SARS‐CoV‐2 infection is driven by extensive interactions between viral proteins and host cellular factors, yet the structural properties of host proteins within these interaction networks remain incompletely understood. Intrinsically disordered proteins and regions are key contributors to protein–protein interaction networks due to their conformational flexibility and associated with it multifunctionality, binding promiscuity, and regulatory versatility. In this study, we performed a systematic, proteome‐wide analysis of intrinsic disorder in human proteins interacting with SARS‐CoV‐2 by integrating five experimentally validated interaction datasets comprising 2055 unique host proteins. Using disorder prediction, structural confidence assessment, functional and domain annotation, protein–protein interaction network analysis, phase‐separation propensity estimation, and independent validation with the D2P2 platform on a selected set of proteins, we characterized the structural organization of the SARS‐CoV‐2 human interactome. Our results reveal a balanced distribution of ordered and disordered host proteins, distinct functional and domain signatures across disorder classes, consistent inverse relationships between disorder and structural confidence, and increased network connectivity and phase‐separation propensity among highly disordered interactors. These findings indicate that SARS‐CoV‐2 exploits structural diversity within the host proteome rather than preferentially targeting a single disorder class and highlight intrinsic disorder as a key contributor to interaction plasticity and network organization at the systems level.

This case report presents an updated interpretation of genetic and chronological data from human remains discovered in Bezdanjača Cave, a Bronze Age burial site located in the Lika region of Croatia. The cave contains a complex necropolis with at least 57 graves and up to 200 individuals, which indicates its use as a collective burial site during the Middle and Late Bronze Age. Based on ancient DNA analysis, 13 males were identified among 38 analyzed individuals, with the majority belonging to the Y-chromosome haplogroup R1b, commonly associated with Bronze Age populations. However, two individuals were assigned to haplogroup I2a1a (I-Y3120). The I2a lineage has deep roots in Europe, and its presence has been confirmed in prehistoric contexts in Croatia and the region. However, newly obtained radiocarbon dates from occipital bones reveal that at least one of the two I2a1a individuals from Bezdanjača Cave dates to the Early Modern period (1645-1950 calibrated CE), which indicates that the remains were deposited in the cave much later than previously assumed. At the same time, these new data do not contradict the possible presence of the I2a1a lineage in Bronze Age populations in this area, as Bronze Age I2a1a samples have been reported from other archaeological sites in Croatia and the wider region. These findings, presented here for the first time, highlight the risks of assuming chronological homogeneity based solely on archaeological context and demonstrate the necessity of direct radiocarbon dating when integrating archaeological and genetic data. An interdisciplinary approach and careful chronological verification in ancient DNA research are essential to avoid misinterpretations in broader population genetic studies.

Aim To investigate integrated biochemical and glycomic signatures in humans related to coordinated metabolic and endocrine adaptations during prolonged fasting, which are essential for maintaining systemic homeostasis. Methods This single-arm longitudinal interventional study enrolled five healthy adults who underwent a 72-hour water-only fast. Blood samples were collected at baseline (T0), immediately after fasting (T1), and after 11 days of refeeding (T2). A broad panel of biochemical, hormonal, inflammatory, and glycomic parameters was determined. Time-dependent differences were evaluated using the Friedman test. Results Fasting induced a distinct biphasic response across multiple circulating markers, with significant alterations in total cholesterol, C-reactive protein, thyroid-stimulating hormone, and free triiodothyronine at T1 followed by recovery toward baseline at T2. Insulin and glucose concentrations declined during fasting and increased after refeeding, although these changes did not reach statistical significance. Plasma, immunoglobulin G (IgG), and IgA N-glycosylation profiles were extensively remodeled, which indicated dynamic metabolic and immune system adaptation. Liver enzymes, electrolytes, and most lipid fractions exhibited only minor and reversible fluctuations. Conclusion A 72-hour fast was associated with metabolic and hormonal changes consistent with an environment conducive to autophagy, although autophagy was not measured directly. The protocol appeared feasible and well tolerated in a small cohort of healthy adults. Given the limited sample size, these findings should be considered preliminary and require confirmation in larger studies incorporating direct autophagy markers and additional time points.

