Search Results (1,209)

Alpha2-plasmin inhibitor (α2PI) has a heterogeneous structure due to proteolytic cleavages in the circulation. The C-terminally cleaved form loses the plasminogen binding site and is, therefore, a slow plasmin inhibitor (NPB-α2PI). As FXIII primarily crosslinks the plasminogen-binding intact form (PB-α2PI) to fibrin, the effect of NPB-α2PI on fibrinolysis has been less studied. Herein, we investigated the effect of C-terminal truncation. Total-, PB-, and NPB-α2PI antigen levels and α2PI incorporation were measured by ELISAs from samples of 80 healthy individuals. Clot lysis parameters of the same subjects were investigated using an in vitro clot lysis assay. α2PI incorporation into the clot was demonstrated by Western blotting. Clot lysis and clot structure were also analyzed using an α2PI-deficient plasma substituted with recombinant PB- and NPB-α2PI. Both plasma and clot-bound levels of total- and NPB-α2PI showed a significant positive correlation with clot lysis parameters. NPB-α2PI was detected in the clot due to non-covalent binding. Regardless of the type of binding, both forms affected the clot structure by increasing the thickness of the fibrin fibers and reducing the pore size. In conclusion, we found that NPB-α2PI can bind non-covalently to fibrin, and this binding contributes to changes in clot structure and inhibition of fibrinolysis. Full article

Background/Objectives: The present study aimed to synthesize folate-conjugated poloxamers and develop polymeric micelles for the dermal delivery of irinotecan and alpha-mangostin for the treatment of melanoma using poloxamer 188 and poloxamer 184, which have never been synthesized with folate before. Methods: Poloxamer 188 and poloxamer 184 were synthesized with folate by esterification. The in vitro skin penetration enhancement of irinotecan- and alpha-mangostin-loaded folate-conjugated polymeric micelles was evaluated. The skin penetration pathway of folate-conjugated polymeric micelles was investigated by colocalization of multiple fluorescently labeled particles using confocal laser scanning microscopy (CLSM). Results: Folate-conjugated poloxamer 188 and poloxamer 184 were successfully synthesized. The prepared irinotecan- and alpha-mangostin-loaded folate-conjugated polymeric micelles from poloxamer 188 and poloxamer 184 had particle sizes of approximately 180 and 150 nm, respectively, indicating a positive charge with a narrow size distribution which could be easily taken up into cells. An in vitro skin penetration study revealed that folate-conjugated polymeric micelles from poloxamer 184 significantly enhanced the skin penetration of irinotecan and alpha-mangostin to a greater extent than the solution. CLSM visualization revealed that folate-conjugated polymeric micelles penetrated through the skin by the transfollicular pathway as the major penetration pathway, whereas penetration by the intercluster pathway, transcellular pathway and intercellular pathway constituted a minor pathway. Conclusions: Folate-conjugated poloxamer 184 polymeric micelles are promising candidates for the dermal delivery of anticancer drugs by the transfollicular pathway as the major skin penetration pathway. 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

