Synergistic Effects of Genipin and Alendronate in 3D-Bioprinted Gelatin-Polyvinylpyrrolidone Scaffolds

In this study, we introduce a one-step semi-solid extrusion 3D printing strategy to fabricate gelatin-polyvinylpyrrolidone (GAG-PVP) scaffolds loaded with a low dose (0.5 wt%) of alendronate (ALN) and crosslinked in situ with 1 wt% genipin. The genipin-crosslinked ALN scaffold (GAG-PVP-GEN-ALN) demonstrated enhanced functional performance compared to both non-crosslinked GAG-PVP and GAG-PVP-ALN controls. Its peak swelling reached 462% at 5 h, surpassing the 362% of the unmodified scaffold and preventing the rapid dissolution observed for GAG-PVP-ALN, before gradually deswelling for 96 h. Water contact angle measurements confirmed that genipin fully restored surface hydrophobicity (101.9°), counteracting the pronounced wettability induced by ALN (47.8°) and exceeding the 78.2° of the GAG-PVP matrix, which is consistent with swelling ratio. Differential scanning calorimetry (DSC) indicated enhanced thermal stability of the crosslinked gelatin, with shifts in both glass transition and denaturation temperatures reflecting greater molecular rigidity despite the presence of glycerol as a plasticizer. Mechanical testing showed that while alendronate alone reduced mechanical performance, the combined inclusion of alendronate and genipin significantly enhanced scaffold properties compared to gelatin-polyvinyl pyrrolidone blend: tensile strength increased from 19.7 MPa to 39.8 MPa, elastic modulus rose from 805 MPa to 1174 MPa, and microhardness improved from 9.24 MPa to 22.3 MPa, values nearing those of native cancellous bone. The sustained ALN release profile extended from an abrupt 3 h burst in GAG-PVP-ALN to a controlled 48 h delivery in GAG-PVP-GEN-ALN, following first-order kinetics. Both direct and indirect cytotoxicity assays confirmed high cell viability (> 85%) without morphological abnormalities. These results highlight that embedding low-dose ALN within a genipin-crosslinked gelatin-PVP network results in a mechanically robust, biocompatible scaffold with tunable swelling and prolonged drug release, offering a versatile platform for localized bone tissue engineering

Influence of patient age and sex on drug-induced liver injury caused by antiseizure medications: a disproportionality analysis of VigiBase

Background: While it has been recognised that children may be at a greater risk of developing valproic acid-induced hepatotoxicity, it is less known if age and sex represent risk factors for drug-induced liver injury (DILI) caused by other antiseizure medications (ASMs). Aim: To investigate the influence of patient age and sex on DILI caused by ASMs. Method: To estimate hepatotoxic potential of 24 ASMs, deduplicated safety reports from VigiBase were accessed and disproportionality analysis was conducted by calculating Empirical Bayesian Geometric Mean and its 90% confidence interval (EB05, EB95). For ASMs with identified signals of disproportionate reporting (EB05 > 2, N > 0) for at least one hepatotoxicity-related event, we compared reporting frequency of overall hepatotoxicity among different age groups and between males and females by combining all events of interest into one custom term “overall DILI”. The same analysis was conducted using the custom term “liver failure”. Results: Signals of disproportionate reporting were identified for 14 (58.33%) ASMs. Valproic acid and carbamazepine had the highest number of identified signals (25 and 17, respectively). Among the 14 ASMs with identified signals, seven (50%) had disproportionally higher reporting of overall DILI in children, one (7.14%) in older people, and nine (64.29%) in females. Further, six (42.86%) ASMs had disproportionally higher reporting of liver failure in children and three (21.43%) in females. Conclusion: Our study suggests the importance of considering patient age and sex when assessing the risk of DILI from specific ASMs. However, the results are mainly hypothesis-generating and the identified signals require further investigation

Fumaria officinalis Dust as a Source of Bioactives for Potential Dermal Application: Optimization of Extraction Procedures, Phytochemical Profiling, and Effects Related to Skin Health Benefits

