Search Results (4,530)

This study aimed to evaluate the impact of liquid–solid (LS) systems on the dissolution profiles of a poorly soluble drug—suvorexant (SUV). In the first stage of this study, LS systems were prepared by using two different non-volatile solvents: ethylene glycol diethyl ether and polyethylene glycol 400 (PEG 400). To compare the properties of different types of LS systems, formulations were prepared that differed in the content of SUV (10 and 20 mg) as well as in the ratio of excipients (microcrystalline cellulose and colloidal silica), which was 10:1 or 1:1. The physicochemical properties of the prepared formulations were characterized by X-ray diffractometry (XRD), thermogravimetry (TGA) and differential scanning calorimetry (DSC). This was followed by a dissolution study of SUV from prepared LS systems, using a 0.4% sodium lauryl sulfate solution as the medium to maintain sink conditions. Results showed that the LS systems change the crystalline structure of SUV to an amorphous one and improve the dissolution rate of SUV. The greatest improvement was achieved by using the microcrystalline cellulose and colloidal silica in a 10:1 ratio for the preparation of the system (CCA variant). It was observed that the type of solvent used and the order of combining excipients during the preparation of LS systems are also important for the properties. The main point was that physicochemical characterization of the prepared formulations lead to a loss of crystallinity of SUV associated with its introduction into liquid–solid systems. Full article

Thermoplastic polyurethane properties are governed by the interplay between soft-segment mobility, hard-segment interactions, and segmented morphology, yet the extent to which atomistic predictions of their thermal and mechanical behavior depend on force-field choice remains insufficiently benchmarked. Here, we combine FTIR, DSC, TGA, and tensile testing with all-atom molecular dynamics simulations to investigate HDI–PEG polyurethane systems across a controlled soft-segment series. Experimentally, films with PEG molecular weights of 400, 1000, and 1500 g/mol were characterized, while simulations were extended to 400–2000 g/mol to compare two complementary force-field frameworks under a consistent protocol: OPLS-AA, a conventional atom-type-based force field, and OpenFF/Sage, a direct-chemical-perception framework augmented here with bespoke torsional refinements. Both force fields reproduce the composition-driven decrease in Tg and density with increasing PEG length, but differ systematically in absolute values, with OPLS-AA predicting higher densities and Tg values than OpenFF. Tensile experiments show the highest elastic modulus for PEG400, a marked decrease at PEG1000, and a partial recovery at PEG1500. Although nanosecond-scale deformation simulations overestimate absolute moduli because they probe high-rate elastic response, they recover composition-dependent stiffness differences, with OpenFF yielding a more pronounced non-monotonic trend than OPLS-AA. Overall, this work provides an experimentally anchored benchmark for assessing which composition-driven trends in HDI–PEG polyurethanes are robust across force-field families, and which observables remain sensitive to model assumptions and simulation scale. Full article

Microbial bio-inoculants have been proposed as management tools to enhance crop performance under variable environmental conditions; however, their effectiveness is often influenced by site-specific factors. This study evaluated the effects of bio-inoculation on seed germination and seedling vigor of pea under osmotic stress induced by polyethylene glycol (PEG 6000), and its interaction with two fertilization levels (75% and 100% of the recommended dose) under field conditions in the Amazonas region of Peru. Under laboratory conditions, germination percentage remained high across all treatments (93.3–100%) and was not affected by bio-inoculation or osmotic potential; however, osmotic stress altered germination dynamics, increasing mean germination time from 1.85–2.09 days at 0 MPa to 2.26–2.43 days at −0.8 MPa, while germination synchrony and seedling vigor decreased as stress increased. The seedling vigor index reached maximum values at −0.2 MPa (4.47–5.29) and declined at −0.8 MPa (1.50–2.00), and multivariate analyses showed that variation in germination responses was mainly associated with germination timing and vigor rather than seed viability. Under field conditions, no significant effects of fertilization level, microbial bio-inoculation, or their interaction were detected on agronomic traits or yield, although variability between locations was observed; plant height ranged from 38.5–46.3 cm in Lamud and from 100.6–108.3 cm in Molinopampa, while grain yield varied from 698–1846 kg/ha and 8771–9919 kg/ha, respectively. Overall, environmental conditions exerted a stronger influence than microbial bio-inoculation on germination dynamics and field productivity, while the findings provide practical guidance for improving pea production with bio-inoculants and optimized fertilization. Full article

