This study aimed to characterize the metabolomic profile alterations in patients with liver cirrhosis resulting from alcoholic liver disease (ALD, n=31) and non-alcoholic fatty liver disease (NAFLD, n=8) compared to matched controls.

Materials and methods: We conducted a study of 39 cirrhotic patients and age-, gender-, and BMI-matched controls (n=59) (p>0.05 for all), which included pre-cirrhotic patients and healthy individuals. Metabolite level in the blood serum was determined by high-performance liquid chromatography and tandem mass spectrometry. In order to find out the relationship between certain metabolites and the presence of liver cirrhosis, a binary logistic regression model was used to determine the odds ratio (OR) and 95% confidence interval (CI).

Results: This study employed targeted metabolomic profiling to analyze 96 serum metabolites in patients with NAFLD and ALD. The analysis of the obtained data allowed us to identify a spectrum of metabolites associated with liver cirrhosis in the outcome of ALD and NAFLD (p < 0.05) and multidirectional shifts in the concentrations of biologically active molecules. Fibrosis progression was associated with significant metabolite alterations: increased levels of kynurenine (OR 2.93 [1.56-5.51]), acetyl-carnitine (OR 1.08 [1.01-1.17]), epinephrine OR 26.5 [5.7-123.4], dimethylarginine OR 20.2 [2.5-164.8], and methionine OR 1.10 [1.05-1.16], along with decreased indole-3-carboxaldehyde OR 0.04 [0.003-0.45]), serotonin (OR 0.038 [0.01-0.15]), norepinephrine (OR 0.25 [0.11-0.56]), taurine (OR 0.98 [0.97-0.99]), and carnosine OR 0.55 [0.34-0.90]).

Conclusion: Comprehensive metabolomics studies remain challenging due to their high cost, leading most liver disease studies to focus on individual metabolite groups. In contrast, our targeted analysis of 96 serum metabolites provides a broad, multifactorial view of the metabolic alterations that occur in the context of ALD and NAFLD cirrhosis. The obtained data, which suggest enhanced oxidative stress, sympathetic activation, and mitochondrial dysfunction in progressive fibrosis, not only confirm previous findings but also uncover novel, previously invisible metabolic pathways and interactions.

This work explores the bioanalytical methods and green procedures for the synthesis of small molecules–ionic liquids with N-glycans (GLc-O-SMol-HmILs), good applicable materials with bioactive properties as potential drugs anti-SARS-CoV-2. Methylimidazolium N-ethylamine has good applicability and is an active ionic liquid that acts with oligosaccharides very easily. Particularly, this ionic liquid is employed in the modification of oligosacharides obtained from human milk, lactoferrin (Hm), to produce a novel small molecule (Gl-SM-o-IL-HmLf). The advanced analytical techniques of UPLC/ESI-QTOF and MALDI-TOF mass spectrometry illustrated the identification and investigation of the modified small molecules–glycoproteins–ionic liquids (Glc-SM-IL-HmLf). Small molecules may be provided as significant potential drug candidates for inhibiting the selected targets of main protease (Mpro), RNA-dependent RNA polymerase (RdRp), Papain-like protease (PLpro) and trans-membrane protease serine 2 (TMPRSS2). The molecule had a slightly stronger interaction than remdesivir with Mpro and TMPRSS2. Similarly, the modified compound interacted with RdRp at a comparable level to the standard drug, which had slightly lower binding energy than remdesivir and its derivatives, and the novel small molecule-IL-HmLf has strong 10 H-bonding with the structures of four targets of main protease. The characterization and quantification of small molecules–ionic liquids linked to oligosaccharides were investigated by LCMS, GCMS, NMR, and UV-vis spectrometer. Molecular docking disclosed the bio-activity of modifying glycoprotein–small Glc-IL-HmLf-molecules by elucidating their interactions with the inspected targets.

Rabies remains a persistent zoonotic threat, particularly in regions where domestic dogs are the main source of human and animal infections. This mathematical model studies the dynamics of rabies transmission between canine populations (dog-to-dog) and from canines to humans (dog-to-human). The model incorporates susceptible, infected, and vaccinated compartments for both species, with pre-exposure vaccination as the key control strategy. Processes such as encapsulation, stability enhancement, and controlled release are modelled as parameters influencing vaccination rates in both dogs and humans. Specifically, the model introduces processing-dependent vaccination functions that reflect improved bioavailability, immunogenicity, and delivery efficiency due to advanced formulation techniques.

