries (LMICs). This review explores next-generation veterinary vaccines, emphasizing innovations in thermostability and delivery platforms to overcome these barriers. Recent advances in vaccine drying technologies, such as lyophilization and spray drying, have improved antigen stability and storage resilience, facilitating effective immunization in remote settings. Additionally, novel delivery systems, including nanoparticle-based formulations, microneedles, and mucosal routes (intranasal, aerosol, and oral), enhance vaccine efficacy, targeting immune responses at mucosal surfaces while minimizing invasiveness and cost. These approaches reduce reliance on cold-chain logistics, improve vaccine uptake, and enable large-scale deployment in field conditions. The integration of thermostable formulations with innovative delivery technologies offers scalable solutions to immunize livestock and aquatic species against major pathogens. Moreover, these strategies contribute significantly to One Health objectives by mitigating zoonotic spillovers, reducing antibiotic reliance, and supporting sustainable development through improved animal productivity. The emerging role of artificial intelligence (AI) in vaccine design—facilitating epitope prediction, formulation optimization, and rapid diagnostics—further accelerates vaccine innovation, particularly in resource-constrained environments. Collectively, the convergence of thermostability, advanced delivery systems, and AI-driven tools represents a transformative shift in veterinary vaccinology, with profound implications for public health, food systems, and global pandemic preparedness. Full article

A facile and cost-effective sol–gel method for the synthesis of uniformly porous alumina (Al₂O₃) was developed using stable CTAB/hexanol/water microemulsions as soft templates. The phase behavior of the ternary system was investigated to identify compositions that form kinetically stable microemulsions, with an optimal ratio of 7.5 wt.% CTAB, 5 wt.% hexanol, and 87.5 wt.% water exhibiting minimal droplet size variation over one week. Gelation was induced by partial neutralization to pH 4.2 with ammonium carbonate, promoting the formation of polynuclear Al species and enabling the uniform entrapment of hexanol droplets. Lyophilization preserved the porous network, and calcination at 500 °C yielded η-Al₂O₃ with a large specific surface area (~225 m²·g⁻¹) and a narrow mesopore size distribution centered around 100 nm, consistent with the original droplet size. Mercury porosimetry and SEM analyses confirmed a highly porous, low-density material (0.75 g·cm⁻³) with an interconnected pore morphology. This scalable synthesis method, supported by the high kinetic stability of the microemulsion, provides sufficient processing time and eliminates the need for post-synthesis purification. It shows strong potential for producing advanced alumina materials for use in energy storage, catalysis, and sensor applications. Full article

The instability of mRNA vaccines presents significant challenges for their storage, transportation, and large-scale distribution, particularly in resource-limited countries. Recently, freeze-drying (lyophilization) has been considered as a promising approach for preserving mRNA vaccine efficacy. This formulation technique enhances the long-term stability of mRNA vaccines by converting them into a stable dry powder. The purpose of this review is to provide an overview of the current knowledge on the progress of freeze-drying techniques for mRNA vaccines, with emphasis on the associated challenges. This review highlights the factors influencing the stability of freeze-dried mRNA vaccines and provides a comprehensive overview of the formulation components, including excipients, buffers, and surfactants, as well as the process parameters and storage conditions that aim to improve stability and shelf-life. By providing these insights, this review supports the advancement of more robust, scalable, and efficient lyophilization protocols, ultimately addressing the stability limitations of mRNA vaccines and enhancing their global accessibility. Full article

Anthrax remains a formidable bioterrorism threat for which new, optimized and thermostable vaccines are needed. We previously demonstrated that five immunizations of rabbits with a multiple-antigenic-peptide (MAP) vaccine in either Freund’s adjuvant or human-use adjuvants can elicit antibody (Ab) against the loop-neutralizing determinant (LND), a cryptic neutralizing epitope in the 2β2-2β3 loop of protective antigen from Bacillus anthracis (B. anthracis), which mediates complete protection of rabbits from inhalation spore challenge with the B. anthracis Ames strain. To develop a more immunogenic vaccine, we molecularly constructed a virus-like particle (VLP) vaccine, comprising the Woodchuck hepatitis core antigen capsid (WHcAg) displaying 240 copies of the LND epitope on each nanoparticle. Initial studies showed that the LND-VLP was immunogenic in rabbits following two immunizations, and passive transfer of the rabbit sera into A/J mice conferred complete protection from aerosol challenge with B. anthracis. Further optimization of the vaccine revealed that the lyophilized LND-VLP vaccine was capable of eliciting highly protective levels of neutralizing antibody with two immunizations, and in some rabbits, a single immunization, using human-use adjuvants. A lyophilized LND-VLP nanoparticle vaccine may be an effective stand-alone vaccine or may complement PA-based vaccines as a future pre- or post-exposure vaccine for anthrax. Full article

