Plasma proteomics technologies are advancing rapidly, offering new opportunities for biomarker discovery and precision medicine. Direct comparisons of available technologies are needed to understand how platform selection affects downstream findings. We compared the performance of a peptide fractionation-based mass spectrometry method (HiRIEF LC-MS/MS) and the Olink Explore 3072 proximity extension assays on 88 plasma samples, analyzing 1129 proteins with both methods. The platforms exhibited complementary proteome coverage, high precision, and concordance in estimating sex differences in protein levels. Quantitative agreement between platforms was moderate (median correlation 0.59, interquartile range 0.33-0.75), mainly influenced by technical factors. Finally, we present a publicly available tool for peptide-level analysis of platform agreement and demonstrate its utility in clarifying cross-platform discrepancies in protein and proteoform measurements. Our findings provide insights for platform selection and study design, and highlight the value of combining mass spectrometry and affinity-based approaches for more comprehensive and reliable plasma proteome profiling. Advancements in plasma proteomics have opened new avenues for biomarker discovery, necessitating a clear understanding of technological capabilities. Here, the authors compare HiRIEF LC-MS/MS and Olink Explore 3072, revealing complementary strengths and moderate quantitative agreement, and introduce PeptAffinity, a resource facilitating detailed peptide-level exploration of differences in protein quantification between platforms.

The aviation industry operates as a complex, dynamic system generating vast volumes of data from aircraft sensors, flight schedules, and external sources. Managing this data is critical for mitigating disruptive and costly events such as mechanical failures and flight delays. This paper presents a comprehensive application of predictive analytics and machine learning to enhance aviation safety and operational efficiency. We address two core challenges: predictive maintenance of aircraft engines and forecasting flight delays. For maintenance, we utilise NASA’s C-MAPSS simulation dataset to develop and compare models, including one-dimensional convolutional neural networks (1D CNNs) and long short-term memory networks (LSTMs), for classifying engine health status and predicting the Remaining Useful Life (RUL), achieving classification accuracy up to 97%. For operational efficiency, we analyse historical flight data to build regression models for predicting departure delays, identifying key contributing factors such as airline, origin airport, and scheduled time. Our methodology highlights the critical role of Exploratory Data Analysis (EDA), feature selection, and data preprocessing in managing high-volume, heterogeneous data sources. The results demonstrate the significant potential of integrating these predictive models into aviation Business Intelligence (BI) systems to transition from reactive to proactive decision-making. The study concludes by discussing the integration challenges within existing data architectures and the future potential of these approaches for optimising complex, networked transportation systems.

Cervical cancer is one of the leading causes of cancer in women, worldwide. Infection with humanpapillomavirus (HPV) has been accepted as the primary cause for the development of invasive cervicalcancer and its precursor lesions. Despite HPV infection has been proposed as an indispensable factor forcervical cancer development, only a subset of neoplastic lesions with HPV infection persist and progress toinvasive cancer. This suggests us that other molecular events are also involved in cancer progression. Aimof this study was to extract mRNA from cytobrush-collected healthy and HPV infected cervical epithelialcells and investigate various RNA extraction and purification protocols for assessment of RNA yield andquality. Taking into consideration that cervical cancer screening is based on the cytology basedPapanicolaou test (Pap test), main challenge is to investigate whether the samples obtained by regular Paptesting can be used for gene expression analysis. For this purpose, a total of 68 cervical specimens werepreviously tested for HPV infection. Following HPV testing, samples were submitted to RNA extractionand compared to the products after additional purification step involving DNase I. Products obtained afterdifferent RNA extraction and purification methods were visualized using 2% agarose gel electrophoresis.In conclusion, DNase I based RNA purification represents a necessary step for the assurance of a high-quality extracted RNA used for gene expression analysis studies. Reliance on commercial kits for RNAextraction only, without performing additional purification step can lead to errors in drawing finalconclusions and/or to false negative gene expression profiling, affecting the overall diagnostic procedure.According to obtained results, the type of sampling used in this study was not suitable for the subsequentgene expression analysis.

Abstract To preserve resources for future generations and promote rural development, supporting ecotourism is essential. This paper provides guidelines for developing ecotourism, highlighting its role in environmental conservation. While mass tourism benefits rural communities, it can cause significant environmental harm. Therefore, this research promotes ecotourism as a sustainable alternative. In rural areas, ecotourism supports development by responsibly using natural resources. The study focuses on the potential of rural settlements in the Semberija region of Bosnia and Herzegovina, assessing their capacity for ecotourism to aid local development. A decision model was developed, considering four main criteria - natural, infrastructure, socio-cultural, and economic - and their sub-criteria. This model evaluates six rural communities’ ecotourism potential. To determine the importance of each criterion, a fuzzy weighting method with the Bonferroni mean operator was used, revealing economic factors as the most influential. The fuzzy ranking method then ranked the settlements, with Amajlije identified as having the highest ecotourism potential. The findings suggest that promoting ecotourism in Amajlije and similar communities can support sustainable rural development, balancing environmental preservation with economic growth.