In the realm of preventive medicine, reinforcement of a zero-tolerance stance on cigarette smoking is imperative. The use of heat-not-burn (HnB) technology by individuals who are unable to quit smoking has prompted strong discussions in both the public health and clinical domains. This raises the question of whether exposure to cigarettes or HnB products has a different effect on blood pressure (BP) values and whether this effect depends on the concentration of heavy metals. A total of 33 participants were divided into four exposure groups: passive HnB exposure (n=7), active HnB users (n=4), passive cigarette smokers (n=11), and active cigarette smokers (n=9). Blood concentrations of cadmium (Cd) and lead (Pb) were determined by atomic absorption spectroscopy (AAS) using a Graphite Furnace Atomic Absorption Spectrometer, Agilent 240Z AA (Agilent Technologies, Santa Clara, CA, USA). Samples were prepared by wet digestion with nitric acid solution. After centrifugation, the supernatant was transferred into vials for AAS analysis. Concentrations of Cd and Pb were measured before and after exposure, and changes were analyzed using generalized linear models (GLM) adjusted for sex, age, and body mass index (BMI). All participants were smokers who had stopped smoking five days prior to the experiment. None of the participants had any chronic diseases. After a five-day abstinence (washout period), they were exposed to active and passive consumption of HnB or traditional cigarettes. BP was measured before and after exposure to tobacco smoke. Average Cd levels were similar across all groups (p = 0.55): passive HnB 1.59 µg/L, active HnB 1.29 µg/L, passive cigarette smokers 1.65 µg/L, and active cigarette smokers 1.64 µg/L. Pb levels varied significantly by exposure type (p = 0.006), as follows: 35.95 µg/L with passive HnB exposure, 26.98 µg/L with active HnB exposure, 52.45 µg/L and 36.50 µg/L in passive and active cigarette smokers, respectively. Upon monitoring, an elevation in Cd levels was significantly associated with a rise in systolic pressure (p = 0.026) and a decrease in diastolic pressure (p = 0.006), indicating an increase in pulse pressure and potential arterial stiffness. Pb concentration showed no direct effect on hemodynamic parameters. These findings suggest that Cd and Pb levels vary based on type of exposure. There was a significant difference between traditional cigarettes and HNB products, with the latter exhibiting lower concentrations, as well as between active and passive exposure. Changes in BP tend to be more closely linked to Cd, as higher levels correlate with increased systolic and decreased diastolic pressures, indicating its role in arterial stiffness and early vascular dysfunction.

Atherosclerosis is a progressive vascular disease characterized by lipid-rich plaque accumulation, oxidative stress, and chronic inflammation, contributing to coronary heart disease, stroke, and peripheral arterial disease. This study investigated the impact of inflammation, vascular calcification, and statin therapy on redox balance in blood and carotid artery plaques, aiming to identify potential biomarkers for disease assessment. Thirty-two patients undergoing carotid endarterectomy provided 34 plaque samples. Enzyme activities in plaque/erythrocytes and –SH group concentration in plasma/plaque were measured. Pathological analysis was performed to determine inflammation/calcification grade, the presence of mast cells and plaque composition. The results showed that mast cells were associated with reduced non-protein –SH groups, indicating selective thiol consumption and serving as a qualitative marker of oxidative burden. Reduced catalase activity in erythrocytes was associated with advanced calcification, pointing to long-standing systemic oxidative stress. Statin therapy enhanced systemic superoxide-dismutase 1 activity, increased –SH groups, and modulated plaque-specific glutathione reductase activity, attenuating sex-related differences in redox regulation. These findings highlight the complex interplay between systemic and local oxidative processes in atherosclerosis through alterations in redox-related biomarkers such as plasma –SH group concentrations and catalase activity.