Titanium–aluminum–vanadium (Ti6Al4V) is a material chosen for spine, orthopedic, and dental implants due to its combination of desirable mechanical and biological properties. Lasers have been used to modify metal surfaces, enabling the generation of a surface on Ti6Al4V with distinct micro- and nano-scale structures. Studies indicate that topography with micro/nano features of osteoclast resorption pits causes bone marrow stromal cells (MSCs) and osteoprogenitor cells to favor differentiation into an osteoblastic phenotype. This study examined whether the biological response of human MSCs to Ti6Al4V surfaces is sensitive to laser treatment-controlled micro/nano-topography. First, 15 mm diameter Ti6Al4V discs (Spine Wave Inc., Shelton, CT, USA) were either machined (M) or additively manufactured (AM). Surface treatments included no laser treatment (NT), nanosecond laser (Ns), femtosecond laser (Fs), or nanosecond followed by femtosecond laser (Ns+Fs). Surface wettability, roughness, and surface chemistry were determined using sessile drop contact angle, laser confocal microscopy, X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM). Human MSCs were cultured in growth media on tissue culture polystyrene (TCPS) or test surfaces. On day 7, the levels of osteocalcin (OCN), osteopontin (OPN), osteoprotegerin (OPG), and vascular endothelial growth factor 165 (VEGF) in the conditioned media were measured. M NT, Fs, and Ns+Fs surfaces were hydrophilic; Ns was hydrophobic. AM NT and Fs surfaces were hydrophilic; AM Ns and Ns+Fs were hydrophobic. Roughness (Sa and Sz) increased after Ns and Ns+Fs treatment for both M and AM disks. All surfaces primarily consisted of oxygen, titanium, and carbon; Fs had increased levels of aluminum for both M and AM. SEM images showed that M NT discs had a smooth surface, whereas AM surfaces appeared rough at a higher magnification. Fs surfaces had a similar morphology to their respective NT disc at low magnification, but higher magnification revealed nano-scale bumps not seen on NT surfaces. AM Fs surfaces also had regular interval ridges that were not seen on non-femto laser-ablated surfaces. Surface roughness was increased on M and AM Ns and Ns+Fs disks compared to NT and Fs disks. OCN was enhanced, and DNA was reduced on Ns and Ns+Fs, with no difference between them. OPN, OPG, and VEGF levels for laser-treated M surfaces were unchanged compared to NT, apart from an increase in OPG on Fs. MSCs grown on AM Ns and Ns+Fs surfaces had increased levels of OCN per DNA. These results indicate that MSCs cultured on AM Ns and AM Ns+Fs surfaces, which exhibited unique roughness at the microscale and nanoscale, had enhanced differentiation to an osteoblastic phenotype. The laser treatments of the surface mediated this enhancement of MSC differentiation and warrant further clinical investigation. Full article

Polymicrobial biofilms involving fungal and bacterial species are increasingly recognized as contributors to persistent infections, particularly in the oral cavity. Candida albicans and Aggregatibacter actinomycetemcomitans are two commensals that can turn into opportunistic pathogens and are able to form robust biofilms. Objectives: This study aimed to assess the interaction dynamics between these two microorganisms and to evaluate their susceptibility to fluconazole and azithromycin in single- and mixed-species forms. Methods: Biofilm biomass was quantified using crystal violet assays, while biofilm cell viability was assessed through CFU enumeration (biofilm viability assay). To assess the resistance properties of single versus mixed-species coincubations, we applied the antimicrobial susceptibility test (AST) to each drug, and analysed spatial organization with confocal laser scanning microscopy, using PNA-FISH. Results: The results indicated that both species can coexist without significant mutual inhibition. However, a non-reciprocal synergism was also observed, whereby mixed-species biofilm conditions promoted the growth of A. actinomycetemcomitans, while C. albicans growth remained stable. As expected, antimicrobial tolerance was elevated in mixed cultures, likely due to enhanced extracellular matrix production and potential quorum-sensing interactions, contributing to increased resistance against azithromycin and fluconazole. Conclusions: This study provides novel insights into previously rarely explored interactions between C. albicans and A. actinomycetemcomitans. These findings underscore the importance of investigating interspecies interactions within polymicrobial biofilms, as understanding their mechanisms, such as quorum-sensing molecules and metabolic cooperation, can contribute to improved diagnostics and more effective targeted therapeutic strategies against polymicrobial infections. Full article

Biofilms cause a variety of problems, such as food spoilage, food poisoning, infection, tooth decay, periodontal disease, and metal corrosion, so knowledge on biofilm prevention and removal is important. A detailed observation of the three-dimensional structure of biofilms on the nanoscale is expected to provide insight into this. In this study, we report on the successful in situ nanoscale observations of a marine bacterial biofilm on glass in phosphate buffer solution (PBS) using both scanning ion conductance microscopy (SICM) and confocal laser scanning microscopy (CLSM) over the same area. By observing the same area by SICM and CLSM, we were able to clarify the three-dimensional morphology of the biofilm, the arrangement of bacteria within the biofilm, and the difference in local ion conductivity within the biofilm simultaneously, which could not be achieved by observation using a microscope alone. Full article

