Life doi: 10.3390/life16081323

Authors: Miguel Jiménez-Gómez Beatriz Montull-Veiga Víctor Manuel Mora-Cuesta Eva Revilla-López Myriam Aguilar-Pérez Alicia de-Pablo-Gafas Juan Margallo-Iribarnegaray Carlos Andrés Quezada-Loaiza Ana Hernández-Voth Francisco López-Medrano María Ruiz-Rodríguez Rodrigo Alonso-Moralejo

Respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) are clinically relevant pathogens in lung transplant (LT) recipients, but their impact on lung function and the benefit of ribavirin remains uncertain. We conducted a prospective multicenter observational cohort study of adult LT recipients with RSV or hMPV infection diagnosed between 2021 and 2024 at five centers, with follow-up for up to 12 months. Ribavirin was prescribed at the treating physician’s discretion. Multivariable analysis assessed the association between ribavirin and percentage change in forced expiratory volume in the first second (FEV1) at 90 days, using FEV1 three months before infection as baseline and adjusting for baseline FEV1, pre-existing chronic lung allograft dysfunction, and time since transplantation. Seventy-six patients were included; 60 (78.9%) had RSV and 16 (21.1%) hMPV. Lower respiratory tract infection occurred in 48.7%, and an acute ≥10% FEV1 decline at 14 days was observed in 29.3%. Among patients with lower respiratory tract infection, 45.9% received corticosteroids alone and 37.8% corticosteroids plus ribavirin. No statistically significant between-group differences in allograft dysfunction or mortality were observed. RSV and hMPV infections after LT were frequently associated with lower respiratory involvement and lung function decline. In this observational cohort, ribavirin use was not independently associated with 90-day FEV1 change; however, residual confounding and limited statistical power preclude conclusions regarding treatment efficacy.

Life doi: 10.3390/life16081322

Authors: Antonio L. García-Cordero Ehira Romero-Castelán Juana Fernández-López Fernando Díaz-Sánchez Jose M. Lorenzo Jose A. Rodriguez Raquel Lucas-González Eva M. Santos

This study evaluated the effect of chickpea and rice incorporation to replace corn in extruded snacks on protein digestibility, mineral bioaccessibility, and estimated glycemic index (eGI) using a standardized in vitro digestion model (INFOGEST 2.0). Four formulations were analyzed: a control (100% corn, CF) and chickpea-enriched snacks (F30, F60, and the commercial flavored F60 formulation, CCP). Chickpea replacement increased protein, dietary fiber, and mineral contents while reducing total starch. Protein digestibility remained high across all samples (89.5–96.6%), although slightly lower in chickpea/rice-enriched formulations. However, these formulations provided a higher amount of digestible protein because of their increased protein content. Mineral bioaccessibility varied depending on the element, but chickpea and rice incorporation generally enhanced mineral concentrations in the fraction potentially available for absorption despite the presence of phytic acid and insoluble dietary fiber. Starch hydrolysis was rapid during the early stages of digestion, leading to high estimated glycemic index (eGI) values in all samples. Nevertheless, chickpea/rice-enriched formulations exhibited lower eGI values compared to the corn control, likely due to reduced starch content and matrix effects. Overall, chickpea and rice replacement improved the nutritional profile of extruded snacks by enhancing protein quality and mineral availability, despite its impact on glycemic response being limited, highlighting opportunities for further product optimization.

Life doi: 10.3390/life16081321

Authors: Stoyan Markov

Introduction: Transnasal penetrating intracranial injuries represent an exceptionally rare form of traumatic brain injury, characterized by high mortality and a substantial risk of severe neurosurgical and infectious complications. Self-inflicted injuries involving the penetration of a foreign body through the nasal cavity into the brain are exceedingly uncommon and require prompt diagnosis and a multidisciplinary treatment approach. Objective: This study aimed to present a rare clinical case of transnasal penetration of a foreign body (a ballpoint pen refill) into the brain parenchyma during a suicide attempt, and to describe the diagnostic and therapeutic approach. Case Report: A 46-year-old woman with an acute psychotic disorder was admitted following a suicide attempt involving the insertion of a ballpoint pen refill through the right nasal cavity. Computed tomography revealed a retained intracranial foreign body embedded within the brain parenchyma, traversing the cribriform plate and associated with pneumocephalus. The foreign body was successfully removed via an endoscopic endonasal approach. Owing to the cerebrospinal fluid leak that developed after removal of the foreign body, reconstruction of the anterior skull base defect was performed using fascia lata and tissue adhesive. The postoperative course was uneventful, with no neurological complications and gradual resolution of the intracerebral hemorrhagic changes and pneumocephalus. The patient was discharged without neurological deficits and was referred for psychiatric treatment with a diagnosis of catatonia accompanied by episodes of psychomotor agitation. Conclusions: Transnasal penetrating brain injuries are rare but potentially fatal. Early radiological diagnosis, meticulous preoperative planning, and an endoscopic endonasal approach enable safe removal of the foreign body and reliable reconstruction of the anterior skull base defect. Multidisciplinary collaboration among neurosurgeons, otorhinolaryngologists, radiologists, and psychiatrists is essential for achieving favorable outcomes in these patients.

Life doi: 10.3390/life16081320

Authors: Emil-Tiberius Trasca Dumitru Radulescu Eleonora Daniela Ciupeanu-Calugaru Razvan Mercut Vlad Dumitru Baleanu Elena-Irina Caluianu Alexandru-Marian Vieru Patricia-Mihaela Radulescu

Background/Objectives: Comparisons of abdominal Surgical-Site Infection (SSI) rates during COVID-19 often treat recorded incidence as a direct measure of biological risk, although the pandemic altered operative volume and case mix, prevention practices, and post-discharge detection. This review examines when reported SSI changes are interpretable. Methods: We conducted a critical narrative review, informed by SANRA and critical interpretive synthesis, of MEDLINE/PubMed, Embase, Scopus, and Europe PMC (January 2018–25 June 2026). Comparative SSI studies formed the core evidence set, supported by contextual studies on emergency presentation, biological risk, prevention, and surveillance. Results: Studies reported lower, unchanged, non-monotonic, and higher SSI rates. These differences became more coherent when the recorded rate was separated into operative composition, true infection probability, and detection probability. No core study measured all domains required to isolate a biological pandemic effect. Evidence from acute pancreatitis and abdominal trauma showed that disease severity, care access, inflammatory status, and prognostic relationships varied with health-system context. Conclusions: We propose the Surgical-Site Infection Signal-Validity Framework (SSI-SVF). It classifies findings as probable true improvement, apparent improvement, masked deterioration, mixed deterioration, or indeterminate and requires joint assessment of operative composition, true infection risk, and detection probability before interpreting a calendar-period change as altered surgical safety.

Life doi: 10.3390/life16081319

Authors: Keyla Nunes Farias Gomes Raíssa Maria dos Santos Galvão Natalia Lidmar von Ranke Carlos Rangel Rodrigues Caroline de Souza Ferreira Pereira Julianne Soares Pereira Matheus Amorim Rosa e Silva Brenda Bairral Queiroz Ornellas Jonathas Albertino de Souza Oliveira Carneiro Geovana Espindola Jardim André Lopes Fuly José Augusto Albuquerque dos Santos Robson Xavier Faria

Rheumatoid arthritis (RA) is a chronic autoimmune disease that affects mainly peripheral joints because of inflammation of the synovial membrane. Current treatments, such as nonsteroidal anti-inflammatory drugs (NSAIDs) and glucocorticoids, although effective, are associated with high costs and several adverse effects. In this context, natural products have emerged as promising alternatives because of their potential therapeutic effects and lower toxicity. The objective of this review was to identify and evaluate natural substances with potential applications in RA treatment on the basis of studies published between 2015 and 2020. A literature search was conducted in SciELO, PubMed, and Google Scholar using the keywords “rheumatoid arthritis”, “treatment”, “toxicity”, and “natural products”. Additionally, we applied in silico methods to predict pharmacokinetic and toxicological parameters using ADMET Predictor® (Simulation Plus) and compared the results with those of commercial drugs such as diclofenac, ibuprofen, and naproxen. Target fishing (reverse docking) was also performed to identify possible molecular targets related to RA. Seven natural compounds were identified, mostly evaluated through in vivo studies. Among them, paeoniflorin, quercetin, resveratrol, and celastrol are in clinical phases and present potential as RA treatments. In silico analysis highlighted curcumin, tetramethylpyrazine, and resveratrol as the most promising candidates, with ADMET profiles comparable or superior to those of current NSAIDs. In conclusion, natural products represent viable alternatives for RA therapy. However, further studies are essential to better understand their safety, pharmacokinetics, and drug interactions to ensure their clinical applicability.

Life doi: 10.3390/life16081317

Authors: Abdulellah Saad Alharbi Abdulaziz Arif A. Alshammari Mai B. Alwesmi Vasudevan Mani

Type 2 diabetes mellitus (T2DM) is a widely distributed dysmetabolic condition increasingly linked to neurocognitive decline, neuroinflammation, oxidative stress, and neuronal apoptosis. Chronic hyperglycemia and insulin resistance disrupt homeostasis in the central nervous system, contributing to neurodegenerative processes. This study evaluated the effects of semaglutide (SMG), a glucagon-like peptide-1 receptor agonist, on T2DM-associated neurobehavioral and biochemical alterations using a rat model. T2DM was induced by nicotinamide–streptozotocin, followed by oral administration of SMG at a dose of 1.44 mg/kg for one month. Cognitive function was evaluated using the elevated plus maze (EPM) and novel object recognition (NOR) paradigms. Fasting blood glucose and body weight were monitored throughout the experiment. Neuroinflammatory markers (COX-2, TNF-α, and IL-6), oxidative stress biomarkers (MDA, GSH, and catalase), and apoptosis-associated proteins (Caspase-3, Bax, and Bcl-2) were measured in brain tissue homogenates using ELISA. Diabetic rats exhibited marked cognitive deficits, hyperglycemia, increased neuroinflammatory markers, and altered apoptosis-related biomarkers, with reduced Bcl-2 expression. Treatment with SMG significantly improved learning and memory performance, fasting blood glucose, and body weight. At the molecular level, SMG treatment was associated with lower levels of MDA, TNF-α, Bax, IL-6, COX-2, and Caspase-3, together with higher levels of Bcl-2, catalase, and GSH compared with untreated diabetic rats. Overall, oral SMG treatment was associated with improved cognitive performance and metabolic control, accompanied by attenuation of diabetes-associated neuroinflammatory, oxidative stress, and apoptosis-related alterations in brain tissue. However, the present study was not designed to distinguish direct neuroprotective effects from changes secondary to improved metabolic control. Further studies are required to clarify the underlying mechanisms and determine the translational relevance of these findings.

Life doi: 10.3390/life16081318

Authors: Emanuel Maitz Reinhard Domanyi Gerold Schwantzer Gerald Sendlhofer Alexandros Andrianakis Dietmar Thurnher Thomas Weiland

Objectives: Chronic rhinosinusitis (CRS) is a prevalent chronic inflammatory upper airway disease. Despite a higher radiological disease burden in males, clinical CRS diagnoses are disproportionately registered in females, suggesting systematic underdiagnosis in young men. The 22-item Sinonasal Outcome Test (SNOT-22) represents the reference patient-reported outcome measure (PROM) for sinonasal symptom burden. This study aimed to determine the prevalence of sinonasal symptom burden and to identify associated clinical, laboratory, and lifestyle parameters in a large cohort of young male adults. Methods: In this prospective cross-sectional study, 500 young male adults underwent, as part of the conscription examination, standardized assessment including the SNOT-22, spirometry (FEV1%), whole blood count (eosinophils), and serum CRP. SNOT-22 scores were categorized as none (0–20), moderate (21–50), and severe (>50). Associations were analyzed using Spearman correlation, Mann–Whitney U, and Chi2 tests. Results: The mean SNOT-22 score was 18.8 (SD 15.2; range 0–73). A total of 38.2% of participants scored ≥21, indicating at least moderate sinonasal symptom burden. SNOT-22 correlated significantly with CRP (r = 0.117, p = 0.009) and inversely with relative FEV1% (r = −0.103, p = 0.022). Eosinophils showed a numeric gradient across SNOT-22 categories (3.08% vs. 3.31% vs. 3.56%) without reaching significance. Conclusions: More than one third of young male adults reported significant self-reported sinonasal symptom burden. Symptom severity was associated with systemic inflammatory markers and reduced pulmonary function, supporting the unified airway concept. These findings support the utility of SNOT-22 as a low-threshold assessment tool for self-reported sinonasal symptom burden in preventive health examinations.

Life doi: 10.3390/life16081316

Authors: Bünyamin Arı Fatmagül Can Umur Batak Turan Cihan Dülgeroğlu

Serial presepsin measurements were evaluated for their prognostic value compared with conventional laboratory markers in predicting in-hospital mortality among older adults with hip fractures. This prospective observational study included 85 patients aged ≥65 years admitted with acute hip fractures between December 2025 and May 2026. Residual serum remaining after routine clinical laboratory sampling was obtained on admission (Day 1), Day 3, and Day 5; serum presepsin was measured by commercial ELISA, and routine parameters (C-reactive protein [CRP], white blood cell count, lymphocytes, monocytes, platelets, and liver enzymes) were analysed in the hospital laboratory. Renal function was assessed by serial creatinine and estimated glomerular filtration rate (eGFR). Patients were classified as survivors or non-survivors according to in-hospital outcome. Temporal trajectories were modelled with a linear mixed-effects model fitted to log-transformed presepsin, receiver operating characteristic (ROC) analysis assessed predictive performance, and internal validity was examined by bootstrap resampling. Of the 85 patients, 68 survived and 17 died during hospitalization; all deaths occurred between hospital days 5 and 12. Presepsin diverged progressively between groups, reaching significantly higher concentrations in non-survivors by Day 5 (median 207.70 vs. 147.21 ng/L; p < 0.001), with a Day 5 group-by-time interaction ratio of 1.76 (95% CI 1.38–2.25; p < 0.001). Day 5 presepsin showed the highest predictive accuracy (AUC = 0.868, 95% CI 0.774–0.945; optimism-corrected AUC 0.866), significantly outperforming CRP (AUC = 0.606; p = 0.003) and platelet count (AUC = 0.485; p < 0.001). The association persisted after adjustment for age, ASA class, fracture type, and eGFR, and Day 5 presepsin added discrimination to a baseline clinical model (ΔAUC 0.125; p = 0.015). The Youden-derived cutoff of 169.54 ng/L was unstable across bootstrap resamples (95% range 169.5–207.7 ng/L). Serial presepsin measurement, particularly on Day 5, is associated with in-hospital mortality in this population; these exploratory findings require external validation before any clinical application can be considered.

Life doi: 10.3390/life16081315

Authors: Eugen-Toma Radu Nastaca Alina Palade Crina Cristina Solomon Oana Gabriela Somcutian Manuela Pia Pumnea Maria Totan Claudia Maria Mihuț Cecilia Georgescu Adina Frum Carmen Maximiliana Dobrea Felicia Gabriela Gligor

Background/Objectives: Microplastics (MPs) are emerging environmental contaminants with potential implications for human health. Experimental studies suggest that MPs accumulate in the liver and promote inflammatory and fibrotic changes, but evidence from human tissues remains limited. This study investigated the presence of suspected MP-like particles in post-mortem human liver tissue and their associations with clinical, biochemical, hematological and histopathological parameters. Methods: Post-mortem liver tissue samples were collected from 55 adults. Suspected MP-like particles were extracted using hydrogen peroxide digestion, filtration, Nile Red staining and image-based quantification. Histopathological evaluation assessed inflammation, fibrosis, necrosis and fatty liver degeneration. Polarized light microscopy was used as a supportive morphological assessment method. Statistical analyses included Mann–Whitney U tests, Fisher’s exact tests and Spearman correlation analysis. Results: Detectable hepatic suspected MP-like particles were identified in 9 of 55 individuals (16.4%). Individuals with detectable suspected MP-like particles had significantly lower alanine aminotransferase (ALT) levels and leukocyte counts. The hepatic suspected MP-like particle burden showed weak positive correlation with liver fibrosis and inflammation and weak inverse correlation with ALT levels and leukocyte count. Fisher’s exact test showed that liver fibrosis and liver inflammation were significantly associated with detectable hepatic suspected MP-like particles, with higher unadjusted odds observed in the corresponding 2 × 2 contingency tables. No statistically significant associations were found for liver necrosis or fatty liver degeneration. Conclusions: Detectable suspected MP-like particles were identified in post-mortem human liver tissues and were more closely associated with fibrotic and inflammatory histopathological changes than with routine biochemical abnormalities. Larger studies using standardized detection methods are needed to confirm these results.

Life doi: 10.3390/life16081314

Authors: Katarina Dunjic Momir Dunjic Marina Gazdic Jankovic Marina Miletic Kovacevic Nikolina Kastratovic Biljana Ljujic Tatjana Novakovic Milan Filipovic Tatjana Filipovic Zhao Jing Marija Dunjic Miroslav M. Sovrlić Sandra S. Konstantinović Jelena S. Stanojević Milan D. Kostić Stefano Turini

Background: Previous studies have examined bioactive constituents of Prunus dulcis oil, Pinus sylvestris essential oil, and related phytochemical matrices separately; however, their combined chemical profile, comparator-context molecular-docking landscape, and comparative cellular response in melanoma and non-malignant fibroblast models remain insufficiently characterized. Methods: The analyzed study samples were profiled by GC/MS and GC/FID-based fatty-acid methyl ester analysis. An archived molecular-docking dataset was used to summarize structurally plausible interactions of six formulation-associated markers with eight melanoma-relevant or melanoma-adjacent targets and to present representative three-dimensional complexes. Because complete protocol metadata and co-crystallized-ligand redocking records were unavailable, the computational findings were interpreted descriptively. Neutral-red uptake assays assessed the 24 h viability of B16F10 melanoma cells and MRC-5 fibroblasts after exposure to each oil, the dual-oil formulation, or cisplatin. Four-parameter logistic models provided estimated IC50 values, and exact two-sided permutation tests were used for Spearman rank analyses. Results: The dual-oil formulation yielded estimated IC50 values of 1.42% v/v in B16F10 cells and 4.85% v/v in MRC-5 cells, corresponding to an apparent selectivity index of 3.41. The complete eight-concentration series was non-monotonic and showed no significant rank association (B16F10: ρs = −0.4286, exact p = 0.2992; MRC-5: ρs = −0.3810, exact p = 0.3599). In a post hoc analysis of the descending branch (≥0.37% v/v), the association was perfect in B16F10 cells (ρs = −1.0000, exact p = 0.0167) but did not reach significance in MRC-5 cells (ρs = −0.9000, exact p = 0.0833). The docking matrix identified target-dependent numerical prioritization patterns; however, small score differences were not interpreted as evidence of stronger binding or cellular target engagement. Conclusions: The findings define an exploratory chemical–computational–cellular framework and a preliminary formulation-level viability phenotype. They do not establish synergy, apoptosis, pathway modulation, therapeutic efficacy, or clinical safety. Independent multi-batch chemical confirmation, a fully documented and redocking-validated computational protocol, expanded melanoma and normal-skin cell panels, orthogonal functional assays, longer exposure periods, mechanistic validation, and subsequent in vivo studies are required.