Hybrid supramolecular–nanocomposite hydrogels based on polyethylene glycol (PEG), β-cyclodextrin–adamantane host–guest interactions, and silica nanoparticles represent an important class of hierarchical soft materials with tunable viscoelastic and transport properties. This review critically analyzes recent progress in cyclodextrin–silica hybrid PEG hydrogels, focusing on the mechanistic coupling between stiffness, stress relaxation, and molecular transport arising from the interplay between reversible supramolecular crosslinks and nanoparticle-induced confinement effects. Particular attention is given to how host–guest exchange kinetics regulate dynamic bond rearrangement and affinity-mediated retention of hydrophobic cargo, while silica nanoparticles enhance mechanical reinforcement and modify diffusion pathways through tortuosity and interfacial polymer–particle interactions. The analysis highlights how nanoparticle size, loading level, and surface functionalization influence relaxation spectra and network topology, as well as how environmental stimuli may affect supramolecular bond stability and overall material performance. Comparison with alternative inorganic fillers and mesoporous silica architectures further clarifies the specific advantages of silica in achieving balanced mechanical stability and controlled transport behavior. Overall, current evidence indicates that hybrid CD–silica networks enable partial decoupling of stiffness, relaxation dynamics, and diffusion, although complete independence remains constrained by fundamental polymer physics relationships. These insights support the development of predictive structure–property frameworks for advanced biomedical and controlled release applications. Full article

Lettuce (Lactuca sativa L.) is an important leafy vegetable crop, yet the efficiency and reliability of genome editing platforms in lettuce remain limited, particularly for precision base editing applications. In this study, we established an optimized PEG-mediated protoplast transformation system for lettuce through systematic evaluation of key parameters, including protoplast density, incubation time, plasmid size, and transformation method. Under optimized conditions, a maximum transient transformation efficiency of up to 81% was achieved. Using this optimized protoplast platform, we comparatively evaluated the performance of three single-base editing systems—adenosine base editor (ABE), glycosylase-based guanine base editor (gGBE), and rice alkylpurine DNA glycosylase-mediated A-to-K base editor (rAKBE)—targeting the LsALS gene, encoding acetolactate synthetase as a herbicide target with great value in weed control. Among the tested editors, ABE exhibited the highest A-to-G editing efficiency, reaching 9.3%. In contrast, gGBE and rAKBE showed lower editing efficiencies. Together, this study established a robust and reproducible protoplast-based platform for transient genome editing in lettuce and provides a practical framework for the rapid evaluation of base editing tools and target sites, firstly for gGBE and rAKBE evaluation in lettuce. The optimized system facilitates functional genomics studies and supports the development of precision breeding strategies in lettuce. Full article

Persea americana (avocado) is a fruit rich in nutrients; however, its industry is facing major threats from pathogen infection. Here, we clearly identified Colletotrichum fructicola as the pathogen causing avocado diseases in Pu’er City, Yunnan Province. However, the biological characteristics, genetic transformation system, and early cell cycle regulation of this pathogen remained unclear. In this study, C. fructicola exhibited a maximum growth rate on complete medium (CM), with the conidial yield reaching 2 × 10⁵ conidia/mL after 24 h in liquid CM. Conidia of C. fructicola had nearly fully germinated at 4 h post-inoculation (hpi), with the appressorium formation rate exceeding 95% at 12 hpi. We also established a PEG-CaCl₂-mediated genetic transformation system. The GFP-tagged transformants showed no significant differences in core biological function from the wild type. Using eGFP labeling, we visually elucidated the early cell cycle regulation of C. fructicola. Furthermore, cell cycle inhibitor assays demonstrated that C. fructicola conidial germination is independent of nuclear division and relies on cytoskeletal modulation, whereas appressorium formation and mycelial expansion require functional cell cycle regulation. This is probably the first study to systematically elucidate the cell cycle regulatory characteristics of C. fructicola isolated from avocado, and to successfully develop its genetic transformation system. These results provide important theoretical and technical support for the formulation of integrated control strategies against C. fructicola, as well as facilitating the sustainable development of the avocado industry. Full article