Fumaria officinalis (fumitory), in the form of dust, was employed as a source of bioactive extracts whose chemical profile and biological potential were investigated. According to the results of the optimization of the extraction protocol, the extract with the highest polyphenol yield was prepared using fumitory dust under the optimal conditions determined using the statistical tool, 23 full factorial design: 50% ethanol and a 30:1 mL/g ratio during 120 s of microwave extraction (22.56 mg gallic acid equivalent/g of plant material). LC-MS and spectrophotometric/gravimetric analyses quantified the polyphenol, flavonoid, tannin, alkaloid, and protein contents. Caffeoylmalic acid, quercetin dihexoside, quercetin pentoside hexoside, rutin, and methylquercetin dihexoside were the most dominant compounds. The highest total flavonoid, condensed tannin, alkaloid, and protein yields were determined in the extract prepared using microwaves. In addition to the proven antioxidant potential, in the present study, the anti-inflammatory activity of fumitory extracts is also proven in the keratinocyte model, as well as a significant reduction of H2O2-induced reactive oxygen species production in cells and the absence of keratinocyte cytotoxicity. Thus, detailed chemical profiles and investigated biological effects related to skin health benefits encourage the potential application of fumitory dust extracts in dermo-cosmetic and pharmaceutical preparations for dermatological circumstances

Associations of excessive gestational weight gain with changes in components of maternal reverse cholesterol transport and neonatal outcomes

Excessive gestational weight gain may be associated with unfavorable pregnancy outcomes. We explored the impact of excessive weight gain on components of HDL metabolism in maternal plasma: sterol composition of HDL particles, distribution of HDL subclasses and SCARB1, ABCA1 and ABCG1 genes expressions and their associations with newborns' characteristics. The study included 124 pregnant women, 58 with recommended and 66 with excessive weight gain. Concentrations of cholesterol synthesis marker, desmosterol, within HDL increased during pregnancy in both groups of participants. In women with excessive weight gain, levels of cholesterol absorption marker, campesterol, within HDL were significantly lower in the 3rd trimester compared to the 1st and 2nd trimesters. Relative proportions of large HDL 2b subclasses increased during pregnancy in women with recommended weight gain. Women with high pre-pregnancy BMI and excessive gestational weight gain had the lowest levels of β-sitosterol within HDL and the highest relative proportions of HDL 3a and HDL 3b subclasses in the 2nd trimester. Large HDL 2b particles were in positive correlation, while smaller HDL 3 subclasses and SCARB1 gene expressions were in negative correlation with APGAR scores. In conclusion, excessive weight gain could contribute to altered metabolism of HDL, and subsequently to poorer neonatal outcomes

Strawberry and Drupe Fruit Wines Antioxidant Activity and Protective Effect Against Induced Oxidative Stress in Rat Synaptosomes

The aim of this study was to investigate the antioxidant capacity of fruit wines and their protective effects against hydrogen peroxide-induced oxidative stress in rat synaptosomes in vitro. The wines were produced from strawberries and drupe fruits (i.e., plum, sweet cherry, peach, and apricot) through microvinification with a pure S. cerevisiae yeast culture. Fruit wines were produced with and without added sugar before the start of fermentation, whereas subvariants with and without pits were only applied to drupe fruit wines. First, synaptosomes were treated with the wines, while oxidative stress was induced with H2O2. Subsequently, the activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)) and the content of malondialdehyde (MDA), an indicator of membrane injury, were determined. In addition, the Briggs–Rauscher reaction (BR) was used to evaluate the inhibition capacity against free radicals. All investigated fruit wines increased the activity of the studied antioxidant enzymes and decreased MDA content compared to the corresponding controls (synaptosomes treated with H2O2). After synaptosomal treatment with plum wine, the highest activities were observed for SOD (5.57 U/mg protein) and GPx (0.015 U/mg protein). Strawberry wine induced the highest CAT activity (0.047 U/mg protein) and showed the best ability to reduce lipid peroxidation, yielding the lowest MDA level (2.68 nmol/mg). Strawberry, plum, and sweet cherry wines were identified as samples with higher antioxidant activity in both principal component analysis (PCA) and hierarchical cluster analysis (HCA). Finally, plum wine exhibited the highest inhibitory activity in the BR reaction (397 s). The results suggest that fruit wines could be considered potential functional food due to their protective effects against oxidative stress

Parenteral Nanoemulsion for Optimized Delivery of GL-II-73 to the Brain—Comparative In Vitro Blood–Brain Barrier and In Vivo Neuropharmacokinetic Evaluation

