Search Results (1,056)

Background: Antibacterial resistance (ABR) poses a major challenge to global health, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the prominent multidrug-resistant strains. MRSA has developed resistance through the expression of Penicillin-Binding Protein 2a (PBP2a), a key transpeptidase enzyme involved in bacterial cell wall biosynthesis. Objectives: The objective was to design and characterize a novel small-molecule inhibitor targeting PBP2a as a strategy to combat MRSA. Methods: We synthesized a new indole triazole conjugate (ITC) using eco-friendly and click chemistry approaches. In vitro antibacterial tests were performed against a panel of strains to evaluate the ITC antibacterial potential. Further, a series of in silico evaluations like molecular docking, MD simulations, free energy landscape (FEL), and principal component analysis (PCA) using the crystal structure of PBP2a (PDB ID: 4CJN), in order to predict the mechanism of action, binding mode, structural stability, and energetic profile of the 4CJN-ITC complex. Results: The compound ITC exhibited noteworthy antibacterial activity, which effectively inhibited the selected strains. Binding score and energy calculations demonstrated high affinity of ITC for the allosteric site of PBP2a and significant interactions responsible for complex stability during MD simulations. Further, FEL and PCA provided insights into the conformational behavior of ITC. These results gave the structural clues for the inhibitory action of ITC on the PBP2a. Conclusions: The integrated in vitro and in silico studies corroborate the potential of ITC as a promising developmental lead targeting PBP2a in MRSA. This study demonstrates the potential usage of rational drug design approaches in addressing therapeutic needs related to ABR. Full article

Antioxidative neuroprotection is effective at preventing ischemic stroke (IS). Ferulic acid (FA) offers benefits in the treatment of many diseases, mostly due to its antioxidant activities. In this study, a glycosylated ferulic acid conjugate (FA-Glu), with 1,2,3-triazole as a linker and bioisostere between glucose at the C6 position and FA at the C4 position, was designed and synthesized. The hydrophilicity and chemical stability of FA-Glu were tested. FA-Glu’s protection against DNA oxidative cleavage was tested using pBR322 plasmid DNA under the Fenton reaction. The cytotoxicity of FA-Glu was examined via the PC12 cell and bEnd.3 cell tests. Antioxidative neuroprotection was evaluated, in vitro, via a H₂O₂-induced PC12 cell test, measuring cell viability and ROS levels. Antioxidative alleviation of cerebral ischemia–reperfusion injury (CIRI), in vivo, was evaluated using a rat middle cerebral artery occlusion (MCAO) model. The results indicated that FA-Glu was water-soluble (LogP −1.16 ± 0.01) and chemically stable. FA-Glu prevented pBR322 plasmid DNA cleavage induced via •OH radicals (SC% 88.00%). It was a non-toxic agent based on PC12 cell and bEnd.3 cell tests results. FA-Glu significantly protected against H₂O₂-induced oxidative damage in the PC12 cell (cell viability 88.12%, 100 μM) and inhibited excessive cell ROS generation (45.67% at 100 μM). FA-Glu significantly reduced the infarcted brain areas measured using TTC stain observation, quantification (FA-Glu 21.79%, FA 28.49%, I/R model 43.42%), and H&E stain histological observation. It sharply reduced the MDA level (3.26 nmol/mg protein) and significantly increased the GSH level (139.6 nmol/mg protein) and SOD level (265.19 U/mg protein). With superior performance to FA, FA-Glu is a safe agent with effective antioxidative DNA and neuronal protective actions and an ability to alleviate CIRI, which should help in the prevention of IS. Full article