Life doi: 10.3390/life16081313

Authors: Alejandro Gugliucci

Recent research establishes carbohydrate response element-binding protein (ChREBP) as a central regulator of fructose-induced hepatic lipogenesis, underscoring its critical role in mediating the effects of excessive sugar intake on liver metabolism. Consequently, this narrative review presents a perspective of hepatic sugar metabolism and ChREBP signaling, highlighting areas for further exploration to better understand and address the metabolic consequences of fructose-driven activation of ChREBP. The discussion encompasses five principal domains: (1) fructose metabolism; (2) the structure and function of ChREBP; (3) interactions between glucose and fructose metabolism with ChREBP activity; (4) mechanisms that intensify fructose’s influence on ChREBP pathways; and (5) future research opportunities. Under physiological conditions, ChREBP mitigates accumulation of glycolytic intermediates; however, excessive sugar consumption overwhelms these capacities, thereby increasing lipogenesis. Fructose demonstrates notably stronger activation of ChREBP in vivo relative to glucose. Given ChREBP’s dual role in maintaining metabolic homeostasis and promoting insulin resistance under high fructose intake, therapeutic targeting poses considerable challenges. We propose the testable hypothesis that the MG generated from excessive fructose metabolism may bind to ChREBP’s lysine residues, stabilizing the protein and reducing degradation by impeding ubiquitination. This stabilization of ChREBP would prolong its activity, thereby further promoting sustained hepatic lipogenesis and exacerbating the risk of MASLD. Experimental and clinical research is needed to confirm its validity and define the molecular pathways involved. Future studies should aim to identify glucose metabolites involved in ChREBP regulation and clarify its beneficial versus adverse effects. The specific functions and regulation of ChREBPβ vs. ChREBPα remain to be elucidated.

Life doi: 10.3390/life16081312

Authors: Sebastiano Ettore Spoto

Astrobiological observations usually return atmospheric, mineralogical, molecular, isotopic, or textural states rather than organisms. The interval between environmental habitability and confirmed life detection is therefore an evidential problem. This article develops planetary geochemical metabolism (PGM) as a scale-explicit description of abiotic water–rock–fluid reactions, including atmospheric or ice-shell boundary conditions where relevant, that sustain redox disequilibria, catalytic mineral interfaces, prebiotic molecular fluxes, and preservable mineral products. Proto-biosignatures are defined as contextual, non-diagnostic signatures of this interval: signals that do not demonstrate life, but increase the plausibility of prebiotic network organization relative to low-organization abiotic chemistry. A nondimensional Geochemical Metabolic Potential, ΦPGM, is formalized as a target-specific screening index describing redox exergy, catalytic-interface density, reaction-network closure, environmental cycling efficacy, and preservation potential. The index is not calibrated as a probability of life. A reproducible Latin-hypercube experiment across Msim=5000 synthetic environments examines internal model behavior rather than ranking planets. Higher ΦPGM values mark hypotheses to be tested under declared scale, uncertainty, and observability constraints. Applications to early Earth, Noachian Mars, ocean worlds, and terrestrial exoplanets illustrate that habitability, prebiotic organization, biological inference, and preservation are distinct quantities.

Life doi: 10.3390/life16081311

Authors: Siti Hajar Adam Nor Syaza Syahirah Amat Junaidi Shariff Halim Mohd Helmy Mokhtar

Ficus deltoidea Jack (Moraceae), locally known as Mas Cotek, is a medicinal plant traditionally used throughout Southeast Asia for the management of diabetes mellitus. This review summarises the available evidence on the antidiabetic properties of F. deltoidea based on eleven in vitro, nine in vivo and one clinical study identified through a structured literature search. In vitro investigations show that F. deltoidea inhibits α-glucosidase and α-amylase, stimulates insulin secretion in pancreatic β-cells via both K+-ATP channel-dependent and -independent pathways, enhances glucose uptake in hepatocytes and adipocytes, promotes adiponectin secretion and inhibits protein tyrosine phosphatase 1B (PTP1B). Vitexin and isovitexin, the predominant C-glycosyl flavonoids in F. deltoidea leaves, appear to be the main bioactive compounds responsible for these effects. Meanwhile, in vivo studies in streptozotocin-induced diabetic rodents report dose-dependent reductions in fasting blood glucose, improved glucose tolerance, restoration of pancreatic islet architecture, modulation of hepatic gluconeogenic and glucose-metabolic genes, and protection against diabetic nephropathy and bone loss. Inter-varietal differences in chemical composition and biological activity were observed, with var. trengganuensis and var. intermedia reported as the most active. The only available clinical trial in adults with prediabetes (1000 mg/day for 8 weeks) showed a reduction in LDL and total cholesterol but no significant change in fasting blood glucose or insulin. The discrepancy between preclinical and clinical findings highlights the need for standardised extracts, pharmacokinetic studies and adequately powered clinical trials in patients with established type 2 diabetes mellitus.

Life doi: 10.3390/life16081310

Authors: Su-Min Cha

Generative artificial intelligence (AI) and large language models (LLMs) are increasingly evaluated in rehabilitation, yet their clinical validity, reproducibility, and safety remain uncertain. This scoping review mapped peer-reviewed studies of generative AI/LLMs across rehabilitation assessment, clinical reasoning, decision support, planning, education, and functional classification. Following JBI methodology and PRISMA-ScR, five databases were searched for English-language studies published from 1 January 2015 to 24 July 2026. Two reviewers independently conducted study selection, data extraction, methodological appraisal, and application-domain coding. Of 2126 records, 43 publications representing 42 unique studies were included, predominantly from 2025–2026 and involving GPT/ChatGPT/OpenAI-family systems. At the unique-study level, six application domains were identified: clinical reasoning and decision support (n = 13), rehabilitation education, simulation, and feedback (n = 10), rehabilitation planning and prescription (n = 9), guideline adherence and clinical-question support (n = 6), adaptive feedback and rehabilitation support (n = 2), and assessment and functional classification (n = 2). Evidence was concentrated in benchmark, scenario-based, and educational evaluations, with limited patient-level outcomes. Heterogeneous methods, incomplete reporting of reproducibility, inconsistent safety assessment, and possible selective publication limited comparability and clinical generalizability. Generative AI/LLMs should therefore be used primarily as clinician-supervised assistive tools, with prospective validation, standardized reporting, and active safety evaluation prioritized.

Life doi: 10.3390/life16081309

Authors: Lucian Mihai Florescu Cosmin Vasile Obleagă Mădălin Mămuleanu Ioana Andreea Cîrlig Alesandra Florescu Mihai Alexandru Ene Aurelia Ștefania Domenco Alexandru Marian Olaru Alexandra Gabriela Cosmina Țâru Raluca Elena Nica Rossy Vlăduț Teică Ioana Andreea Gheonea

Automated colorectal tumor segmentation on routine computed tomography (CT) remains technically challenging because of limited soft-tissue contrast, heterogeneous tumor morphology, complex bowel anatomy, and marked imbalance between tumor and background pixels. This retrospective technical feasibility study aimed to develop and evaluate an attention-enhanced ResNet50–U-Net architecture for automated segmentation of primary colorectal tumors on contrast-enhanced abdominopelvic CT images. The dataset comprised 493 axial CT image–mask pairs containing visible colorectal tumors, including 70 institutional images obtained from 25 patients and 423 images from publicly available colorectal cancer imaging resources. Reference masks were generated by manual tumor delineation performed by an experienced radiologist and reviewed by a second abdominal radiologist, with uncertain contours resolved by consensus. The proposed model combined an ImageNet-pretrained ResNet50 encoder with an attention-guided U-Net decoder and was trained using a composite Binary Cross-Entropy and Focal Tversky loss function to address foreground–background class imbalance. Performance was assessed using five-fold image-level cross-validation. Predicted probability maps were binarized using a threshold of 0.5, and segmentation metrics were calculated through global micro-averaging within each validation fold. The model achieved a mean Intersection over Union of 0.7406 ± 0.0276, a Dice similarity coefficient of 0.8507 ± 0.0182, a pixel-level sensitivity of 0.9559 ± 0.0266, and a pixel-level background specificity of 0.9956 ± 0.0003. Qualitative assessment demonstrated substantial spatial agreement between predicted masks and expert annotations, although minor boundary discrepancies and small false-positive components were observed. These findings support the technical feasibility of attention-enhanced encoder–decoder architectures for colorectal tumor segmentation on routine CT images. However, because validation was performed at the image level, entirely tumor-negative images were not included (preventing the evaluation of clinical specificity and false-positive rates), and no independent external test cohort was available, further patient-level, multicenter, and DICOM-based validation is required before clinical implementation. While the model demonstrated robust internal performance, future studies must prioritize large-scale external validation and patient-level cross-validation to rigorously assess generalizability and specificity.

Life doi: 10.3390/life16081308

Authors: Marta Ferrer-Cornet Marvin Garcia-Reyes Mireia Bragulat-Arévalo Andrea Guala Juan Garrido-Oliver Alba Catala-Santarrufina Pere Lopez-Gutierrez Ruperto Oliveró-Soldevila Gisela Teixidó-Turà Ignacio Ferreira-Gonzalez Jose Rodriguez-Palomares Sergi Bellmunt-Montoya Lydia Dux-Santoy

As abdominal aortic aneurysms (AAA) rupture results in a high mortality rate, early detection and risk stratification are critical for timely elective intervention. Although the maximum diameter is the sole imaging metric used to assess risk, other factors may provide complementary information. This study investigated the role of inflammation and hemodynamics in AAA progression. Methods: Patients with AAA underwent hybrid 18F-fluorodeoxyglucose ([18F]FDG) PET/CMR imaging, including a four-dimensional flow sequence and contrast-enhanced magnetic resonance angiography, and were followed to assess AAA diameter growth rate. Abdominal aorta hemodynamics and [18F]FDG PET target-to-background (TBR) were quantified at 28 standardized regions. Correlations between AAA diameter, hemodynamics, PET and growth rate were assessed. Results: Thirty-two AAA patients (84.4% men) with a median age of 73 and an interquartile range of [68; 77] years were prospectively recruited. The AAA maximum diameter was 48.1 [44.8; 55.0] mm, and the growth rate was 2.7 [1.3; 4.0] mm/year during a follow-up of 2 [1.1; 3.5] years. The AAA maximum diameter (R = 0.443, p = 0.011) and flow vorticity (R = 0.453, p = 0.009) showed significant bivariate correlation with the growth rate. When correcting for the aneurysm diameter, the helicity density was significantly correlated with the growth rate (p = 0.045), while the AAA thrombus volume was inversely associated with it (p = 0.042). Aneurysms with negative helicity density (counterclockwise rotation) grew significantly faster (p = 0.042) than those with positive helicity. In multivariable analysis, beyond the AAA diameter, the thrombus volume, helicity density and TBR were inversely related to the growth rate. Conclusions: Integrating flow, inflammatory and thrombus information may improve AAA growth rate prediction beyond the maximum aneurysm diameter.

Life doi: 10.3390/life16081307

Authors: Hao Zou Yuanjie Zhou Suiyin Lin Tingting Pang Linxi Zhang Jing Zhou Renzhi Wu

Sugarcane bagasse is an important renewable lignocellulosic resource, yet its bioconversion efficiency remains low, primarily because wild-type Saccharomyces cerevisiae cannot utilize xylose, which limits the industrial production of cellulosic ethanol. In this study, a high-ethanol-yielding strain 31.1 was obtained from Scheffersomyces stipitis (formerly known as Pichia stipitis) 1960 through 60Co mutagenesis and long-term domestication. Strain 31.1 exhibited an ethanol productivity of 0.78 g/(L·h), a sugar-to-ethanol conversion rate of 0.38 g/g, and a fermentation efficiency of 82.61%. Using the wild-type strain 1960 and a low-yielding strain 12.1 as controls, comparative genomics and transcriptomics were employed to elucidate the mechanism underlying the high ethanol production. Our findings are as follows: at the genomic level, there were 271 genomic structural variations. At the transcriptomic level, most genes involved in secondary metabolite synthesis, antibiotic synthesis, ribosomal pathways, amino acid biosynthesis, and oxidative phosphorylation pathways were down-regulated. Additionally, two key genes—XYL1 (xylose reductase gene) and XUT4 (high-affinity xylose transporter gene)—were significantly up-regulated. Through comprehensive integration of phenotypic comparison (e.g., fermentation performance of the high-yield strain 31.1 in yeast propagation and ethanol fermentation), comparative genomics, transcriptomics, and bioinformatics analyses of pathways involved in oxidative phosphorylation and the cell cycle (related to yeast cell growth), we identified 60 candidate key genes associated with high xylose-to-ethanol yield in S. stipitis. These genes are predominantly involved in the cell cycle pathway, including CDC15 and PHO81. In conclusion, our study preliminarily reveals the mechanisms underlying the high xylose ethanol production of the high-yield strain at the genomic and transcriptomic levels.

Life doi: 10.3390/life16081306

Authors: Lampros Fotos Aris Pourlis Eirini Valasi Georgios I. Papakonstantinou Konstantinos Kousenidis Athanasios Sougias George Tsegas Eleftherios Chourdakis Zisis Tsiropoulos Alexandra V. Michailidou Christos Vlachocostas Vasileios G. Papatsiros

Heat stress (HS) suppresses voluntary feed intake in lactating sows, yet individual-level characterization of indoor and outdoor Temperature–Humidity Index (THI) linked to feed intake is rare in Mediterranean commercial settings. This prospective observational study enrolled 272 F1-crossbred sows (parities 1–7) farrowing from May–October 2025 at a commercial Greek farm. Outdoor THI was computed from 8760 hourly records and indoor THI from calibrated dataloggers; individual daily lactation feed intake (days 1–30) was recorded via RFID transponder-feeders. Building cooling (ΔTHI = 3.7–14.3) attenuated but did not eliminate mild-to-moderate indoor stress from June–September. Only parity and stillbirth count were Bonferroni-significant correlates of feed intake (parity: β = +0.142, p = 0.002). A sow-day Gradient Boosting model, evaluated by grouped cross-validation to prevent within-sow leakage, achieved CV-R2 = 0.406 ± 0.045, providing a validated, herd-tested framework for parity-stratified feeding management, pending external validation across farms, genetics, and climates.

Life doi: 10.3390/life16081305

Authors: Igor Djordjević Ana Miletić Filip Ivanjac Danica Popović Antić Minja Miličić Lazić Vitomir Konstantinović

Background: Temporomandibular disorders (TMD) are associated with musculoskeletal pain affecting the masticatory muscles and temporomandibular joint (TMJ). Low-level laser therapy (LLLT) is a therapeutic modality with analgesic, anti-inflammatory, and biostimulatory effects. Protocol and laser parameter settings for TMD management have not yet been established. The aim of this prospective study was to evaluate the effectiveness of low-level laser therapy in reducing pain in TMD patients. Methods: Sixty-three participants treated at the Department of Prosthodontics, School of Dental Medicine, University of Belgrade, were sorted to the LLLT group and the control splint-medication group (SMG), which received occlusal splint therapy, with nonsteroidal anti-inflammatory drug treatment (NSAID; Ibuprofen). Results: Both therapeutic approaches resulted in significant pain reduction and improvement in patients’ quality of life. No statistically significant differences were observed between the groups, LLLT patients demonstrated a higher rate of successful treatment outcomes, reduction in pain intensity during the treatment. Conclusion: The applied LLLT was associated with significant reduction in pain intensity and improvement in oral health-related quality of life in patients with TMD. Although no statistically significant differences between the groups were demonstrated at the individual post-treatment assessment points, these findings support LLLT as a potentially useful non-invasive conservative treatment option.

Life doi: 10.3390/life16081304

Authors: Liudmila Mikhaleva Maria Martynova Zarina Gioeva Nikolay Shakhpazyan Nikita Chizhikov Valentina Pechnikova Mikhail Gushchin Alexander Ilyichev

Prostate cancer (PCa) is one of the most prevalent malignancies in men worldwide. Established clinicopathological parameters do not fully describe the biological heterogeneity of PCa or consistently predict its clinical course after radical prostatectomy, supporting the investigation of additional prognostic markers. Stroma AReactive Invasion Front Areas (SARIFA) is a recently described histomorphological pattern characterized by direct contact between tumor cells and adipocytes without intervening desmoplastic or inflammatory stroma at the invasion front. Limited retrospective evidence from prostatectomy specimens indicates that SARIFA positivity is associated with adverse pathological features. However, its independent prognostic value and clinical utility remain to be established. Because SARIFA can potentially be assessed on routinely prepared hematoxylin and eosin-stained sections without additional molecular assays or immunohistochemical staining, it represents a potentially accessible candidate marker, although its reproducibility, standardization, and cost-effectiveness require formal evaluation. This review summarizes current evidence concerning lipid metabolic remodeling and tumor–adipocyte interactions that may contribute to the SARIFA phenotype in PCa. Androgen receptor-regulated lipid metabolism, uptake of adipocyte-derived fatty acids, adipokine signaling, and extracellular vesicle-mediated communication provide a plausible mechanistic framework. Nevertheless, direct experimental evidence linking several of these processes specifically to SARIFA in PCa remains limited, and some proposed relationships are based on indirect evidence or findings from other tumor types. Further studies should establish standardized scoring criteria, intra- and interobserver reproducibility, external validation, and incremental prognostic value before the clinical implementation of SARIFA can be considered.

Life doi: 10.3390/life16081303

Authors: Valerica Creanga Zarnescu Liliana Mititelu-Tartau Ilie Onu Daniel Andrei Iordan Liliana-Lăcrămioara Pavel

Background: Non-steroidal anti-inflammatory drugs (NSAIDs) remain the cornerstone of pharmacological treatment for osteoarthritis (OA) and are recommended by international guidelines for the management of symptomatic pain. Despite their well-established efficacy, substantial interindividual variability exists in treatment response, with some patients experiencing meaningful pain relief while others derive little clinical benefit despite appropriate drug selection and dosing. This variability reflects, at least in part, the biological heterogeneity of OA pain. Objective: To review the mechanisms underlying variability in NSAID responsiveness in OA and to propose a practical framework for personalized NSAID prescribing based on pain phenotype, individual safety profile, and treatment timing. Methods: A narrative review of the contemporary scientific literature was conducted using major biomedical databases to summarize the current evidence on phenotype-guided NSAID therapy in OA. Particular attention was given to the emerging concepts of nociceptive, nociplastic, and neuropathic-like pain phenotypes and their implications for personalized anti-inflammatory therapy. Results: Current evidence indicates that OA pain is a heterogeneous and dynamic condition in which inflammatory nociceptive, nociplastic, and neuropathic-like mechanisms coexist to varying degrees. NSAIDs primarily target inflammatory nociceptive pain by inhibiting cyclooxygenase (COX)-mediated prostaglandin synthesis and reducing peripheral sensitization. Consequently, patients with predominantly inflammatory nociceptive pain are the most likely to benefit from NSAID therapy, whereas those with predominant nociplastic or neuropathic-like pain mechanisms may require alternative or multimodal treatment strategies. Beyond pain phenotype, optimal NSAID selection should integrate cardiovascular, gastrointestinal, and renal risk assessment, recognizing the important pharmacological and safety differences among individual agents. Treatment timing is also clinically relevant, as anti-inflammatory therapy appears most effective when initiated during periods of active inflammatory nociceptive pain. Based on the available evidence, we propose a conceptual clinical decision framework integrating patient selection, NSAID choice, and treatment timing. Conclusions: Personalized NSAID prescribing should move beyond a diagnosis-based approach toward a mechanism-based strategy that integrates pain phenotyping, individualized safety assessment, and appropriate treatment timing. The proposed framework is built upon three complementary principles, identifying the right patient, selecting the right treatment, and initiating therapy at the right time. It provides a practical foundation for implementing precision medicine in the pharmacological management of OA.