Modern football is characterized by playing both in defense and attack, which requires greater energy demands from players, and the amount and intensity of movement increases from year to year. The aim of the study was to determine differences in the distance covered and movement intensities of players in relation to their position in the team. The sample was players who played all 90 minutes of the knockout phase of the 2022 World Cup (N=224), according to positions: goalkeepers (n=31), defense (n=101), midfielders (n=61), and attackers (n=31). Data were taken from the official FIFA website (www.fifa.com): distance covered (m), distance covered in zone 1 (speed 0-7 km/h), in zone 2 (7-15 km/h), in zone 3 (15-20 km/h), in zone 4 (20-25 km/h) and in zone 5 (>25 km/h), number of runs in zone 4, number of sprints in zone 5, maximum achieved speed (km/h). Differences between positions were determined by discriminant analysis. Three discriminant functions were isolated that are statistically significant at the 99% level (sig.=0.000) (Can. Cor.=0.934; Can. Cor =0.492 and Can. Cor.=0.356). The highest correlations with the first function, which maximally differentiates positions (Wilks Lambda =0.085; sig.=0.000), have the variables: intensity of movement in zone 2, number of runs in zone 4 and total distance covered. The second function (Wilks Lambda =0.662; sig.=0.000) is determined by the maximum achieved speed and distance covered in zone 3. The third function (Wilks Lambda =0.873; sig.0.000) is determined by intensity of movement in zone 4, number of sprints, distance covered in zone 5 and in zone 1. Excluding the goalkeeper position, it is evident that positions in the team are approaching each other in relation to distance covered, intensity of movement and maximum speed of movement, which supports the thesis that in modern football, polyvalent football players who can be used in multiple positions in the team are increasingly profiled

Unergative verbs assign the agent theta role to subjects whereas unaccusative verbs occur with theme subjects. Theory suggests that, unlike subjects of unergative verbs, theme subjects of unaccusative verbs are merged in the post-verbal internal argument position and moved to the pre-verbal external argument position. In a cross-modal lexical priming experiment, we tracked (re‑)activation patterns of the subject in sentences with imperfective and perfective unergative and unaccusative verbs in Bosnian/Croatian/Serbian (BCS). We investigated the interplay between unaccusativity and verbal aspect. Our findings are that the subject of perfective unaccusative verbs is (re‑)activated post-verbally, at the gap position, whilst this is not the case for unergative verbs and imperfective unaccusative verbs.

Background/Objectives: Antimicrobial resistance (AMR) is a major public health problem driven partly by inappropriate antibiotic use. Students of health studies represent future healthcare professionals with an important role in patient education, infection prevention, and antimicrobial stewardship. This study assessed knowledge, attitudes, and behaviours regarding antibiotic use and AMR among students of the Faculty of Health Studies, University of Mostar. Methods: An anonymous cross-sectional online survey was conducted in March 2025 using a self-selected convenience sample. The questionnaire was adapted from a previously published survey among Cypriot university students and distributed through student WhatsApp groups and by e-mail. Of 1113 invited students, 220 completed the survey, yielding a response rate of 19.8%. Results: During the previous 12 months, 39.5% of respondents reported antibiotic use. Most respondents reported adherence to medical instructions regarding dosage and duration of therapy, while 20.5% reported self-medication with antibiotics and 29.5% reported keeping unused antibiotics at home. Approximately 42% perceived antibiotics as easy or very easy to obtain without a prescription. Only 36.4% of respondents correctly distinguished antibiotics from other medications. Although most respondents recognised that bacteria can develop resistance, misconceptions persisted regarding humans and viruses. Differences between study programmes were observed for some attitudes and perceptions, whereas gender and year of study were not significantly associated with most responses. Conclusions: Health studies students demonstrated partial knowledge of antibiotics and AMR, together with behaviours that may contribute to inappropriate antibiotic use. Strengthened curricular content on rational antibiotic use, infection management, infection prevention, and antimicrobial stewardship appears justified. The findings are also consistent with the need to consider broader stewardship measures, including better enforcement of existing prescription-only dispensing requirements in Bosnia and Herzegovina.

