Search Results (85)

The problem of spreading harmful infections through contaminated surfaces has become more acute during the recent coronavirus pandemic. The design of self-cleaning materials, which can continuously decompose biological contaminants, is an urgent task for environmental protection and human health care. In this study, the surface of blended cotton/polyester fabric was functionalized with N-doped TiO₂ (TiO₂-N) nanoparticles using titanium(IV) isopropoxide as a binder to form durable photoactive coating and additionally decorated with Cu species to promote its self-cleaning properties. The photocatalytic ability of the material with photoactive coating was investigated in oxidation of acetone vapor, degradation of deoxyribonucleic acid (DNA) fragments of various lengths, and inactivation of PA136 bacteriophage virus and Candida albicans fungi under visible light and ultraviolet A (UVA) radiation. The kinetic aspects of inactivation and degradation processes were studied using the methods of infrared (IR) spectroscopy, polymerase chain reaction (PCR), double-layer plaque assay, and ten-fold dilution. The results of experiments showed that the textile fabric modified with TiO₂-N photocatalyst exhibited photoinduced self-cleaning properties and provided efficient degradation of all studied contaminants under exposure to both UVA and visible light. Additional modification of the material with Cu species substantially improved its self-cleaning properties, even in the absence of light. Full article

Waste and chemicals generated from industry have been a major source of pollution and a prominent threat to human health via the food chain; hence, an efficient and durable material that can be used to detoxify polluted soil and water bodies is necessary to attain ecosystem equity and security. This study hypothesized that biochar (BC) made from poultry manure (PM) through pyrolysis and fortification with iron (Fe–BC) can be used to remove methyl orange dye from aqueous solution. Furthermore, this study evaluated the effect of solution pH on the sorption of methyl orange through batch sorption studies. The similarity in the modeled data and experimental data was measured by the standard error of estimate, whereas sorption isotherms were examined using nonlinear forms of different sorption equations. With the use of Langmuir models, a maximum sorption capacity of 136.25 mg·g⁻¹ and 98.23 mg·g⁻¹ was recorded for Fe–BC and BC, respectively. Fe–BC possessed a higher adsorption ability in comparison to BC. The pseudo-second-order best described the sorption kinetics of both adsorbents at R² = 0.9973 and 0.9999, indicating a strong interaction between MO and Fe–BC. Furthermore, the efficiency with which MO was removed by the absorbent was highest at lower pH (pH = 4). It is therefore concluded that Fe–BC can be used as an effective and environmentally friendly material for detoxification of wastewater; however, further research on the application and usage of biochar modified techniques for enhancing adsorption efficacy on a large scale should be encouraged. Full article

Chronic pain affects a significant portion of the population, with fewer than 30% achieving adequate relief from existing treatments. This study describes the humanization methodology and characterization of an effective non-opioid single-chain fragment variable (scFv) biologic that reverses pain-related behaviors, in this case by targeting P2X4. After nerve injury, ATP release activates/upregulates P2X4 receptors (P2X4R) sequestered in late endosomes, triggering a cascade of chronic pain-related events. Nine humanized scFv (hscFv) variants targeting a specific extracellular 13-amino-acid peptide fragment of human P2X4R were generated via CDR grafting. ELISA analysis revealed nanomolar binding affinities, with most humanized molecules exhibiting comparable or superior affinity compared to the original murine antibody. Octet measurements confirmed that the lead, HC3-LC3, exhibited nanomolar binding kinetics (KD = 2.5 × 10⁻⁹ M). In vivo functional validation with P2X4R hscFv reversed nerve injury-induced chronic pain-related behaviors with a single dose (0.4 mg/kg, intraperitoneal) within two weeks. The return to naïve baseline remained durably reduced > 100 days. In independent confirmation, the spared nerve injury (SNI) model was similarly reduced. This constitutes an original method whereby durable reversals of chronic nerve injury pain, anxiety and depression measures are accomplished. Full article