Life doi: 10.3390/life16081302

Authors: Liliana Cepoi Tatiana Chiriac Ludmila Rudi Svetlana Codreanu Ana Valuța Svetlana Djur Tudor Trifan Vera Potopová

The chemical form of copper is a key determinant of metal bioavailability and may profoundly influence cyanobacterial metabolism and the biological activity of culture-derived products. In this study, Nostoc linckia was cultivated in the presence of metallic copper nanoparticles (CuNPs), copper oxide nanoparticles (CuONPs), or CuSO4·5H2O (15 mg Cu L−1). Biomass productivity, biochemical composition, and antioxidant capacity, together with the physicochemical and biochemical characteristics of the culture liquid, were determined. The culture liquid was subsequently evaluated through the seed priming of four maize (Zea mays L.) hybrids. The different copper forms induced distinct metabolic responses in Nostoc linckia. CuONPs promoted biomass accumulation while maintaining protein and photosynthetic pigment contents and inducing an adaptive oxidative response, whereas CuNPs, and particularly CuSO4, impaired biomass production and primary metabolism. These metabolic changes were reflected in the culture liquid properties and were associated with genotype-dependent differences in maize germination dynamics, seedling vigor, and biochemical composition. Integrated correlation, hierarchical clustering, and principal component analyses revealed coordinated relationships linking cyanobacterial physiology, culture liquid profiles, and plant responses. Collectively, these findings demonstrate that the chemical form of copper drives metabolic remodeling in Nostoc linckia, thereby shaping the biochemical composition and biostimulant properties of its culture liquid.

Life doi: 10.3390/life16081301

Authors: Jawaria Areeba Noor Yusra Zarlashat Muhammad Ebad Asif Khan Enrique Mandado Loureiro Edit Dósa

Hypertension remains a major global health burden, and control rates remain suboptimal because of poor medication adherence and limitations of existing therapies, including escape within the renin–angiotensin–aldosterone system. Zilebesiran, a first-in-class, subcutaneously administered small interfering RNA therapeutic, represents a promising advance in hypertension management. Through N-acetylgalactosamine-mediated hepatic delivery, zilebesiran selectively silences angiotensinogen (AGT) messenger RNA, the transcript encoding the common precursor of all angiotensin peptides. This upstream intervention reduces AGT production and produces durable blood pressure lowering that can persist for up to 6 months after a single dose. This review summarizes the mechanism of action of zilebesiran, its pharmacokinetic and pharmacodynamic properties, and the available phase 1 and phase 2 clinical evidence, including the KARDIA program. We also place zilebesiran within the evolving antihypertensive landscape by comparing it with aldosterone synthase inhibitors, dual endothelin receptor antagonists, and brain aminopeptidase A inhibitors. Finally, we discuss translational challenges, including reversal strategies for emergency situations, monitoring considerations, and potential roles for personalized dosing. Early-phase and phase 2 trials show dose-dependent and durable reductions in serum AGT and blood pressure with infrequent dosing; however, long-term safety, cardiovascular outcome benefit, and generalizability in diverse high-risk populations remain to be established, and zilebesiran remains investigational while phase 3 outcome testing is underway.

Life doi: 10.3390/life16081300

Authors: Konstantin Gunbin Zamart Ramazanova Bakhyt Matkarimov Murat Saparbaev Sergey Nikolaev Andrey Yurchenko

Cutaneous melanoma and basal cell carcinoma (BCC) represent the two primary malignancies driven by solar ultraviolet (UV) radiation. To map the spatial topology and distance dependence of their mutational signatures, we deployed new analytical bioinformatics workflow utilizing two convergent strategies: a data-driven read-level phasing framework to discriminate between collateral mutational events versus independent ones and a hypothesis-driven spatial permutational simulation model to capture density-dependent local deviations. A whole-genome analysis employing this framework unexpectedly reveals two fundamentally distinct lesion-processing landscapes, present in both BCC and melanoma. While independent mutations consistently reproduce canonical UV signatures (SBS7a–c) in both tumor types, collateral mutations tell a distinct and more varied narrative between BCC and melanoma. These collateral mutations are unusually abundant for non-UV characteristics, such as age-related SBS1 and SBS5, and display a notable 3′ to 5′ asymmetry near UV-induced lesions in pyrimidine dimers. We also demonstrated that BCC displays a pronounced enrichment of dinucleotide base substitutions flanking UV-signature sites on the 3′ side, particularly CA>TG and CG>TA changes. Ultimately, these topological patterns in both tumors indicate that a primary photoproduct seeds secondary mutagenesis within its local chromatin environment, dramatically altering our understanding of UV-induced lesion processing.

Life doi: 10.3390/life16081299

Authors: Andrea Marzullo Cecilia Salzillo

Myocardial infarction is a major cause of cardiovascular death and a key substrate for sudden cardiac death. Traditionally, histopathological analysis of infarction has focused on the temporal sequence of morphological changes, from coagulative necrosis to inflammatory infiltrate and cicatricial fibrosis. However, recent molecular studies have highlighted how these processes are tightly regulated by cell death pathways, including apoptosis, autophagy, and ferroptosis, and by electrical and microvascular remodeling mechanisms that contribute to cardiac instability. This review integrates histopathological and molecular evidence relating to post-infarction evolution, with particular attention to the infarct border zone, the privileged substrate for arrhythmogenesis. Key molecular markers and cells involved in the inflammatory response and wound healing are discussed, as well as implications for ventricular reentry circuit formation and sudden cardiac death risk. An integrated understanding of these mechanisms offers innovative perspectives for the identification of predictive biomarkers and the development of therapeutic strategies aimed at reducing post-infarction arrhythmic outcomes.

Life doi: 10.3390/life16081298

Authors: Roko Šantić Marko Kumrić Lovre Martinović Nikola Pavlović Azer Rizikalo Marino Vilović Josip Vrdoljak Joško Božić

Chronic low-grade inflammation accompanies cardiometabolic disease, while routine risk assessment is based mainly on fasting measurements. This structured narrative review summarises human evidence for discrete postprandial changes in triglyceride-rich lipoproteins and remnants, glucose and insulin, endotoxin-related markers, innate immune cells and endothelial function. The evidence is comparatively consistent for postprandial lipaemia, glycaemic excursions and acute flow-mediated dilation responses, whereas endotoxin, cytokine and cellular findings are smaller, assay-sensitive and less consistently replicated. Based on this evidence, we introduce PRISM-CM (Postprandial Inflammatory Stress Modules in CardioMetabolic disease) as an author-developed, hypothesis-generating evidence map. It comprises five candidate biological domains—lipid–remnant burden, glucose–insulin stress, endotoxin handling, innate immune-cell activation and endothelial response—with recovery kinetics treated as a cross-domain analytic dimension. The framework was not derived by clustering, consensus methods or predictive modelling; its illustrative response patterns are not validated endotypes. A composite score is not proposed because the independence, reproducibility and incremental predictive value of the candidate measurements have not been established. Potential future diagnostic and prognostic applications require prospective validation. Reference ranges, age- and sex-specific norms, within-person reproducibility, reproducible response patterns and outcome-linked thresholds remain unknown. PRISM-CM is therefore not a clinical algorithm, risk score or treatment-selection tool.

Life doi: 10.3390/life16081297

Authors: Virginie Philippe Alexis E. Laurent André Berchtold Nathalie Hirt-Burri Lee Ann Applegate Robin Martin

Purpose: To determine whether specific in vitro biologic characteristics and manufacturing kinetics of cultured human articular chondrocytes (HACs) can serve as predictive biomarkers for patient-reported and structural outcomes following second-generation autologous chondrocyte implantation (ACI) in the knee. Materials and Methods: This prospective cohort study evaluated 67 patients (mean age 25.1 ± 8.3 years) treated with second-generation ACI for large focal cartilage defects (mean size 5.4 ± 2.5 cm2) between 2017 and 2024. HACs were expanded under Good Manufacturing Practice (GMP) conditions using human platelet lysate-supplemented media. Four biological determinants were analyzed: (1) initial chondrocyte yield (ICY) isolated from the cartilage biopsy; (2) chondrogenic activity (relative ACAN and COL2A1 expression); (3) final culture confluence level; and (4) HAC population doubling time (PDT) pre- and post-cryopreservation. Clinical outcomes (KOOS and IKDC) and MRI outcomes (MOCART) were assessed at 2 years (T1) and at a mean final follow-up of 4.2 ± 1.7 years (T2) via univariate and multiple regression analyses. Results: Static biosynthetic markers (ACAN/COL2A1 expression) and final culture confluence did not significantly correlate with longitudinal clinical (KOOS/IKDC) or structural (MOCART) outcomes. Conversely, the retained kinetic parameters served as strong prognostic indicators. Multiple regression revealed that higher ICY values significantly predicted improvements across most KOOS subscales from baseline to T2, including KOOS Symptoms (β = 63.56; p < 0.01), KOOS Pain (β = 88.83; p < 0.001), KOOS ADL (β = 107.84; p < 0.001), and KOOS Sport and Recreation Function (β = 92.3; p < 0.05). Furthermore, a prolonged PDT, indicative of diminished in vitro proliferative vigor, inversely correlated with functional recovery in IKDC (β = −26.12; p < 0.01), KOOS Pain (β = −23.09; p < 0.05), and KOOS ADL (β = −21.71; p < 0.05) scores. Conclusions: The intrinsic proliferative vigor of the HAC cellular payload (shorter PDT) and higher initial cell yields are robust kinetic biomarkers associated with markedly superior mid-term clinical outcomes following second-generation ACI for large focal chondral defects in the knee. In contrast, standard morphological and gene expression metrics failed to predict in vivo functional success. These findings advocate for a risk-based paradigm shift in GMP quality control methodologies, emphasizing dynamic HAC growth kinetics over static cellular features to optimize patient-specific regenerative potential.

Life doi: 10.3390/life16081296

Authors: Christos Eliopoulos Ioanna Langousi Matthaios Tselios Eleni Kougia Vasileios G. Papatsiros Giorgos Markou Dimitrios Arapoglou

The present study examined the supplementation of Spirulina platensis (SP) in various ratios in the basal diet of laying hens targeting the determination of the optimum amount that is required for the improvement of egg quality parameters and oxidative stress. A total of 300 24-week-old Lohmann Brown laying hens were selected and kept at a commercial farm (Vasileios Kompoulis Ltd. farm, Megara, Attiki, Greece). The employed farm was supplied with 25 similar cages. The control group was fed the basal diet, whereas the experimental feedstuff was fortified with SP by 0.1, 0.2, 2 and 5% w/w. At the end of the trial the addition by 5% w/w improved the egg weight, the yolk color, carotenoids, protein content and enhanced the eggshell strength. Additionally, the specific treatment improved the oxidative stress of the employed laying hens by recording the lowest values in TBARS and CARBs content. On the contrary the fortification with 5% w/w enhanced TAC, GSH and CAT activity. Overall, SP addition forms an intriguing approach for the improvement of the performance, physiology, and health of laying hens, with a parallel contribution to the development of more sustainable and viable feed ingredients for poultry production systems.

Life doi: 10.3390/life16081295

Authors: Ekaterina Otlyga Dmitry Otlyga Yuliya Krivova Alexandra Proshchina Olga Junemann Marina Shahina Sergey Saveliev

The development of the human adrenal gland involves dramatic structural reorganization, including migration of adrenal medulla cells, changes in cell shape, and complex dynamics of distinct cell populations. These processes require dynamic remodeling of the cytoskeleton. In this study, we investigated vimentin—a key regulator of cell shape changes—in a population of developing adrenal medulla cells, which we call large cells (LCs). We performed double immunofluorescence for tyrosine hydroxylase and vimentin on adrenal glands from human fetuses aged from 8 gestational weeks to 11 days postnatally and compared our findings with recently published single-cell transcriptomics data. We found that the ratio of vimentin-positive LCs did not change significantly with age, and staining intensity was even higher in later fetuses, indicating that vimentin immunoreactivity does not decline with age. In contrast, analysis of the scRNA-seq dataset revealed a decrease in the proportion of vimentin-positive cells within the subpopulation of late chromaffin cells. We attribute these differences to both the high stability of vimentin filaments relative to their short-lived mRNA and the physiological heterogeneity of developing chromaffin cells, the precise causes of which remain to be determined.

Life doi: 10.3390/life16081294

Authors: Ioana-Madalina Bilha Stefana Catalina Bilha Nada Akad Adrian Covic Simona Hogaș Mihai Marian Hogaș Ioana Alina Halip Calina Anda Sandu-Boz Camelia Margareta Bogdanici Irina Draga Caruntu

Background: Chronic kidney disease (CKD) is characterized by persistent inflammation, metabolic dysregulation, and mineral and bone disorder (CKD-MBD), all of which may affect ocular tissues. While retinal manifestations of CKD have been increasingly investigated, data regarding clinical anterior segment modifications and their relationship with systemic biochemical parameters remain limited. Methods: We performed a cross-sectional study including 130 participants: 53 non-CKD subjects, 29 patients undergoing maintenance hemodialysis (HD), and 48 kidney transplant recipients (KTRs). All participants underwent comprehensive ophthalmic examination, including best-corrected visual acuity (BCVA), intraocular pressure (IOP) measurement using Goldmann applanation tonometry, Schirmer I test, tear break-up time (TBUT) and assessment of cataract and blepharitis prevalence. Serum creatinine estimated glomerular filtration rate (eGFR), calcium, phosphate, parathyroid hormone (PTH), magnesium, 25-hydroxyvitamin D, and Activin A were recorded. Associations between ocular and systemic parameters were evaluated using correlation and regression analyses. Results: TBUT, Schirmer I and IOP did not differ significantly among groups (all p > 0.05). After adjustment for age, sex, BMI and diabetes mellitus (DM), HD patients showed worse visual acuity than the non-CKD reference group, reflected by higher logMAR BCVA values (adjusted B = +0.128, p = 0.024). In KTRs, higher corrected calcium was independently associated with reduced tear film parameters, correlating with both shorter TBUT (B = −0.903, p = 0.037) and lower Schirmer I values (B = −5.947, p = 0.008). Circulating Activin A increased progressively from controls to KTR to HD patients (median 189.5 vs. 273.0 vs. 314.0 pg/mL, p < 0.001) but showed no independent association with any anterior segment parameter; it was instead associated with cumulative glucocorticoid exposure (B = +1.602, p = 0.024). Conclusions: In CKD, clinical anterior segment and ocular surface differences were largely explained by age, sex, and comorbidity rather than renal replacement modality, with visual acuity deficit persisting in HD after adjusting for these covariates. At the individual level, corrected calcium was the parameter most consistently associated with tear film parameters in KTRs, whereas elevated Activin A was not independently associated with ocular involvement and likely reflects systemic disease activity.

Life doi: 10.3390/life16081293

Authors: Víctor Juan Vera-Ponce Jhosmer Ballena-Caicedo Marcos García-Rodríguez Witre Omar Padilla José Celso Paredes Carranza Fiorella E. Zuzunaga-Montoya

Household biomass combustion remains a source of household air pollution and unequal exposure among women. Its association with hypertension is biologically plausible, but evidence from Latin America remains limited, particularly when the analysis is restricted to women without a prior hypertension diagnosis, which represents the population of greatest interest for early screening. This study aimed to estimate the association between biomass cooking fuel use and screen-detected hypertension among Peruvian women aged 15–49 years without known hypertension. We conducted an analytical cross-sectional study using pooled DHS 2014–2024 microdata. Nonpregnant women aged 15–49 years with valid blood pressure measurements, cooking fuel information, complete covariates, and no self-reported diagnosis or current antihypertensive treatment were included. Screen-detected hypertension was defined as mean systolic blood pressure ≥ 140 mmHg or mean diastolic blood pressure ≥ 90 mmHg. Poisson regression with robust variance based on the complex sampling design was used to estimate prevalence ratios (PRs). The main model was adjusted for age, altitude, area of residence, natural region, education, wealth quintile, and survey year. Sensitivity, fuel-specific, effect-modification, positivity, sequential-adjustment, extended-adjustment, and continuous blood-pressure analyses were performed. The analytical sample included 301,575 women, of whom 78,499 used biomass/solid fuel and 18,571 had screen-detected hypertension. The weighted prevalence of screen-detected hypertension was 8.5% among clean-fuel users and 6.4% among biomass/solid-fuel users. The crude association was inverse (crude PR: 0.75; 95% CI: 0.71–0.79), but it reversed after adjustment (adjusted PR: 1.20; 95% CI: 1.10–1.31). Wood was associated with a higher adjusted prevalence (adjusted PR: 1.25; 95% CI: 1.14–1.36). Coal/charcoal showed an imprecise null estimate (adjusted PR: 1.00; 95% CI: 0.76–1.31). The association was stronger in urban than in rural areas (adjusted PR: 1.28 vs. 1.09; interaction p = 0.005). Among Peruvian women without known hypertension, biomass/solid cooking fuel use was associated with a higher adjusted prevalence of screen-detected hypertension, supporting the inclusion of clean-fuel transition within integrated cardiovascular prevention strategies.

Life doi: 10.3390/life16081292

Authors: Giorgia Sforza Carmen Maritato Gaia Anzini Alessia Carboni Claudia Ruscitto Laura Papetti Massimiliano Valeriani

Marfan syndrome (MFS) is a connective tissue disorder classically associated with cardiovascular, musculoskeletal, and ocular manifestations. Neurological involvement is increasingly recognized and is most commonly related to spontaneous intracranial hypotension secondary to dural ectasia and cerebrospinal fluid (CSF) leakage. By contrast, idiopathic intracranial hypertension (IIH) is exceptionally rare in pediatric patients with MFS. We report the case of an 8-year-old girl with Marfan syndrome presenting with neck pain, diplopia, bilateral papilledema, and bilateral sixth cranial nerve palsy. Brain MRI demonstrated optic nerve tortuosity, distension of the perioptic subarachnoid spaces, and partial empty sella, while venous sinus thrombosis and spinal CSF leakage were excluded. Lumbar puncture confirmed markedly elevated CSF opening pressure (48 cmH2O), consistent with IIH. CSF and plasma neurofilament light chain levels were elevated, whereas anti-MOG antibodies and autoimmune investigations were negative. The patient showed rapid clinical improvement following therapeutic CSF drainage and acetazolamide treatment. To the best of our knowledge, this represents only the second report of pediatric IIH associated with Marfan syndrome. Our patient developed idiopathic intracranial hypertension, an uncommon neurological manifestation in this condition. Through this case, we aim to highlight the diagnostic challenges, discuss the possible pathophysiological mechanisms underlying this rare association and emphasize the importance of considering intracranial hypertension in the differential diagnosis of children with Marfan syndrome presenting with neuro-ophthalmological symptoms.

