Abstract: This research describes the development and thorough characterization of a novel, versatile, and biocompatible hybrid nanocarrier of the NO-releasing agent NOC-18, with a specific focus on optimizing the purification process. In this study, we focused on the sustained release of NO using the biocompatible and diagnostic hybrid magnetic nanoparticles (hMNPs) containing cerium-doped maghemite (CM) NPs, embedded within human serum albumin (HSA) protein. A comprehensive study was conducted using Electron Paramagnetic Resonance (EPR) alongside the Griess assay to evaluate the NO release from the chosen NO donor NOC-18 and to assess the limitations of the molecule under various reaction conditions, identifying the optimal conditions for binding NOC-18 with minimal NO loss. Two types of particles were designed: In-hMNPs, where NOC-18 is encapsulated within the particles, and Out-hMNPs, where NOC-18 is attached onto the surface. Our results demonstrated that In-hMNPs provided a sustained and prolonged release of NO (half-life 50 h) compared to rapid release for the Out-hMNPs, likely due to the strong bonds formed with cerium, which helped stabilize the NO molecules. These results represent a promising approach to the design of the dual-function agent that combines the contrasting properties for tumor MRI with the possibility of increasing the permeability of tumor vasculature. Employment of such a dual-function agent in combination with nanotherapeutics may augment their efficacy by facilitating their access to the tumor.

Abstract: L-Carnosine (Car) is an endogenous dipeptide with high potential for drug discovery approaches in neurogenerative diseases, whereas TAT1 is a small arginine-rich peptide derived from the HIV-1 trans-activator protein (TAT) known to stimulate proteasome activity. Three isomeric peptides were synthetised by adding the Car moiety to the TAT1 sequence at the C- and N-termini and in the middle of the peptide sequence. High-resolution and energy-resolved CID MS/MS experiments were performed to differentiate the three isomeric peptides. At first glance, the obtained MS/MS spectra showed a high degree of similarity between the peptides. A wealth of low-intensity fragment ions peaks resulting mainly from arginine-specific neutral losses, which are useless for structural elucidation, or peaks that cannot be easily assigned were observed. Energetic study was also non-conclusive with that respect. However, Principal Component Analysis (PCA) showed that the three peptides could be clearly distinguished when the entire MS/MS spectra were considered rather than just the intensity of the precursor ions peak. Interestingly, the PCA loadings revealed the characteristic fragment ions of each peptide (although with smaller intensities) providing hints on consecutive fragmentation patterns. Some of these specific peaks could also be assigned to scrambling during fragmentation. These results demonstrate the potential of PCA as a simple chemometric tool for semi-automated peak identification in complex MS/MS spectra.

Abstract: The annually wasted amount of food had surpassed 1 billion metric tons. Food waste is considered as an important source for recovery of bioactive compounds, such as carotenoids. There is a demand for antioxidants, nutraceuticals and natural colorants in various industries and carotenoids are one of the commonly used compounds which fit with this description. Pumpkin and spinach waste, whose combined amount is over 2 million metric tons, contains bioactive compounds and these wasted foods could be utilized for recovery of carotenoids. Carotenoids are hydrophobic molecules; therefore, commercial extraction processes often use highly non-polar solvents, and these are rarely environmentally friendly. The aim of this research was to develop effective extraction processes for carotenoids from pumpkin and spinach using environmentally friendly green chemicals. A series of deep eutectic solvents (DESs) composed with L-menthol and carboxylic aliphatic acids were made for extraction of carotenoids from pumpkin (Cucurbita moschata) and spinach (Spinacia oleracea) via mechanical mixing assisted extraction (MMAE) and homogenization assisted extraction (HAE). Response surface methodology (RSM) and analysis of variance (ANOVA) analysis were used to analyze the data and optimization. The DESs composed from L-Menthol and Propionic Acid had the best effect on the extraction of total carotenoid content (TCC) (represented as ß-Carotene) from pumpkin and spinach via solutions with 1:2 and 1:4 molar ratios, respectively. The yield of carotenoid extraction is expressed in μg-ß-Carotene/g of pumpkin or spinach. Under the calculated optimum conditions, the yields are estimated to be 11.528 μg-ß-Carotene/g-pumpkin for MMAE method, 8.966 μg-ß-Carotene/g-pumpkin for HAE method, 16.924 μg-ß-Carotene/g-spinach for MMAE method and 18.870 μg-ß-Carotene/g-spinach for HAE method.