Background and Objectives: Vitamin D signaling plays critical roles in immune regulation, bone metabolism, and cellular differentiation across multiple tissues. However, the spatial and temporal expression patterns of key vitamin D signaling components—the vitamin D receptor (VDR) and the enzyme 1α-hydroxylase (encoded by CYP27B1)—during human nephrogenesis have not been mapped at the protein level. The primary objective of this study was to characterize VDR and 1α-hydroxylase expression across critical stages of human kidney development, complementing prior transcriptomic and single-cell descriptions, and to contextualize these developmental observations against the dysregulation of vitamin D pathway genes in adult renal and urothelial malignancies. Materials and Methods: Immunofluorescence analysis was performed on FFPE kidney tissue from 12 specimens (3 per stage) at 10, 22 and 38 gestational weeks and postnatally at 1.5 years. For each specimen, at least three non-adjacent sections were stained and 6 non-overlapping cortical fields were imaged at ×40 (18 fields per stage). Fluorescence-area percentages were quantified in ImageJ 1.54g, and group differences were assessed by one-way ANOVA with Tukey’s post hoc test at both field- and specimen-level. An accompanying bioinformatic analysis evaluated the differential expression of VDR, CYP27B1, and CYP24A1 in adult renal and urothelial malignancies (TCGA cohorts: KICH, KIRC, KIRP, BLCA) using unpaired Welch’s t-test, with Benjamini–Hochberg FDR correction applied across all 16 tumor-versus-normal comparisons (12 gene-wise + 4 post hoc log2(CYP24A1/CYP27B1) ratios). Results: VDR showed its highest mean fluorescence area at 10 weeks (3.40% (95% CI 3.24–3.56); field-level Tukey p < 0.0001 versus other stages) and its lowest at 22 weeks (0.69% (0.64–0.74)). 1α-hydroxylase was also highest at 10 weeks (5.44% (5.29–5.60); p < 0.0001) and stabilized at lower levels thereafter (3.04–4.26%). Co-expression of both proteins was observed throughout development except in 22-week glomeruli. In TCGA, all 12 significant gene-wise comparisons retained significance after BH FDR correction (q < 0.05). VDR showed cohort-specific dysregulation: reduced in KICH (q = 2 × 10−4) but increased in KIRC and KIRP (q = 2 × 10−4 for both). CYP24A1 was reduced in all three renal cohorts (q ≤ 0.029) and unchanged in BLCA. CYP27B1 showed cohort-specific direction (reduced in KIRC; increased in KICH, KIRP, and BLCA). Conclusions: This study provides an initial immunofluorescence-based spatial description of VDR and 1α-hydroxylase across human kidney development, revealing a coordinated redistribution from immature glomeruli at 10 weeks to mature tubular segments at later stages. The TCGA analysis demonstrates that vitamin D pathway dysregulation in renal carcinoma is cohort-specific and is not abolished by multiple-testing correction. Together, these results indicate that the developmentally engaged vitamin D pathway retains kidney-specific functional relevance in adult renal pathology and provide a baseline reference for future mechanistic studies.

Pomegranate peel, an abundant agro-industrial by-product, represents a sustainable source of bioactive polyphenols, particularly punicalagin, which has been associated with antioxidant and photoprotective potential. This study aimed to develop microemulsions (MEs) containing pomegranate peel extract for dermal delivery of punicalagin using biocompatible surfactant systems. Three MEs differing in surfactant–cosurfactant composition (ME-A, ME-P, and ME-E) were prepared. Each formulation solubilized 1% (w/w) of pomegranate peel extract and was evaluated regarding in vitro release behavior, skin permeation/retention, antioxidant activity, and in vitro sun protection factor (SPF). All investigated MEs provided sustained release of punicalagin (≈10–17% of the applied dose in 8 h). ME-A, based on an alkyl polyglucoside surfactant, showed a significantly higher cumulative release of punicalagin (60.4 µg/cm2) compared with ME-E and ME-P. In skin penetration/permeation studies, ME-A also exhibited the highest numerical total delivery of punicalagin (≈48.2 µg/cm2 after 24 h), although differences among formulations were not statistically significant. All formulations demonstrated high antioxidant activity in the DPPH assay and measurable in vitro photoprotective potential, with SPF values ranging from approximately 11 to 14. Overall, pomegranate peel extract-loaded MEs showed potential as dermal delivery systems capable of improving solubilization and modulating skin delivery of punicalagin. The combination of agro-waste-derived bioactives with biocompatible surfactants highlights the potential of these systems as sustainable approaches for skincare formulations.