High welfare standards for animals used in research is as much an ethical issue as it is a cornerstone of high-quality science. Researchers can improve both animal welfare and data reliability by implementing strategies that reduce stress in experimental animals. One modern and effective approach is to monitor animals within their familiar home-cage environment. Home-cage monitoring (HCM) systems integrate multiple approaches to automatically, continuously, and non-invasively monitor the physiology and behaviour of laboratory animals within their home environments. HCM favours the animals’ natural rhythms and behaviours while reducing stress from various sources and the need for human intervention. In this article, we explore how HCM contributes to the 3Rs framework introduced by Russell and Burch and focus particularly on how to select the most appropriate HCM system for specific research needs. We discuss available resources and practical limitations for system choice, and provide a brief outlook on the evolving role of artificial intelligence to analyse HCM data. We also discuss the opportunities and barriers to HCM adoption, particularly in relation to countries with developing research structure and limited funding in Europe. Our central message is clear: use of HCM technologies supports 3Rs and promotes both better science and better animal welfare. Pametne kletke, večja dobrobit: Podpora načelom 3R v raziskavah na živalih s spremljanjem v domači kletki in ustrezno izbiro sistema Izvleček: Visoki standardi dobrobiti živali v raziskavah niso zgolj etična obveznost, temveč tudi temelj visokokakovostne znanosti. Raziskovalci lahko izboljšajo tako dobrobit živali kot tudi zanesljivost podatkov z uvedbo strategij, ki zmanjšujejo stres pri poskusnih živalih. Eden izmed sodobnih in učinkovitih pristopov je spremljanje živali v njihovem domačem okolju. Sistemi za spremljanje v domači kletki (HCM, angl. home-cage monitoring) združujejo več pristopov za samodejno, neprekinjeno in neinvazivno spremljanje fiziologije in vedenja laboratorijskih živali v njihovem domačem okolju. HCM podpira naravne ritme in vedenja živali ter zmanjšuje stres iz različnih virov in potrebo po posegih človeka. V članku opisujemo, kako HCM prispeva k načelom 3R, ki sta ga uvedla Russell in Burch, s posebnim poudarkom na izbiri najprimernejšega sistema HCM za specifične raziskovalne potrebe. Obravnavamo razpoložljive vire in praktične omejitve pri izviri sistema ter podajamo kratek pogled na razvijajočo se vlogo umetne inteligence pri analizi podatkov HCM. Prispevek obravnava tudi priložnosti in ovire pri uvajanju HCM, zlasti v povezavi z državami z manj razvito raziskovalno infrastrukturo in omejenimi sredstvi v Evropi. Naše osrednje sporočilo je jasno: uporaba tehnologij HCM podpira načela 3R ter spodbuja boljšo dobrobit živali in boljšo znanost. Ključne besede: laboratorijske živali; avtomatsko spremljanje vedenja; izboljšave; kontinuirno zbiranje podatkov; stres

In the Paschen minimum region, the mechanisms of avalanche vacuum breakdown, the anomalous Paschen breakdown mechanism, and the Townsend breakdown mechanism alternate. The boundaries between these mechanisms are not sharply defined, instead, they intermingle and influence each other. Since these electrical breakdown mechanisms are based on gas discharge, this study critically examines the mathematical model of electrical discharge mechanisms based on ionization expressions and coefficients according to Townsend and Takeishi. Based on this analysis, new expressions for the ionization coefficient are proposed, assuming a Maxwellian spectrum of free electron gas. Experimental verification has confirmed that such an ionization coefficient provides the best results in the Paschen effect region. By applying the derived algorithm for calculating breakdown voltage values and comparing them with corresponding experimental values, an explanation of the electrical breakdown mechanism of noble gases in the Paschen region is provided. Experiments were conducted under well-controlled laboratory conditions with minimal measurement uncertainty.