This interdisciplinary approach bridges mathematical epidemiology and pharmaceutical technology. Earlier rabies models focus on transmission and static vaccination, often ignoring vaccine formulation and delivery. Our current work fills this gap by incorporating pharmaceutical and particle engineering parameters into the vaccination terms of the model, thereby providing a more comprehensive framework for optimizing rabies control strategies in endemic regions.

Positivity and boundedness analyses confirm that all model variables remain biologically feasible and bounded over time. Stability analysis identifies thresholds for disease elimination or persistence. Numerical simulations show that enhancing pharmaceutical parameters increases vaccination impact, reducing peak infection prevalence in dogs from 18% to 5% and in humans from 4% to 0.8%, and shortening elimination time from 8 years to 3 years. Formulations with controlled release and improved stability maintain over 90% reduction in transmission for more than 5 years, compared to 60% over 3 years for conventional vaccines. This will ensure that the model’s predictions are validated against realistic conditions and can effectively guide rabies control strategies.

Endometrial cancer (EC) incidence is rising globally, creating an urgent need for novel and effective therapeutic strategies. This study aimed to investigate the anticancer effects and underlying molecular mechanisms of neferine (Nef) in EC. Through mRNA sequencing analysis, we identified the MAPK signaling pathway as a key target influenced by Nef treatment. In vitro experiments using Ishikawa cells demonstrated that Nef significantly inhibited cell proliferation, migration, and invasion abilities. Mechanistic studies revealed that Nef downregulated critical proteins in the MAPK pathway, including MEK, ERK, and JNK1, as well as metastasis-related markers such as MMP2, MMP9, and Vimentin. The MAPK activator C16-PAF was able to reverse Nef-induced suppression of MMP2 expression, further confirming the involvement of the MAPK pathway. In vivo, administration of Nef at 20 mg/kg effectively suppressed tumor growth and inhibited MAPK pathway activation in a xenograft mouse model. Compared with current therapies, which often have considerable side effects and limited efficacy especially in advanced stages, Nef’s multi-targeted mechanism of action and natural origin suggest its potential as a promising candidate for EC treatment. Future studies should focus on evaluating its therapeutic efficacy in combination with standard treatments, as well as its pharmacokinetic properties and safety profile in clinical settings.

The main aim of this research was to provide a complete and new ultraviolet (UV) analysis of acetylsalicylic acid, a non-steroidal drug, in acetonitrile as a solvent. It is a low-cost method which is accurate, precise, easy to apply, and needs low reagent consumption. Using this method, we can precisely find the amount of pure acetylsalicylic acid from various pharmaceuticals in any specialized laboratories. A statistical validation process was successfully applied. The linearity of the method, system precision, method precision, method robustness, and method accuracy were studied and successfully determined. To evaluate the linearity of the method and exactly quantify pure acid acetylsalicylic acid from two pharmaceuticals by the proposed method, the absorption spectrum was plotted for a standard solution of 16.83 µg/mL in acetonitrile p.a. as a pure spectral solvent. The maximum absorption wavelength was found to be λ = 213 nm. The ultraviolet spectrophotometric method was linear over the entire concentration range between 1.20 µg/mL and 16.00 µg/mL, and the linear regression value was R² = 0.999. Acetylsalicylic acid pure calculated content was found to be 497.46 mg for the first pharmaceutical and 494,67 mg for the second product. These amounts were very close to the officially stated content of 500 mg, with very low average percentage deviations from this official content, (+) 0.51 % and (+) 1.06 %, respectively. The calculated average percentage deviations were below the maximum percentage limit (± 5%) stated by the European and International Pharmacopoeias Rules. This method can be easily applied for the Technical Quality Control of pharmaceutical products containing acetylsalicylic acid as an active substance.

Introduction:
Chronic Obstructive Pulmonary Disease (COPD) is a progressive inflammatory lung disorder characterized by irreversible airflow limitation and systemic involvement. While previous studies have implicated microRNA-34a (miR-34a) and Notch1 in COPD pathogenesis, most were restricted to in vitro models, specific exposure types, or genetic association studies lacking clinical correlation. This study explores the diagnostic and therapeutic relevance of the miR-34a–Notch1 axis in clinically phenotyped COPD patients. Modulation of this axis using targeted oligonucleotides such as antagomir-34a, an inhibitor of miR-34a, offers potential for therapeutic intervention.