Nephrogenic diabetes insipidus (NDI) is a rare hereditary disorder characterized by renal resistance to arginine vasopressin (AVP), resulting in the kidneys’ inability to concentrate urine. Approximately 90% of NDI cases follow an X-linked inheritance pattern and are associated with pathogenic variants in the AVPR2 gene, which encodes the vasopressin receptor type 2. The remaining 10% are attributed to mutations in the AQP2 gene, which encodes aquaporin-2, and may follow either autosomal dominant or recessive inheritance patterns. We present the case of a male infant, younger than nine months of age, who was clinically diagnosed with NDI at six months. The patient presented recurrent episodes of polydipsia, polyuria, dehydration, hypernatremia, and persistently low urine osmolality. Despite adjustments in pharmacologic treatment and strict monitoring of urinary output, the clinical response remained suboptimal. Given the lack of improvement and the radiological finding of an absent posterior pituitary (neurohypophysis), the possibility of coexistent central diabetes insipidus (CDI) was raised, prompting a therapeutic trial with desmopressin. Nevertheless, in the absence of clinical improvement, desmopressin was discontinued. The patient’s management was continued with hydrochlorothiazide, ibuprofen, and a high-calorie diet restricted in sodium and protein, resulting in progressive clinical stabilization. Whole-exome sequencing identified a novel homozygous missense variant in the AQP2 gene (c.398T > A; p.Val133Glu), classified as likely pathogenic according to the American College of Medical Genetics and Genomics (ACMG) criteria: PM2 (absent from population databases), PP2 (missense variant in a gene with a low rate of benign missense variation), and PP3 (multiple lines of computational evidence supporting a deleterious effect)]. NDI is typically diagnosed during early infancy due to the early onset of symptoms and the potential for severe complications if left untreated. In this case, although initial clinical suspicion included concomitant CDI, the timely initiation of supportive management and the subsequent incorporation of molecular diagnostics facilitated a definitive diagnosis. The identification of a previously unreported homozygous variant in AQP2 contributed to diagnostic confirmation and therapeutic decision-making. The diagnosis and comprehensive management of NDI within the context of polyuria-polydipsia syndrome necessitates a multidisciplinary approach, integrating clinical evaluation with advanced molecular diagnostics. The novel AQP2 c.398T > A (p.Val133Glu) variant described herein was associated with early and severe clinical manifestations, underscoring the importance of genetic testing in atypical or treatment-refractory presentations of diabetes insipidus. Full article

Increasing the discharge of saline wastewater from an industrial field poses a challenge for applicable Anammox-based technologies. This study established the integrated partial sulfur-driven denitrification and Anammox (SPDA) system to explore the effects of different salinity levels on nitrogen conversion features. The results of batch tests suggested that sulfur-driven denitrification exhibited progressive suppression of nitrate reduction (97.7% → 12.3% efficiency at 0% → 4% salinity) and significant nitrite accumulation (56.4% accumulation rate at 2% salinity). Anammox showed higher salinity tolerance but still experienced drastic TN removal decline (97.6% → 17.3% at 0% → 4% salinity). Long-term operation demonstrated that the SPDA process could be rapidly established at 0% salinity and stabilize with TN removal efficiencies of 98.1% (1% salinity), 72.8% (2% salinity), and 70.2% (4% salinity). The robustness of the system was attributed to the appropriate strategy of gradual salinity elevation, the promoted secretion of protein-dominated EPS, the salinity-responsive enrichment of Sulfurimonas (replacing Thiobacillus and Ferritrophicum) as sulfur-oxidizing bacteria (SOB), and the sustained retention and activity of Brocadia as AnAOB. The findings in this study deepen the understanding of the inhibitory effects of salinity on the SPDA system, providing a feasible solution for saline wastewater treatment with low cost and high efficiency. Full article

The covalent cross-linking of caseins by the enzyme transglutaminase (Tgase) stabilizes the structure of casein micelles. In our study, the effects of a pretreatment of skim milk (SM) by Tgase on milk protein fractionation by microfiltration were tested. Tgase was found to induce amount-dependent modifications of all milk proteins in SM and a reduction in deposit resistance for laboratory dead-end filtrations of up to 20%. This improvement in process performance could partially be confirmed in pilot-scale cross-flow filtrations of Tgase-pretreated SM and micellar casein solutions (MCC). These comparative trials with untreated retentates under a variation of ΔpTM (0.5–2 bar) at 10 and 50° revealed distinct differences in deposit behavior and achieved the reduction in deposit resistance in a range of 0–20%. The possibility of pre-fouling with enzymatically pretreated MCC prior to SM filtration was also investigated. Under different pre-fouling conditions, practical modes of retentate change, and pre-foulant compositions, a switch to untreated SM consistently resulted in an immediate and major increase in deposit resistance by 50–150%. This was partially related to the change in the ionic environment and the protein fraction. Nevertheless, our results underline the potential of Tgase pretreatment and pre-fouling approaches to alter filtration performance for different 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