Abstract: Precise binding free energy predictions for ligands targeting metalloproteins, especially zinc-containing histone deacetylase (HDAC) enzymes, require specialized computational approaches due to the unique interactions at metal-binding sites. This study evaluated a docking algorithm optimized for zinc coordination to determine whether it could accurately differentiate between protonated and deprotonated states of hydroxamic acid ligands, a key functional group in HDAC inhibitors. By systematically analyzing both protonation states, we sought to identify which state produced docking poses and binding energy estimates most closely aligned with experimental values. The docking algorithm was applied across HDAC 2, 4, and 8, comparing protonated and deprotonated ligand correlations to experimental data. Results demonstrated that the deprotonated state consistently yielded stronger correlations with experimental data, with R² values for deprotonated ligands outperforming protonated counterparts in all HDAC targets (average R2 = 0.80 compared to the protonated form where R2 = 0.67). These findings emphasize the significance of proper ligand protonation in molecular docking studies of zinc-binding enzymes, particularly HDACs, and suggest that deprotonation enhances predictive accuracy. The study’s methodology provides a robust foundation for improved virtual screening protocols to evaluate large ligand libraries efficiently. This approach supports the streamlined discovery of high-affinity, zinc-binding HDAC inhibitors, advancing therapeutic exploration of metalloprotein targets. A comprehensive, step-by-step tutorial has been provided to facilitate a thorough understanding of the methodology and enable reproducibility of the results.

Abstract:

(1) Aim of this in-vitro study was to investigate the handling of proximal-cervical undermined enamel margins on adhesive performance of differently fabricated and differently cemented ceramic inlays and partial crowns (2) Methods: 192 extracted third molars received mod (n=96) and partial crown (n=96) preparations. A mesial 2x2x4 mm cervical groove was created in dentin to simulate a deeper (dentin) caries excavation. This dentin groove was either left (G/groove), filled with composite (F/filling) or completely removed (D/dentin). Labside (e.max Press) and chairside (e.max CAD) inlays and partial crowns were adhesively luted with Syntac/Variolink Esthetic (SV) or Adhese Universal/Variolink Esthetic (AV). Initially, and again after thermomechanical loading (TMB: 1 million cycles at 50 N, 25,000 thermocycles at 5°C/55°C), epoxy replicas were examined for marginal gaps using scanning electron microscopy (200x magnification). Light microscopy (10x magnification) was used to evaluate proximal cervical crack propagation. (3) Results: Regardless of the adhesive system, D groups generally showed significantly lower marginal quality (p<0.05), with the universal adhesive performing better than the multi-step adhesive system (p<0.05). Subgroups G and F were similar in marginal quality (p>0.05) and not worse than the controls (p>0.05) regardless of the adhesive system, but showed less cracking in F than in G (p<0.05). In general, fewer cracks were observed in CAD/CAM restorations than in laboratory-fabricated restorations (p<0.05). Partial crowns showed better marginal quality and less cracking than inlays (p<0.05). (4) Conclusions: If the dentin level is lower than the enamel level in ceramic preparations after caries excavation in the proximal box, the resulting undermined enamel should not be removed.