Methods:
A total of 46 participants were enrolled, comprising 23 clinically diagnosed COPD patients and 23 healthy controls, following institutional ethical approval and written informed consent. The mean age of COPD patients was 62.26 ± 9.17 years, and that of the control group was 47.05 ± 12.67 years. Group comparisons were adjusted for age using multivariate analysis to reduce confounding. Pulmonary function tests (PFTs)—including Forced Expiratory Volume in 1 second (FEV1), Forced Vital Capacity (FVC), and the FEV1/FVC ratio—were conducted alongside Diffusing Capacity of the Lung for Carbon Monoxide per unit Alveolar Volume (DLCO/VA) and Arterial Blood Gas (ABG) analysis. The serum marker Blood Urea Nitrogen (BUN) was also assessed. Expression levels of miR-34a and Notch1 were quantified using qRT-PCR.

Results:
COPD patients exhibited significant upregulation of miR-34a and downregulation of Notch1 (p < 0.001). These alterations were correlated with reduced pulmonary function and abnormal ABG parameters. Receiver Operating Characteristic (ROC) analysis revealed high diagnostic accuracy for Notch1 (AUC = 0.974) and FEV1/FVC (AUC = 0.867). ANOVA demonstrated significant variation in Notch1 expression across ABE-defined COPD subtypes, suggesting its potential utility for severity stratification.

Conclusion:
These findings support the miR-34a–Notch1 axis as a promising biomarker and therapeutic target. Antagomir-34a shows potential as a precision medicine approach for COPD management.

The growing challenge of antibiotic resistance has prompted the search for cost-effective and efficient alternatives. Developing new synthetic or natural antimicrobial drugs is often resource-intensive and time-consuming, leading current research to focus on plant-based extracts and nanoparticle synthesis capable of generating reactive oxygen species or forming metal complexes. This study aims to optimize the eco-friendly synthesis of silver (AgNPs) and zinc oxide nanoparticles (ZnO NPs) with antibacterial properties for topical application. A standardized ethanolic extract of Paeonia anomala was used as a reducing and stabilizing agent. Phytocomplex formation with zinc ions under various pH conditions was modeled using Medusa software. The effects of microwave, UV irradiation, and ultrasound on nanoparticle synthesis and stability were assessed. Physicochemical characteristics and colloidal stability were monitored via UV-Vis spectrophotometry, with surface plasmon resonance (SPR) peaks observed at 408 nm for AgNPs and 320 nm for ZnO NPs, confirming nanoscale formation. Particle size and zeta potential were also evaluated. Storage at room temperature led to slightly more aggregation compared to refrigeration at 4 °C, though the effect was minimal. Neither the Paeonia anomala tincture nor the AgNPs derived from it exhibited antioxidant activity. However, AgNPs synthesized with the extract and stabilized via ultrasound demonstrated strong antimicrobial activity against Staphylococcus aureus and Escherichia coli at concentrations as low as 0.015 mM, while ZnO NPs required at least 2 mM to achieve similar effects.

Twin-screw melt granulation (TSMG) is a promising continuous process for producing solid pharmaceutical dosage forms. However, the application of Near-Infrared (NIR) spectroscopy as a Process Analytical Technology (PAT) tool in this context is still underexplored. This study evaluates the feasibility of using in-line NIR spectroscopy combined with chemometric modeling to monitor key formulation and process variables in TSMG, including binder content, screw configuration, and sample temperature. Experiments used lactose monohydrate and polyethylene glycol (PEG) 6000 as a model formulation. Blends were processed under four conditions, varying PEG content (10–20 wt%), screw design (kneading or conveying), and screw speed (100–175 rpm). Granules were analyzed in-line using NIR spectroscopy (1100–2300 nm), collecting 23 spectra per condition. Spectral data were preprocessed (SNV, derivatives) and analyzed via Principal Component Analysis (PCA) and Partial Least Squares (PLS) regression. These conditions reflected a design space supported by prior studies¹. Additionally, ungranulated mixtures with 15 wt% PEG were conveyed through the granulator and their spectra were compared to granulated samples. Measurements were taken immediately after production and after cooling to ambient temperature to assess sample temperature effects. PLS models showed excellent predictive performance for PEG content, with R² values of 0.9998 (raw) and 0.9983 (SNV). PCA graphically discriminated between granulated and non-granulated samples, and between screw configurations, especially using raw and SNV data. Temperature effects on spectral profiles were evident: granules measured post-processing and at room temperature formed separated PCA clusters, especially with derivative preprocessing. These findings confirm that NIR spectroscopy, combined with multivariate data analysis, enables non-destructive, real-time monitoring of formulation and process parameters in TSMG, supporting its integration as a robust PAT tool for enhancing control and product quality in pharmaceutical continuous manufacturing.