The heart’s relentless contractile activity depends critically on mitochondrial function to meet its extraordinary bioenergetic demands. Mitochondria, through oxidative phosphorylation, not only supply ATP but also regulate metabolism, calcium homeostasis, and apoptotic signaling, ensuring cardiomyocyte viability and cardiac function. Mitochondrial dysfunction is a hallmark of cardiomyopathies and heart failure, characterized by impaired oxidative phosphorylation, excessive production of reactive oxygen species (ROS), dysregulated calcium handling, and disturbances in mitochondrial dynamics and mitophagy. These defects culminate in energetic insufficiency, cellular injury, and cardiomyocyte death, driving heart disease progression. Diverse cardiomyopathy phenotypes exhibit distinct mitochondrial pathologies, from acute ischemia-induced mitochondrial collapse to chronic remodeling seen in dilated, hypertrophic, restrictive, and primary mitochondrial cardiomyopathies. Mitochondria also orchestrate cell death and inflammatory pathways that worsen cardiac dysfunction. Therapeutic strategies targeting mitochondrial dysfunction, including antioxidants, modulators of mitochondrial biogenesis, metabolic therapies, and innovative approaches such as mitochondrial transplantation, show promise but face challenges in clinical translation. Advances in biomarker discovery and personalized medicine approaches hold promise for optimizing mitochondrial-targeted therapies. Unlike previous reviews that examined these pathways or interventions individually, this work summarizes insights into mechanisms with emerging therapeutic strategies, such as SGLT2 inhibition in HFpEF, NAD+ repletion, mitochondrial transplantation, and biomarker-driven precision medicine, into a unified synthesis. This framework underscores the novel contribution of linking basic mitochondrial biology to translational and clinical opportunities in cardiomyopathy and heart failure. This review synthesizes the current understanding of mitochondrial biology in cardiac health and disease, delineates the molecular mechanisms underpinning mitochondrial dysfunction in cardiomyopathy and heart failure, and explores emerging therapeutic avenues aimed at restoring mitochondrial integrity and improving clinical outcomes in cardiac patients.

Heavy metals are persistent environmental pollutants with well-documented toxic, genotoxic, and bioaccumulative effects across ecosystems. This study evaluates blood cell morphology as a potential biomarker of environmental pollution in Miniopterus schreibersii (Kuhl, 1819), a migratory bat species. Individuals were captured from a contaminated site (Dardagani underground quarry) and a reference site (Mokra Megara Cave) in Bosnia and Herzegovina. Environmental sampling included guano, soil, and water. Guano was analyzed for nine heavy metals (Cr, Cu, Mn, Fe, Co, Ni, Cd, Pb, Zn) to assess site-specific contamination. Distinct spatial differences in metal accumulation were observed. Elevated Cu and Mn concentrations in guano from the contaminated site indicated anthropogenic input, whereas higher Fe, Ni, and Pb at the reference site reflected natural lithogenic enrichment. Hematological and cytological examinations revealed morphological alterations in blood cells of bats from the contaminated site, including neutrophil hypersegmentation, polychromatophilia, atypical granulation, and nuclear abnormalities in lymphocytes. The total lymphocyte count differed significantly, and these qualitative changes suggest early physiological adjustments or potential indicators of sublethal toxic exposure. By integrating environmental (soil and guano) and biological (blood morphology) parameters, this study demonstrates a non-lethal and ecologically relevant approach to biomonitoring. Blood cell alterations, combined with metal analyses in guano, provide a sensitive tool for detecting potential chronic environmental stress. M. schreibersii is reaffirmed as a valuable sentinel species for ecological monitoring in karst and other vulnerable habitats; however, the limited number of high-quality blood smears obtainable under field conditions still remains a constraint to broader generalization of the findings.

Simple Summary African swine fever is a deadly viral disease of pigs and wild boar that causes major losses for farmers and threatens food security. The disease does not affect people, but its rapid spread and high fatality in pigs make it one of the most serious challenges for animal health in Europe. Since 2019, the disease has been present in Serbia, and in 2023, it was first reported in Bosnia and Herzegovina. In this study, we examined virus samples collected from pigs and wild boar during outbreaks between 2023 and 2025 to better understand how the virus is spreading in the region. By looking at several important parts of the virus genome, we found that all the samples belonged to the same group, known as cluster 19. This shows that the same type of virus has been circulating for several years without major changes. The results suggest that the disease is being maintained locally, mainly through contact between wild boar and pigs kept on small farms with little or no protection. The discovery of the same virus type in Bosnia and Herzegovina highlights that the disease crosses borders, making regional cooperation and continued monitoring essential for controlling its spread.