Background/Objectives: The objective assessment of natural tooth mobility remains challenging in clinical practice. This pilot study aimed to investigate the feasibility, repeatability, and agreement of a modified implant stability measurement system adapted for natural teeth using a custom-fabricated titanium bracket and a modified SmartPeg. Methods: Sixteen systemically healthy patients (10 males, six females) and 94 single-rooted permanent teeth with varying mobility grades were included. The tooth mobility was assessed using the Miller Mobility Index, Periotest M, and resonance frequency analysis (RFA) with the Osstell Beacon device. For the Osstell measurements, a custom titanium bracket bonded to the buccal tooth surface allowed for the placement of a modified SmartPeg. Each tooth was measured twice under standardized conditions, and mean values were recorded. The statistical analyses included Spearman correlation analysis, Cohen’s kappa for agreement with Miller categories, and intraclass correlation coefficients (ICCs) to assess the measurement repeatability. Results: The mean Periotest value was 12.70 ± 13.69, and the mean ISQ (implant stability quotient) value was 69.45 ± 19.37. The repeated measurements demonstrated excellent intra-examiner repeatability for both devices (ICC > 0.95). The Periotest values showed substantial agreement with the Miller mobility grades (κ = 0.763; p < 0.001), whereas the Osstell values demonstrated weak agreement with these ordinal categories (κ = 0.094; p = 0.048). A strong negative correlation was observed between the Periotest and Osstell measurements irrespective of the scales (r = −0.865; p < 0.001). Conclusions: In natural dentition, the resonance frequency analysis demonstrated reproducible measurements under controlled experimental conditions and showed measurable associations with conventional mobility assessments. However, the method remains investigational. The findings do not establish clinical validity for the routine assessment of natural tooth mobility. Further studies with larger sample sizes and statistical models accounting for patient-level clustering are required before clinical implementation can be considered. This study is registered at ClinicalTrials.gov (NCT07188168). Full article

A warm-up is a critical procedure in sports science for enhancing muscular performance and optimizing subsequent athletic activities. However, the physiological and athletic performance effects of a warm-up are often transient, diminishing rapidly during the period of inactivity after the warm-up, which is known as the warm-up transition phase. In this study, a multi-functional thermoregulation wearable composite film of graphene–MXene–bacterial cellulose/polyethylene glycol (G-M-BC/PEG) was developed by integrating MXene (a two-dimensional material with good photothermal conversion performance) and graphene into a bacterial cellulose aerogel framework, subsequently impregnated with polyethylene glycol (PEG-2000). The film showed stable structure, efficient solar photothermal conversion and storage (SPCS), and improved mechanical properties. Under 1 sun irradiation, the optimized G-M-BC/PEG wearable film showed excellent SPCS performance, sustaining a temperature plateau of 38–40 °C for 10 min after the xenon lamp was switched off under 1 sun irradiation, with a leakage rate of only 5.32% after five cycles. By constructing a biomimetic sports human body model, the composite aerogel was shown to significantly elevate muscle surface temperature and effectively mitigate heat loss during the transition phase. In the warm-up effectiveness and sports performance tests, the wearable film improved 200 m sprint performance by 0.8% ± 0.4% (p = 0.039). It also maintained subjective thermal sensation during the warm-up transition phase, with no significant decline at 5 or 10 min after the warm-up and a significant decrease only at 15 min (p = 0.02), while thermal comfort remained stable, suggesting improved neuromuscular readiness. This research provided a novel strategy for the fabrication of advanced aerogel-based wearable devices aimed at precision thermal management and athletic performance optimization. Full article