The spread of per- and polyfluoroalkyl substances (PFAS) in the environment poses a severe threat to soil organisms, aquatic life, and human health. Many PFAS compounds are mobile and easily transported from soils to groundwater and further to surface waters. Leaching tests are valuable tools for assessing the site-specific leaching behaviour of contaminants. Here, we report the results of an evaluation of two standardized leaching tests for PFAS-contaminated soil materials: the batch test (ISO 21268-2:2019) using either demineralized water or 1 mM CaCl₂ as leachants (liquid-to-solid (L/S) ratio of 10) and the up-flow percolation test (ISO 21268-3:2019) using 1 mM CaCl₂ as leachant. One field-contaminated soil and three spiked (12 PFAS compounds) soils (aged 5 months) were included in the study. Desorption kinetics in the batch test were fast and equilibrium was obtained for all PFAS compounds within 24 h, the prescribed equilibration time. The same solubility was obtained for short-chain PFAS (PFBA, PFHxA, PFHpA, PFBS) in demineralized water and 1 mM CaCl₂, whereas significantly lower solubility was often observed for long-chain PFAS in CaCl₂ than in water, probably due to decreased charge repulsion between soil surfaces and PFAS compounds. In the up-flow percolation test, concentrations of short-chain PFAS in leachates decreased rapidly with increasing L/S, in contrast to long-chain PFAS, where concentrations decreased gradually or remained constant. Solid–solution partitioning coefficients (K_d), calculated from the data of the batch and percolation tests (1 mM CaCl₂), were generally in agreement, although differing by more than three orders of magnitude between different PFAS compounds. Uncertainties and pitfalls when calculating K_d values from leaching test data are also explored. Full article

There are two types of noninvasive biomarkers of human embryo developmental potential: those based on a direct assessment of embryo morphology over time and those using spent media after embryo in vitro culture as source of information. Both are derived from previously acquired knowledge on different aspects of pre-implantation embryo development. These aspects include embryo morphology and kinetics, chromosomal ploidy status, metabolism, and embryonic gene transcription, translation, and expression. As to the direct assessment of morphology and kinetics, pertinent data can be obtained by analyzing sequential microscopic images of in vitro cultured embryos. Spent media can serve a source of genomic, metabolomic, transcriptomic and proteomic markers. Methods used in the early pioneering studies, such as microscopy, fluorescence in situ hybridization, autoradiography, electrophoresis and immunoblotting, or enzyme-linked immunosorbent assay, are too subjective, invasive, and/or time-consuming. As such, they are unsuitable for the current in vitro fertilization (IVF) practice, which needs objective, rapid, and noninvasive selection of the best embryo for uterine transfer or cryopreservation. This has been made possible by the use of high-throughput techniques such as time-lapse (for direct embryo evaluation), next-generation sequencing, quantitative real-time polymerase chain reaction, high-performance liquid chromatography, nanoparticle tracking analysis, flow cytometry, mass spectroscopy, Raman spectroscopy, near-infrared spectroscopy, and nuclear magnetic resonance spectroscopy (for spent culture media analysis). In this review, individual markers are presented systematically, with each marker’s history and current status, including available methodologies, strengths, and limitations, so as to make the essential information accessible to all health professionals, even those whose expertise in the matter is limited. Full article

Background/Objectives: The AXL receptor tyrosine kinase is a promising therapeutic target in solid tumors, yet conventional viral vector-engineered CAR-T cells face critical limitations, including risks of insertional mutagenesis and immunogenicity from murine-derived single-chain variable fragments (scFvs). This study aimed to develop and evaluate mRNA-engineered fully human AXL CAR-T (^mfhAXL CAR-T) cells as a safer, scalable alternative for solid tumor immunotherapy. Methods:^mfhAXL CAR-T cells were generated via electroporation-mediated delivery of in vitro transcribed mRNA encoding a fully human AXL-specific CAR. CAR expression kinetics and T-cell viability were quantified by flow cytometry. Antitumor activity was assessed through in vitro co-cultures with AXL-positive lung and pancreatic cancer cells, measuring cytotoxicity, cytokine secretion, and specificity. In vivo efficacy was evaluated in a lung cancer xenograft mouse model, with tumor volume and body weight monitored over 14 days. Results: Flow cytometry confirmed transient but high CAR expression (>90% at 24 h) with preserved T-cell viability (>90%). In vitro, ^mfhAXL CAR-T cells exhibited dose-dependent cytotoxicity and antigen-specific cytokine secretion. In vivo, four administrations of ^mfhAXL CAR-T cells suppressed tumor growth without body weight loss. Conclusions: The mRNA-electroporated ^mfhAXL CAR-T platform enables cost-effective, large-scale production, offering a safer alternative to viral vector-based approaches by eliminating risks of insertional mutagenesis and immunogenicity. Full article