Life doi: 10.3390/life16081291

Authors: Kuo-Chin Hung Min-Tser Liao Cai-Mei Zheng Shih-Feng Chen Li-Ting Lu Chia-Ter Chao Kuo-Cheng Lu

Background/Objectives: Uremic pruritus and sleep disturbance commonly coexist in patients receiving maintenance hemodialysis. This analysis examined whether supplementation with Lactobacillus casei var. rhamnosus LCR35 was associated with changes in pruritus and sleep outcomes. Methods: This 12-week randomized, open-label, controlled study included 78 patients receiving maintenance hemodialysis who were assigned to LCR35 supplementation (n = 38; three sachets daily) or usual care without probiotic supplementation (n = 40). No placebo was used. Outcomes were Pittsburgh Sleep Quality Index (PSQI) and 5-D Itch Scale results. We used analysis of covariance (ANCOVA) for between-group analyses, with the week-12 score as the dependent variable and the corresponding baseline score as a covariate. Results: In the LCR35 group, descriptive improvements were observed in the 5-D Itch Scale and selected PSQI outcomes. However, none of the baseline-adjusted between-group differences was statistically significant: for 5-D Itch Scale, −1.48 (95% confidence interval (CI), −3.64 to 0.68; p = 0.176); for global PSQI, −0.80 (95% CI, −2.09 to 0.50; p = 0.225); for PSQI sleep disturbance, −0.07 (95% CI, −0.27 to 0.13; p = 0.482); and use of sleep medications, −0.28 (95% CI, −0.67 to 0.12; p = 0.163). In the LCR35 group, the exploratory association between changes in pruritus and the PSQI sleep-disturbances component was significant before, but not after, false-discovery-rate (FDR) correction (p = 0.007; FDR q = 0.055). Conclusions: Our preliminary findings raised the possibility that LCR35 might modestly influence pruritus and specific sleep disturbances. However, our results do not firmly establish a treatment effect, nor a patient phenotype most likely to benefit.

Life doi: 10.3390/life16081290

Authors: Wanessa Michelin Joana O. Pinto Bruno Peixoto

Background: The interaction between sex, APOE ε4 status, and clinical progression in Alzheimer’s Disease (AD) remains a subject of debate. While females are often considered at higher risk for AD, the underlying structural neuroanatomical trajectories and how they are modulated by genotype are not fully elucidated. This study aims to evaluate how sex and the APOE ε4 genotype interact to influence longitudinal brain atrophy across three clinical groups. Methods: We analyzed longitudinal data from 2400 participants from the Alzheimer’s Disease Neuroimaging Initiative (ADNI), stratified by clinical group (i.e., cognitively normal, mild cognitive impairment, and AD), sex, and APOE ε4 carrier status. Using Type III Sum of Squares ANCOVA, we modeled the longitudinal variation in brain volume, controlling for baseline brain volume, and baseline severity of neurocognitive impairment and age at entry. Results: While main effects of sex and APOE genotype were not significant, the triple interaction (APOE * Sex * Clinical Group) was marginally significant (p = 0.051). Post hoc analysis revealed a distinct pattern of structural dimorphism within the AD cohort among APOE ε4-negative individuals with females exhibiting significantly greater structural preservation compared to males (Mean difference = 11.32, p = 0.051). Among APOE ε4 carriers, atrophy trajectories for males and females were statistically indistinguishable (p = 0.922), potentially suggesting that the ε4 allele exerts a dominant neurodegenerative influence that overrides sex-specific physiological differences. Conclusions: These emerging findings highlight the importance of jointly considering biological sex and APOE ε4 status to improve the characterization of Alzheimer’s disease heterogeneity and support precision medicine approaches.

Life doi: 10.3390/life16081289

Authors: Najeeb Qadi Amaal Aldakheel Eslam Shosha

Dementia is a growing global health challenge, yet many determinants of late-life cognitive decline emerge decades before symptoms appear. Midlife is a practical window for prevention because hypertension, diabetes, obesity, dyslipidemia, smoking, physical inactivity, unhealthy diet, sleep disturbance, and social isolation can be identified and modified before substantial brain injury becomes apparent. This narrative review synthesizes evidence linking midlife vascular and lifestyle exposures to late-life cognitive impairment and dementia. The most consistent data support a life-course model in which cumulative vascular, metabolic, inflammatory, and behavioral risks interact with neurodegenerative pathology and cognitive reserve. Vascular and metabolic factors act largely through small-vessel disease, endothelial dysfunction, and inflammation, whereas physical activity, healthy diet, sleep, and social engagement may strengthen resilience. Although observational evidence is vulnerable to confounding and single-risk trials may underestimate cumulative benefit, multidomain prevention remains biologically plausible and clinically actionable, as recent trials reaffirm. These priorities are especially salient in the rapidly transitioning Gulf Cooperation Council (GCC) countries, where midlife cardiometabolic risk is high and local evidence is limited. Dementia prevention should be embedded in routine midlife care, with vascular risk management and sustained lifestyle support treated as core elements of lifelong brain health.

Life doi: 10.3390/life16081288

Authors: Abdulaziz A. Albalwi Hamad S. Al Amer Rashid Mir Shahul Hameed Pakkir Mohamed Jamsheed Javid Mohammad Muzaffar Mir Waad Alamri Yousef M. Alshehre Ahmad A. Alharbi

Background: Chronic low back pain (CLBP) is a leading cause of global disability, linked to elevated IL-6 and vitamin D deficiency or insufficiency. While interferential current (IFC) therapy and exercise therapy are established treatments, their combined effect with vitamin D supplementation on inflammatory biomarkers, pain, and functional disability remains underexplored. Objective: The aim of this study was to evaluate the combined effects of physiotherapy and vitamin D supplementation on pain, disability, IL-6 expression, and vitamin D levels in CLBP patients. Methods: This two-arm, parallel-group, randomized controlled trial enrolled 60 CLBP patients, randomized into Group A (physiotherapy alone: IFC therapy and exercise therapy; n = 30) or Group B (physiotherapy plus vitamin D supplementation; n = 30). Outcomes included pain intensity (11-point Numeric Pain Rating Scale (NPRS)), disability (Arabic Oswestry Disability Index (ODI), 0–100%), and serum IL-6 and 25-hydroxyvitamin D (both by ELISA; pg/mL and ng/mL, respectively), which were assessed at baseline and 6 weeks. All 60 participants enrolled had laboratory-confirmed vitamin D deficiency or insufficiency (serum 25(OH)D < 20 ng/mL) at screening. Independent samples t-tests compared continuous baseline characteristics and between-group differences in 6-week change scores; paired t-tests assessed within-group change from baseline to 6 weeks; the chi-square test compared the categorical variable of gender (α = 0.05). Results: Both groups showed significant improvements in NPRS and ODI at six weeks (p < 0.001). Group B demonstrated significant reductions in NPRS (−3.01 vs. −1.71) and ODI (21.83% vs. 16.33%). IL-6 decreased significantly only in Group B (18.24 pg/mL, p < 0.001), with no significant change in Group A (25.96 pg/mL, p = 0.626). Vitamin D levels increased significantly in Group B (31.68 ng/mL, p < 0.001) compared with Group A (19.68 ng/mL, p = 0.115), a between-group difference that was also statistically significant. Conclusions: Combining physiotherapy with vitamin D supplementation improves outcomes in the management of CLBP. This approach effectively reduces pain, functional disability, inflammation, and vitamin D deficiency or insufficiency, offering a comprehensive, evidence-based treatment strategy.

Life doi: 10.3390/life16081287

Authors: Minglun Liang Chunwu Yu Hongyu Jiang Wentao Ma

The “RNA World” hypothesis posits a reasonable scenario for the origin of life. A question central to this scenario is: which RNA species may have emerged as the first ribozyme, thereby initiating the earliest stage of life? To date, experimental and computational studies have focused on ribozymes playing a constructive role in RNA synthesis—such as those facilitating template-directed copying or nucleotide synthesis. However, the answer remains unconvincing, primarily because their functional complexity likely demands lengthy sequences, making de novo emergence improbable. Here, we propose an alternative scenario: a destructive ribozyme, which is potentially quite short, may have emerged first. This ribozyme could cleave other RNA molecules to generate building blocks for its own replication. Notably, the dilemma for a destructive ribozyme is that it may degrade molecules of its own type, thus making it difficult to thrive as an RNA species. Using computational modeling, we demonstrate that an exonuclease ribozyme encoded within a circular RNA genome (thus evading self-attack) can spread within an RNA pool and may subsequently give rise to those constructive ribozymes. Our findings offer a new perspective on the initiation of the RNA world, thereby enriching both theoretical and experimental frameworks for understanding the origin of life.

Life doi: 10.3390/life16081286

Authors: Magdalena Chomczyńska Andrzej Kucharski Anna Szymańska Agnieszka Korolczuk Marcin Szymański

Middle ear neuroendocrine tumours (MeNETs) are rare epithelial neoplasms with neuroendocrine differentiation that pose significant diagnostic and therapeutic challenges. The clinical presentation of MeNETs is often nonspecific and can mimic other middle ear pathologies, such as chronic otitis media, cholesteatoma, or paraganglioma Common symptoms include conductive hearing loss, otalgia, intermittent or persistent tinnitus, ear fullness, and dizziness. We present five patients who underwent surgery in our University Otolaryngology Centre between 2019 and 2025, in whom histopathological examination confirmed the diagnosis of MeNET. Although MeNET is typically considered an indolent tumour, rare cases of locally aggressive behaviour and distant metastases have been reported in the literature. Metastatic potential appears to correlate with histopathological features such as increased mitotic activity, Ki-67 proliferation index > 5%. The treatment of choice for MeNET is surgical resection of the tumour, with the choice of surgical technique depending on the stage of the tumour, its relationship to surrounding anatomical structures, and the possibility of hearing preservation. In our study, we highlighted the importance of radical tumour excision to minimize the risk of recurrence. Given the risk of recurrence and the risk of potential metastases, long-term follow-up is necessary, particularly in patients with advanced-stage tumours.

Life doi: 10.3390/life16081285

Authors: Anna Biazik Magdalena Sobuś Radomir Henklewski Michał Wieteska Paweł Kordowitzki

In this case series, four equines were diagnosed with cystitis, resulting in divergent clinical outcomes. Three animals achieved complete clinical resolution without recurrence, whereas one required euthanasia due to progressive clinical deterioration. All diagnoses were established through a dual-modality investigative protocol comprising cystoscopic evaluation and histopathological examination of bladder mucosal biopsies. The cohort exhibited considerable etiological heterogeneity. One case was classified as chronic idiopathic cystitis, while another was attributed to an opportunistic bacterial infection involving Facklamia spp. A third case demonstrated histopathological features consistent with mild chronic-active inflammation, characterized by mucosal hyperplasia and focal ulceration. Of particular pathological significance was one equine presented with marked epithelial denudation accompanied by pronounced submucosal edema, notably in the absence of appreciable inflammatory cellular infiltration. These characteristic morphological features and lesions observed in the horses share certain histopathological features with inflammatory bladder lesions described as interstitial cystitis in other species. However, these conclusions should be interpreted with caution, as the findings are based on only four horses.

Life doi: 10.3390/life16081284

Authors: Cristian-Norbert Ionescu Gergő Ráduly Marian Pop Karin Ursula Horváth Dan Iovănescu Gheorghe Mühlfay

Skull base pneumatization is anatomically variable and clinically relevant to temporal bone and transsphenoidal surgical corridors, but manual volumetric segmentation is time-consuming. This retrospective pilot study evaluated a clinician-guided deep learning workflow for mastoid and sphenoid sinus compartment segmentation on bone computed tomography. Images were curated and annotated in 3D Slicer using MONAI Label and separate three-dimensional SegResNet models. The mastoid development dataset comprised 122 side-cases, with 28 reserved side-cases; the sphenoid dataset comprised 117 development and 17 reserved examinations. The best internal-validation Dice scores were 0.8660 for the mastoid model and 0.8750 for the sphenoid model. In AI-assisted correction cohorts, mean Dice ranged from 0.9539 to 0.9691 for mastoid and from 0.9347 to 0.9426 for sphenoid. In independently annotated subsets, AI-to-expert Dice was 0.8083–0.8097 for mastoid and 0.8339–0.8473 for sphenoid, while interobserver Dice was 0.8003 and 0.9136, respectively. AI assistance reduced mean segmentation time by 86.5% for mastoid and 81.4% for sphenoid. These findings support clinician-supervised AI segmentation as an efficient starting point for volumetric assessment.

Life doi: 10.3390/life16081283

Authors: Zhigang Zhao Dong Wang Wei Gao Chun-Hsien Su

Exercise training is frequently examined in relation to cognitive and brain health in aging populations. Yet many exercise–cognition studies remain difficult to interpret mechanistically because cognitive outcomes are often assessed without sufficient alignment with exercise dose, biological endpoints, or participant vulnerability. This mechanism-informed narrative review argues that future research in cognitive aging should move beyond outcome-based interpretation alone and more explicitly connect the exercise stimulus with cerebrovascular endpoints, supportive biological markers, baseline vulnerability, and cognitive or translational outcomes. The review treats cerebrovascular endpoints, including cerebral blood flow, cerebrovascular reactivity, endothelial function, vascular compliance, and neurovascular coupling, as primary mechanistic anchors because they are closely related to vascular cognitive vulnerability and are more proximal to biological adaptation than cognitive test scores alone. Immune–inflammatory markers, blood–brain barrier-related indicators, myokines, neurotrophic factors, and exercise-induced circulating factors are considered supportive endpoints because they may clarify broader biological responsiveness when selected according to a clear hypothesis. The review also considers how exercise modality, delivered dose, adherence, baseline fitness, vascular risk, cognitive status, sex, comorbidity, and recovery capacity may shape responder heterogeneity. The purpose of this review is to develop a mechanism-informed framework for interpreting exercise–cognition studies in aging by aligning the delivered exercise stimulus, participant vulnerability, biological responsiveness, and cognitive or translational outcomes. Based on this synthesis, we propose a five-layer endpoint framework that positions cerebrovascular measures as primary mechanistic anchors and supportive biological markers as hypothesis-dependent contextual endpoints. Its distinct contribution is to integrate these elements within a single interpretive structure rather than considering exercise exposure, biological responses, participant characteristics, and cognitive outcomes as separate domains. Rather than serving as a clinical prescription model or formal reporting guideline, the framework is intended to improve hypothesis–endpoint alignment and the mechanistic interpretation of exercise–cognition studies in aging.

Life doi: 10.3390/life16081282

Authors: Anna Podlasek

Stroke remains one of the leading causes of death and acquired disability worldwide [...]

Life doi: 10.3390/life16081281

Authors: George Triantafyllou Ioannis Paschopoulos Daniel Gondorf Niki Lama Evangelos Oikonomou Juan Jose Valenzuela-Fuenzalida Juan Sanchis-Gimeno Jose E. León-Rojas Charalambos Vlachopoulos Gerasimos Siasos Maria Piagkou

Persistent left superior vena cava (PLSVC) is the most frequently reported anatomical variant of the thoracic systemic venous system. While its association with congenital heart disease (CHD) is well recognized, its prevalence within CHD populations and its relative occurrence compared with non-CHD cohorts remain incompletely characterized. This study aimed to estimate the pooled prevalence of PLSVC among patients with CHD and to assess its relative frequency compared with that in control populations. A systematic review of PubMed, Scopus, and Web of Science was conducted from database inception to January 2026. Studies reporting the prevalence of PLSVC in CHD cohorts were included. A random-effects meta-analysis was performed, and risk ratios (RRs) were calculated for studies that included both CHD and non-CHD groups. Sixteen studies (n = 37,370) were included. The pooled prevalence of PLSVC in CHD patients was 6.36% (95% CI: 4.70–8.24). Significant heterogeneity was identified (I2 = 97.6%). An increased likelihood of PLSVC was observed in CHD populations (RR = 15.34; 95% CI: 5.72–41.14). However, this relative risk is based on a highly limited number of comparative studies (n = 4) and exhibits substantial heterogeneity (I2 = 92.47%); thus, it must be interpreted with significant caution. Subgroup analyses did not demonstrate statistically significant differences based on nationality (p = 0.59), study type (p = 0.32) and age group (p = 0.95), while imaging modality was statistically significant (p = 0.01). PLSVC is observed in a notable proportion of patients with CHD, although variability in reported prevalence is substantial and likely influenced by methodological differences across studies. Given its potential implications for catheter-based and surgical procedures, systematic evaluation of thoracic venous anatomy should be considered during the diagnostic and preoperative assessment of patients with CHD.

Life doi: 10.3390/life16081280

Authors: Gorana Nedin Ranković Dane Krtinić Aleksandar Nikolić Ana Cvetanović Mirjana Todorović Mitić Milica Mihajlović Irena Conić Nikola Milenković Nemanja Dimić Nada Pejčić Iva Binić

Background: Patients with malignant diseases receiving chronic opioid therapy are particularly susceptible to drug–drug interactions because of extensive polypharmacy, multimodal anticancer treatment, supportive care, and comorbidities. This study aimed to determine the prevalence of potentially clinically relevant drug–drug interactions and to identify factors associated with their occurrence. Methods: This exploratory observational cross-sectional pilot study included 49 adult oncology patients receiving chronic opioid therapy. Complete medication regimens were screened using the Medscape Drug Interaction Checker and Lexicomp. Lexicomp category D and X interactions were analysed descriptively. Because category D interactions were nearly universal and category X interactions were rare, the presence of at least one Medscape “Serious—Use Alternative” interaction was used pragmatically as the binary outcome for regression modelling. Potential predictors were assessed using univariable binary logistic regression and a parsimonious adjusted model with covariates selected using a clinically informed approach. Because of quasi-complete separation, Firth’s penalized-likelihood logistic regression was used as the primary adjusted analysis. Results: Lexicomp category D interactions were identified in 48 of 49 patients (98.0%), whereas category X interactions were present in 2 patients (4.1%). Medscape serious interactions were detected in 33 patients (67.3%). In the Firth-adjusted model, female sex was associated with lower odds of a Medscape serious interaction (adjusted OR 0.12, 95% CI 0.02–0.52), while cardiovascular disease was associated with higher odds (adjusted OR 5.16, 95% CI 1.27–26.31). Stage IV disease showed a positive but statistically non-significant association, and total medication count was not independently associated with the outcome in sensitivity analysis. Because of the small pilot sample and the resulting wide confidence intervals, these findings should be interpreted as exploratory and hypothesis-generating. Conclusions: Oncology patients receiving chronic opioid therapy had a high burden of potentially clinically relevant drug–drug interactions, predominantly Lexicomp category D interactions, warranting consideration of therapy modification and individualized monitoring rather than absolute avoidance. Regular medication review, use of complementary interaction databases, and individualized clinical assessment may improve pharmacotherapy safety in this vulnerable population.

Life doi: 10.3390/life16081279

Authors: Zuzana Šufliarska Katarína Vdoviaková Lenka Krešáková Ján Danko Pavol Rusnák Ľubomír Medvecký Mária Giretová Jozef Bíreš Filip Humeník Jana Pobehová

Background: Surgical wound dehiscence (SWD) is defined as a complete or partial separation of the surgical incision site after surgery and represents a serious complication in soft tissue surgery. Case presentation: We present a clinical case of wound dehiscence in a 7-month-old dachshund. The primary cause was an attack by another dog, leading to the development of a deep bite wound in the back area perforating the anus and numerous minor injuries in the same area. After initial treatment and apparently good general health, numerous necrotic foci were observed during wound cleaning on the 3rd day after treatment. The wound was subsequently revised and an experimentally developed biomaterial based on chitosan in combination with manuka honey was applied. The use of this combination resulted in an accelerated healing process and almost complete closure of the wound already on the 12th day after application. Conclusions: This case demonstrates and simultaneously points to the promising possibilities of using chitosan-based biomaterials and honey as natural products for the successful treatment of wounds and management of postoperative complications.

