PubMed Central

In this study, we evaluated the impact of different in vitro 3D culture modelling methods on the activity of doxorubicin (DOX) and 5-fluorouracil (5-FU) in human melanoma spheroids. Human melanoma A375 and IGR39 spheroids were generated using the hanging drop and non-adhesive surface methods. Spheroid growth dynamics were assessed by measuring changes in spheroid diameter. To compare the effects of anticancer drugs in spheroids of different sizes, spheroids of approximately 200 and 400 µm were formed. Drug activity was evaluated based on spheroid growth and cell viability using the MTT assay. A375 spheroids formed using the non-adhesive surface method were more sensitive to DOX than spheroids formed using the hanging drop method. In smaller A375 spheroids, 10 µM 5-FU reduced cell viability more effectively in spheroids formed using the hanging drop method. In contrast, IGR39 spheroids formed by the hanging drop method were more resistant than those formed on a non-adhesive surface. However, in IGR39 spheroids, the effects of DOX and 5-FU on growth and viability did not significantly differ between formation methods. In conclusion, A375 spheroid growth was not significantly influenced by the formation method, whereas IGR39 spheroid growth depended on the method used. A375 spheroids formed on non-adhesive surfaces were more sensitive to DOX, whereas 5-FU activity depended on drug concentration and spheroid size. In IGR39 spheroids, the effects of DOX and 5-FU on growth and viability were largely independent of the spheroid formation method.

In view of the importance of texture and colour characteristics for consumer acceptance of fermented dairy products, this study aimed to evaluate the associations between selected whey proteins, fat-soluble vitamins, and instrumental quality traits of organic plain yoghurts. Physicochemical analyses included acidity, total protein, fat, whey proteins (β-lactoglobulin, α-lactalbumin, bovine serum albumin, lactoferrin, and lysozyme), and fat-soluble compounds (vitamins A, D3, and E, and β-carotene). Selected instrumental texture and colour attributes were also determined. Whey protein fractions were associated mainly with texture characteristics, whereas lipophilic vitamins and β-carotene were more closely related to colour attributes. Total protein content was positively associated with firmness (r = 0.510, p ≤ 0.05), while β-lactoglobulin was related to apparent viscosity (r = 0.705, p ≤ 0.05). In contrast, vitamin A, vitamin D3, β-carotene, and fat content were negatively associated with lightness and whiteness, but positively related to yellowness and chroma. Lactoferrin and lysozyme showed stronger relationships with selected secondary texture attributes, particularly gumminess and cohesiveness. Multiple regression analysis indicated that firmness and apparent viscosity were only moderately explained by the selected compositional predictors, whereas colour attributes were more strongly associated with fat-fraction components. The remaining variability likely reflected additional microstructural and processing-related effects. Although the results should be interpreted as associations rather than direct causal relationships, the findings support the concept of protein-driven texture and fat-driven colour development in plain yoghurts and improve understanding of the relationships between bioactive milk components and instrumental plain yoghurt quality.

The postpartum period in beef cows is characterized by complex physiological and reproductive changes that may influence the postpartum reproductive changes and estrus expression. This exploratory pilot study evaluated postpartum reproductive responses in Angus cows receiving grape extract supplementation while investigating associations among thermographic, behavioral, hormonal, and reticulorumen temperature indicators associated with estrus expression. Nineteen Angus cows were assigned to a control group (C; n = 10) or a treatment group receiving slow-release grape extract boluses (T; n = 9). From calving until artificial insemination, ocular and vulvar thermographic images, blood samples, and physiological measurements were obtained weekly between 10:00 and 11:00 a.m. Reticulorumen temperature and activity data were continuously recorded using intraruminal sensors. Kaplan–Meier survival analysis demonstrated an earlier onset of postpartum estrus in supplemented cows compared with controls (p = 0.010). Mean time to first estrus was 23.88 ± 1.86 days in the T group and 39.82 ± 5.05 days in the C group. No significant differences were observed between groups for most individual physiological or hormonal variables. Exploratory correlation analysis revealed moderate associations among vulvar temperature, ocular temperature, activity, estrus index, and reticulorumen temperature indicators. However, because diagnostic accuracy was not evaluated, these findings should not be interpreted as validation of estrus detection performance. The results suggest that multimodal physiological monitoring may provide complementary information related to postpartum estrus expression in beef cows, while grape extract supplementation may be associated with earlier postpartum reproductive recovery. Due to the exploratory study design and limited sample size, further studies are required to validate these preliminary observations.