Acute coronary syndrome (ACS) is increasingly prevalent among young patients. The prevalence of ACS among young women has also increased. It is well known that coronary artery disease (CAD) follows a different pattern in women and men. In order to provide timely and appropriate treatment, it is necessary to understand the gender differences in the risk factors, clinical profile, and early outcome in the young ACS patient population. Understanding and addressing the risk factors linked to the early onset of the disease is therefore essential to lowering the burden of premature CAD (PCAD). This study aimed to analyse the gender differences in the presence of major coronary risk factors, clinical presentation, diagnosis and immediate outcomes in patients with PCAD. We evaluated 218 consecutive patients who were hospitalised in the ICCU of cardiology department with diagnosis of PCAD between November 2023 and May 2025, after satisfying the inclusion criteria. All included patients were 45 years of age or younger, were admitted for their first acute coronary syndrome and underwent primary PCI with optimal revascularization results. The study included 72 female and 146 male patients. The mean age of females was 42.5 ± 2.22 years and males was 39.6 ± 3.85 years (p<0.001). Young women had significantly more risk burden of hypertension (70.2% vs 36.5% p<0.001) diabetes (38.9% vs 28.7%, p<0.001) and dyslipidaemia (61.4% vs. 38.4%, p<0.001) compared to young men. Among young males, smoking was significantly higher (76.5% vs. 25.2%, p<0.001). NSTEMI was more common presentation among females (22.5% vs 8.4%, p<0.001) with more prevalent non obstructive CAD (13.1% vs. 8.2%, p=0.15). Young male patients showed angiographically more severe CAD and greater frequency of multivessel CAD (22.8% vs 17.9%, p=0.58). Young female patients had significantly better mean ejection fraction (EF) (48.1±7.9% vs 39.2±9.8%, p=0.13). Young women had more often acute heart failure (Kilip class ≥ 2) and longer length of stay in the intensive care unit (12.4% vs.6.29%, P=0.04; 5.5 vs 2.7 days, p <0.001 respectively). Mortality rate in the first 48 hours of admission was higher in man than in woman (1.65% vs.1.13%, p=0.65). A distinct gender difference in the risk factor profile for PCAD supports an early, comprehensive, but gender-specific approach to prevention. Young patients with premature CAD may benefit from these findings in terms of risk factor modification and gender-based management approach.

This study aimed to examine the influence of effective playing time (EPT) on match running performance (MRP) in elite soccer players competing in the UEFA Champions League (UCL), while controlling for contextual factors. In total, 1371 individual observations, involving 437 outfield players, participating in 32 teams, were collected from 122 matches using an optical tracking system. The MRP variables included total distance (TD), low- (LIR), moderate- (MIR), and high-intensity running (HIR). Linear mixed models were applied, with match outcome, match location, and opponent quality included as fixed effects, and player and team identity as random effects. The results showed that EPT significantly influenced TD (f=5.01–14.20, all p<0.01) and LIR (f=3.23–21.93, all p<0.05) across most playing positions, with players covering 9–10% greater distances in matches with very long EPT compared to very short EPT. In contrast, no significant effects of EPT were observed for MIR and HIR across any playing position. These findings indicated that longer EPT primarily increased overall activity, mainly through low-intensity activities, while moderate- and high-intensity activities remained relatively stable. Importantly, this pattern was consistent across players from multiple national contexts. From a practical perspective, EPT should be considered when planning and monitoring overall load, whereas moderate- and high-intensity activities should be systematically developed and maintained through training independent of EPT.

Compounds from olive (Olea europaea L.) and pomegranate (Punica granatum L.) have many beneficial effects on human health. This review paper considers the inhibitory potential, under in vitro conditions, of bioactive components of olive and pomegranate on different enzyme systems. Research shows that olive polyphenols (oleuropein, hydroxytyrosol, luteolin, and oleocanthal), as well as pomegranate polyphenols (punicalagin, urolithin A, ellagic acid), inhibit cyclooxygenase and lipoxygenase enzymes, which are associated with inflammatory processes. They also show an inhibitory effect on acetylcholinesterase, butyrylcholinesterase, and β-secretase, which opens up the possibility of a strong neuroprotective effect. Olive and pomegranate polyphenols also have an inhibitory effect on enzymes involved in carbohydrate metabolism, such as amylase and glucosidase, and can help fight diabetes and regulate human metabolism. In addition, polyphenols and extracts of both plants showed an inhibitory effect on cytochrome P450 enzymes, which metabolize most drugs. These data open up the possibility of interactions with certain groups of drugs. The current evidence supports the view that olive and pomegranate polyphenols act as biologically versatile compounds with considerable pharmaceutical and nutraceutical potential. Future investigations integrating enzymology, metabolomics, molecular docking, and clinical validation will be essential for translating these promising in vitro findings into evidence-based therapeutic applications.