Life doi: 10.3390/life16081278

Authors: Alexander Martens Markus Schauer Susanne Mair Mohamad Motevalli Brigitte König

Microbial contamination in hospitality settings remains understudied despite the high density of human contact and diverse operational activities that characterize hotel environments. This study aimed to characterize microbial contamination patterns and environmental hygiene risks across multiple functional areas of family-oriented hotels. A cross-sectional environmental microbiological investigation was conducted in family hotels in Bavaria, Germany. A total of 225 environmental surface samples were collected from guest rooms, child areas, food-related areas, service environments, and water-exposed locations. Bacterial isolates were identified using culture-based microbiology and MALDI-TOF mass spectrometry. Microbial prevalence, contamination severity, microbial richness, and pathogen prevalence were assessed using mixed-effects ordinal logistic regression, generalized additive model (GAM), and integrated hygiene profile, all performed in R (version 4.5.1). Microbial occurrence patterns differed markedly across functional areas. Contamination category distributions differed significantly among areas, with food-related environments showing the strongest enrichment in the highest contamination category (72.7%). Food-related areas showed significantly greater odds of severe contamination than guest rooms (OR = 8.37, 95% CI: 1.89–37.00), child areas (OR = 6.16, 95% CI: 1.20–31.46), and water-exposed environments (OR = 12.34, 95% CI: 2.45–62.10). Microbial richness differed across operational zones (p = 0.048) and was positively associated with contamination severity (β = 0.141, p < 0.001). GAM revealed a significant non-linear richness–contamination relationship (p < 0.001). Integrated hygiene profile consistently identified food-related and service areas as the highest risk environments. Environmental hygiene risks in hospitality settings display functional-area heterogeneity, highlighting the need for targeted, area-specific hygiene management strategies.

Life doi: 10.3390/life16081277

Authors: Wolfgang Nitschke Simon Duval Kilian Zuchan Jostin Monge-Ruiz Frauke Baymann Barbara Schoepp-Cothenet

Across the diverse family of flavoenzymes, the isoalloxazine cofactor was found to display extremely diverse redox properties, both with respect to the absolute value of the potential regime wherein it operates and to its redox cooperativity, that is, the relative positioning of its individual 1-electron transitions. Taking together electrochemical data and 3D structural information reported for selected representatives of the flavoenzyme family, we assessed the contribution of pK value modifications at the three protonatable nitrogens of the isoalloxazine moiety. While the absolute value of the redox regime appears only weakly dependent on such pK modifications, the diversity of redox cooperativity is readily rationalized by (protein-induced) stabilization/destabilization of the proton primarily on N5 and to lesser degrees on N1 and N3. The mathematical formalism underlying the interdependence of pK values and redox midpoint potentials is subsequently extended to representatives of the family featuring extremely positive redox cooperativity (i.e., the electron bi/confurcating flavoenzymes). Observed structural idiosyncrasies of these cases were found to rationalize the extremely strong inversion (ΔE ≪ −800 mV) of 1-electron midpoint potentials in the framework of this formalism.

Life doi: 10.3390/life16081276

Authors: Antonella Cinzia Marra

The Pleistocene hippopotamuses of the Siculo-Maltese Archipelago are re-examined through the study of historical collections and newly recovered material. Fossils attributed to Hippopotamus pentlandi and Hippopotamus melitensis were described anatomically and compared morphologically and morphometrically with the continental species Hippopotamus antiquus and Hippopotamus amphibius. Comparison with the continental taxa confirms the mosaic of characters that has long complicated the identification of the ancestral taxon of Hippopotamus pentlandi. At the same time, the Sicilian hippopotamus is shown to possess a distinctive skeletal organization, combining a shortened, more gracile skull with shortened, robust limbs. Most of the Maltese material falls within the range of Hippopotamus pentlandi, although it shows a general trend towards smaller size. The available record remains insufficient to determine the characters of Hippopotamus melitensis as a distinct endemic taxon, while the occurrence of Hippopotamus pentlandi in Malta cannot be excluded. The anatomical pattern documented here provides the morphological basis for future phylogenetic analyses of the Mediterranean insular hippopotamuses.

Life doi: 10.3390/life16081275

Authors: Naiara Díaz-Marín Carmen López-de-la-Fuente Yolanda Marcén Jorge Pérez-Rey Carmen Bilbao María José López-de-la-Fuente

Acquired brain injury (ABI) frequently results in visual field loss, a disabling condition that negatively affects visual exploration, reading, mobility, activities of daily living, and quality of life. This systematic review aimed to synthesise current evidence on the effectiveness of rehabilitation interventions for visual field loss following ABI. The review was conducted according to PRISMA 2020 guidelines and prospectively registered in PROSPERO (registration number: CRD420261359820). Four electronic databases (PubMed, Scopus, Web of Science, and ProQuest) were searched for studies published between January 2015 and May 2026. Twenty-eight studies met the eligibility criteria, including randomised controlled trials, quasi-experimental studies, and case series. Interventions were classified as compensatory, substitutive, or restorative, the latter including neuromodulation techniques. Compensatory interventions provided the most consistent evidence of benefit, improving visual exploration, reading, mobility, activities of daily living, and vision-related quality of life. Restorative interventions produced more variable findings, with some studies reporting improvements in visual field sensitivity and stimulus detection, whereas evidence for substitutive approaches was limited because few eligible studies were identified. Overall, functional improvements were reported more consistently than objective visual field expansion. Current evidence supports visual rehabilitation as an important component of ABI management, although further high-quality research using standardised protocols and outcome measures is needed to establish the most effective interventions for different patient populations.

Life doi: 10.3390/life16081274

Authors: Piotr Listos Krzysztof Fronc Antonina Mazur Mateusz Bosiacki Jolanta Kotlińska Tymoteusz Słowik Joanna Listos

Levetiracetam (LEV) is an antiepileptic drug and is used in the management of various types of seizures and other clinical conditions, including anxiety and neuropathic pain. Its unique mechanism primarily involves selective interaction with synaptic vesicle protein 2A (SV2A), which stabilizes synaptic function and the release of neurotransmitters. The objective of the present study was to evaluate LEV activity in morphine (MPH) dependence. In mice, LEV was used at doses ranging from 31.25 to 125 mg/kg, depending on the experimental paradigm. It was studied in three schedules: (1) MPH tolerance to antinociceptive effects (the hot plate test—HPT); (2) MPH withdrawal signs, manifested as jumps, induced by naloxone (NAL)—2 mg/kg, i.p.; (3) MPH sensitization to locomotor activity. In this study, LEV reduced MPH tolerance to analgesic effects. Moreover, LEV diminished the intensity of NAL-precipitated MPH withdrawal signs. This drug was also effective in attenuating MPH-induced sensitization. These findings suggest that LEV may modulate behavioral adaptations associated with repeated MPH exposure. However, further mechanistic studies are required to identify the molecular and neurochemical pathways involved in these effects.

Life doi: 10.3390/life16081273

Authors: Yuanhao Li Xiaobin Li Huiguo Yang Haiying Li Yafei Liang Mingcong Ding Aikemu Maimaitijiang Jie Ren Honglei Sun Jiajia Liu

Early-weaned squabs are sensitive to dietary energy–protein supply, but combined effects remain unclear. We tested metabolizable energy (ME; 11.9, 12.2, and 12.5 MJ/kg) and crude protein (CP; 18, 19, and 20%) in 216 15-day-old gray-feathered King pigeon (Columba livia domestica) squabs using a 3 × 3 factorial design. Higher ME increased abdominal fat percentage, whereas CP affected slaughter traits. ME × CP interactions occurred for live weight, liver index, breast and leg muscle quality, and most digestive enzyme activities. At 18% CP, live weight was significantly higher at 12.5 than at 12.2 MJ/kg ME, whereas the opposite numerical trend occurred at 19% CP. Liver index was lower at 18% CP + 11.9 MJ/kg ME than at 18% CP + 12.2 MJ/kg ME. At 12.5 MJ/kg ME, breast muscle a* was lower at 20% CP than at 18% or 19% CP. Enzyme activities were generally highest at 20% CP + 11.9 MJ/kg ME. Targeted 16S rRNA sequencing associated the highest-energy/lowest-protein treatment with Lactobacillus-related taxa, whereas the opposite treatment showed higher diversity and greater contributions of Pseudomonadota- and Enterobacterales-related taxa. Responses were trait-specific; microbial findings represent associations between two contrasting diets.

Life doi: 10.3390/life16081272

Authors: Shivankar Agrawal Laurent Dufossé Sunil Kumar Deshmukh Shilpa A. Verekar

The rapid emergence and dissemination of antimicrobial resistance (AMR) among bacterial pathogens, particularly the ESKAPE group (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.), represents one of the most pressing global public health challenges, contributing to increased morbidity, mortality, and healthcare costs. The limited development of new antibiotic classes over the past two decades has intensified the search for structurally novel antimicrobial agents and adjuvants capable of overcoming multidrug resistance. Natural products continue to serve as an invaluable source of anti-infective drug leads owing to their remarkable structural diversity and broad spectrum of biological activities. Mangrove ecosystems, located at the interface of terrestrial and marine environments, harbor highly diverse microbial communities, including fungi that have evolved under extreme environmental conditions and produce a wide range of unique secondary metabolites. Beyond their ecological significance, mangrove-associated fungi have emerged as prolific producers of bioactive compounds with promising antibacterial activity against multidrug-resistant pathogens. This review comprehensively summarizes recent advances (2018–2026) in the discovery of antibacterial metabolites from mangrove-associated fungi active against ESKAPE pathogens, which discusses their structural diversity, reported antimicrobial activities, and emerging strategies for accelerating natural product discovery, including genome mining, metabolomics, OSMAC, adaptive laboratory evolution, and artificial intelligence-assisted approaches. A total of 139 chemically distinct metabolites (Compounds 1–139) isolated from mangrove-associated fungi are critically reviewed, highlighting their potential as promising leads for the development of next-generation antimicrobial agents against ESKAPE pathogens.

Life doi: 10.3390/life16081271

Authors: Emanuela Elena Mihai Matei Teodorescu Eugenio Moita Gonçalves Rajiv Reebye Mihai Berteanu

Background: Stroke remains a leading cause of long-term disability worldwide. Post-stroke spasticity (PSS) is increasingly recognized as a complex complication resulting from the interaction between neural hyperexcitability and non-neural muscular adaptations, including fibrosis, altered muscle architecture, and increased stiffness. Consequently, effective management requires a comprehensive approach targeting both components. Musculoskeletal ultrasound (MSK US) has emerged as a practical tool for muscle characterization, treatment targeting, and longitudinal monitoring within neurorehabilitation programs. Aim: To propose a clinical practice framework integrating serial MSK US into multimodal PSS management and to illustrate its application through a proof-of-concept clinical case. Methods: The proposed framework combines standardized clinical assessment, serial MSK US evaluation, BoNT-A injections, physical modalities (neuromuscular electrical stimulation and extracorporeal shock wave therapy), and task-oriented rehabilitation. Ultrasound findings, including muscle echogenicity, architecture, and Modified Heckmatt Scale (MHS) characteristics, were integrated with clinical outcomes and rehabilitation goals to support individualized decision-making. Results: Serial MSK US provided complementary information regarding muscle quality and potential non-neural contributors to spasticity syndrome, supporting treatment planning and follow-up. Although only modest morphological changes were observed over time, the illustrative case showed favorable changes in clinical assessments including spasticity, motor performance, gait, and functional independence following the multimodal intervention. Conclusions: This proof-of-concept framework highlights the potential role of serial MSK US as a practical decision-support and monitoring tool within multimodal PSS management. Integrating clinical assessment with ultrasound-informed evaluation of both neural and non-neural components may facilitate more individualized rehabilitation strategies and optimize functional outcomes. Further studies are warranted to validate and standardize this approach.

Life doi: 10.3390/life16081270

Authors: Chih-Ya Chang Yen-Sheng Lin Po-Yin Chen Li-Wei Chou

Adhesive capsulitis (AC) is a common fibroinflammatory shoulder disorder, affecting 2–5% of the general population and up to 20% of people with diabetes mellitus. Although historically considered self-limiting, the course frequently exceeds two years and imposes a substantial functional burden. Intra-articular corticosteroids remain the predominant first-line option, but their benefit is often short-lived and is offset by transient hyperglycemia in diabetic patients, potential tissue toxicity with repeated use, and limited durability. These limitations have prompted interest in non-steroidal, ultrasound-guided alternatives. This narrative review appraises two complementary strategies: ultrasound-guided regenerative injectates (hyaluronic acid, hypertonic dextrose prolotherapy, and platelet-rich plasma) and ultrasound-guided perineural hydrodissection of the suprascapular, axillary, and subscapular nerves. We summarize comparative efficacy with corticosteroids, durability, injection anatomy, and the subgroups most likely to benefit. We then propose—explicitly as a clinically defensible framework rather than a validated guideline—a phase-specific paradigm in which a timely non-steroidal injection provides early pain control while stage-appropriate rehabilitation drives functional recovery. The hypothesis that this sequence may compress the symptomatic course to approximately three to six months is supported by preliminary evidence and requires prospective validation. Because no head-to-head trial has compared the proposed paradigm with the corticosteroid-first standard, these recommendations warrant corresponding caution.

Life doi: 10.3390/life16081269

Authors: Tsair-Fwu Lee Guang-Zhi Lin Wen-Ping Yun Yi-Lun Liao Liyun Chang Chao-Hong Liu Cheng-Shie Wuu Yu-Chang Hu Yu-Wei Lin Pei-Ju Chao Yang-Wei Hsieh

Objective: This study aims to develop a preliminary explainable image-based auxiliary assessment framework for radiation pneumonitis (RP) by integrating a generative adversarial network (GAN), Gradient-weighted Class Activation Mapping (Grad-CAM), and ensemble learning to improve image-based classification performance, interpretability, and computational efficiency. Methods: Chest CT images from 46 lung cancer patients who underwent VMAT were retrospectively collected, yielding 542 RP and 1857 non-RP images. Images were preprocessed using Otsu-based segmentation and standardized before being input into the RP-GAN for feature extraction. Grad-CAM was applied to qualitatively visualize attention patterns across convolutional layers and guide feature-layer selection, while PCA reduced the extracted features from 12,288 to 730 dimensions. An ensemble stacking classifier combining RF, SVM, KNN, and XGBoost with logistic regression as a meta-learner was constructed. Model performance was evaluated using AUC, accuracy, PPV, NPV, specificity, recall, and F1-score. Results: Grad-CAM highlighted conv2d_4 and conv2d_5 as the most informative layers. PCA reduced training time from 49 min to 49 s with minimal performance loss. In the internal hold-out test set, the ensemble model achieved the highest point estimates for AUC and accuracy among the evaluated classifiers (AUC = 0.921; accuracy = 87.1%). The model identified RP-positive CT slices and provided qualitative visual cues for suspected RP-related regions. Conclusions: The proposed GAN–Grad-CAM ensemble framework showed promising internal classification performance for post-treatment CT-based RP image assessment with substantially improved computational efficiency. Its qualitative visual outputs may support clinical image review, although further external validation and quantitative localization assessment are required before clinical application as an auxiliary image-review tool.

Life doi: 10.3390/life16081268

Authors: Kai Wang Haojia Zhang Chang Zhou Zijin Sun Wei Shao Kunjing Liu Wenyuan Ma Chunyu Wang Xueqian Wang Qingguo Wang Yanyan Jiang Fafeng Cheng

Insomnia is associated with disrupted sleep architecture, hippocampal pathological alterations, neurotransmitter imbalance, and neuroinflammatory responses. This study evaluated the effects of the Prunella vulgaris compound (PVC) on sleep architecture and hippocampal alterations in p-chlorophenylalanine (PCPA)-induced insomnia model mice and examined their association with NLRP3-related pyroptotic signaling. PVC was characterized using UPLC-Q-Exactive Orbitrap MS and targeted HPLC quantification. PCPA-induced insomnia models were established in ICR and C57BL/6N mice for pharmacodynamic and molecular assessments and EEG/EMG recording, respectively. Sleep outcomes were assessed using the pentobarbital-induced sleep test and EEG/EMG; hippocampal histomorphology, neurotransmitters, and inflammasome- and pyroptosis-related markers were evaluated using histological, biochemical, RT-qPCR, and Western blot analyses. UPLC-MS tentatively characterized 198 constituents, and salviaflaside, rosmarinic acid, and linarin were quantified by HPLC. PVC shortened sleep latency, prolonged sleep duration, reduced wakefulness, increased NREM and REM sleep, and improved sleep continuity. It also attenuated hippocampal histomorphological alterations; modulated 5-HT, GABA, glutamate, and dopamine levels; and reduced NLRP3 inflammasome- and pyroptosis-related markers together with IL-1β and IL-18. PVC improved insomnia-like phenotypes and attenuated hippocampal pathological alterations in PCPA-induced insomnia model mice. These effects were associated with improved sleep architecture and neurotransmitter homeostasis and with reduced NLRP3/caspase-1/GSDMD-related signaling.

Life doi: 10.3390/life16081267

Authors: Silvijus Abramavičius Dalia Akramienė Tashfeen Tashfeen Dovilė Abramavičienė Edgaras Stankevičius

Pseudomonas aeruginosa is an important opportunistic pathogen associated with diverse healthcare-associated infections and increasing antimicrobial resistance. This systematic review evaluated antimicrobial treatment regimens, clinical outcomes, and pharmacokinetic/pharmacodynamic (PK/PD) considerations in P. aeruginosa infections. A systematic review was conducted according to PRISMA guidelines. PubMed/MEDLINE and the Cochrane Library were searched for studies published between January 2000 and March 2026. Data on antimicrobial therapies, clinical and microbiological outcomes, and PK/PD parameters were extracted. Study quality was assessed using the NHLBI quality assessment tool. Narrative synthesis was performed for all eligible studies, and a quantitative meta-analysis was conducted for studies with standardized outcome data. Of the 982 records identified, 249 studies met the inclusion criteria, and 34 were included in the quantitative meta-analysis. The evidence base was highly heterogeneous regarding study design, patient populations, infection types, and treatment regimens. Respiratory infections, particularly in cystic fibrosis patients, accounted for most studies. Quantitative synthesis showed no consistent superiority of any antimicrobial regimen (or combination of them), while treatment outcomes were influenced by infection characteristics, antimicrobial resistance patterns, and PK/PD factors. No universally superior antimicrobial regimen was identified for Pseudomonas aeruginosa infections. Some novel and promising treatment options have been identified, such as anti-Pseudomonas aeruginosa LPS monoclonal antibody panobacumab and inhaled Clostridium butyricum delivered via oxygen-driven nebulization. Two novel treatment modalities did not yield a clinically relevant effect, namely, the bispecific monoclonal antibody MEDI3902 (gremubamab) and the IC43 Pseudomonas aeruginosa vaccine. Treatment effectiveness appears to depend on clinical context, antimicrobial susceptibility, and PK/PD considerations, highlighting the need for further standardized research.