This study presents the structural and compositional characterisation of a newly developed Ti55Al27Mo13 alloy synthesised via aluminothermic reaction. The alloy was designed to overcome the limitations of conventional processing routes for high–melting–point elements such as Ti and Mo, enabling the formation of a complex, multi–phase microstructure in a single high–temperature step. The aim was to develop and characterise a material with microstructural features expected to enhance wear resistance, oxidation behaviour, and thermal stability in future applications. The alloy is intended as a precursor for composite nanopowders and surface coatings applied to aluminium–, magnesium–, and iron–based substrates subjected to mechanical and thermal loading. Elemental analysis (XRF, EDS) confirmed the presence of Ti, Al, Mo, and minor elements such as Si, Fe, and C. Microstructural investigations using laser confocal and scanning electron microscopy revealed a heterogeneous structure comprising solid solutions, eutectic regions, and dispersed oxide and carbide phases. Notably, the alloy exhibits high hardness values, reaching >2400 HV in Al₂O₃ regions and ~1300 HV in Mo– and Si–enriched solid solutions. These results suggest the material’s substantial potential for protective surface engineering. Further tribological, thermal, and corrosion testing, conducted with meticulous attention to detail, will follow to validate its functional performance in target applications. Full article

Although corn germ meal is a rich source of dietary fiber, it contains a relatively low proportion of soluble dietary fiber (SDF) and is frequently contaminated with high levels of zearalenone (ZEN). Solid-state fermentation has the dual effects of modifying dietary fiber (DF) and degrading mycotoxins. This study optimized the solid-state fermentation process of corn germ meal using Bacillus subtilis K6 through response surface methodology (RSM) to enhance SDF yield while efficiently degrading ZEN. Results indicated that fermentation solid-to-liquid ratio and time had greater impacts on SDF yield and ZEN degradation rate than fermentation temperature. The optimal conditions were determined as temperature 36.5 °C, time 65 h, and solid-to-liquid ratio 1:0.82 (w/v). Under these conditions, the ZEN degradation rate reached 96.27 ± 0.53%, while the SDF yield increased from 9.47 ± 0.68% to 20.11 ± 1.87% (optimizing the SDF/DF ratio from 1:7 to 1:3). Scanning electron microscopy (SEM) and confocal laser scanning microscope (CLSM) revealed the structural transformation of dietary fiber from smooth to loose and porous forms. This structural modification resulted in a significant improvement in the physicochemical properties of dietary fiber, with water-holding capacity (WHC), oil-holding capacity (OHC), and water-swelling capacity (WSC) increasing by 34.8%, 16.4%, and 15.2%, respectively. Additionally, the protein and total phenolic contents increased by 23.0% and 82.61%, respectively. This research has achieved efficient detoxification and dietary fiber modification of corn germ meal, significantly enhancing the resource utilization rate of corn by-products and providing technical and theoretical support for industrial production applications. Full article

Background/Objectives: Magnetic iron oxide nanoparticles (IONPs), human serum albumin (HSA) and folic acid (FA) are prospective components for hybrid nanosystems for various biomedical applications. The magnetic nanosystems FA-HSA@IONPs (FAMs) containing IONPs, HSA, and FA residue are engineered in the study. Methods: Composition, stability and integrity of the coating, and peroxidase-like activity of FAMs are characterized using UV/Vis spectrophotometry (colorimetric test using o-phenylenediamine (OPD), Bradford protein assay, etc.), spectrofluorimetry, dynamic light scattering (DLS) and electron magnetic resonance (EMR). The selectivity of the FAMs accumulation in cancer cells is analyzed using flow cytometry and confocal laser scanning microscopy. Results: FAMs (d_N~55 nm by DLS) as a drug delivery platform have been administered to cancer cells (human breast adenocarcinoma MCF-7 and MDA-MB-231 cell lines) in vitro. Methylene blue, as a model photosensitizer, has been non-covalently bound to FAMs. An increase in photoinduced cytotoxicity has been found upon excitation of the photosensitizer bound to the coating of FAMs compared to the single photosensitizer at equivalent concentrations. The suitability of the nanosystems for photodynamic therapy has been confirmed. Conclusions: FAMs are able to effectively enter cells with increased folate receptor expression and thus allow antitumor photosensitizers to be delivered to cells without any loss of their in vitro photodynamic efficiency. Therapeutic and diagnostic applications of FAMs in oncology are discussed. Full article