Background/Objectives: Radiolabeled interleukin-2 (IL2) could allow for imaging activated T-lymphocytes in the tumor microenvironment (TME). The aims of this study were to assess the shelf life of a lyophilized kit containing THP-desIL2 to allow for the labeling of IL2 with ⁶⁸Ga at room temperature and to test the in vitro binding of ⁶⁸Ga-THP-desIL2 on different T-cell populations in order to determine which specific T-cell subset expresses the CD25 subunit of the IL2 receptor (IL2R). Methods: desIL2 was conjugated with THP and lyophilized. ⁶⁸Ga labeling was performed and several quality controls, including HPLC, iTLC and SDS-PAGE, were carried out at different storage times (1, 3 and 6 months) and temperatures (4 °C and −80 °C). Moreover, flow cytometric analysis on different T-cell populations and the in vitro and competitive binding of ⁶⁸Ga-THP-desIL2 were performed. Results: The lyophilized kit of THP-desIL2 was stable up to 6 months at −80 °C, preserving its sterility, integrity and acceptable values of labeling yield (51.80 ± 3.74%), radiochemical purity (>96%) and specific activity (5.59 ± 0.40 MBq/µg). Binding of ⁶⁸Ga-THP-desIL2 on activated lymphocytes was specific and exhibited a low dissociation constant from IL2R on stimulated Tregs (Kd: 10⁻⁹–10⁻¹⁰ mol/L). Conclusions: We assessed the shelf life of a lyophilized kit containing THP-desIL2 for the easy labeling of IL2 with ⁶⁸Ga at room temperature. The kit can be stored at −80 °C up to 6 months, thus facilitating the adoption of ⁶⁸Ga-THP-desIL2 into clinical practice. ⁶⁸Ga-THP-desIL2 showed high affinity and specificity for CD25 on activated T-lymphocytes, particularly Tregs, thus opening new opportunities for imaging immune cells trafficking in the TME. Full article

Amelanchier alnifolia is a plant known for its nutritional and bioactive properties. Its leaves contain a high concentration of active compounds with significant antioxidant and antidiabetic effects, including strong α-glucosidase inhibitory potential. The combination of these bioactive leaf extracts with prebiotic substances, such as fructooligosaccharides (FOS), galactooligosaccharides (GOS), and chitooligosaccharides (COS), enables the development of functional systems with enhanced beneficial properties. In this study, process optimization for leaves extraction was performed using a Plackett–Burman screening design, which identified key parameters for further optimization using the Box–Behnken design. The optimal extraction conditions were determined as follows: methanol content 58.06%, solid-to-solvent ratio 26.03 m/v, and extraction time 73.56 min. These conditions yielded the highest the total phenolic content (TPC). A comparative analysis of different cultivars revealed significant variations in polyphenol content among them. The formulated lyophilized systems with GOS, FOS and COS positively influenced the chlorogenic acid release profile, while maintaining the extract’s antidiabetic and antioxidant properties. FT-IR analysis confirmed the molecular interactions responsible for these effects. The prebiotic effectiveness of the systems was quantitatively evaluated using two key parameters: the prebiotic index (PI), and the prebiotic activity score (PAS). Microbiological analyses demonstrated the beneficial effects of prebiotic-enriched systems characterized by better prebiotic action on Bifidobacterium strains than the pure extract. These findings suggest that A. alnifolia leaf extracts, in combination with prebiotics, could serve as promising functional ingredients with potential applications in health-promoting and antidiabetic formulations. Full article

Astragalus sinicus, a key leguminous green manure widely cultivated in Southern China’s rice-based cropping systems, plays a pivotal role in sustainable agriculture by enhancing soil organic matter sequestration, improving rice yield, and elevating grain quality. The symbiotic nitrogen-fixing association between A. sinicus and its matching rhizobia is fundamental to its agronomic value; however, suboptimal inoculant efficiency and field application methodologies constrain its full potential. To address these limitations, we conducted a multi-phase study involving (1) rhizobial strain screening under controlled greenhouse conditions, (2) an optimized lyophilization protocol evaluating cryoprotectant (trehalose, skimmed milk powder and others), and (3) seed pelleting trails with rhizobial viability and nodulation assessments over different storage periods. Our results demonstrate that Mesorhizobium huakuii CCBAU 33470 exhibits a superior nitrogen-fixing efficacy, significantly enhancing key traits in A. sinicus, including leaf chlorophyll content, tiller number, and aboveground biomass. Lyophilized inoculants prepared with cryoprotectants (20% trehalose or 20% skimmed milk powder) maintained >90% bacterial viability for 60 days and markedly improved nodulation capacity relative to unprotected formulations. The optimized seed pellets sustained high rhizobial loads (5.5 × 10³ cells/seed) with an undiminished viability after 15 days of storage and nodulation ability after 40 days of storage. This integrated approach of rhizobial selection, inoculant formulation, and seed coating overcomes cultivation bottlenecks, boosting symbiotic nitrogen fixation for A. sinicus cultivation. Full article