A nanoparticle formulation was generated from distiller dried grains with solubles (DDGS), and its effect on the production of anthraquinones (AQs) was evaluated on Rubia tinctorum hairy roots. The DDGS material was washed with water and ethyl acetate to remove mainly the soluble organic/inorganic molecules and reduce the fat content, respectively, followed by an alkaline treatment to remove the polysaccharides. The resulting alkaline solutions were then lyophilized and redispersed in deionized water to generate a monodispersed nanoparticulate formulation (DDGS-NP) with a hydrodynamic diameter and zeta potential of 227 ± 42 nm and −53 ± 7 mV, respectively. The formulation demonstrated good colloidal stability over time, and sterilized DDGS-NPs maintained comparable physicochemical properties. The nanoparticles were enriched in protein fractions, unsaturated fatty acids, and orthophosphate anion components from DDGS, as determined by solid-state Nuclear Magnetic Resonance (NMR), X-ray photoelectron spectroscopy (XPS), organic elemental analysis (OEA), and inductively coupled plasma optical emission spectrometry (ICP-OES) techniques. The DDGS-NPs were tested at different concentrations on Rubia tinctorum hairy roots, in comparison to or in combination with methyl jasmonate (MeJ), for their capacity to induce the production of AQs. All DDGS-NP concentrations increased the production of specific AQs to 7.7 (100 mg L⁻¹), 7.8 (200 mg L⁻¹), and 9.3 µmol/gFW (500 mg L⁻¹), with an extracellular AQ accumulation of 18 µM for the highest DDGS-NP concentration, in comparison with the control hairy roots (~2 µM AQ). The plant growth was not affected at any of the tested nanoparticle concentrations. Interestingly, the combination of DDGS-NPs and MeJ resulted in the highest extracellular AQ accumulation in R. tinctorum root cultures. Full article

Background/Objectives: Gallic acid, a natural phenolic compound, was used as a crosslinking agent to achieve protein–polyphenol conjugation under alkaline conditions, presenting an innovative approach to stabilize gelatin. Methods: The formulated inks were evaluated for their rheological properties and 3D printing performance. Once the scaffolds were printed, physicochemical properties were assessed by color changes and FTIR. Additionally, three different post-processing methods were studied to avoid toxicity: incubation in PBS, incubation in NaOH followed by PBS neutralization, and incubation in HCl followed by PBS neutralization. Results: The inks exhibited shear-thinning behavior with self-supporting capacity after extrusion, indicating their suitability for use as inks in 3D printing. After printing, changes in color and in the amide I band/amide II band ratio were observed due to alkaline oxidation, confirming the gelatin crosslinking. Among the tested treatments, incubation in PBS or NaOH followed by neutralizing with PBS proved to be the most suitable for obtaining cytocompatible scaffolds. The mechanical properties demonstrated the suitability of the proposed crosslinking systems for creating scaffolds. Conclusions: This strategy confirms that gallic acid-mediated crosslinking under alkaline conditions enables the fabrication of cytocompatible and mechanically stable gelatin-based scaffolds, making them suitable for tissue engineering. Full article

A high intake of saturated fats and cholesterol from processed meats is associated with increased cardiovascular disease risk. This study aimed to develop a nutritionally enhanced Bologna-type sausage by partially replacing the beef content with a structured emulsion gel (EG) formulated from pine nut oil, inulin, carrageenan, and whey protein concentrate. The objective was to improve its lipid quality and functional performance while maintaining product integrity and consumer acceptability. Three sausage formulations were prepared: a control and two variants with 7% and 10% EG, which substituted for the beef content. The emulsion gel was characterized regarding its physical and thermal stability. Sausages were evaluated for their proximate composition, fatty acid profile, cholesterol content, pH, cooking yield, water-holding capacity, emulsion stability, instrumental texture, microstructure (via SEM), oxidative stability (TBARSs), and sensory attributes. Data were analyzed using a one-way and two-way ANOVA with Duncan’s test (p < 0.05). The EG’s inclusion significantly reduced the total and saturated fat and cholesterol, while increasing protein and unsaturated fatty acids. The 10% EG sample achieved a PUFA/SFA ratio of 1.00 and an over 80% reduction in atherogenic and thrombogenic indices. Functional improvements were observed in emulsion stability, cooking yield, and water retention. Textural and visual characteristics remained within acceptable sensory thresholds. SEM images showed more homogenous matrix structures in the EG samples. TBARS values increased slightly over 18 days of refrigeration but remained below rancidity thresholds. This period was considered a pilot-scale evaluation of oxidative trends. Sensory testing confirmed that product acceptability was not negatively affected. The partial substitution of beef content with pine nut oil-based emulsion gel offers a clean-label strategy to enhance the nutritional quality of Bologna-type sausages while preserving functional and sensory performance. This approach may support the development of health-conscious processed meat products aligned with consumer and regulatory demands. Full article