A biochar (BC) generated by the pyrogasification of wood chips from authorized forestry cuts was extensively characterized and evaluated for its efficacy in retaining/releasing two agrochemicals, namely the fungicide penconazole (PEN), the herbicide S-metolachlor (S-MET), and the xenoestrogen bisphenol A (BPA) widely present in industrial effluents. The elemental composition of BC was evaluated using CN elemental analysis and total reflection X-ray fluorescence (TXRF) spectroscopy which showed the abundance of elements typically found in BCs (Ca, K, P) along with essential trace elements such as Fe and Mn. Scanning electron microscopy coupled with energy-dispersive X-ray analysis (SEM-EDX) described the surface features of BC along with the major surface elements, while Brunauer–Emmett–Teller (BET) analysis revealed, as expected, a large specific surface area (366 m² g⁻¹). High porosity (0.07 cm³ g⁻¹) was demonstrated by the density functional theory (DFT) method, while Fourier transform infrared (FT-IR) spectroscopy highlighted the presence of a prominent aromatic structure and the abundance of reactive functional groups responsible for the binding of the compounds. The sorption/desorption capacity of BC was studied by means of sorption kinetics and isotherms in batch trials, and by modeling the experimental data with various theoretical equations. All compounds reached sorption equilibrium on BC very rapidly, following preferentially pseudo-second-order kinetics. Freundlich adsorption constants of PEN, S-MET, and BPA were 37.3, 13.2, and 11.6 L g⁻¹, respectively, thus demonstrating the great affinity of BC for hydrophobic pollutants. The adsorption process was hysteretic as only a small fraction of each compound was slowly desorbed from BC. The overall results obtained highlighted the great potential of BC of acting as a biosorbent of contaminants, which is of great importance for the containment of pollution in agricultural soils and for limiting the entry of toxic compounds into the human and animal food chain. Full article

Diesel contamination in water bodies poses a significant environmental challenge due to the toxic effects of its water-soluble fraction (WSF) on aquatic ecosystems and human health. The aim of this work was the design of a new technological procedure for the purification of water contaminated with the WSF of diesel. The procedure is based on the adsorption of organic pollution on an organozeolite, after which the biodegradation of the adsorbed pollutant takes place. The material for obtaining organozeolite was a natural zeolite from the Zlatokop deposit (Vranje, Serbia). The zeolitic surface was modified with hexadecyltrimethylammonium bromide (HDTMA-Br), a cationic quaternary ammonium salt. The adsorption experiments, with initial WSF concentrations of 2.5–25 mg/L, at pH 6 and at 20 °C, were performed in a batch system using organozeolite, and the results showed that more than 90% of the WSF of diesel was removed, reaching equilibrium after 1 h. The maximum adsorbed capacity of organozeolite for the removal of the WSF of diesel fuel from water under the tested conditions was 22.2 mg/g. Equilibrium data were well fitted by a linear isotherm model, while a pseudo-second-order equation well fitted the kinetic data. After adsorption, a 15-day biodegradation experiment was carried out under batch conditions. The results showed that the examined consortium of microorganisms degraded 80% of the adsorbed contaminant. Additional respirometric analyses showed that, in parallel with the degradation of the contaminant, the degradation of the long-chain HDTMA ions at the surface of the organozeolite also occurred. To the best of our knowledge, this is the first study combining adsorption and biodegradation to remove the WSF of diesel from water. Full article

Hyaluronan (HA) is one of the crucial components of the extracellular matrix in vertebrates and is synthesized by three hyaluronan synthases (HASs), namely HAS1, HAS2, and HAS3. The low expression level of HASs in normal keratinocytes and other various types of cells presents a recognized challenge, impeding biological and pathological research on their localization. In this study, the human proteins HAS1, HAS2, and HAS3 with fused maltose-binding protein (MBP) tags were successfully expressed at high levels and purified for the first time in HEK293F cells. The enzymatic properties of the three HAS proteins were further characterized and compared. A pulse-field gel electrophoresis analysis of the size distribution of the hyaluronan generated in vitro by the membrane proteins demonstrated that the three HAS isoforms generate HA polymer chains at different molecular masses. Kinetic studies demonstrated that the three HAS proteins differed in their catalytic efficiency and apparent K_m values for the two substrates, UDP-GlcA and UDP-GlcNAc. Furthermore, the cellular hyaluronan secretion by the three isoenzymes was evaluated and quantified in the HEK 293T cells transfected with GFP-tagged HAS1-GFP, HAS2-GFP, and HAS3-GFP using an ELISA assay. These findings enhance our understanding of the membrane protein HASs in mammalian cells. Full article