Abstract: This study evaluates the chemical, physical, mechanical, and biological properties of untreated and heat-treated Cryptomeria japonica wood from the Azores, Portugal. Heat treatment was performed at 212°C for 2 hours following the Thermo-D class protocol. Chemical analysis revealed an increase in ethanol extractives and lignin content after heat treatment, attributed to hemicellulose degradation and condensation reactions. Dimensional stability improved significantly, as indicated by reduced swelling coefficients and higher anti-swelling efficiency (ASE), particularly in the tangential direction. Heat-treated wood demonstrated reduced water absorption and increased density, enhancing its suitability for applications requiring dimensional stability. Mechanical tests showed a decrease in bending strength by 19.6% but an increase in the modulus of elasticity (MOE) by 49%, reflecting changes in the wood's structural integrity. Surface analysis revealed significant color changes, with darkening, reddening and yellowing, aligning with trends observed in other heat-treated woods. Biological durability tests indicated that both untreated and treated samples were susceptible to subterranean termite attack, although heat-treated wood exhibited a higher termite mortality rate, suggesting potential long-term advantages. This study highlights the impact of heat treatment on Cryptomeria japonica wood, emphasizing its potential for enhanced stability and durability in various applications.

Abstract: Pancreatitis is a prominent and severe type of inflammatory disorder with has grabbed a lot of scientific and clinical interest to prevent its onset. It should be detected early to avoid the development of serious complications, which occur due to long term damage. The accurate measurement of biomarkers that are released from the pancreas during inflammation is essential for the detection and early treatment of patients with severe acute and chronic pancreatitis, and this is sub-optimally performed in clinically relevant practices mainly due to the complexity of the procedure and the cost of the treatment. Clinically available tests for early detection of pancreatitis are often time consuming. Early detection of pancreatitis also relates to disorders of the exocrine pancreas, such as cystic fibrosis in the hereditary form and the cystic fibrosis-like syndrome in the acquired form of pancreatitis which are genetic disorders whose symptoms can be correlated to the presence of over expression of specific markers such as creatinine in biological fluids like urine. In this review we have studied how to develop a minimally invasive system using hydrogel-based biosensor, which are highly absorbent and biocompatible polymers which can respond to specific stimuli such as enzymes, pH, temperature, or the presence of biomarkers. These biosensors are helpful for real-time health monitoring, environmental monitoring, and medical diagnostics since they translate biological reactions into quantifiable data. These analytical devices can be used to improve early detection of severe pancreatitis in real time.

Abstract: Cannabidiol (CBD), isolated from the Cannabis plant, has been gaining increased attention in recreational purposes but has been less explored for applications in dentistry. CBD has shown to be effective against many strains of oral bacteria. Bacteria-associated denture infections is a recurring disease affecting most denture wearers. Poly(methyl methacrylate) (PMMA) remains the main material employed in the fabrication of dentures due to its desirable physical, mechanical, and aesthetic properties. However, the improvement of its antimicrobial properties remains a challenge. To address this need, we have developed a denture surface synthesized from PMMA loaded with CBD nanoparticles for bactericidal effects. The antimicrobial agent CBD was purified, and then synthesized into denture coatings by UV curing and the curing time being investigated. The coatings were characterized by 1H NMR, SEM and FTIR spectroscopies. The antimicrobial activity of the CBD was evaluated before synthesizing it into the PMMA material on Staphylococcus aureus, Escherichia coli and S. agalactiae. Anti biofilm studies were conducted against Staphylococcus aureus, Escherichia coli and S. agalactiae biofilms. Dissolution and leaching studies of the synthesized coatings were conducted to determine the release pattern of CBD molecules from the coatings. The results showed that CBD exhibited a significant bactericidal effect on gram positive bacteria with the MIC value of 2- 2.5 µg/mL and MBC of 10- 20 µg/mL but remained was ineffective on planktonic gram -negative bacteria. The results for biofilms studies showed a significant reduction in the biofilm formation in the CBD added coatings as compared to only PMMA coatings. The PMMA/CBD coatings significantly reduced 99 % of biofilm growth not only in gram-positive but also in gram-negative as well. The coated MMA/CBD surface has antifouling properties which targets adhesin proteins and blocked biofilm formation. The CBD molecule leached out to the bacteria and disrupted bacterial cytoplasmic cell walls causing cell lysis. The effectiveness of such antimicrobial agents in coatings relies on the drug dissolving in the fluids prior to absorption into the systemic circulation. The rate of dissolution of the CBD from the coating is crucial and the experimental results show excellent release of CBD molecules in vitro. This study showed that CBD can be incorporated into denture materials, enabling antibiotic-free dentures for preventing dental biofilms and reducing dental plaques.