Background/Objectives: GL-II-73 is a positive allosteric modulator that is selective for α5GABAA receptors and has physicochemical properties that favor nanocarrier formulations when parenteral delivery to the central nervous system is desired. Our aim was to develop an optimized nanoemulsion containing GL-II-73 and subsequently test whether this would improve permeation across the blood–brain barrier (BBB) and availability in the brain. Methods: The nanoemulsions were formulated and subjected to detailed physiochemical characterization. The optimized formulation was tested in comparison to a solution of GL-II-73 in the appropriate solvent in an in vitro model of the blood–brain barrier based on human induced pluripotent stem cell-derived microvascular endothelial cells, astrocytes, and pericytes. Plasma and brain exposure to GL-II-73 and its metabolite MP-III-022 was investigated in an in vivo neuropharmacokinetic study in rats exposed to the selected nanoemulsion and the conventional solution formulation. Results: The selected biocompatible nanoemulsion exhibited satisfactory physicochemical properties for parenteral administration, with a Z-ave of 122.0 ± 1.5, PDI of 0.123 ± 0.009 and zeta potential of −40.7 ± 1.5, pH of 5.16 ± 0.04, and adequate stability after one year of storage, and allowed the localization of GL-II-73 in the stabilization layer. The permeability of GL-II-73 through the BBB was twice as high with the selected nanoemulsion as with the solution. The availability of GL-II-73 and MP-III-022 (also a positive allosteric modulator selective for α5GABAA receptors) in the brain was 24% and 61% higher, respectively, after intraperitoneal administration of the nanoemulsion compared to the solution; the former increase was statistically significant. Conclusions: The increased permeability in vitro proved to be a good predictor for the improved availability of GL-II-73 in brain tissue in vivo from the formulation obtained by encapsulation in a nanoemulsion. The putative additive effect of the parent molecule and its metabolite MP-III-022 could lead to enhanced and/or prolonged modulation of α5GABAA receptors in the brain

Urinary biomarkers in prediction of subclinical acute kidney injury in pediatric oncology patients treated with nephrotoxic agents

Background: Acute kidney injury (AKI) is a common complication in pediatric oncology patients, most often caused by nephrotoxic drugs. We aimed to assess whether levels of urinary kidney injury molecule-1 (uKIM-1), neutrophil gelatinase-associated lipocalin (uNGAL), liver fatty acid binding protein (uL-FABP) and Vanin-1 (uVNN-1), individually and in combination-integrated could be early markers for cytotoxic treatment induced AKI. Methods: Children with different malignant diseases treated with cisplatin (CIS) or ifosfamide (IFO) were included. AKI was defined using pediatric KDIGO (Kidney Disease Improving Global Outcomes) criteria by comparing pretreatment serum creatinine (sCr) values with those acquired at 48 h after the first or second chemotherapy cycle. Five serum (at baseline, 2, 6, 24 and 48 h after treatment) and four urine samples (at baseline, 2, 6 and 24 h after treatment) were obtained. Urinary biomarkers (uBm) were normalized to urine creatinine. Results: Thirty-eight patients were assessed. Within 48 h following chemotherapy 6 (15.79%) patients experienced AKI. Patients with AKI were younger and tend to have lower baseline sCr values than patients without AKI, but these differences were not statistically significant. Compared to baselines, all uBm were significantly increased during the first 6 h while sCr concentrations did not change significantly during the study period. The median increases in uBm during the first 6 h after treatment were 529.8% (interquartile range – IQR, 63.9-1835.2%) – 2194.0% (IQR, 255.3-4695.5%) in AKI vs. 302.2% (IQR 114.6-561.2%) -429.8% (156.5–1467.0%) in non-AKI group depending of tested uBm. The magnitude of these changes over time didn’t differ significantly between groups. The area under receiver operator curve (AUC) for uL-FABP and uNGAL at 24 h after chemotherapy were 0.81 and 0.72, respectively. The ROC analysis revealed that the other individual biomarkers’ performance at any time-point wasn’t statistically significant (AUC < 0.7). A model of integrated-combined uBm, 2 h (AUC 0.78), 6 h (AUC 0.85) and 24 h after (AUC 0.92) treatment with CIS and/or IFO showed good utility for early AKI prediction. Conclusions: The results of this study support that the use of the uBm to improves early AKI prediction in patients receiving CIS and/or IFO containing chemotherapy. Further studies on larger comparable groups of patients are needed

Antibiofilm Effects of N-Acetyl Cysteine on Staphylococcal Biofilm in Patients with Chronic Rhinosinusitis