Background: Existing therapies for xerostomia are primarily symptomatic, providing temporary mucosal hydration without addressing underlying pathological changes in the oral cavity. In this context, medicated chewing gums containing ascorbic acid and lysozyme hydrochloride offer a promising approach, combining antimicrobial, antioxidant, and trophic effects with physiological salivary stimulation and prolonged local delivery. Methods: For the development of compressed chewing gum formulation, the physicochemical (particle size distribution, moisture absorption capacity, and microscopic characteristics) and technological (flowability, angle of repose, bulk and tapped density, Carr’s index (CI), and Hausner ratio (HR)) properties of the active substances and their formulations with excipients were evaluated. Pharmacological activity was assessed in an atropine-induced xerostomia rat model. Results: The physical mixture of all components showed inferior flow properties compared with the formulation containing pre-granulated lysozyme hydrochloride, as evidenced by higher Carr’s index and Hausner ratio values (CI = 17, HR = 1.20 vs. CI = 13, HR = 1.14), indicating improved processability after pre-granulation. The effect of relative humidity during formulation was also assessed, with an optimal level of 40% required to ensure process stability due to the hygroscopic nature of the components. Based on these data, technological approaches ensuring processability were established, including wet pre-granulation of lysozyme hydrochloride and premixing of ascorbic acid to reduce oxidation risk. These approaches resulted in an optimized compression mass with excellent flowability (CI = 8, HR = 1.09), suitable for the preparation of medicated chewing gum. An optimal compression force (7 kN) ensured suitable rheological and textural properties, resulting in rapid and nearly complete release of the active ingredients from the medicated chewing gum, consistent with kinetic analysis. In vivo studies using an atropine-induced xerostomia rat model demonstrated that the combination of ascorbic acid and lysozyme hydrochloride significantly increased salivary secretion (2.17-fold vs. control pathology group) and reduced salivary gland mass coefficients (by 13–18% compared with the control pathology group and groups receiving individual active ingredients), alongside improvement of oxidative stress markers, including a reduction in TBA-reactants (by 51.6%) and an increase in catalase activity (by 51.0%). Conclusions: The developed medicated chewing gum showed favorable technological properties, efficient release of active ingredients, and anti-xerostomic activity in vivo, indicating its potential for xerostomia relief and oral health support.

Preterm birth remains a major global health concern, affecting approximately one in ten neonates, with an estimated 15 million infants born prematurely each year. Prematurity and clinical factors such as antibiotics, cesarean delivery, and limited access to mother’s own milk disrupt microbiota development in VLBW infants; although human milk supplies nutrients and a microbial community, its composition and clinical role are not yet well understood. However, the composition and clinical significance of the human milk microbiota (HMM) in VLBW infants remain insufficiently characterized. Background: This review aims to summarize recent evidence (2021–2025) on the microbiome of MOM in mothers of VLBW (<1500 g) preterm infants and to evaluate its potential role in neonatal health. Methods: The study used a systematic literature review, searching PubMed and Google Scholar with predefined criteria and keywords. Results and Conclusions: MOM microbiota of VLBW in infants is dominated by Staphylococcus, Enterococcus, Streptococcus, Enterobacteriaceae, and Acinetobacter, with lower levels of Veillonella, Clostridium sensu stricto, Pseudomonas, Haemophilus, and Bifidobacterium; its diversity increases over lactation, and feeding type influences infant gut colonization and immune development, though links to necrotising enterocolitis (NEC) remain limited. Further research using multi-omic approaches is needed to clarify these mechanisms and their clinical implications.