Abstract:

Medicinal plants have been used extensively as sources of a wide variety of biologically active compounds for many centuries and as crude materials or pure compounds for treating various disease conditions. The leaves of the plant have been applied in treating snakebite, stomach ache, cough and so on. The plant leaves were extracted with hexane and methanol using the soxhlet extraction process. In this study, leaf extracts of G. senegalensis were profiled and evaluated for their phospholipase A2 inhibitory potential via experimental and computational approaches. Characterization of the extracts was done using GC-MS analysis, Antisnake venom screening was conducted using PLA2 acidimetric assay while Insilco molecular docking studies was performed using AutoDock vina in PyRx and ADMET was predicted using swiiADME and protox-II online servers. GC-MS analysis revealed the presence of 50 compounds from which 14, 4, 15, 13 and 13 were for hexane, chloroform, ethyl acetate, butanol and aqueous fractions respectively. The PLA2 acidimetric assay was used to screen the fractions for inhibitory activity against N. nigricollis venom in vitro. The results showed that the aqueous fraction was the most active, with PLA2 inhibition ranging from 66.18 to 74.67% at 1.0 to 0.125 mg/cm3, respectively. The fractions inhibited the hydrolytic effects of the N. nigricollis PLA2 enzyme, exhibiting considerable (p<0.05) antisnake venom activity. In comparison to the standards, four compounds exhibited a higher docking score (-8.7 to -8.4 kcal/mol). Insilico ADME and Drug-likeness revealed the compounds have passed absorptivity test for oral medication as well as indicating lower likelihood of interacting with other drugs. The results also showed the compounds to be slightly toxic. The results of this study supported the use of G. senegalensis in traditional medicine by demonstrating that its leaves contains phytoconstituents with antisnake properties.

Abstract: With the growth of global energy demand and the reduction of fossil fuels, the search for clean energy has become particularly urgent. Hydrogen energy, due to its cleanliness and high efficiency, is regarded as an ideal alternative energy source. Methane catalytic cracking technology, as an effective way to produce hydrogen, has significant economic and environmental value.This study focuses on the application of vanadium-based catalysts in biogas cracking for hydrogen production. Vanadium-based catalysts have shown great potential in the dehydrogenation of hydrocarbons due to their cost-effectiveness, environmental compatibility, high catalytic activity, and stability. An innovative one-step method was used to prepare Ni-VC/TiO2 composite catalysts, and the impact of vanadium carbide (VC) content on the performance of the catalyst was studied. The addition of VC not only optimized the catalyst's active surface area but also enhanced its conductivity and electron transfer capability, thereby improving catalytic efficiency.Experimental results indicate that an increase in VC content leads to a decrease in the specific surface area and pore volume of the catalyst, but an increase in pore size, which is beneficial for the adsorption and diffusion of gas molecules. Moreover, the reaction temperature has a significant effect on the performance of the catalyst: at 500°C, the 10Ni-5VC/TiO2 catalyst exhibited the highest stability and hydrogen production rate. However, at higher temperatures, the methane conversion rate and hydrogen yield will drop sharply due to the increased graphitization of coke, which accelerates the deactivation of the catalyst.Through regeneration experiments, we found that using CO2 as an activator at 600°C can effectively restore the activity of the catalyst. However, as the number of regenerations increases, the performance of the catalyst gradually decreases, which is related to the increase in Ni grain size and the formation of surface amorphous coke. SEM and TEM analysis showed that coke mainly exists in the form of filamentous carbon, which affects the long-term stability of the catalyst.