The rise in multidrug resistance in Klebsiella pneumoniae is an alarming issue, especially in invasive infections among patients with co-morbidities. With the gain of hypervirulence traits, multidrug-resistant K. pneumoniae has led to a significant increase in chronic infections and associated mortality. This study aims to explore the distribution of multidrug-resistant and hypervirulent (hv) K. pneumoniae in invasive infections in a tertiary care hospital. A total of 231 K. pneumoniae isolates were collected over a period of six months from invasive infections. These isolates were tested phenotypically and genotypically for the presence of antimicrobial resistance, along with molecular detection of hypervirulence determinants (iucA, rmpA, rmpA2, peg344, iroB). High levels of resistance to β-lactams, fluoroquinolones, and aminoglycosides were observed. Carbapenemase-encoding genes were widely distributed, and 22% showed the presence of at least one hypervirulence gene, most commonly iucA and rmpA. Co-carriage of resistance and hypervirulence determinants in K. pneumoniae was observed in nearly 20% of the isolates, indicating the emergence of MDR-hvKP phenotypes in the hospital setting. Mortality was significantly higher among patients infected with MDR isolates, whereas hypervirulence markers were not independently associated with mortality. The presence of MDR–hypervirulent strains remains clinically concerning and underscores the need for continued genomic surveillance. Full article

Background: Percutaneous endoscopic gastrostomy (PEG) and percutaneous radiologic gastrostomy (PRG) are established techniques for long-term enteral access. Contemporary comparisons of complication patterns, length of stay (LOS), and utilization trends remain limited. Methods: We conducted a retrospective cohort study of adult patients undergoing initial gastrostomy placement at a single academic center between 2021 and 2024 (n = 341). The primary outcome was any 30-day procedure-related complication. Secondary outcomes included complication subtypes, LOS, and procedural volume trends. Multivariable regression analyses were performed to adjust for potential confounders. Results: Among 341 patients, 195 underwent PEG and 146 PRG. Overall complication rates were similar (PEG 16.4% vs. PRG 14.4%, p = 0.31). Infectious complications were numerically higher with PEG (4.1% vs. 1.4%), though not statistically significant. Mean LOS was 3.2 days for PEG and 2.8 days for PRG (p = 0.12). On multivariable analysis, gastrostomy technique was not associated with complications (aOR 0.88, 95% CI 0.48–1.61) or LOS. PRG utilization increased substantially over the study period, comprising 60.7% of procedures by 2024. Conclusions: PEG and PRG demonstrated no statistically significant differences in safety outcomes, with no statistically significant differences in complications or LOS. A marked shift toward PRG utilization was observed over time. These findings support individualized, patient-centered selection of gastrostomy technique, while acknowledging limited power to detect small but clinically meaningful differences. Full article

Background: Despite surgical repair, large paraesophageal hernias (PEHs) often recur. To minimize recurrence, adjunctive measures, such as mesh and gastropexy, have been explored, but their impact on recurrence and reoperation rates remains unclear. Therefore, we analyzed our single-institution case series, where absorbable hiatal overlay mesh and percutaneous endoscopic gastrostomy (PEG) placement were utilized systematically. Methods: Patients undergoing laparoscopic large PEH repair by a single surgeon between 1 January 2006 and 31 May 2021 were identified. Demographic data, hernia size, number of hiatal sutures used, use of mesh and/or PEG, fundoplication type, and complications were extracted by retrospective chart review. Hernia recurrence was assessed though postoperative radiographic and endoscopic studies or need for reoperation. Fisher’s exact, chi-square, Mood’s two-median and t-tests were used for between-group comparisons. Generalized linear models were used to assess associations between mesh and PEG placement and number of hiatal sutures and to compare risk differences for recurrence between treatment types (partial versus complete fundoplication; mesh versus no mesh; and PEG versus no PEG). Kaplan–Meier estimator with log-rank test was used to assess time to recurrence. Results: Overall, 413 patients (median age 66 years) underwent laparoscopic large PEH repair and fundoplication (51% partial and 49% complete fundoplications). Of these, 78% had overlay absorbable mesh and 51% had a PEG. With an average follow-up time of over 5 years, we found 19.9% had radiographic or endoscopic recurrence. Although cohort stratification based on mesh implementation and fundoplication type did not identify differences in recurrence rates, significantly lower recurrence rates were noted in patients with PEG compared to no-PEG (14.8% vs. 23.5%, p = 0.01). Notably, of five reoperations, all were in complete fundoplication patients, and one occurred in a patient with PEG placement at the index operation. Conclusions: PEG placement during laparoscopic PEH repair may lead to fewer recurrences in high-risk patients. Future prospective studies are warranted. Full article