This study aimed to develop a functional powder using whey and milk matrices, leveraging the protective capacity of chia–alginate hydrogels and the advantages of electrohydrodynamic spraying (EHDA), a non-thermal technique suitable for encapsulating probiotic cells under stress conditions commonly encountered in food processing. A hydrogel matrix composed of chia seed mucilage and sodium alginate was used to form a biopolymeric network that protected probiotic cells during processing. The encapsulation efficiency reached 99.0 ± 0.01%, and bacterial viability remained above 9.9 log₁₀ CFU/mL after lyophilization, demonstrating the excellent protective capacity of the hydrogel matrix. Microstructural analysis using confocal laser scanning microscopy (CLSM) revealed well-retained cell morphology and homogeneous distribution within the hydrogel matrix while, in contrast, scanning electron microscopy (SEM) showed spherical, porous microcapsules with distinct surface characteristics influenced by the encapsulation method. Encapsulates were incorporated into beverages flavored with red fruits and pear and subsequently freeze-dried. The resulting powders were analyzed for moisture, protein, lipids, carbohydrates, fiber, and color determinations. The results were statistically analyzed using ANOVA and response surface methodology, highlighting the impact of ingredient ratios on nutritional composition. Raman spectroscopy identified molecular features associated with casein, lactose, pectins, anthocyanins, and other functional compounds, confirming the contribution of both matrix and encapsulants maintaining the structural characteristics of the product. The presence of antioxidant bands supported the functional potential of the powder formulations. Chia–alginate hydrogels effectively encapsulated L. reuteri, maintaining cell viability and enabling their incorporation into freeze-dried beverage powders. This approach offers a promising strategy for the development of next-generation functional food gels with enhanced probiotic stability, nutritional properties, and potential application in health-promoting dairy systems. Full article

Cell-free biosensors harness the selectivity of cellular machinery without living cells’ constraints, offering advantages in environmental monitoring, medical diagnostics, and biotechnological applications. This review examines recent advances in cell-free biosensor development, highlighting their ability to detect diverse analytes including heavy metals, organic pollutants, pathogens, and clinical biomarkers with high sensitivity and specificity. We analyze technological innovations in cell-free protein synthesis optimization, preservation strategies, and field deployment methods that have enhanced sensitivity, and practical applicability. The integration of synthetic biology approaches has enabled complex signal processing, multiplexed detection, and novel sensor designs including riboswitches, split reporter systems, and metabolic sensing modules. Emerging materials such as supported lipid bilayers, hydrogels, and artificial cells are expanding biosensor capabilities through microcompartmentalization and electronic integration. Despite significant progress, challenges remain in standardization, sample interference mitigation, and cost reduction. Future opportunities include smartphone integration, enhanced preservation methods, and hybrid sensing platforms. Cell-free biosensors hold particular promise for point-of-care diagnostics in resource-limited settings, environmental monitoring applications, and food safety testing, representing essential tools for addressing global challenges in healthcare, environmental protection, and biosecurity. Full article

The use of methane as a carbon source for producing bacterial single-cell protein (SCP) has been one of the most interesting developments in recent years. Most of these upcoming industries are using a methanotroph, Methylococcus capsulatus Bath, for SCP production using natural gas as the substrate. In the present study, we have explored the possibility of using an indigenously isolated methanotroph from a rice field in India, Methylomagnum ishizawai strain KRF4, for producing SCP from biogas [derived from cow dung]. The process was eco-friendly, required minimal instruments and chemicals, and was carried out under semi-sterile conditions in a tabletop fish tank. As the name suggests, Methylomagnum is a genus of large methanotrophs, and the strain KRF4 had elliptical to rectangular size and dimensions of ~4–5 µm × 1–2 µm. In static cultures, when biogas and air were supplied in the upper part of the growing tank, the culture grew as a thick pellicle/biofilm that could be easily scooped. The grown culture was mostly pure, from the microscopic observations where the large size of the cells, with rectangular-shaped cells and dark granules, could easily help identify any smaller contaminants. Additionally, the large cell size could be advantageous for separating biomass during downstream processing. The amino acid composition of the lyophilized biomass was analyzed using HPLC, and it was seen that the amino acid composition was comparable to commercial fish meal, soymeal, Pruteen, and the methanotroph-derived SCP-UniProtein^®. The only difference was that a slightly lower percentage of lysine, tryptophan, and methionine was observed in Methylomagnum-derived SCP. Methylomagnum ishizawai could be looked at as an alternative for SCP derived from methane or biogas due to the comparable SCP produced, on the qualitative level. Further intensive research is needed to develop a continuous, sustainable, and economical process to maximize biomass production and downstream processing. Full article