Coatings that are tolerant of poor surface preparation are often used for rapid, real-time maintenance of aging steel surfaces. In this study, a modified epoxy (EP) anti-rust coating was proposed, utilizing methyl gallate (MG) as a rust conversion agent, graphene oxide (GO) as an active functional material, and epoxy resin as the film-forming material. The anti-rust mechanism was investigated using potentiodynamic polarization (PDP), electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM), laser scanning confocal microscopy (LSCM), and the scanning vibration electrode technique (SVET). The results demonstrated that over a period of 21 days, the impedance of the coating increases while the corrosion current density decreases with prolonged soaking time. The coating exhibited a maximum impedance of 2259 kΩ, and a lower corrosion current density of 8.316 × 10⁻³ A/m², which demonstrated a three-order magnitude reduction compared to the corrosion current density observed in mild steel without coating. LSCM demonstrated that MG can not only penetrate the tiny gap between the rust particles, but also effectively convert harmful rust into a complex. SVET showed a much more uniform current density distribution in the micro-zones of mild steel with the anti-rust coating compared to uncoated mild steel, indicating that the presence of GO not only enhanced the electrical conductivity of the coating, but also improved the structure of the coating, which contributed to the high performance of the modified epoxy anti-rust coating. This work highlights the potential application of anti-rust coating in the protection of metal structures in coastal engineering. Full article

Bio-based polyurethane asphalt binder (PUAB) derived from castor oil (CO) is environmentally friendly and exhibits extended allowable construction time. However, CO imparts inherently poor mechanical performance to bio-based PUAB. To address this limitation, attapulgite (ATT) with fibrous nanostructures was incorporated. The effects of ATT on bio-based PUAB were systematically investigated, including cure kinetics, rotational viscosity (RV) evolution, phase-separation microstructures, dynamic mechanical properties, thermal stability, and mechanical performance. Experimental characterization employed Fourier transform infrared spectroscopy, Brookfield viscometry, laser scanning confocal microscopy, dynamic mechanical analysis, thermogravimetry, and tensile testing. ATT incorporation accelerated the polyaddition reaction conversion between isocyanate groups in polyurethane (PU) and hydroxyl groups in ATT. Paradoxically, it reduced RV during curing, prolonging allowable construction time proportionally with clay content. Additionally, ATT’s compatibilizing effect decreased bitumen particle size in PUAB, with scaling proportionally with clay loading. While enhancing thermal stability, ATT lowered the glass transition temperature and damping properties. Crucially, 1 wt% ATT increased tensile strength by 71% and toughness by 62%, while maintaining high elongation at break (>400%). The cost-effectiveness and significant reinforcement capability of ATT make it a promising candidate for producing high-performance bio-based PUAB composites. Full article

The continuous casting of Ti-Nb microalloyed steel was simulated with high temperature confocal laser scanning microscopy (HTCLSM). Evolution of the sample surface morphology was observed in-situ, during cooling conditions chosen to represent different locations in a cast slab. Calculations with a thermodynamics model of carbonitride precipitate formation agreed with the transmission electron microscopy (TEM) analysis that fine reliefs observed on the sample surface were actually caused by interior precipitation of (Ti,Nb)(C,N). Precipitation and the resulting reliefs changed with location beneath the slab surface, simulated casting speed, and steel composition. With the same casting speed and steel composition, reliefs in the simulated slab surface sample appeared earlier and were larger than in the slab center. With increased casting speed, reliefs were observed later and decreased in size. With increased titanium or niobium content, reliefs appeared earlier and increased in number. TEM measurement showed that the precipitate diameters were mainly smaller than 4 nm, with a few between 4 and 8 nm. The property of surface reliefs observed via HTCLSM correlated qualitatively with the number and size of internal precipitates measured with TEM, showing this to be an effective tool for indirectly characterizing nanoscale secondary phase precipitation inside the sample. Full article