Background: Rabies is a highly fatal zoonotic disease that causes approximately 59,000 human deaths worldwide each year. Current inactivated rabies vaccines require multiple doses and are associated with high costs. The full-length rabies virus glycoprotein (RVG), a membrane protein, exhibits substantial instability in its trimeric structure during recombinant expression. This instability makes it difficult to obtain high-purity, correctly folded antigens. Objectives: This study focuses on the preparation of a full-length recombinant RVG subunit vaccine candidate expressed in a glycoengineered Pichia pastoris system with mammalian-like glycosylation. Methods: The full-length RVG gene (including the transmembrane domain and cytoplasmic tail) from the Challenge Virus Standard-11 (CVS-11) strain was codon-optimized and inserted into the pPICZαA vector to construct the recombinant expression plasmid pPICZαA-RVG. The plasmid was transformed into glycoengineered Pichia pastoris X33-7 (low-mannose type) by electroporation for inducible expression. The target protein was purified by nickel affinity chromatography, anion-exchange chromatography, and Superdex-200 size-exclusion chromatography. The structural characteristics of the purified protein were analyzed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The purified antigen was formulated with the adjuvants AS03 or MF59. BALB/c mice (n = 5 per group) were immunized intramuscularly following a four-dose schedule (days 0, 7, 14, and 28). Antigen-specific IgG antibody titers were measured by ELISA, and neutralizing antibody titers were determined using the rapid fluorescent focus inhibition test (RFFIT). Results: Glycoengineered Pichia pastoris yeast strains expressing wild-type RVG (RVG-WT) or a mutant variant (RVG-M6: R84S, R199S, H270P, R279S, K300S, and R463S) were successfully constructed. The purified RVG antigen formed nanoparticles with an average particle size of approximately 75 nm. Immunized mice generated robust RVG-specific IgG responses, with titers reaching approximately 6.31 × 10⁵ for RVG-WT after the fourth immunization, compared to 3.16 × 10³ for RVG-M6 and 5.62 × 10³ for the RVG-WT-PEG control. Two weeks after the fourth immunization, RVG-WT formulated with AS03 or MF59 induced significant neutralizing antibody responses compared with the control group (p < 0.0001 and p < 0.01, respectively). The neutralizing antibody titers reached 1:79.43 in the AS03 group and 1:33.11 in the MF59 group, whereas the WT-PEG + AS03 control group showed a low titer of 1:3.72. In contrast, RVG-M6 formulated with MF59 failed to induce detectable neutralizing antibodies (1:3.02). Furthermore, RVG-WT + AS03 induced significantly higher neutralizing antibody responses than the WT-PEG + AS03 control group (p < 0.0001), and a significant difference was also observed between the RVG-WT + MF59 and RVG-M6 + MF59 groups (p < 0.01). Conclusions: The glycoengineered Pichia pastoris expression system successfully produced uniform full-length rabies virus glycoprotein nanoparticles with high purity. When formulated with the AS03 adjuvant, RVG-WT induced high-titer neutralizing antibodies in mice, suggesting a promising strategy for the development of recombinant subunit vaccines against rabies. However, this study is limited by the absence of challenge studies and validation in target animal species, which will be further investigated in future work. Full article