Atherosclerosis-driven arterial occlusions remain a major cause of ischemic complications, including stroke, myocardial infarction, and limb loss. Although surgical interventions have advanced, the absence of targeted pharmacological strategies continues to limit therapeutic success, particularly due to the systemic toxicity of many promising drugs. Localized vascular drug delivery offers a new frontier, allowing for the modulation of inflammation at early stages of atherosclerotic plaque formation. In this study, we developed a nanocarrier-based delivery platform consisting of liposomes (LPs) and polymeric nanoparticles (PNPs) designed to encapsulate therapeutic monoclonal antibodies (mAbs) and polyphenols. To enable site-specific delivery, the nanocarriers were functionalized with immunouteroglobin-1 (IUG-1), a recombinant protein that selectively binds to the extra-domain B of fibronectin, which is overexpressed in atherosclerotic lesions. LPs were produced via a thin-film hydration technique, while PNPs were obtained using a water/oil/water double-emulsion solvent evaporation method. IUG-1 conjugation was achieved through carbodiimide/N-hydroxysuccinimide chemistry. Nanocarrier characterization included scanning electron microscopy, nanoparticle tracking analysis, and dynamic light scattering to determine morphology, size distribution, and ζ-potential. Both LPs and PNPs showed average diameters of approximately 200 nm and a stable negative surface charge. IUG-1 functionalization led to an increased particle size and an altered ζ-potential, confirming successful surface modification. LP and PNP exposure to human serum for 24 or 48 hours revealed good stability. Drug release profiles were evaluated to confirm controlled delivery over time. A sustained release of mAbs was observed in human serum over time, and biocompatibility tests on endothelial cells, macrophages, and red blood cells demonstrated high tolerance across all concentrations without cytotoxicity or hemolytic effects. These results suggest that the developed IUG-1-functionalized nanocarriers represent a promising and versatile platform for the targeted therapy of atherosclerosis, offering enhanced site-specific efficacy and reduced systemic toxicity. These achievements lay the groundwork for preclinical evaluation in vascular medicine.

Abstract

Introduction:
Zolpidem, a widely prescribed non-benzodiazepine hypnotic from the imidazopyridine class, is used for the short-term treatment of insomnia due to its sedative and sleep-inducing properties. However, its rapid onset, strong sedative effects, and amnesic properties make it a substance of choice in drug-facilitated crimes (DFCs). Increasing reports of zolpidem misuse emphasize the need for novel detection methods in forensic toxicological investigations.

Method:
To address the need for a robust and eco-friendly analytical approach, a miniaturized Vortex-Assisted Ultrasonication-based Cellulose Paper Sorptive extraction in combination with Liquid chromatography mass spectrometry/mass spectrometry (VAUS-CPSE-LC-MS/MS) method was developed and validated for the quantification of Zolpidem in different beverages, including alcoholic, non-alcoholic, and water samples. Isopropanol alcohol (IPA) was selected as the elution solvent and a “2×2” cm sized filter paper was selected for the experiment. To optimize the method, the statistical tool “Minitab Statistica 22.0” was employed where the Placket Burman Design and Central Composite Design were formed to identify influential parameters.

Results:
Optimal conditions for the extraction were a vortex time of 90 seconds, ultrasonication time of 12.5 minutes, pH of 7.5, extraction time of 6 minutes, back-extraction solvent volume of 1.5 mL, and 6.5 pieces of filter paper. Chromatographic separation was achieved using UHPLC with a binary mobile phase of 0.1% formic acid in Milli-Q water (Solvent A) and 0.1% formic acid in acetonitrile (Solvent B). The calibration curve for target analytes was linear in the range of 1.325–1000 ng/mL with a regression coefficient (R²) of 0.999. The method demonstrated excellent sensitivity with limits of detection (LODs) and limits of quantification (LOQs).

Conclusion:
The developed VAUS-CPSE-LC-MS/MS method proved to be a sensitive, eco-friendly, and cost-effective approach for the detection and quantification of Zolpidem in complex beverage matrices. This method offers strong potential for application in forensic toxicology investigations involving drug-facilitated crimes, robberies, and suspicious deaths.