Life doi: 10.3390/life16081266

Authors: Liviu Vasile Laurențiu Augustus Barbu Nicolae-Dragoș Mărgăritescu Stelian-Stefaniță Mogoantă Tiberiu Stefăniță Țenea Cojan Gabriel Florin Răzvan Mogoș

Background: Early identification of complicated appendicitis remains challenging, particularly in elderly patients who often present with atypical symptoms and delayed diagnosis. This study evaluated the association between age, inflammatory markers, and the severity of acute appendicitis. Methods: This retrospective comparative study included 311 adult patients (182 males, 58.5%, and 129 females, 41.5%) who underwent appendectomy between January 2020 and December 2024. Demographic, clinical, and laboratory data were analyzed. Logistic regression and receiver operating characteristic (ROC) analyses were performed to assess the association between age and complicated appendicitis. Results: Complicated appendicitis was identified in 120 patients (38.6%). Patients with complicated appendicitis were significantly older than those with uncomplicated disease (49.53 ± 16.85 vs. 39.53 ± 17.58 years, p < 0.001). In the prespecified multivariable logistic regression model including age, sex, leukocyte count, and C-reactive protein (CRP), increasing age remained significantly associated with complicated appendicitis (adjusted OR 1.25 per 10-year increase, 95% CI 1.10–1.45, p = 0.002). Leukocyte count (adjusted OR 1.05 per 1000/mm3 increase, 95% CI 1.02–1.08, p < 0.001) and CRP (adjusted OR 1.12 per 10 mg/L increase, 95% CI 1.08–1.16, p < 0.001) were also significantly associated with complicated appendicitis in the multivariable model, whereas male sex was not. ROC analysis showed poor-to-modest discriminatory ability of age for distinguishing uncomplicated from complicated appendicitis (AUC = 0.669, 95% CI: 0.609–0.731), indicating that age alone has limited value as a standalone predictor. Conclusions: Although increasing age remained significantly associated with complicated appendicitis after adjustment for sex, leukocyte count, and C-reactive protein (CRP), the adjustment was limited to variables consistently available in this retrospective dataset. Therefore, residual confounding cannot be excluded, and the observed association should not be interpreted as evidence of a fully independent effect. Furthermore, the discriminatory performance of age was only modest (AUC = 0.669), indicating that age should not be used as a standalone predictor and should be interpreted within the overall clinical context.

Life doi: 10.3390/life16081265

Authors: Niki Tombolesi Emanuele Francini Gretta Veronica Badillo-Pazmay Carlo Fortunato Francesca Marchegiani Fabiola Olivieri Stefania Fumarola Rosanna Maniscalco Giulia Matacchione

Ageing is characterized by progressive metabolic and inflammatory dysregulation, in which methylglyoxal (MGO), a highly reactive dicarbonyl by-product of glycolysis, is involved in the formation of advanced glycation end products (AGEs). This review provides an integrated overview of the bidirectional relationship between MGO-derived carbonyl stress and the gut microbiota, focusing on its implications for ageing and age-related diseases. We summarize current evidence on MGO production, clearance, tissue distribution, and reactivity, with particular attention to the intestinal lumen as a site where dietary compounds, host metabolism, and microbial activity converge. Age-related dysbiosis may impair intestinal barrier integrity, alter microbial metabolite production, and promote chronic low-grade inflammation, thereby reinforcing metabolic dysfunction and favoring free MGO accumulation. Conversely, MGO and AGEs can reshape microbial communities, compromise epithelial tight junctions, and amplify inflammatory signalling through receptor-dependent and independent mechanisms. Evidence from in vitro, animal, and clinical studies supports a role for the MGO–microbiota axis in metabolic, cardiovascular, gastrointestinal, neurodegenerative, and frailty-related conditions. Targeting microbiota composition, intestinal barrier function, and MGO scavenging pathways may therefore represent a promising strategy to mitigate carbonyl stress and preserve health during ageing.

Life doi: 10.3390/life16081264

Authors: Natalia Starodubtseva Alina Poluektova Alisa Tokareva Alexey Kononikhin Alexander Brzhozovskiy Anna Bugrova Evgenii Kukaev Zulfiya Khodzhaeva Evgeny Nikolaev Gennady Sukhikh

First-trimester prediction of preeclampsia (PE) remains a major clinical challenge, particularly outside specialized fetal medicine centers. This study aimed to identify and validate serum protein biomarkers for early PE prediction using an integrated proteomic approach. A prospective cohort of 64 first-trimester singleton pregnancies (32 future PE cases, 32 matched controls) was analyzed. Untargeted proteomics was performed using DIA-PASEF-MS, followed by targeted cross-platform verification with MRM-MS. Machine learning classifiers (support vector machines, SVM, and random forest) were trained on differentially abundant proteins (FDR < 0.01, VIP > 1.5). DIA-MS identified 33 protein markers associated with complement activation, IGF transport regulation, and platelet degranulation. An SVM model with a linear kernel achieved 95% accuracy (AUC = 0.95, sensitivity = 95%, specificity = 97%). Four markers (AFM, AHSG, C8A, IGHG1) were confirmed across platforms, confirming the discovery findings. Cross-platform correlation was high: 71% of overlapping proteins showed r > 0.5 (p < 0.001), with the highest concordance observed for potential PE marker AHSG (r = 0.8, p < 0.001). PRSS1, IGHV1-4, and SERPINC1 showed a strong correlation with proteinuria (|r| > 0.5, p < 0.05), linking the proteomic signature to clinical severity. Integrated DIA-MS and MRM-MS proteomics yields a reproducible, high-performance serum signature for first-trimester PE prediction. The identified markers reflect core pathophysiological pathways and offer potential to augment current FMF-based screening algorithms.

Life doi: 10.3390/life16081263

Authors: Lihua Liu Yuhang Zhang Feiming Huang Yusheng Bao Jian Zhang

Choroidal neovascularization (CNV) is a common and serious complication in various retinal diseases that involves the growth of new blood vessels from the choroid into the subretinal space, leading to a visual threat. Its underlying mechanism has not been fully uncovered. Discovering new genes related to CNV is an important way to reveal its molecular mechanisms. In this study, we used DisGeNET as the initial data source to identify genes related to CNV. The gene cluster method and additional screening tests were designed to explore the genetic landscape related to CNV. Our comprehensive analysis revealed many genes with high confidence, specifically identifying novel candidates that traditional topological methods missed. Notably, we identified SERPINE1, COL1A1, and ANGPT1 as unique gene signatures using our network embedding approach. Functionally, SERPINE1 regulates the plasminogen activation system to control proteolytic balance, COL1A1 maintains the structural integrity of the extracellular matrix during invasion, and ANGPT1 is critical for the maturation and stabilization of nascent vessels. Meanwhile, some genes were also identified by other methods, such as MMP2, a regulator of extracellular matrix degradation. The newly found genes are essential for understanding the process that initiates CNV.

Life doi: 10.3390/life16081262

Authors: Oana-Bogdana Barboi Constantin Simiras Radu-Alexandru Vulpoi Diana-Elena Floria Vadim Rosca Delia Gabriela Ciobanu-Apostol Corina Hincu Vasile-Liviu Drug

Background: Eosinophilic jejuno-duodenitis is a rare inflammatory disorder characterized by eosinophilic infiltration of the gastrointestinal tract, with highly variable clinical presentations. Mucosal eosinophilic duodenitis and jejunitis associated with ascites is uncommon and may mimic inflammatory or neoplastic conditions. Case Presentation: A 26-year-old male presented with persistent left-sided abdominal pain without bowel habit changes. Laboratory investigations revealed only slight inflammatory syndrome and peripheral eosinophilia. Contrast-enhanced abdominopelvic computed tomography demonstrated minimal ascites and circumferential wall thickening with contrast enhancement of duodenal and jejunal loops in the left flank. Given the nonspecific imaging findings, differential diagnoses included Crohn’s disease, intestinal lymphoma, and infectious enteritis. Endoscopic evaluation with histopathological analysis confirmed dense eosinophilic infiltration of the duodenal and jejunal mucosa, establishing the diagnosis of eosinophilic jejuno-duodenitis after exclusion of secondary causes. Management and Outcome: The patient was treated with budesonide, with rapid clinical improvement and resolution of inflammatory markers. Follow-up magnetic resonance enterography performed three months after initiation of treatment demonstrated complete resolution of bowel wall thickening and disappearance of the minimal ascites. Conclusions: Eosinophilic jejuno-duodenitis should be considered in young patients presenting with small bowel thickening, ascites, and peripheral eosinophilia. Awareness of this entity is essential to avoid misdiagnosis and unnecessary invasive procedures.

Life doi: 10.3390/life16081261

Authors: Selma Hamimed Rayane Merazka Amel Kamah Fatima Zohra Kamah Mouna Keroui

CRISPR (clustered regularly interspaced short palindromic repeats)-based genome engineering is reshaping how environmental contamination can be interrogated and remediated, offering a level of programmability and specificity that conventional physicochemical workflows seldom match. Microplastics polymer fragments below 5 mm that now pervade virtually every ecosystem are especially difficult to monitor and remove because of their chemical heterogeneity, sub-millimeter size, and capacity to adsorb co-pollutants. This review examines how the molecular logic of CRISPR-Cas systems is being repurposed for two complementary goals: sensitive analytical detection and microbially driven degradation of plastic particles. We first outline the biochemistry of Cas-mediated cis- and trans-cleavage that underpins isothermal, amplification-free biosensing, and then survey direct strategies, in which polymer-binding DNA (deoxyribonucleic acid) aptamers are coupled to Cas12a (CRISPR-associated protein 12a), alongside indirect strategies that read out the molecular stress signatures provoked by microplastic exposure in sentinel organisms and plastisphere communities. On the remediation side, we discuss how targeted editing, CRISPR interference, and rationally assembled microbial consortia enhance enzymatic depolymerization and redirect carbon flux toward valuable bioproducts. By integrating detection and remediation within a single conceptual framework, we identify the principal bottlenecks, aptamer selectivity in complex matrices, reagent stability under field conditions, and host metabolic burden, and outline research priorities for translating these tools from proof of concept toward deployable environmental technologies.

Life doi: 10.3390/life16081260

Authors: Jun Lao Nannan Wang Chuyu Yao Xiangmin Piao

Background: Flower color is a well-defined trait in Platycodon grandiflorus, whereas little is known about the effects of flower color variation on the metabolic profiles of reproductive organs. Triterpenoid saponins and flavonoids have common precursors upstream, suggesting a potential carbon flux trade-off. Methods: Untargeted UPLC-MS/MS metabolomics was performed on the reproductive organs of purple- and white-flowered P. grandiflorus at six stages of flower development. Mfuzz time-series clustering, PCA, and metabolite correlation networks were used for data analysis. Results: Six clusters were assigned to 25 metabolites (17 triterpenoid saponins and 8 flavonoids). Flavonoid glycosides were found to possess a conserved inverted V-shaped accumulation pattern in both germplasms. By contrast, saponins derived from triterpenoids showed strong germplasm-dependent accumulation patterns. White-flowered plants showed sustained accumulation, with a peak at the young fruit stage. In purple-flowered plants, accumulation peaked transiently at the withering stage, followed by a decline. PCA validated that metabolic divergence increased with developmental progression. Conclusions: Based on the metabolomic profiles, we hypothesize a putative trade-off model in which floral color divergence may change upstream carbon flux allocation between the triterpenoid saponin and flavonoid pathways. The young fruit stage of white-flowered plants is a candidate harvesting period for bioactive saponins. These conclusions are based only on the pattern of metabolite accumulation and need to be validated by multi-omics.

Life doi: 10.3390/life16081259

Authors: Suliman Aladhadh

Predictive models used for the assessment of cardiovascular disease (CVD) risk must not only be highly accurate but also explainable and reliable for clinical decision-making. In this paper, we present a methodological framework for explainable machine learning and deep learning approaches to CVD risk prediction that uses clinically derived electrocardiogram (ECG) features and conventional CVD risk factors. A large synthetic dataset containing 30,000 patient cases was created according to a clinically validated distribution to provide reproducible and privacy-preserving benchmarking. A series of interpretability tests were performed on multiple models of machine learning and deep learning categories within a unified experimental setup. Out of the tested models, XGBoost and BiLSTM provided the highest discrimination capabilities in the machine learning and deep learning categories, respectively. Explaining the predictions via the SHapley Additive explanations (SHAP) approach, calibration of probability estimations, uncertainty quantification, and ablation studies were applied to assess the explainability, reliability, and robustness of the models. We show that the presented framework allows for the production of consistent and interpretable predictions along with the evaluation of the models’ explainability and predictive reliability. Instead of focusing on the single-feature importance, this paper presents a reproducible methodological framework for the explainable modeling of CVD risk.

Life doi: 10.3390/life16081258

Authors: Rogier Schipperheijn Frederik G. Dikkers Bernadette S. de Bakker

Introduction: The ecto-endodermal boundary in the human oral cavity remains debated. Rather than a distinct line, this transition forms a complex interface, especially in the developing mouth and pharynx. This ambiguity is clinically relevant, as, for example, HPV-induced tumors often arise where ecto- and endoderm meet. The boundary is generally placed at the posterior third of the tongue and behind the uvula, though its developmental basis remains unclear. This review summarizes the literature on the embryological development of the mouth and pharynx to trace origins and interactions of ectodermal and endodermal derivatives and identify potential tumor initiation regions. Methods: PubMed articles on oral structures were reviewed to approximate the ecto-endodermal border. Results: Teeth originate from ectoderm. The origin of salivary glands and papillae depends on their location. No study reports were found for the tonsils, incisive papilla, tubarial glands, and faucial pillars. Conclusions: A clear educational image of the origin of the structures of the human mouth is made to help identify regions of risk for oral cancer.

Life doi: 10.3390/life16081256

Authors: Mejdi Snoussi Emira Noumi Manal Mohammed Alzahrani Khulood Fahad Alabbosh Qusai Alsenani Mamdouh Alshammari Mohd Adnan Arif Jamal Siddiqui Riadh Ben Salah Naourez Ktari Karim Hosni Vincenzo De Feo Adel Kadri

Background/Objectives: Allium atroviolaceum Boiss. is a functional food rich in phenolic compounds; however, the contribution of different plant organs to its bioactivity remains insufficiently understood. Methods: An integrated approach combining UHPLC-QTOF-IMS based metabolite profiling, network pharmacology, molecular docking, and experimental antioxidant assays was employed. Results: Metabolite profiling revealed that the flowers were rich in anthocyanins and flavonoids; leaves in flavonol glycosides; bulbs in flavonoids, phenolics, and lipids; and flower stalks in flavonoid glycosides and sterols. Interestingly, flowers exhibited the strongest antioxidant scavenging activity, with the lowest IC50 values of 3.08 ± 0.21 µg/mL and 89 ± 2.8 µg/mL in the DPPH and ABTS assays, respectively. They also showed the highest FRAP response at 2 mg/mL (1.367 ± 0.010 at 2 mg/mL). Moreover, flower stalks were particularly effective in limiting lipid oxidation, displaying the greatest activity in the β-carotene bleaching assay, with an IC50 value of 14.75 ± 0.29 µg/mL. Network-based target prioritization highlighted several oxidative stress/inflammation-associated proteins, including EGFR, HRAS, TGFB1, BCL2, JUN, TNF, TP53, MAPK1, and PTGS2. Additionally, Caffeoyl pinoresinol showed the strongest predicted interaction with PTGS2 and was therefore selected as a candidate ligand for further computational evaluation. Conclusions: A. atroviolaceum exhibits organ-specific phytochemical diversity and potent antioxidant activity mediated through multiple molecular targets. The in silico analyses identified candidate compound–target associations related to oxidative stress and inflammatory pathways, but these findings remain predictive and require biochemical and cell-based validation.

Life doi: 10.3390/life16081257

Authors: Sahil Ajit Saraf Wai Po Kevin Teng Kolangara Veetil Santosh Sencer Karakaya Amala Abbas Tony Kiat Hon Lim Kankanamage Malinda Amesh Karasinghe Zachariah Chowdhury Priti Singh Anubhav Narwal Samrat Bhattacharjee Bidyut Bikash Gogoi Bahoran Singh Mohamid Afroz Khan Daljeet Kaur Kaveh Taghipour Li Yan Khor

Introduction: Prostate needle biopsy reporting is time-consuming because each case typically includes 12–18 cores that require detailed assessment before a final case-level diagnosis is issued. Reporting is also influenced by pathologist expertise, particularly for tumour detection and ISUP Grade Group assignment. This study evaluated an artificial intelligence (AI)-based system designed to identify tumour and provide segmentation-based visual outputs highlighting Gleason patterns and suggesting an ISUP Grade Group. Methods: A strongly supervised approach was used to develop the algorithm. Twenty-seven pathologists annotated 2115 prostate needle biopsy whole-slide images, followed by two levels of senior pathologist reviews. An independent external test dataset of 150 prostate needle biopsy whole-slide images was then evaluated. Ground truth (GT) was established by two genitourinary pathologists, with disagreements resolved by consensus. The same slides were independently reviewed by 11 pathologists without AI assistance, while the AI system analysed the slides in parallel. AI performance and pathologist consensus were compared to GT. Results: For benign versus malignant classification, the AI identified all 109 malignant slides, with no false-negative predictions, while the pathologist consensus missed two malignant slides. The AI correctly classified 38/41 benign slides, compared to 39/41 for the pathologist consensus. For ISUP Grade Group assignment, the AI showed exact agreement with GT in 57/109 malignant cases. The AI more frequently assigned a higher Grade Group than GT. Conclusions: In this retrospective evaluation, the AI system showed high sensitivity for tumour detection and promising ordinal agreement for ISUP Grade Group assignment. These findings support further evaluation of the system within supervised prostate biopsy workflows.

Life doi: 10.3390/life16081255

Authors: Annunziata Raimondo Annunziata Nigro Mara Corbisieri Valentina Giudice Bianca Serio Serena Lembo Carmine Selleri

Cutaneous graft-versus-host disease (GVHD) is a common complication of allogeneic haematopoietic stem cell transplantation (allo-HSCT). Despite established guidelines, discrepancies persist between clinical trial evidence and real-world practice, particularly for newer agents such as ruxolitinib. To characterize the phenotypic spectrum of cutaneous GVHD, evaluate real-world treatment strategies—with a specific focus on systemic and topical ruxolitinib—and assess adherence to national and international guidelines, we conducted a retrospective observational study of patients undergoing allo-HSCT between 2015 and 2025 who were referred to a dedicated dermato-haematology clinic. Clinical, histological, and therapeutic data were collected. Cutaneous manifestations were classified according to National Institutes of Health (NIH) and Italian Group for Blood and Marrow Transplantation (GITMO) criteria. Of 62 transplanted patients, 44 (71%) developed GVHD, with the skin as the most frequently involved organ. Acute GVHD predominantly presented with maculopapular eruptions, whereas chronic GVHD showed heterogeneous phenotypes, including sclerotic variants. First-line management was largely guideline-concordant. Systemic ruxolitinib was administered to 3% of patients in the aGVHD group and 3% in the cGVHD group. Steroid-refractory and steroid-dependent status was not systematically recorded; therefore, treatment eligibility could not be reliably determined, and the observed frequencies should not be interpreted as evidence of underuse. Topical ruxolitinib was not used and remains investigational for cutaneous GVHD. Interpretation should also consider that the study period encompassed changes in regulatory approval, reimbursement, and access to targeted therapies. Structured multidisciplinary assessment may support the management of complex cutaneous GVHD, although its effects on treatment decisions and patient outcomes require prospective evaluation.

Life doi: 10.3390/life16081254

Authors: Bianca Liana Grigorescu Leonard Azamfirei Sânziana Bora Dorin Bica Irina Săplăcan Raduly Gergo Mihaly Veres

Intensive care medicine represents one of the most challenging setting in modern healthcare, where specific mechanisms intertwine and form a dynamic biological model, where organ dysfunction can easily evolve to multi-organ dysfunction, continuously reshaping the patient’s clinical course. The critically ill patient represents a biological system resulted from interaction between maladaptive and adaptative mechanisms, therefore generates a large volume of data that can exceeds human cognitive capacity. Artificial intelligence can integrate multimodal physiological, laboratory, and clinical data into a dynamic representation of the patient’s biological trajectory. AI-tools and machine learning technologies have evolved to potential clinical support tools, with great perspectives for future implementation, but currently with limited use in clinical practice. This article is a narrative review of artificial intelligence in ICU, aiming to present current evidence and limitations.