With the rapid development of nuclear energy, the contamination of environmental water systems by uranium has become a significant threat to human health. To efficiently remove uranium from these systems, three types of silica-based polyamine resins—SiPMA-DETA (SiPMA: silica/poly methyl acrylate; DETA: diethylenetriamine), SiPMA-TETA (TETA: triethylenetetramine), and SiPMA-TEPA (TEPA: tetraethylenepentamine)—were successfully prepared, characterized, and evaluated in batch experiments. Characterization results showed that the silica-based polyamine resins were successfully prepared, and they exhibited a uniform shape and high specific surface area. SiPMA-DETA, SiPMA-TETA, and SiPMA-TEPA had nitrogen contents of 4.08%, 3.72%, and 4.26%, respectively. Batch experiments indicated that these adsorbents could efficiently remove uranium from aqueous solutions with a pH of 5–9. The adsorption kinetics of U(VI) were consistent with the pseudo-second-order model, indicating that the adsorption process was chemisorption and that adsorption equilibrium was achieved within 10 min. SiPMA-TEPA, with the longest polyamine chain, exhibited the highest adsorption capacity (>198.95 mg/g), while SiPMA-DETA, with the shortest polyamine chain, demonstrated the highest U(VI) adsorption efficiency (83%) with 100 mM Na₂SO₄. SiPMA-TEPA still removed over 90% of U(VI) from river water and tap water. The spectral analysis revealed that the N-containing functional groups on the ligand were bound to anionic uranium–carbonate species and possibly contributed to the adsorption efficiency. In general, this work presents three effective adsorbents for removing uranium from environmental water systems and thus significantly contributes to the field of environmental protection. Full article

The uptake of four liposomal formulations was tested with the murine endothelial cell line bEnd.3 and the human glioblastoma cell line U-87 MG. All formulations were composed of DPPC, cholesterol, 5 mol% of mPEG (2000 Da, conjugated to DSPE), and the dye DiD. Three of the formulations had an additional PEG chain (nominally 5000 Da, conjugated to DSPE) with either succinimide (NHS), glucose (PEG-bound at C-6), or 4-aminophenyl β-D-glucopyranoside (bound at C-1) as ligands at the distal end. Measuring the uptake kinetics at 1 h and 3 h for liposomal incubation concentrations of 100 µM, 500 µM, and 1000 µM, we calculated the liposomal uptake saturation S and the saturation half-time t_1/2. We show that only succinimide has an elevated uptake in bEnd.3 cells, which makes it a very promising and so far largely unexplored candidate for BBB transfer and brain cancer therapies. Half-times are uniform at low concentrations but diversify for high concentrations for bEnd.3 cells. Contrary, U-87 MG cells show almost identical saturations for all three ligands, making a uniform uptake mechanism likely. Only mPEG liposomes stay at 60% of the saturation for ligand-coated liposomes. Half-times are diverse at low concentrations but unify at high concentrations for U-87 MG cells. Full article

Skin bioprinting has the potential to revolutionize treatment approaches for injuries and surgical procedures, while also providing a valuable platform for assessing and screening cosmetic and pharmaceutical products. This technology offers key advantages, including flexibility and reproducibility, which enable the creation of complex, multilayered scaffolds that closely mimic the intricate microenvironment of native skin tissue. The development of an ideal hydrogel is critical for the successful bioprinting of these scaffolds with incorporated cells. In this study, we used a hydrogel formulation developed in our laboratory to fabricate a 3D-bioprinted skin model. The hydrogel composition was carefully selected based on its high compatibility with human skin cells, incorporating alginate, methyl cellulose, and nanofibrillated cellulose. One of the critical challenges in this process, particularly for its commercialization and large-scale production, is ensuring consistency with minimal batch-to-batch variations. To address this, we explored methods with which to preserve the physicochemical properties of the hydrogels, with a focus on freezing techniques. We validated the pre-frozen hydrogels’ printability, rheology, and mechanical and surface properties. Our results revealed that extended freezing times significantly reduced the viscosity of the formulations due to ice crystal formation, leading to a redistribution of the polymer chains. This reduction in viscosity resulted in a more challenging extrusion and increased macro- and microporosity of the hydrogels, as confirmed by nanoCT imaging. The increased porosity led to greater water uptake, swelling, compromised scaffold integrity, and altered degradation kinetics. The insights gained from this study lay a solid foundation for advancing the development of an in vitro skin model with promising applications in preclinical and clinical research. Full article