Background The aim of this study was to examine the prevalence of malnutrition and its association with the frequency of falls and respiratory infections among older adults residing in Croatian nursing homes. Materials and methods The study included 148 participants, 112 (75.7%) women and 36 (24.3%) men, aged 65 years and older, living in the nursing homes in Rijeka and Opatija. The Mini Nutritional Assessment-Short Form (MNA-SF) and data from the medical records were used for data collection. Results The study showed that seven (4.7%) of the older adults living in nursing homes were malnourished, while 53 (35.8%) were at risk of malnutrition. It was found that participants with malnutrition and nutritional risk were more likely to develop respiratory infections (r=-0.37). No correlation was found between malnutrition and the frequency of falls in older adults (r=0.01). Conclusion Malnutrition and the risk of malnutrition are common problems in older adults in nursing homes, requiring regular monitoring and timely intervention. The results confirmed the association between malnutrition and respiratory tract infections, while also highlighting the possibility of co-occurrence of obesity and malnutrition, which is often overlooked.

Simple Summary Advanced gastric cancer is generally associated with a poor prognosis. Stroma AReactive Invasive Front Area (SARIFA) is a recently recognized aggressive histological feature, defined as five tumor cells in direct contact with adipocytes within perigastric, submucosal, or perivascular adipose tissue. The aim of our retrospective study was to evaluate the correlation of SARIFA with pathohistological variables and its impact on overall survival. A cohort of 102 Croatian patients with locally advanced gastric cancer was analyzed, and a significant association between SARIFA and nodal metastases as well as perineural invasion was observed. Patients with both lymphovascular invasion and SARIFA had a significantly higher proportion of affected lymph nodes. They also exhibited a shorter, though not statistically significant, overall survival compared with patients with one or neither of these factors (median 9.2 vs. 16.1 months). A positive SARIFA status may serve as a biomarker of invasiveness and an additional prognostic risk factor. Abstract Background/Objectives: Advanced gastric cancer usually has an unfavorable prognosis. Stroma AReactive Invasion Front Area (SARIFA) is a newly recognized biomarker of aggressiveness, easily recognized as five tumor cells in direct contact with adipocytes in perigastric, submucosal, and perivascular adipose tissue. We investigated this phenomenon and correlated it with other pathohistological variables. Material and Methods: The sample includes 102 Croatian patients with locally advanced gastric cancer, who underwent total gastrectomy/lymphadenectomy between 2012–2018 and in 2023 at University Hospital Split, Croatia, and had pathological stage pT3 or pT4. Representative histological specimens were analyzed for SARIFA, and results were compared with other variables and overall survival. External validation and gene expression analysis of CD36 and FABP4 were performed using the TCGA-STAD cohort. Results: SARIFA was significantly associated with positive pN status (p = 0.009) and perineural invasion (p = 0.043). Patients with SARIFA had a more than fivefold increased risk of nodal involvement (OR = 6.35; 95% CI: 1.35–29.84; p = 0.019). Lymphovascular invasion (LVI) was associated with nodal disease (OR = 4.39; 95% CI: 1.194–16.143; p = 0.026), and SARIFA was marginally associated (OR = 4.886; 95% CI: 0.985–24.241; p = 0.052). Patients who had both LVI and SARIFA had a higher proportion of affected lymph nodes (p = 0.009). SARIFA status did not significantly affect overall survival. Gene expression analysis showed a significant increase in CD36 expression, while FABP4 expression was elevated but not statistically significant, in SARIFA-positive cases. Conclusions: SARIFA could be used as a marker for invasiveness and further investigated due to its predictive potential.

Background/Objectives: Hip osteoarthritis (HOA) is a progressive joint disease characterized by cartilage loss, subchondral bone changes, and synovial inflammation. While tumor necrosis factor receptor 1 (TNFR1), interleukin-6 (IL-6), and transforming growth factor-beta 1 (TGF-β1) are recognized as key mediators of joint pathology, their compartment-specific expression in the human hip synovium remains insufficiently characterized. Therefore, we aimed to investigate their localization and expression in the intimal and subintimal compartments of synovial tissue in patients with HOA compared to controls (CTRL). Methods: Synovial membrane samples were obtained from 19 patients with primary HOA undergoing total hip arthroplasty and 10 CTRL subjects undergoing arthroplasty for acute femoral neck fracture without HOA. Specimens were processed for hematoxylin and eosin (H&E) and immunofluorescence staining. Expression of TNFR1, IL-6, and TGF-β1 was quantified in the intima and subintima using ImageJ analysis. Group differences were assessed using two-way Analysis of variance (ANOVA) with Tukey’s test when assumptions were met; for heteroscedastic outcomes we applied Brown–Forsythe ANOVA with Dunnett’s T3 multiple comparisons. Results: Histological analysis confirmed synovitis in HOA samples, with intimal hyperplasia and mononuclear infiltration. IL-6 was significantly upregulated in the intima of HOA synovium compared with CTRLs, while subintimal expression remained unchanged. In contrast, TGF-β1 expression was reduced in the HOA intima, eliminating the normal intima–subintima gradient. For TNFR1, the within-HOA contrast (int > sub) was significant, whereas the intimal HOA vs. CTRL comparison showed a non-significant trend. Transcriptomic analysis supported IL-6 upregulation, while TNFR1 and TGF-β1 did not reach statistical significance at the mRNA level in an orthogonal, non-hip (knee-predominant) dataset. Conclusions: These findings demonstrate compartment-specific cytokine dysregulation in HOA, with increased intimal TNFR1 and IL-6 alongside reduced intimal TGF-β1. The synovial lining emerges as a dominant site of inflammatory signaling, underscoring its importance in disease progression.