Life doi: 10.3390/life16081253

Authors: Samuel Crowley George-Abraam Tawfik Richard Villa Claire Larson Isaac Yeung Sergio D. Bergese

Patients with heart failure require special considerations in the perioperative setting given the challenges caused by heart failure pathophysiology. This pathophysiology is complex and involves several mechanisms that contribute to both disease progression and the clinical syndrome, including cardiac remodeling, neurohormonal activation, and resultant hemodynamic derangements. Anesthetic management of these patients requires diligence at all stages of care. Preoperative assessment includes risk stratification, management of comorbid conditions, and heart failure medication management. Anesthetic agents have differing effects on hemodynamics and selection of these agents must be aimed at preventing hemodynamic collapse in patients with heart failure. In addition, the increased risk of intraoperative complications among these patients often necessitates means of invasive hemodynamic monitoring and use of vasopressor or inotropic support. Finally, the postoperative care of heart failure patients requires attention to the volume status of these patients, as well as considerations for pain management and modulation of the surgical stress response. In this review article, we aim to review how the pathophysiologic mechanisms and changes that occur with heart failure impact anesthetic management and patient-centered perioperative care.

Life doi: 10.3390/life16081252

Authors: Junda Wu Qingyun Huang Xin Wang Xiaoping Wei Bo Yu Rong Yang Longxin Xu Yongcai Zhu

Guizhou cattle are important indigenous bovine genetic resources for regional beef production and germplasm conservation. This study characterized the transcriptome-wide N6-methyladenosine (m6A) enrichment landscape in longissimus dorsi muscle from three representative indigenous Guizhou cattle breeds—Guanling (GL), Wuchuan (WC), and Weining (WN)—and compared these profiles with those of Simmental (XM) cattle as a commercial reference. MeRIP-seq was used to detect group-level m6A-enriched regions and regions with differential m6A enrichment, followed by Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and database-inferred protein–protein interaction analyses. qRT-PCR was used to examine transcript abundance of selected network-prioritized candidate genes, rather than to validate m6A enrichment. MeRIP-seq detected 17,659, 19,530, 17,501, and 16,271 m6A-enriched regions in GL, WC, WN, and XM cattle, respectively. Compared with XM cattle, 2822, 5914, and 3655 regions with differential m6A enrichment were detected in GL, WC, and WN cattle, respectively. ACTB, CTNNB1, and AKT2 were prioritized for follow-up, and qRT-PCR showed breed-associated differences in their transcript abundance. These exploratory findings provide an epitranscriptomic resource and candidate targets for future MeRIP-qPCR, phenotypic, and functional studies.

Life doi: 10.3390/life16081251

Authors: Lukas S. Fiedler Tobias Meyer Fabian Burk Fynn Roters

Background: Obesity and diabetes mellitus (DM) are established risk factors for impaired wound healing, but their associations with early postoperative complications following surgical tracheostomy in critically ill patients remain insufficiently characterized. This study evaluated the associations of obesity and DM with clinically documented early peristomal wound infection following bedside surgical tracheostomy using either the Visor or Björk technique. Methods: This retrospective single-centre cohort study included 92 ICU patients who underwent bedside surgical tracheostomy between 2022 and 2023. Postoperative complications were retrospectively assessed from clinical documentation at postoperative days 5–7 and 10–14. Documentation at days 5–7 was available for all 92 patients, whereas documentation at days 10–14 was available for 78 patients. The analyses therefore evaluated complications documented during the available postoperative follow-up period of up to 14 days rather than complete 14-day cumulative incidence. Associations of obesity and DM with clinically documented peristomal wound infection were assessed using exploratory univariable analyses. Results: Clinically documented peristomal wound infection occurred in 18 of 92 patients (19.6%) during the available postoperative follow-up period. Infection was documented in 10 of 28 patients with obesity (35.7%) compared with 8 of 64 patients without obesity (12.5%; crude OR 3.89, 95% CI 1.33–11.35; Fisher’s exact p = 0.020). Infection was also more frequent in patients with DM (12/31, 38.7%) than in those without DM (6/61, 9.8%; crude OR 5.79, 95% CI 1.91–17.57; Fisher’s exact p = 0.002). Postoperative haemorrhage was numerically more frequent in patients with obesity (14.3% vs. 6.3%), but this association was not statistically significant (crude OR 2.50, 95% CI 0.58–10.78; Fisher’s exact p = 0.242). Conclusions: Obesity and DM were associated with higher crude odds of clinically documented early peristomal wound infection following bedside surgical tracheostomy. Given the exploratory, unadjusted analyses and incomplete follow-up through days 10–14, these findings should be considered hypothesis-generating and require confirmation in larger prospective cohorts.

Life doi: 10.3390/life16081250

Authors: Simon Wagner Anne Schroeder Sara Martina Steinmann Claudia Kunst Kirstin Pollinger Manuela Gunckel Elisabeth Aschenbrenner Martina Müller Karsten Gülow

Colorectal cancer (CRC) is a leading cause of cancer death, with resistance and apoptosis evasion—often via Bcl-2—representing major challenges. The redox-modulating drug dimethyl fumarate (DMF) has demonstrated efficacy in hematologic malignancies; however, its potential in solid tumors remains unclear. Here, we show that DMF, especially in combination with the Bcl-2 inhibitor venetoclax (ABT-199), induces apoptosis in HCT-116 CRC cells. DMF impairs mitochondrial respiration, causing membrane hyperpolarization, ATP depletion, autophagy, and cell cycle arrest. Combined treatment increases metabolic stress, reduces proliferation, and induces sustained G2 arrest with downregulation of cyclins and CDKs. These findings highlight a combined effect targeting redox balance and apoptosis in CRC. Given their clinical availability, DMF and ABT-199 represent a promising combination for further preclinical evaluation.

Life doi: 10.3390/life16081249

Authors: Elena Bischoff Stoyanka Vladeva Fabian Bischoff Mira Hristova Nikola Kirilov

Pregnancy is associated with physiological adaptations in calcium metabolism and skeletal homeostasis that may result in temporary reductions in bone mineral density (BMD). However, routine skeletal assessment during pregnancy remains limited because dual-energy X-ray absorptiometry (DXA) involves ionizing radiation. This study aimed to compare bilateral femoral neck BMD and Z-scores in pregnant and non-pregnant women using Radiofrequency Echographic Multi-Spectrometry (REMS), a radiation-free ultrasound-based technology, and to assess correlation between bilateral measurements. In this cross-sectional study, 100 women were enrolled, including 40 pregnant women and 60 non-pregnant women with comparable age and body mass index (BMI). Bilateral femoral neck BMD and age-adjusted Z-scores were evaluated using REMS. Pregnant women had a mean age of 33 ± 5 years, pre-pregnancy BMI of 25.8 ± 6.5 kg/m2, and gestational age of 21 ± 5 weeks, with no significant differences in age or pre-pregnancy BMI compared with controls. Pregnant women demonstrated significantly lower left femoral neck BMD than controls (0.805 vs. 0.862 g/cm2; p = 0.0018) and lower left femoral neck Z-scores (−0.08 vs. 1.18 SD; p = 0.003). A strong positive correlation between left and right femoral neck BMD was observed (R = 0.78). Similarly, right femoral neck BMD was significantly lower in pregnant women compared with controls (0.835 g/cm2 vs. 0.892 g/cm2; p = 0.0026). The right femoral neck Z-score was 0.15 ± 1.15 SD in pregnant women and 1.45 ± 1.10 SD in controls (p = 0.002). REMS demonstrated the ability to detect differences in femoral neck bone parameters between pregnant and non-pregnant women while providing bilateral measurement consistency. These findings support the feasibility of REMS as a safe, radiation-free approach for assessing maternal skeletal status during pregnancy. However, the cross-sectional design and lack of longitudinal follow-up limit conclusions regarding the progression and reversibility of pregnancy-associated bone changes. Future prospective studies with larger cohorts and postpartum monitoring are needed to further define the clinical role of REMS in maternal bone health assessment.

Life doi: 10.3390/life16081248

Authors: Luis Romero-Vera David Ulloa-Díaz Jonathan Concha-Poblete Misael Sandoval-Cárcamo Claudio Carvajal-Parodi Francisco Guede-Rojas Carlos Jorquera-Aguilera Cristobal Ulloa-Barboza Claudio Hernández-Mosqueira Gustavo Pavez-Adasme Eduardo Guzmán-Muñoz Héctor Fuentes-Barría Jorge Leschot-Gatica

Background: Professional women’s football requires precise neuromuscular characterisation because of the high mechanical demands associated with acceleration, deceleration, jumping, braking, landing, and rapid force production. The aim was to characterise the multivariate neuromuscular profile derived from the countermovement jump (CMJ) and isometric mid-thigh pull (IMTP), and to examine neuromuscular components according to tactical positions in professional female football players. Methods: Twenty-three professional female football players were assessed using the CMJ and IMTP. Force-time, impulse, rate of force development, jump performance, landing, and inter-limb imbalance variables were extracted. Principal component analysis (PCA) was applied to the standardised variables, and individual component scores were compared according to tactical positions using one-way analysis of variance (ANOVA). Results: The first five principal components explained 79.19% of the total variance. PC1 represented the propulsive force and the rate of force development; PC2 reflected total force and mechanical impulse; PC3 represented early explosiveness; PC4 grouped indicators of inter-limb imbalance; and PC5 integrated jump performance and landing control. No statistically significant differences were observed between tactical positions. Conclusions: The combination of CMJ, IMTP, and PCA enabled multiple muscle-force variables to be synthesised into interpretable neuromuscular dimensions. The findings provide preliminary and exploratory evidence to support the individual monitoring of female football players. Given the low participant-to-variable ratio, the PCA findings should be considered exploratory.

Life doi: 10.3390/life16081247

Authors: Zhixia Zhao Huizi Liu Sheng Yang Xing Liu Qiuxia Chen Jinwang Wang

Background/Objectives: bHLH transcription factors regulate key plant processes including development, stress adaptation, and metabolism. In Kandelia obovata, a mangrove species well known for its high tolerance to harsh environments, the bHLH gene family has not been systematically investigated. Here, this study aims to perform genome-wide identification and expression profiling of KobHLH genes under normal and saline conditions. Methods: bHLH members were identified by homology searches (HMMER and BLASTp) against the K. obovata protein dataset and verified with SMART-based domain analysis. Subsequent analyses covered physicochemical features, conserved motif composition, and phylogenetic relationships. Transcript abundance was quantified via RNA-seq in various organs (roots, stems, leaves, flowers, and fruits) as well as under saline conditions. Differentially expressed transcripts were selected for co-expression network construction. Results: Phylogenetic analysis of the 118 identified KobHLH loci assigned them into 18 distinct clades. Organ-preferential expression patterns were observed, leading to their classification into three putative functional categories. Upon salinity exposure (10–30‰), 61 of the 118 KobHLH genes exhibited differential expression, grouped into ten significant clusters. A co-expression network comprising 20 hub genes and 198 edges was constructed. Conclusions: This study provides the first genome-wide characterization of the bHLH family in K. obovata, revealing its evolutionary divergence and identifying 61 salt-responsive candidates, notably KobHLH108 and KobHLH5 associated with secondary metabolism, for further functional validation. The findings provide a foundation for future functional studies on stress adaptation mechanisms in this ecologically valuable mangrove species.

Life doi: 10.3390/life16081246

Authors: Cristina Galisteo Dáša Straková Alicia García-Roldán Rafael R. de la Haba Cristina Sánchez-Porro Antonio Ventosa

Hypersaline soils are poly-extreme terrestrial habitats characterized by high salinity, in some cases heavy-metal contamination, temperature fluctuations, and nutrient limitation. These conditions impose strong selective pressures, and many prokaryotic inhabitants still remain uncultured. Here, we conducted an extensive culturomics study of 549 isolates from the hypersaline soils of the Odiel Saltmarshes Natural Area (Southwest Spain) and compared the results with previously generated shotgun metagenomic datasets from the same environment in order to evaluate taxonomic composition, functional potential, and ecological representativeness. Cultivation across media containing 7.5%, 15%, and 25% (w/v) total salts yielded microorganisms belonging to three major phyla: Pseudomonadota, Bacillota (Bacteria) and Halobacteriota (Archaea). At the genus level, bacterial isolates were dominated by Marinobacter, Halomonas, and Aquibacillus at 7.5% (w/v) salinity, whereas extremely halophilic archaea, including Halorubrum, Halogeometricum, and Haloarcula, were predominantly recovered from media containing 25% (w/v) salts. Among the isolates, 57 strains displayed identity values < 98.65% for 16S rRNA gene sequence comparison, suggesting their putative status as new taxa. Comparison with metagenomic datasets showed that culture-dependent approaches successfully recovered the dominant haloarchaeal groups but missed some abundant bacterial phyla, such as Gemmatimonadota. Conversely, culturomics enabled the isolation of unknown species from the rare biosphere, including representatives of the novel genus Terrihalobacillus, which are typically detected at low abundance in metagenomic datasets. Together, these results demonstrate the complementarity of culturomics and metagenomics and provide an insight into the microbial communities inhabiting the hypersaline soils of the Odiel Saltmarshes.

Life doi: 10.3390/life16081245

Authors: Ioannis Ilias Chrysi Keskinidou Georgios Poupouzas Vasileios Issaris Nikolaos S. Lotsios Efthymia Botoula Marinella Tzanela Dimitra A. Vassiliadi Stelios Kokkoris Charikleia S. Vrettou Alice G. Vassiliou Ioanna Dimopoulou

Background: The growth hormone (GH)–insulin-like growth factor (IGF) axis is profoundly dysregulated in critical illness. GDF-15 and individual IGF-binding proteins (IGFBPs) have separately been proposed as prognostic biomarkers, but to our knowledge, no prior study has simultaneously characterized all major GH–IGF axis components and GDF-15 in the same critically ill cohort, precluding assessment of their joint intercorrelation structure. Aim: To provide the first simultaneous characterization of the intercorrelation structure among ten GH–IGF axis components and GDF-15 in a single ICU cohort, testing whether this structure is robust to adjustment for illness severity; and, secondarily, to describe admission discriminatory performance relative to APACHE II and SOFA. Methods: This was a prospective observational pilot study of 43 critically ill adults with admission (T01) measurement of ten GH–IGF axis biomarkers and longitudinal follow-up to day 15. Spearman correlations and hierarchical clustering characterized the admission intercorrelation structure; partial correlations adjusting for APACHE II and SOFA, and bootstrap confidence intervals, assessed robustness. Secondary analyses included the examination of admission discrimination (ROC/AUC), a leave-one-out cross-validated combined model, and longitudinal trajectories. All analyses were exploratory, hypothesis-generating, and unadjusted for multiple comparisons unless stated. Results: Hierarchical clustering identified a coherent cluster comprising GDF-15, IGFBP-1, IGFBP-2, and growth hormone-binding protein (GHBP), distinct from classical GH-resistance markers (GHR vs. healthy controls, GHR vs. admission) and from GH, IGF-1, acid-labile subunit (ALS), and IGFBP-3. Within this cluster, GDF-15 correlated with IGFBP-1 (ρ = 0.65, 95% bootstrap CI 0.44–0.78), IGFBP-2 (ρ = 0.50, CI 0.19–0.71), and GHBP (ρ = 0.47, CI 0.20–0.68); GDF-15 showed no correlation with classical GH-resistance markers. These correlations were essentially unchanged after adjusting for APACHE II or SOFA (partial ρ within 0.03–0.16 of unadjusted values), indicating the structure is not attributable to shared confounding by illness severity. This robustness extended to further adjustment for IL-6, age, BMI, and mechanical-ventilation duration, and results from all 45 pairwise T01 correlations were re-examined with Benjamini–Hochberg false-discovery-rate correction (9 of 11 nominally significant pairs retained q < 0.05). However, the GDF-15–GHBP correlation, unlike the GDF-15–IGFBP-1/IGFBP-2 correlations, attenuated substantially after adjustment for IL-6 and was not consistent across a brain-injury/non-brain-injury subgroup sensitivity analysis, indicating this specific link is less specific than the others. In secondary exploratory analyses, GDF-15 had the highest individual admission AUC (0.74) among biomarkers but was substantially outperformed by APACHE II (AUC 0.90) and SOFA (AUC 0.81); a combined GDF-15 + IGFBP-2 model did not improve on GDF-15 alone. Conclusions: This study identifies a severity-independent intercorrelation structure linking GDF-15 to inhibitory IGFBPs, distinct from classical GH-resistance signaling, in critically ill patients. Although the findings do not support any clinical application at this stage, further study in adequately powered, multicenter cohorts can be contemplated.

Life doi: 10.3390/life16081244

Authors: Piotr Morasiewicz Monika Zaborska Łukasz Tomczyk Katarzyna Sznajder Igor Kowal Krystian Kazubski

Background: Good Ilizarov treatment outcomes are associated with low complication rates, rapid formation of strong bone regenerate, and relatively early removal of the Ilizarov external fixator. Some authors recommend the use of pharmaceutical agents that facilitate and accelerate bone union. There are no previous studies evaluating Osteogenon in Ilizarov osteotomies. The purpose of our study was to assess radiological and clinical effects of Osteogenon in patients undergoing osteotomy combined with Ilizarov fixation. Methods: In this retrospective study we assessed 35 patients who had undergone lower leg lengthening osteotomies with the Ilizarov method in the years 2021–2022 and received adjunctive Osteogenon. The control group comprised 60 patients matched for age, sex, BMI, and comorbidities, who did not receive Osteogenon. We assessed the following clinical and radiological parameters: the duration of fixation, total initial limb shortening, total limb lengthening, achieving bone union following osteotomy, elongation index, alignment index, complications, analgesic use, and satisfaction with treatment. Results: The median duration of fixation was worse (longer) in the Osteogenon group (183 days) compared to the control group (143 days), p = 0.014. There were no differences between groups in initial limb shortening, median limb lengthening, median elongation index, median alignment index, and complication rate. A proportion of 22.9% of patients in the Osteogenon group and 80% patients in the control group took analgesics on post-treatment follow-up, p < 0.001. In the study, 65.71% of Osteogenon patients and 55% of control patients were very satisfied with the treatment. Conclusions: Taking Osteogenon improves patient’s satisfaction with treatment and limits analgesic use following osteotomies combined with Ilizarov fixation. Osteogenon was associated with a significantly longer external fixation period and failed to demonstrate measurable improvements in radiological outcomes. Any considerations regarding the potential benefits of using Osteogenon should be treated with caution.