Objective: The aim of this study was to optimize the formulation of a C60-modified self-microemulsifying drug delivery system loaded with triptolide (C60-SMEDDS/TP) and evaluate the cytoprotective effect of the C60-SMEDDS/TP on normal human cells. Results: The C60-SMEDDS/TP exhibited rapid emulsification, an optimal particle size distribution of 50 ± 0.19 nm (PDI 0.211 ± 0.049), and a near-neutral zeta potential of −1.60 mV. The release kinetics of TP from the C60-SMEDDS/TP exhibited a sustained release profile and followed pseudo-first-order release kinetics. Cellular proliferation and apoptosis analysis indicated that the C60-SMEDDS/TP (with a mass ratio of TP: DSPE-PEG-C60 = 1:10) exhibited lower toxicity towards L02 and GES-1 cells. This was demonstrated by a higher IC50 (40.88 nM on L02 cells and 17.22 nM on GES-1 cells) compared to free TP (21.3 nM and 11.1 nM), and a lower apoptosis rate (20.8% on L02 cells and 26.3% on GES-1 cells, respectively) compared to free TP (50.5% and 47.0%) at a concentration of 50 nM. In comparison to the free TP group, L02 cells and GES-1 cells exposed to the C60-SMEDDS/TP exhibited a significant decrease in intracellular ROS and an increase in mitochondrial membrane potential (ΔψM). On the other hand, the C60-SMEDDS/TP demonstrated a similar inhibitory effect on BEL-7402 cells (IC50 = 28.9 nM) and HepG2 cells (IC50 = 107.6 nM), comparable to that of the free TP (27.2 nM and 90.4 nM). The C60-SMEDDS/TP group also exhibited a similar intracellular level of ROS and mitochondrial membrane potential compared to the SMEDDS/TP and free TP groups. Method: Fullerenol-Grafted Distearoyl Phosphatidylethanolamine-Polyethylene Glycol (DSPE-PEG-C60) was synthesized and applied in the self-microemulsifying drug delivery system. The C60-SMEDDS/TP was formulated using Cremophor EL, medium-chain triglycerides (MCT), PEG-400, and DSPE-PEG-C60, and loaded with triptolide (TP). The toxicity and bioactivity of the C60-SMEDDS/TP were assessed using normal human liver cell lines (L02 cells), normal human gastric mucosal epithelial cell lines (GES-1 cells), and liver cancer cell lines (BEL-7402 cells and HepG2 cells). The production of reactive oxygen species (ROS) after the C60-SMEDDS/TP treatment was assessed using 2′,7′-dichlorofluorescein diacetate (DCFDA) staining. The alterations in mitochondrial membrane potential (ΔψM) were assessed by measuring JC-1 fluorescence. Conclusions: The cytoprotection provided by the C60-SMEDDS/TP favored normal cells (L02 and GES-1) over tumor cells (BEL-7402 and HepG2 cells) in vitro. This suggests a promising approach for the safe and effective treatment of TP. Full article

A very important class of models widely used nowadays to describe and predict, at least in stochastic terms, the behavior of many-particle systems (where the word “particle” is not meant in the purely mechanical sense: particles can be cells of a living tissue, or cars in a traffic flow, or even members of an animal or human population) is the Kinetic Theory for Active Particles, i.e., a scheme of possible generalizations and re-interpretations of the Boltzmann equation. Now, though in the literature on the subject this point is systematically disregarded, this scheme is based on Markov Chains, which are special stochastic processes with important properties they share with many natural processes. This circumstance is here carefully discussed not only to suggest the different ways in which Markov Chains can intervene in equations describing the stochastic behavior of any many-particle system, but also, as a preliminary methodological step, to point out the way in which the notion of a Markov Chain can be suitably generalized to this aim. As a final result of the discussion, we find how to develop new very plausible and likely ways to take into account possible effects of the external world on a non-isolated many-particle system, with particular attention paid to socio-economic problems. Full article

The interaction between IgM and C1q represents the first step of the classical pathway of the complement system in higher vertebrates. To identify the significance of particular IgM/C1q interactions, recombinant IgMs were used in both hexameric and pentameric configurations and with two different specificities, along with C1q derived from human serum (sC1q) and two recombinant single-chain variants of the trimeric globular region of C1q. Interaction and complement activation assays were performed using the ELISA format, and bio-layer interferometry measurements to study kinetic behavior. The differences between hexameric and pentameric IgM conformations were only slightly visible in the interaction assay, but significant in the complement activation assay. Hexameric IgM requires a lower concentration of sC1q to activate the complement compared to pentameric IgM, leading to an increased release of C4 compared to pentameric IgM. The recombinant C1q mimetics competed with sC1q in interaction assays and were able to inhibit complement activation. The bio-layer interferometry measurements revealed K_D values in the nanomolar range for the IgM/C1q interaction, while the C1q mimetics exhibited rapid on and off binding rates with the IgMs. Our results make C1q mimetics valuable tools for developing recombinant C1q, specifically its variants, for further scientific studies and clinical applications. Full article