BACKGROUND Enteric infectious diseases claim more than 1 million lives annually and are among the top ten causes of death in children younger than 5 years. Remarkable global investment has been dedicated to enteric infectious disease prevention and control; however, the shifting global health landscape is testing the continuance of progress. To evaluate the current status and guide future interventions, we present the latest epidemiological estimates of enteric infectious diseases from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023 and assess progress towards the Global Action Plan for the Prevention and Control of Pneumonia and Diarrhoea (GAPPD) mortality target of fewer than 20 deaths per 100 000 children younger than 5 years by 2025. METHODS We quantified the incidence, mortality, and disability-adjusted life-years (DALYs) of enteric infectious diseases by age, sex, and year across 204 countries and territories from 1990 to 2023. In GBD 2023, the following were considered under the category of enteric infectious diseases: diarrhoeal diseases, enteric fever (typhoid and paratyphoid), invasive non-typhoidal Salmonella spp (iNTS) infections, and other intestinal infectious diseases. We also examined 15 aetiologies contributing to diarrhoeal diseases. Incidence and prevalence were estimated with DisMod-MR (version 2.1), a Bayesian meta-regression tool, drawing on data from systematic reviews, population-based surveys, claims data, and hospital sources. Cause-specific mortality was modelled with Cause of Death Ensemble Modelling based on data from sources including vital registration, mortality surveillance, verbal autopsy, and minimally invasive tissue sampling. Years of life lost and years lived with disability were computed and combined to derive DALYs. For aetiology-specific estimation, population-attributable fractions (PAFs) for 15 pathogens were derived with a counterfactual framework. Point estimates and 95% uncertainty intervals (UIs) were generated from 250 draws from the posterior distribution. FINDINGS In 2023, enteric infectious diseases resulted in an estimated 1·27 million (95% UI 0·963-1·68) deaths globally, declining from 3·69 million (3·04-4·56) in 1990. The global age-standardised mortality rate (ASMR) decreased from 74·1 (62·0-92·9) per 100 000 population to 16·4 (12·6-21·3) per 100 000 population during the same period. Diarrhoeal diseases accounted for most deaths in 2023 (1·11 million [0·811-1·54]), followed by enteric fever and iNTS. South Asia and sub-Saharan Africa remained the most affected regions in 2023, with 599 000 (441 000-882 000) and 501 000 (373 000-648 000) deaths due to enteric infectious diseases, respectively, predominantly from diarrhoeal disease. Rotavirus was the leading cause of all-age diarrhoeal disease deaths (PAF 16·3% [12·0-21·5]), followed by norovirus (10·2% [2·4-17·0]) and Shigella spp (9·3% [5·4-15·2]). Among children younger than 5 years, PAFs of deaths due to diarrhoeal diseases were 40·2% (32·5-48·5) for rotavirus, 24·0% (15·1-36·7) for Shigella spp, and 23·4% (13·7-34·3) for adenovirus. Across 204 countries and territories, 141 met the GAPPD mortality target in 2023. The driving aetiologies among countries that did not meet the target in 2023 varied slightly by GBD super-region, but the highest or second-highest number of deaths in children younger than 5 years were consistently attributed to rotavirus. Astrovirus and sapovirus, newly included in GBD 2023, were responsible for 24 600 (6290-49 000) and 18 800 (4650-44 400) deaths, respectively, in 2023, mainly in children younger than 5 years. INTERPRETATION Our findings show that mortality and ASMRs of enteric infectious diseases declined substantially between 1990 and 2023. This decline is consistent with the expansion of public health measures and broader socioeconomic development. However, the burden in 2023 remains considerably high, with the highest mortality concentrated in sub-Saharan Africa and south Asia. Considering that more than a quarter of all countries had yet to meet the GAPPD mortality target in 2023, sustained efforts are needed to address the persistent burden in affected countries and to adapt to the changing global health landscape. FUNDING Gates Foundation.