Background and aims.
The use of large language models (LLMs), such as ChatGPT, is rapidly expanding in clinical medicine but data in gastroenterology are limited. We aimed to assess the prevalence and patterns of LLM use among European gastroenterologists, evaluate perceived benefits and risks and identify barriers and priorities for future integration.
Methods.
A Europe-wide online survey was conducted between June and August 2025. A 24-item questionnaire was distributed via national gastroenterology societies and social media. Questions on specific applications, reliability and clinical impact were shown only to respondents with prior LLM experience.
Results.
A total of 966 gastroenterologists from 34 European countries responded, 55.3% reported using LLMs, mainly weekly (54.4%) or daily (20.2%). Common uses included education (59.2%), clinical decision support (58.2%) and scientific writing (39.1%). Among academic users, summarisation (80.1%) and grammar refinement (74.4%) were most frequent. Most users rated LLMs as moderately or highly reliable (81.0%), and 84.3% believed they improve outcomes. Nearly two-thirds (64.5%) felt LLMs can support endoscopy practice. Inaccuracies were reported by 42.7% and were associated with lower trust (ρ=–0.40, p<0.001). Although only 7.8% had formal training, 90.0% supported structured education and 85.2% favoured curricular integration. Most respondents would recommend LLM use (91.4%) and expect broader adoption within 5 years (93.4%). Reported barriers included institutional regulations (45.7%), cost (36.1%), complexity (29.4%) and time constraints (27.9%).
Conclusion.
LLMs are increasingly integrated into clinical and academic workflows in European gastroenterology. Structured training, specialty-specific development and clear governance are essential for safe implementation.

Context: Patients with congenital adrenal hyperplasia (CAH) often face challenges in achieving their target adult height. The QEPS growth model, previously used for assessing healthy and pathological growth patterns, has not been applied to individuals with CAH. Objective: To evaluate growth patterns in patients with CAH using the QEPS growth model and to compare their growth characteristics with a healthy reference population. Design: A retrospective longitudinal study analyzing growth data collected from 1986 to 2008.Setting: The study was conducted in a single tertiary care center. Patients: The study included 25 patients (13 girls) with CAH, subtyped into salt wasting (SW; 12 boys, 8 girls) and simple virilizing (SV; 5 girls). Growth data were compared with a healthy reference cohort. Interventions: All patients were treated with hydrocortisone, and patients with CAH-SW received mineralocorticoids. Main outcome measures: Growth patterns were analyzed using the QEPS model, which includes specific early life growth (E-function), basic childhood growth (Q-function), and specific pubertal growth (P-function). Final adult height was compared with the reference population and parental heights. Results: CAH-SW boys and girls were longer at birth, exhibited reduced early-life growth, and had reduced puberty-specific growth (lower Pmax), resulting in shorter adult height (1.7 SDS). CAH-SV girls had earlier pubertal growth onset, also leading to reduced adult height (−1.6 SDS). Both groups showed similar basic childhood growth. Conclusions: Patients with CAH-SW displayed distinct growth patterns, including longer birth length but reduced specific early and pubertal growth, resulting in shorter adult height compared with reference populations and parental heights.

To obtain genetic parameters relevant for the management of animal genetic resources, this study provides a comparative characterization of the Trakehner (TRAK) horse population in relation to four other horse populations maintained in Lithuania – Arabian (ARAB), Baltic Warmblood (BW), Lithuanian Warmblood (LW), and Zemaitukai (ZEM). These populations represent diverse histories, selection goals, and breeding practices. Pedigree data (n = 22 666) and parentage verification genetic data based on blood group markers (n = 1293) were used to evaluate genetic diversity, population structure, and inter-population relationships. Despite a 25 % population decline in recent years, the TRAK population maintained moderate inbreeding (F = 0.037) and a balanced genetic structure. The effective population size (Ne) was lower than in BW and LW but higher than in the endangered ZEM population. TRAK retained a high number of alleles (Na = 3.83), with observed heterozygosity (Ho = 0.460) slightly lower than that observed in open sport horse populations. Comparison of all individuals and the Lithuanian reference populations revealed stable or increased heterozygosity in currently breeding individuals, particularly within TRAK. Genetic distances and assignment tests confirmed close relationships among TRAK, BW, and LW, reflecting shared ancestry. In contrast, ARAB and ZEM populations remained genetically distinct. PCA and heatmap analyses further supported the separation between closed and open populations in the current breeding context. These findings highlight that population structure, historical gene flow, and management practices jointly shape genetic variability. The results can be used to optimize mating plans by balancing sire and dam contributions, monitor inbreeding and effective population size trends, manage gene flow between related populations (TRAK, BW, LW), and develop targeted conservation strategies for genetically distinct breeds such as ZEM and ARAB.