The kidney’s intricate physiology relies on finely tuned gene regulatory networks that coordinate cellular responses to metabolic, inflammatory, and fibrotic stress. Beyond protein-coding transcripts, non-coding RNAs (ncRNAs), including microRNAs (miRNAs), long non-coding RNAs (lncRNAs), and circular RNAs (circRNAs), have emerged as pivotal regulators of renal biology. By modulating transcriptional, post-transcriptional, and epigenetic pathways, ncRNAs govern podocyte integrity, tubular adaptation, intercellular signaling, and immune activation. Dysregulation of these networks is now recognized as a hallmark of major kidney diseases, ranging from diabetic nephropathy and acute kidney injury to chronic kidney disease, glomerulopathies, and polycystic kidney disease. Mechanistic studies have revealed how pathogenic ncRNAs drive apoptosis, inflammation, fibrosis, and cystic remodeling, while protective ncRNAs mitigate these processes, highlighting their dual roles as both disease mediators and therapeutic targets. The exceptional stability of ncRNAs in urine, plasma, and exosomes further positions them as minimally invasive biomarkers with diagnostic and prognostic value. Translational advances include anti-miR and mimic-based therapies (e.g., lademirsen targeting miR-21, miR-29 mimics, anti-miR-17 oligonucleotides), alongside lncRNA silencing strategies, although challenges in delivery, safety, and redundancy remain significant. This review integrates molecular mechanisms with translational perspectives, providing a comprehensive synthesis of how ncRNAs shape renal pathophysiology. By bridging mechanistic insights with emerging diagnostic and therapeutic applications, we highlight the potential of ncRNAs to transform nephrology, paving the way for biomarker-driven precision medicine and novel interventions aimed at intercepting kidney injury at its regulatory roots. In clinical terms, ncRNA-based biomarkers and therapeutics promise earlier detection, more precise risk stratification, and individualized treatment selection within precision nephrology.

Background/Objectives: Postural orthostatic tachycardia syndrome (POTS) is a form of dysautonomia characterized by excessive tachycardia during orthostatic stress. It is frequently observed in patients with syncope, Chronic Fatigue Syndrome (CFS), and post-COVID-19 syndrome (PCS), yet the underlying mechanisms may differ across these conditions. This study aimed to assess autonomic nervous system (ANS) function in patients with syncope, CFS of insidious onset, and CFS post-COVID-19 who presented with POTS, and to compare them with age- and sex-matched patients without POTS. Methods: In this retrospective cross-sectional study, 138 patients over 18 years of age were included following head-up tilt testing (HUTT). Patients were divided into six groups: syncope with and without POTS, CFS with insidious onset with and without POTS, and CFS post-COVID-19 with and without POTS. All participants underwent HUTT, cardiovascular reflex testing (CART) by Ewing, five-minute resting ECG with short-term Heart Rate Variability (HRV) analysis, and 24 h Holter ECG monitoring. Results: The prevalence of POTS across groups ranged from 5% to 7%. Female predominance was consistent across all subgroups. In syncope with POTS, hypertensive responses during HUTT, lower rates of normal Valsalva maneuver results, and reduced HF values in short-term HRV suggested baroreceptor dysfunction with sympathetic overdrive. In both CFS subgroups with POTS, CART revealed higher rates of definite parasympathetic dysfunction, along with more frequent extreme blood pressure variation during HUTT and reduced vagally mediated HRV parameters (rMSSD, pNN50). Across groups, no significant differences were observed with regard to long-term HRV across groups. Conclusions: Distinct autonomic profiles were identified in POTS patients depending on the underlying condition. Syncope-related POTS was associated with baroreceptor dysfunction and sympathetic predominance, whereas CFS-related POTS was characterized by parasympathetic impairment and impaired short-term baroreflex regulation. Evaluating dysautonomia patterns across disease contexts may inform tailored therapeutic strategies and improve management of patients with POTS.