Gardenia jasminoides Ellis is a commercially important medicinal and ornamental plant; however, its functional genomics remain poorly understood because of the lack of efficient cell-based research tools. To address this limitation, we established an optimized method for isolating viable protoplasts from petal and mesophyll tissues of G. jasminoides and developed a polyethylene glycol (PEG)-mediated transient expression system. For petal protoplast isolation, the optimal enzyme combination consisted of 3.0% cellulase R-10 and 1.0% macerozyme R-10 supplemented with 0.5 M D-mannitol, yielding 5.26 × 10⁶ protoplasts per gram fresh weight (FW) with 80.63% viability. For mesophyll protoplast isolation, 1.5% cellulase R-10 and 0.5% macerozyme R-10 supplemented with 0.5 M D-mannitol produced 8.75 × 10⁶ protoplasts g⁻¹ FW with 84.55% viability. PEG-mediated transient transformation was optimized at 20% PEG4000 for petal protoplasts and 40% PEG4000 for mesophyll protoplasts, resulting in efficient GFP expression. This system was successfully applied to subcellular localization analyses of floral regulatory proteins (GjAP3, GjPI, and GjSEP) and defense-related proteins (GjNPR1 and GjTGA2), as well as to the validation of protein–protein interactions between GjSEP and GjPI and between GjNPR1 and GjTGA2 using bimolecular fluorescence complementation and yeast two-hybrid assays. Collectively, these results establish a reliable and species-specific protoplast-based platform for rapid functional characterization of genes in G. jasminoides, providing an effective tool for future studies on gene regulation, metabolic engineering, and molecular breeding in this horticultural plant species. Full article

Granule-bound starch synthase (GBSS) is primarily recognized for its role in amylose production and starch granule formation in plant plastids. While its biochemical function in storage organs has been well documented, its broader contribution to plant growth and stress adaptation remains less defined. To explore these aspects, the maize gene ZmGBSS1 was ectopically expressed in Arabidopsis thaliana and its physiological effects were examined. Subcellular localization assays confirmed that ZmGBSS1 is specifically localized to chloroplasts. Phenotypic analysis of transgenic lines revealed that overexpression of ZmGBSS1 significantly altered early seedling development, promoted root elongation, and accelerated flowering, with flowering occurring approximately four days earlier than in wild-type plants. Changes in development were accompanied by increased starch accumulation, elevated amylose levels, and a higher abundance of enlarged starch granules within chloroplasts. Under drought and PEG-induced osmotic stress, transgenic plants maintained improved growth performance and recovery capacity, together with greater proline accumulation and chlorophyll retention. These physiological advantages coincided with more rapid starch utilization and clear rises in transcripts for proline synthesis enzymes (AtP5CS1, AtP5CS2) and starch-degrading proteins (AtBAM1, AtBAM3, AtDPE1). Collectively, these findings suggest that ZmGBSS1 not only regulates starch biosynthesis but also plays a crucial role in coordinating plant development and drought stress responses, highlighting its potential for improving stress tolerance through metabolic regulation. Full article

Chrysanthemum (Chrysanthemum × morifolium Ramat.) is a globally popular ornamental plant, but most cultivars lack efficient petal-based transient transformation systems, limiting floral trait molecular mechanism exploration. Protoplasts are versatile tools for gene localization, interaction, and functional characterization. Here, we established a petal protoplast isolation and transient transformation system for C. morifolium ‘Wandai Fengguang’ via L₉(3⁴) orthogonal design: optimal isolation (0.6 M mannitol, 8 h enzymatic digestion time, 0.4% macerozyme R-10, 4% cellulase R-10) and transformation (40% PEG4000, 12 μg plasmid, 10 min transfection, a protoplast density of 1 × 10⁶ protoplasts mL⁻¹). Under these conditions, protoplast yield was 5.14 × 10⁶ protoplasts g⁻¹·FW, viability 87.41%, and transformation efficiency 51.50%, with good applicability for six additional germplasms. We further analyzed CmVIT1 protein localization. Compared with the previous system, this system significantly improved protoplast yield and transformation efficiency, facilitating the transient transformation of genes related to floral traits in chrysanthemum and providing a methodological framework for other horticultural crops. Full article