Staphylococcal bacterial biofilm plays an important role in the pathogenesis and bacterial persistence of chronic rhinosinusitis. N-acetyl cysteine (NAC) has an inhibitory role in biofilm formation, suppressing adhesion and matrix production or favoring dispersal of preformed biofilm. The aim of this study was to examine the in vitro effect of NAC on Staphylococcal biofilm formation by bacterial strains isolated from tissue samples of patients with chronic rhinosinusitis with or without nasal polyps (CRSwNP and CRSsNP). Prospective study included 75 patients with CRS. The biofilm-forming capacity of isolated strains was detected by microtiter-plate method and the effects of sub-inhibitory (1/2x, 1/4x, and 1/8x minimal inhibitory concentration, MIC) and supra-inhibitory minimal concentrations (2x, 4x, and 8xMIC) of NAC on biofilm production were investigated. Staphylococcal bacterial strains were isolated in 54 (72%) patients, and the most frequently isolated species were Staphylococcus aureus (40.7%). Coagulase-negative Staphylococci species were weak producers of biofilm, while S. aureus was a strong biofilm producer. Concentration of 3.1 mg/mL (1/2 MIC) was sufficient to completely prevent biofilm formation in 77.8% of the isolates, where 49.6 mg/mL (8xMIC) led to the complete eradication of formed biofilm in 81.5% of the isolates. The subinhibitory and eradication effects were dose- and strain-dependent. There were no significant differences in MIC values between isolates from patients with CRSwNP and CRSsNP isolates. NAC proved to be effective in inhibiting biofilm formation and reducing formed biofilm by Staphylococcal isolates from patients with CRS. A comparable antibiofilm effect was exhibited in both phenotypes of CRS, indicating that NAC’s antibiofilm activity was independent of the underlying clinical phenotype, and more targeted on biofilm matrix components

Assessment of structural and functional characteristics of HDL and LDL in lung cancer patients in order to elucidate mechanisms of cholesterol metabolic pathways disorders

Introduction: Cholesterol metabolism dysregulation is recognized as one of the hallmarks of the cancer with highest mortality rate and the second most common malignancy – lung cancer (LC). LDL and HDL particles, the latter being carriers of the antioxidant paraoxonase-1 (PON1), were already proven to be altered in cancer pa- tients. We have tried to investigate in more depth the cholesterol metabolism perturbances in LC, by analzying content of each of the LDL and HDL subclass and (anti)oxidative activity of each of the HDL subclass separately. Materials and Methods: LDL and HDL subclasses from blood samples of 89 LC patients and 84 healthy subjects were separated and HDL subclasses PON1 activity assessed using Rainwater method and Gugliucci’s zymogram method, respectively. Results: LC patients had higher relative proportion of HDL 2 particles, lower proportion of HDL 3 particles, and significantly lower activity of PON1 compared to control group (CG). Relative proportion of PON1 activity was higher on HDL 2b fraction and lower on all HDL 3 fractions of LC patients compared to CG. Relative proportions of LDL I and LDL II particles were increased, while proportions of LDL IV and small dense LDL particles were decreased in LC patients. Relative proportions of HDL and LDL subfractions and PON1 activities on HDL sub- fractions were found to be dependent on LC type and size, number of comorbidities and sites of progression, and overall response to therapy. Conclusion: PON1 activity and lipoprotein subfractions distribution seem to be indicators of possible metabolic pathways (disorders) in L

Rational design of SIRT2 inhibitors: Integrating 3D-QSAR, molecular docking and machine learning techniques

SIRT2 influences processes such as apoptosis, DNA repair, and the cell cycle, making it an attractive target for therapeutic intervention in cancer treatment. Although potent inhibitors have been developed, the main challenge remains the design of highly selective SIRT2 inhibitors, especially due to the structural similarity of the NAD+ binding site across other members of the sirtuin family. Currently, derivatives of 5 ((3-amidobenzyl)oxy)nicotinamides stand out as some of the most selective and effective SIRT2 inhibitors, paving the way for further research toward the development of new therapeutically relevant molecules. [1] In this study, we developed a 3D-Quantitative Structure-Activity Relationship (3D-QSAR) model based on a dataset of 86 nicotinamide-based SIRT2 inhibitors complemented by GRIND-derived pharmacophore models. [2] External validation confirmed the reliability of the 3D-QSAR model in predicting SIRT2 inhibition within the defined range of application. [3] The model interpretation enabled the design of novel SIRT2 inhibitors. Furthermore, based on molecular docking results for the SIRT1–3 isoforms, we developed two classification machine learning models to predict the selectivity of inhibitors for the SIRT1/2 and SIRT2/3 isoforms, which was confirmed by external validation. The integration of 3D-QSAR, selectivity modeling and ADMET predictions facilitated the identification of promising selective SIRT2 inhibitors (Figure 1). These in silico identified inhibitors will be synthesized and tested in vitro to confirm their efficacy and selectivity and pave the way for novel SIRT2-targeted therapies for cancer and neurodegenerative diseases.33rd Annual GP2A Medicinal Chemistry Conference, XIVth Paul Ehrlich MedChem Euro-PhD Network Meeting & COST Action OneHealthdrugs, Abstract Book Nantes Université – France, 11th – 13th June 202