Keratin, a fibrous structural protein, has been employed as a biomaterial for hemostasis and tissue repair due to its structural stability, mechanical strength, biocompatibility, and biodegradability. While extensive research has focused on developing scaffolds using keratin extracted from various sources, no studies to date have explored the use of keratin derived from human nail clippings. In this study, keratin was extracted from human nail clippings using the Shindai method and used to fabricate and compare two types of scaffolds for bone tissue engineering via the freeze-drying method. The first scaffold consisted of keratin combined with gelatin (KG), while the second combined keratin, gelatin, and hydroxyapatite (HAp) (KGH), the latter synthesized from blood cockle clam shells using the wet precipitation method. Physicochemical characterization and surface morphology analysis of keratin and both scaffolds showed promising results. Tensile strength testing revealed a significant difference in Young’s modulus. The KG scaffold exhibited higher porosity, water uptake, and water retention capacity compared to the KGH scaffold. In vitro biocompatibility studies revealed that the KGH scaffold supported higher cell proliferation compared to the KG scaffold. This study demonstrates the potential of using human nail-derived keratin in composite scaffold fabrication and serves as a foundation for future research on this novel biomaterial source. Full article

Biopolymers, owing to their environmentally friendly and sustainable characteristics, have become a promising alternative for soil stabilization in geotechnical engineering. The application of protein-based biopolymers as binders for soil stabilization is less prevalent in geotechnical engineering compared to polysaccharide-based biopolymers. This study explores the potential of gelatin, a protein-based biopolymer derived from animal collagen, for stabilizing silty sand and improving its geotechnical properties. Gelatin was mixed into the soil at concentrations ranging from 0.25% to 2% of the dry weight of soil, and its effects on various soil characteristics were evaluated. The tests conducted include liquid limit, plastic limit, compaction behavior, and unconfined compressive strength (UCS); the addition of 1% gelatin led to an approximate 1.69 times increase in the strength of the unamended soil. After 28 days of curing, the UCS improved by approximately 5.03 times compared to the untreated soil, and the treated soil exhibited increased resistance to deformation under load. Microstructural analysis using scanning electron microscopy (SEM) revealed that gelatin facilitated the formation of a cohesive matrix, enhancing particle bonding and reducing void spaces within the soil. Carbon footprint analysis (CFA) conducted on an isolated footing stabilized with gelatin showed that the carbon emissions were reduced by 99.8% and 99% compared to traditional stabilizers such as lime and cement. Additionally, the interaction between the biopolymer and the fine-grained soil is distinctly evident in the FTIR and XRD analysis through hydrogen bonding and the formation of cementitious compounds. Full article

Cancer remains one of the most formidable challenges to human health; hence, developing effective treatments is critical for saving lives. An important strategy involves reactivating tumor suppressor genes, particularly p53, by targeting their negative regulator MDM2, which is essential in promoting cell cycle arrest and apoptosis. Leveraging a drug repurposing approach, we screened over 24,000 clinically tested molecules to identify new MDM2 inhibitors. A key innovation of this work is the development and application of a selective cleaning algorithm that systematically filters assay data to mitigate noise and inconsistencies inherent in large-scale bioactivity datasets. This approach significantly improved the predictive accuracy of our machine learning model for pIC₅₀ values, reducing RMSE by 21.6% and achieving state-of-the-art performance (R² = 0.87)—a substantial improvement over standard data preprocessing pipelines. The optimized model was integrated with structure-based virtual screening via molecular docking to prioritize repurposing candidate compounds. We identified two clinical CB1 antagonists, MePPEP and otenabant, and the statin drug atorvastatin as promising repurposing candidates based on their high predicted potency and binding affinity toward MDM2. Interactions with the related proteins MDM4 and BCL2 suggest these compounds may enhance p53 restoration through multi-target mechanisms. Quantum mechanical (ONIOM) optimizations and molecular dynamics simulations confirmed the stability and favorable interaction profiles of the selected protein–ligand complexes, resembling that of navtemadlin, a known MDM2 inhibitor. This multiscale, accuracy-boosted workflow introduces a novel data-curation strategy that substantially enhances AI model performance and enables efficient drug repurposing against challenging cancer targets. Full article