Thylakoid rhodanese-like protein (TROL) serves as a thylakoid membrane hinge linking photosynthetic electron transport chain (PETC) complexes to nicotinamide adenine dinucleotide phosphate (NADPH) synthesis. TROL is the docking site for the flavoenzyme ferredoxin-NADP⁺ oxidoreductase (FNR). Our prior work indicates that the TROL-FNR complex maintains redox equilibrium in chloroplasts and systemically in plant cells. Improvement in the knowledge of redox regulation mechanisms is critical for engineering stress-tolerant plants in times of elevated global drought intensity. To further test this hypothesis and confirm our previous results, we monitored light-independent ROS propagation in the leaves of Arabidopsis wild type (WT), TROL knock-out (KO), and TROL ΔRHO (RHO-domain deletion mutant) mutant plants in situ by using confocal laser scanning microscopy with specific fluorescent probes for the three different ROS: O₂^·−, H₂O₂, and ¹O₂. Plants were grown under the conditions of normal substrate moisture and under drought stress conditions. Under the drought stress conditions, the TROL KO line showed ≈32% less O₂^·− while the TROL ΔRHO line showed ≈49% less H₂O₂ in comparison with the WT. This research confirms the role of dynamical TROL-FNR complex formation in redox equilibrium maintenance by redirecting electrons in alternative sinks under stress and also points it out as promising target for stress-tolerant plant engineering. Full article

Cobboldia elephantis (Oestridae: Gasterophilinae) is an obligate parasite of the alimentary tract of the Asian elephant, causing gastric myiasis. Current knowledge of its first instar larval morphology has been limited to observations under light microscopy, significantly constraining our understanding of morphological evolution within the genus Cobboldia and the broader subfamily Gasterophilinae. In this study, we provided ultrastructural and three-dimensional characterizations of C. elephantis using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) based on newly hatched first instar larvae. Features of the first instar larva of C. elephantis, which are either unique or shared with C. loxodontis Brauer, 1896, include (i) three pairs of mouthhooks, (ii) posterior spiracles bearing peristigmatic tufts, (iii) anal division with two spine-clusters and a row of elongated spines, and (iv) a planed, button-like sensilla and a sensillum placodeum. We also compared the first instar larval morphology of C. elephantis with that of Cobboldia loxodontis, Gyrostigma rhinocerontis (Owen, 1830), Gasterophilus pecorum (Fabricius, 1794), Portschinskia magnifica Pleske, 1926, and Oestrus ovis Linnaeus, 1758. Species of Gasterophilinae share several unique features of the first instar larva, including (i) an anal division composed of three subdivisions, (ii) spiracular slits on the posterior spiracles, and (iii) conserved positioning of thoracic sensilla. These findings fill a key gap in our knowledge of C. elephantis larval morphology and suggest that these distinctive structures play a role in adaptation to its parasitic lifestyle. Full article

Arsenic (As) and cadmium (Cd) in rice grains are major global food safety concerns. Iron (Fe) can help reduce both, but current Fe treatments suffer from poor stability, low leaf absorption, and fast soil immobilization, with unclear underlying mechanisms. To address these issues, an Fe-based metal–organic framework (MIL-88) was modified with sodium alginate (SA) to form MIL-88@SA. Its stability as a foliar inhibitor and its leaf absorption were tested, and its effects on As and Cd accumulation in rice were compared with those of soluble Fe (FeCl₃) and chelating Fe (HA + FeCl₃) in a field study on As–Cd co-contaminated rice paddies. Compared with the control, MIL-88@SA outperformed or matched the other Fe treatments. A single foliar spray during the tillering stage increased the rice yield by 19% and reduced the inorganic As and Cd content in the grains by 22.8% and 67.8%, respectively, while the other Fe treatments required two sprays. Its superior performance was attributed to better leaf affinity and thermal stability. Laser ablation inductively coupled plasma–mass spectrometry (LA–ICP–MS) and confocal laser scanning microscopy (CLSM) analyses revealed that Fe improved photosynthesis and alleviated As–Cd stress in leaves, MIL-88@SA promoted As and Cd redistribution, and Fe–Cd co-accumulation in leaf veins enhanced Cd retention in leaves. Full article