In recent years, the study of interleukins (ILs), crucial cytokines involved in inflammation, has garnered significant attention within coronary artery disease including atherosclerosis. This review provides a detailed overview of anti-inflammatory ILs, elucidating their functions within the pathogenesis of atherosclerosis. We examine aspects of all the known anti-inflammatory ILs role in atherosclerosis, the direct impact of these ILs on the inflammation; endothelial, smooth vascular cells and macrophage’s function; and their interactions with signaling pathways and molecules. The potential for diagnostic possibilities and targeted drug therapy to modulate anti-inflammatory ILs activity in atherosclerosis was explored. Taken together, findings from recent studies suggest that the main pathways through which ILs exerts its anti-inflammatory effects are: (1) taking part in the regulation of cholesterol transport or oxidised low-density lipoprotein (oxLDL) phagocytosis (IL-1Ra and IL-36Ra—indirectly); (2) affecting different blood cells’ participation in the inflammation (monocytes, lymphocytes, macrophages); (3) taking place in the remodelation of the arterial wall (affecting smooth muscle and endothelium cells). Overall, IL-35, IL-37, and IL-38 appear to be the most promising for modulation of signaling pathways in experimental works and could be investigated as treatment targets. Recombinant IL-10 is investigated in experimental models as therapeutic tool. IL-1Ra is started being translated into clinical practice already. IL-13 and IL-19 are the least studied. It turns out that anti-inflammatory ILs are unlikely to serve as diagnostic markers for atherosclerosis due to their limited specificity and inconsistent associations with disease progression, as well as insufficient validation in large human cohorts. Moreover, key challenges related to delivery, dosing, and safety remain unresolved.

Postoperative delirium is a common complication following sub-thalamic nucleus deep brain stimulation surgery in Parkinson's disease patients. Postoperative delirium has been shown to prolong hospital stays, harm cognitive function, and negatively impact outcomes. Utilizing radiomics as a predictive tool for identifying patients at risk of delirium is a novel and personalized approach. This pilot study analyzed preoperative T1-weighted and T2-weighted magnetic resonance images from 34 Parkinson's disease patients, which were used to segment the thalamus, amygdala, and hippocampus, resulting in 10,680 extracted radiomic features. Feature selection using the minimum redundancy maximal relevance method identified the 20 most informative features, which were input into eight different machine learning algorithms. A high predictive accuracy of postoperative delirium was achieved by applying regularized binary logistic regression and linear discriminant analysis and using 10 most informative radiomic features. Regularized logistic regression resulted in 96.97% (±6.20) balanced accuracy, 99.5% (±4.97) sensitivity, 94.43% (±10.70) specificity, and area under the receiver operating characteristic curve of 0.97 (±0.06). Linear discriminant analysis showed 98.42% (±6.57) balanced accuracy, 98.00% (±9.80) sensitivity, 98.83% (±4.63) specificity, and area under the receiver operating characteristic curve of 0.98 (±0.07). The feed-forward neural network also demonstrated strong predictive capacity, achieving 96.17% (±10.40) balanced accuracy, 94.5% (±19.87) sensitivity, 97.83% (±7.87) specificity, and an area under the receiver operating characteristic curve of 0.96 (±0.10). However, when the feature set was extended to 20 features, both logistic regression and linear discriminant analysis showed reduced performance, while the feed-forward neural network achieved the highest predictive accuracy of 99.28% (±2.71), with 100.0% (±0.00) sensitivity, 98.57% (±5.42) specificity, and an area under the receiver operating characteristic curve of 0.99 (±0.03). Selected radiomic features might indicate network dysfunction between thalamic laterodorsal, reuniens medial ventral, and amygdala basal nuclei with hippocampus cornu ammonis 4 in these patients. This finding expands previous research suggesting the importance of the thalamic-hippocampal-amygdala network for postoperative delirium due to alterations in neuronal activity.