Banana (Musa paradisiaca) is a climacteric fruit with high postharvest perishability, limiting its export potential. This study evaluated the effectiveness of a natural protein–polysaccharide edible coating—comprising whey, agar, cassava starch, and glycerol—on maintaining the physicochemical quality of green bananas during 28 days of refrigerated storage (13 °C, 95% RH). Seven formulations were tested, including an uncoated control. Physicochemical parameters such as weight loss, firmness, fruit dimensions, peel color, titratable acidity, pH, and soluble solids (°Brix) were systematically monitored. Significant differences were observed among treatments (ANOVA, p < 0.001). The most effective coating (T5), composed of 16.7% whey, 16.7% agar, 33.3% cassava starch, and 33.3% glycerol (based on 30 g/L solids), reduced weight loss by 58.8%, improved firmness retention by 48.4%, and limited sugar accumulation by 17.0% compared to the control. It also stabilized pH and acidity, preserved peel thickness and color parameters (L*, a*, b*), and delayed ripening. These findings confirm the coating’s capacity to form a cohesive semipermeable barrier that modulates moisture loss and respiration, making it a functional and sustainable alternative for extending banana shelf life in tropical supply chains. Full article

Background: “CFTR modulators” (also named “caftor”) have been developed and introduced into clinical practice to improve the functionality of defective CFTR protein. Therapeutic drug monitoring (TDM) is not currently used for CFTR modulators in routine clinical practice and there is still much to learn about the pharmacokinetic/pharmacodynamic (PK/PD) and the safety profiles of these drugs in a real-world setting. Moreover, therapeutic ranges are not yet available for both pediatric and adult cystic fibrosis (CF) patients. Methods: A new and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method for contemporary quantification of ivacaftor (IVA), tezacaftor (TEZ) and elexacaftor (ELX) in plasma samples has been developed and validated. The clinical performance of our method has been tested on samples collected during the routine clinical practice from n = 25 pediatric patients (aged between 7 and 17 years) affected by cystic fibrosis. This LC-MS/MS method has been validated according to ICH (International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use) guidelines for the validation of bioanalytical methods. Results: Our method fulfilled ICH guidelines in terms of accuracy, precision, selectivity, specificity and carry-over. Intra- and inter-day accuracy and precision were ≤15%. The 9-day autosampler stability was 90–100% for TEZ and ELX; meanwhile, it fell to 76% for IVA. An injection volume of 1 µL and a wider quantification range (0.1–20 µg/mL) represent a novelty of our method in terms of sensitivity and fields of application. Finally, the evaluation of PK exposure parameters for IVA revealed strong agreement with previously published reports and with results from the summary of product characteristics (SmPCs). Conclusions: This method could be adopted to contemporarily measure ELX/TEZ/IVA plasma levels for both PK studies and monitor therapy compliance, especially in case of poor or partial responses to treatment, or to evaluate drug–drug interactions when multiple concomitant medications are required. Considering also the high cost burden of these medications to the health system, a TDM-based approach could facilitate more cost-effective patient management. Full article

In the context of modern dermocosmetic development, multifunctional topical gel-cream formulations must be efficient for both therapeutic efficacy and cosmetic applications. This study presents a comparative physicochemical and pharmacotechnical analysis of three topical gel-cream formulations developed by Brand Chanand^®: Acne Control Cleanser (ACC), Acne Face Cream (AFC), and Gentle Cream Cleanser Serum Control, Regenerating, Hydrating, Calming (IRC). Each formulation is enriched with a specific blend of bioactive compounds, including botanical oils, vitamins, and proteins, designed to treat acne, to support skin regeneration, and to maintain the skin barrier. A multidisciplinary approach was used, including Fourier Transform Infrared Spectroscopy with Attenuated Total Reflectance (FTIR-ATR), differential scanning calorimetry (DSC), rheological evaluation, pH and density determination, spreadability analysis, and oxidative stability testing to evaluate the products. Antioxidant capacity was assessed through multiple in vitro assays. The results demonstrated that all three gel-cream formulations exhibit pseudoplastic rheological behaviour, suitable for topical application. AFC showed the highest oxidative stability and antioxidant activity, while IRC presented superior spreadability and cosmetic efficacy, likely due to its complex composition. ACC displayed faster absorption and was ideal for targeted use on oily or acne-prone skin. The differences observed in the stability and performance suggest that the ingredient synergy, base composition, and solubility profiles show notable variations in dermato-cosmetic formulations. These findings highlight the formulation–performance relationship in topical gel-cream formulations and support the development of new cosmetic products tailored for sensitive and acne-prone skin. Full article