Life doi: 10.3390/life16081243

Authors: Dongze Li Kaiming Chen Shibo Kong Zhengyu Gou Xinmiao Feng Jiezhong Wu

Background: Uncertainty persists regarding the dose–response association between exercise exposure and blood pressure (BP) control in chronic kidney disease (CKD). This dose–response network meta-analysis examined whether different exercise modalities and model-estimated metabolic-equivalent exposure levels were associated with changes in systolic BP (SBP) and diastolic BP (DBP). Methods: Exercise dose was estimated as metabolic equivalents of task minutes per week (MET-min/week) from reported session duration, frequency, and intensity. Primary outcomes were changes in SBP and DBP, with exploratory subgroup analyses by dialysis status, baseline BP, and intervention duration. Results: The analysis included 26 randomized controlled trials (1218 participants). Aerobic exercise was associated with lower SBP, with the greatest model-estimated reduction at 880 MET-min/week (MD, −7.73 mm Hg). Resistance and mind–body exercise also showed model-estimated reductions, whereas combined aerobic-resistance training was not statistically significant. Dose–response estimates differed by dialysis status and intervention length, but several subgroup estimates were based on sparse data and wide uncertainty. Evidence certainty ranged from very low to high (CINeMA). Conclusions: Exercise interventions may be associated with BP reduction in CKD, but the identified MET-min/week ranges should be interpreted as exploratory, model-derived estimates rather than validated therapeutic thresholds. Better reported randomized trials are needed to confirm dose ranges, safety, adherence, and tolerability in dialysis and non-dialysis CKD populations.

Life doi: 10.3390/life16081242

Authors: Alessia Perna Adriano Paolucci Teresa Esposito Giulia Spernanzoni Annalisa Bruno Rosa Maria Russo Maria Pierdomenico Massimo Santoro

(1) Background: Airports workers are exposed to a mixture of airborne pollutants generated by aircraft operations, ground support equipment, road traffic, and indoor microenvironments. Indoor air quality, influenced by outdoor pollutant and ventilation dynamics, represents an important, but overlooked determinant of occupational exposure. Fine and ultrafine particulate matter (PM) can induce oxidative stress, inflammation, and xenobiotic responses, affecting not only the respiratory system, but also other organs. (2) Methods: This review examines the health effects of occupational exposure among airport workers, with emphasis on indoor air pollution besides aircraft engine emissions. Studies addressing exposure characterization, biological effects, biomarkers, and risk management strategies were critically evaluated. (3) Results: Occupational exposure is driven by both combustion-derived pollutants and indoor–outdoor air exchange processes. Ultrafine particles, black carbon, polycyclic aromatic hydrocarbons, and trace metals contribute to oxidative stress, inflammatory responses, and xenobiotic pathway activation. Monitoring indoor microclimatic parameters, including temperature, atmospheric pressure, and humidity, may facilitate the identification of event-related deterioration in indoor air quality. (4) Conclusions: Indoor air pollution should be recognized as a key component of airport occupational exposure. Integrating indoor/outdoor air quality monitoring and biomarker-based surveillance may improve risk assessment and support more effective protection of airport workers, while advanced predictive tools, including AI-based exposure modelling, represent a promising direction that still requires dedicated validation.

Life doi: 10.3390/life16081241

Authors: Shun-Xin Deng Lih-Geeng Chen Shih-Yi Hsiung Vinh Tuyen T. Le Chia-Jung Lee Yves S. Y. Hsieh Ching-Chiung Wang

Wolfiporia extensa (Peck) Ginns (WE) is a fungus widely used in Traditional Chinese Medicine. Its four medicinal parts, Poria (WP), Rubra Poria (RP), Poriae Cutis (PC), and Poria Cum Pini Radix (PPR), are prescribed for distinct therapeutic purposes; however, their quality standards and pharmacological differences remain unclear. This study established reliable quality markers and compared the neuroprotective activities of four medicinal parts of WE. Seven triterpenoids were quantified using validated high-performance liquid chromatography (HPLC) assays, while monosaccharide profiles were analyzed using 1-phenyl-3-methyl-5-pyrazolone-HPLC. The International Commission on Illumination L*a*b* colorimetry and chemometric models were used for authentication. The neuroprotective effects of aqueous extracts were assessed in SH-SY5Y, BV-2, and HOG cell models. Network pharmacology was used to predict compound–target pathway relationships. The content of poricoic acid A (PAA) was the highest in PC; the content of pachymic acid (PA) was the highest in RP; whereas dehydrotumulosic acid (DTUA) was enriched in WP. WP demonstrated the highest mannose and galactose content. The water extract of WP reduced H2O2-induced cytotoxicity in SH-SY5Y cells, whereas the PC extract alleviated L-α-lysophosphatidylcholine-induced injury in human oligodendroglioma cells. Network analysis identified PA, PAA, and poricoic acid B (PAB) as candidate bioactive compounds involved in neuroprotective effects. The PA/PAB ratios of ≥10, 1–9, and ≤1 were used to identify WP, RP, and PC, respectively. L* values of >80, 55–79, and <55 effectively discriminated WP, RP, and PC, respectively. This integrative study identified chemical and functional quality markers and demonstrated distinct neuroprotective profiles in different parts of WE, supporting their standardized and rational application in traditional medicine and nutraceuticals.

Life doi: 10.3390/life16081240

Authors: Ziyang Yu Shuaiwang Huang Xiyang Peng

Background: Exercise rehabilitation (ER) has become an important treatment regimen during the recovery phase of acute ischemic stroke (AIS). Optimal exercise prescriptions for post-stroke motor recovery remain controversial. Methods: PubMed, Embase, the Cochrane Library, and Web of Science were systematically searched through November 2025. The Cochrane risk-of-bias (RoB 1) assessment tool was utilized to assess the risk of bias in the original studies. Results: This study included 66 trials comprising 3675 patients. For balance, whole-body tilting postural training (WTPT), unilateral strength training (UST) and robot-assisted unilateral gait training (RAUGT) showed high SUCRA rankings. Activities of daily living likely improved most with robot-assisted Tai Chi training (RATCT), home rehabilitation guidance (HRG) and robot-assisted gait training (RAGT). Fugl-Meyer Assessment (FMA) gains appeared greatest following robot-assisted hand training with electrical stimulation (RAHT + ES), neural inhibition therapy (NIT) and graded motor imagery with electrical stimulation (GMI + ES). Walking endurance seemed most responsive to visual feedback training (VFT), UST and visual aids training (VAT), while functional mobility showed greatest improvement with tilt sensor-assisted gait training with electrical stimulation (TAGT + ES), gait training (GT) and virtual reality therapy with motor imagery (VRT + MI). Conclusions: This study provides the first comprehensive evidence synthesis evaluating the comparative effectiveness of specific exercise modalities across distinct recovery domains after AIS. Rather than supporting generic interventions, the findings highlight the distinct comparative advantages of various emerging technologies and traditional practices. These results offer an evidence-based framework for clinicians to optimize post-stroke rehabilitation protocols and highlight priority areas for future research on dose–response and long-term outcomes.

Life doi: 10.3390/life16081239

Authors: Aliki Karkala Alexandros Kalkanis Serafeim-Chrysovalantis Kotoulas Şakir Kaan Çetindağ Dries Testelmans Vasileios Paraschou Dionisios Spyratos Dimitrios G. Raptis Paschalis Steiropoulos Athanasia Pataka

Morning headaches (MHs) are a recognized but incompletely understood symptom in obstructive sleep apnea (OSA). Because symptoms and comorbidities that commonly coexist with OSA may influence both headache susceptibility and perceived sleep quality, the respective contributions of respiratory abnormalities and sleep disruption to MHs remain uncertain, particularly in women. We therefore aimed to identify clinical and sleep study factors associated with frequent MHs in patients with OSA. In this retrospective cohort study, 3227 adults with confirmed OSA evaluated between 2013 and 2023 were classified according to MH frequency (<1 episode/week vs. ≥1 episode/week). Demographic, clinical, questionnaire and sleep study variables were analyzed using univariate and multivariable logistic regression. Frequent MHs were reported by 35.2% of patients. On univariate analysis, MHs were associated with female sex, higher body mass index and waist circumference, greater OSA severity, worse nocturnal oxygenation, fatigue, excessive daytime sleepiness, and insomnia symptoms. However, in multivariable analysis restricted to patients with complete polysomnographic data (N = 501), female sex (OR 4.23, 95% CI 1.39–12.85), waist circumference (OR 1.03, 95% CI 1.01–1.06), and arousal index (OR 1.05, 95% CI 1.01–1.08) emerged as independent predictors, whereas conventional respiratory severity indices did not. These findings suggest that sleep fragmentation, rather than OSA severity alone, may be a key determinant of MHs, particularly in women, underscoring the need for sex-specific approaches beyond apnea–hypopnea index-based assessment.

Life doi: 10.3390/life16081238

Authors: Jakub Szrama Piotr Smuszkiewicz Amadeusz Woźniak Ashish Lohani Krzysztof Zwoliński Paweł Sobczyński Nina Łabędź Aleksandra Dańczak-Pazdrowska Paweł Pazdrowski Anna Mikołajczyk-Lorkiewicz Joanna Wojciechowska Marcin Stopa Adriana Polańska

Toxic epidermal necrolysis (TEN) is a rare, life-threatening mucocutaneous adverse drug reaction associated with extensive epidermal necrosis and severe systemic complications. We report the case of a 25-year-old woman who developed severe lamotrigine-induced TEN four weeks after treatment initiation. The disease rapidly progressed to involve approximately 60% of the body surface area, requiring intensive care unit admission, mechanical ventilation, and multidisciplinary management. Treatment included high-dose intravenous methylprednisolone, intravenous immunoglobulins, etanercept, plasmapheresis, and comprehensive supportive care. During hospitalization, the patient developed gastrointestinal complications manifested by pneumatosis intestinalis and portal venous gas, raising suspicion of bowel ischemia and prompting exploratory laparotomy. Following treatment, gradual clinical improvement and complete resolution of the cutaneous lesions were achieved. However, persistent ocular sequelae developed, including meibomian gland dysfunction, punctate epithelial erosions, conjunctival scarring, trichiasis, tear film instability, and early corneal neovascularization, requiring long-term ophthalmological follow-up. To provide clinical context, a narrative review of the literature on the diagnosis, multidisciplinary management, gastrointestinal manifestations, ocular complications, and systemic treatment of TEN was performed. A comprehensive literature search was conducted to identify relevant articles focusing on the intensive care management and specific organ-related manifestations of Toxic Epidermal Necrolysis (TEN). The gathered literature was narratively synthesized to present a cohesive update on current clinical practices and management controversies. This case highlights the potentially devastating multiorgan course of TEN and emphasizes the importance of early recognition, specialized intensive care, and long-term multidisciplinary follow-up.

Life doi: 10.3390/life16081237

Authors: Ahmet Mehmet Somuncu Ahmet Taner Uysal

MicroPulse laser trabeculoplasty (MLT) lowers intraocular pressure (IOP) with subthreshold pulses that spare the surrounding tissue, yet its effect on the corneal endothelium is far less studied than that of selective laser trabeculoplasty, and almost unstudied in treatment-naïve eyes. We retrospectively reviewed 70 treatment-naïve glaucoma patients (one eye each) treated with a 577 nm yellow laser (Easyret®) in micropulse mode (300 µm, 300 ms, 1000 mW, 360°). Endothelial cell density, mean cell area, hexagonality, the coefficient of variation in cell size, and central corneal thickness were recorded by specular microscopy (SP-1P, Topcon), and IOP by Goldmann applanation, at baseline and 1, 3, and 6 months. Linear mixed-effects models used every available observation, so no patient was excluded for a missed visit. IOP fell from 25.9 ± 2.5 mmHg to 20.6 and 21.0 mmHg at one and three months (−5.32 and −5.00 mmHg, roughly 20%; both p < 0.001), then returned to 24.8 mmHg by six months, and the proportion reaching a 20% reduction fell from 62.5% to 4.6%. Endothelial cell density, mean cell area, and central corneal thickness held steady, and every morphological change stayed within the device’s repeatability limits on equivalence testing. MLT lowered IOP early but transiently; over six months no clinically meaningful change in corneal endothelial morphology was detected.

Life doi: 10.3390/life16081236

Authors: Ilker Çermikli Tufan Alatli Salih Kocaoglu

Background: Acute upper gastrointestinal bleeding (UGIB) remains a leading cause of emergency department (ED) admission and carries a mortality of 5–10%. Static hemodynamic parameters perform poorly as predictors of fluid responsiveness, exposing patients either to under-resuscitation or to the harms of fluid overload. The mini fluid challenge (MFC), in which a 100 mL bolus is used to elicit a measurable change in the left ventricular outflow tract velocity–time integral (ΔVTI), is a validated dynamic test in intensive care but has not been evaluated in undifferentiated ED haemorrhagic shock. We aimed to determine whether ΔVTI-guided resuscitation is feasible at the bedside in UGIB and to explore how ΔVTI relates to in-hospital mortality. Methods: Prospective, single-centre, randomised quasi-experimental comparative study conducted between February 2024 and February 2025. Adults presenting with UGIB were allocated by treatment period, and credentialed emergency physicians to an ultrasound-guided protocol group (UGPG) or a standard care protocol group (SCPG). ΔVTI was measured in both groups but was disclosed to the treating team only in UGPG; in SCPG, it was recorded offline and analysed post hoc. The primary outcome was the feasibility of ΔVTI acquisition; the association of ΔVTI with in-hospital mortality was a secondary, exploratory outcome. Results: Eighty-five patients were enrolled (UGPG n = 47; SCPG n = 38). ΔVTI acquisition was completed within the intended time window in all enrolled patients, establishing bedside feasibility. Across the whole cohort, ΔVTI discriminated non-survivors poorly (AUC 0.55, 95% CI 0.44–0.66), and its performance differed markedly by group: AUC 0.83 (95% CI 0.67–0.93) in SCPG versus 0.69 (95% CI 0.54–0.82) in UGPG. In SCPG, non-survivors had substantially higher ΔVTI than survivors (58.6 ± 30.5% vs. 28.4 ± 30.9%; p = 0.030), a pattern consistent with persistent, uncorrected fluid responsiveness. In UGPG, the association ran in the opposite direction, mortality being concentrated among ΔVTI < 10% non-responders (p = 0.020). Conclusions: ΔVTI-guided resuscitation is feasible in the ED management of UGIB. The direction of the ΔVTI–mortality association differed according to whether ΔVTI was visible to clinicians, generating the testable hypothesis that unrecognised fluid responsiveness contributes to death in standard care. These findings are hypothesis-generating only: the small sample and the low number of events preclude any causal inference and require confirmation in an adequately powered, randomised, etiology-stratified trial.

Life doi: 10.3390/life16081235

Authors: Ming Chen Linlin Zhao Liting Yang Mingxia Jin Jiaqi Kong Qin Yang

While the relationship between body mass index (BMI) and physical fitness in youth often follows an inverted U-shape, populations-based data from sub-plateau settings remain scarce. This study characterized the cross-sectional BMI–fitness association among 14,744 students (9931 females) at a single university in Gansu Province, China (altitude ~1483 m). Fitness was assessed across seven components (vital capacity, speed, power, flexibility, endurance, and muscular strength/endurance) and summarized as a standardized Physical Fitness Index (PFI). Overweight/obesity prevalence was 11.2% (males 18.3%, females 7.7%)—lower than the 14.0% national average for same-aged university students. BMI displayed a monotonic inverse association with PFI; underweight and normal-weight groups performed comparably, while overweight and obese groups showed progressively lower PFI, with a stronger association in males (β = −0.46) than females (β = −0.36). The weight-normalized vital capacity index (VCWI)—a derived indicator of pulmonary function—showed the strongest inverse correlation (males r = −0.47; females r = −0.40). Associations were most pronounced for VCWI and endurance, whereas flexibility exhibited a negligible BMI-related gradient. These findings describe a cohort-specific pattern, emphasizing the need for broader investigation of regional variations, and should not be interpreted as evidence of an altitude-mediated effect.

Life doi: 10.3390/life16081234

Authors: Enas N. Morgan Ayman M. Mousa Rasha A. Elmansy Hanan Seleem Marwa M. Fawzi Amany Refaat Mahmoud Reham Abdulla Aboukhalil Hagir H. T. Ahmed Reem A. Younis Tarek Hamdy Abd-ElHamid Asmaa Jabeen Ashwag Alsharidah Samah M. Abozaid Abdullah M. Alnuqaydan Khaled E. A. Soliman Enas Haridy Ahmed

Monosodium glutamate (E621) is a common flavor enhancer in highly processed food. Although it makes food more enjoyable, chronic intake may lead to excitotoxicity in brain areas. The current study investigates histological and biochemical neurodegenerative alterations in the rat hippocampus following E621 administration and evaluates the potential neuroprotective properties of ascorbic acid (AA) against E621-induced adverse effects. Forty adult male albino rats were divided into four groups: control group (G1), AA group (G2), E621 group (G3), and AA + E621 group (G4). All animals received a daily intraperitoneal (IP) injection for 30 days. Hippocampal samples were processed and stained with hematoxylin and eosin (H&E), immunostained for GFAP, synaptophysin (a synaptic protein), and caspase-3, and biochemically analyzed for oxidative markers, including malondialdehyde (MDA) and superoxide dismutase (SOD). G3 exhibited significant neurodegenerative changes, characterized by pyknotic granular cells and cytoplasmic vacuolation, with significantly elevated GFAP, synaptophysin, and caspase-3 immunoreactivity in the dentate gyrus (DG). These structural deficits correlated with elevated MDA levels and reduced SOD levels in G3. In contrast, simultaneous administration of AA with E621 resulted in substantial preservation of neuronal morphology, a reduction in caspase-3 immunoreactivity, and a restoration of synaptic vesicle density in G4. E621 induces hippocampal injury by increasing ROS levels and dysregulating GFAP, synaptophysin, and caspase-3. At the same time, AA preserves neuronal integrity and synaptic homeostasis, suggesting its potential role as a protective dietary supplement against brain injury induced by the E621 flavor enhancer.

Life doi: 10.3390/life16081233

Authors: Derya Çetin Sarışık Ahmet Serhat Aydın Mehmet Söyler Raif Zileli Ali Özkurt Coskun Yılmaz Hamza Küçük Mevlüt Karataş

Background: Aerobic capacity and respiratory efficiency are critical determinants of performance in elite female football, characterized by repeated high-intensity efforts and rapid recovery demands. Halotherapy, involving the inhalation of dry salt aerosol, has been shown to improve pulmonary health; however, its effects on exercise-related outcomes in athletic populations remain insufficiently investigated. This study aimed to examine the effects of a 12-week post-training halotherapy intervention on respiratory muscle strength, pulmonary function, and aerobic performance in elite female football players. Methods: Twenty-eight elite female football players competing in the Turkish Women’s First League were randomly assigned to a halotherapy group (HG, n = 14) or a control group (CG, n = 14). Both groups completed an identical preseason training program, while the HG additionally performed 45 min halotherapy sessions three times per week in a controlled halochamber. Pre- and post-intervention assessments included maximal inspiratory and expiratory pressures (MIP, MEP), spirometric parameters (FVC, FEV1, PEF, MVV), and maximal oxygen uptake (VO2max) estimated using the Yo-Yo Intermittent Recovery Test Level 1 (Yo-Yo IRT1), along with total distance covered. A 2 × 2 mixed-model ANOVA was used to assess time and group × time interaction effects. Results: Significant group × time interactions were observed for MIP, MEP, VO2max, FEV1, PEF, MVV, and Yo-Yo IRT1 performance (p < 0.05), with large effect sizes (ηp2 = 0.269–0.924), favoring the halotherapy group. Forced vital capacity (FVC) increased over time in both groups (p = 0.141), with no significant interaction effect. Changes in the control group were comparatively smaller across most outcomes. Conclusion: Post-training halotherapy was associated with improvements in respiratory muscle strength, ventilatory function, aerobic capacity, and intermittent running performance in elite female football players. These findings suggest that halotherapy may be considered as a complementary, non-invasive strategy to support respiratory and aerobic adaptations. Further research is required to confirm these findings and to clarify the underlying physiological mechanisms. Trial registration: NCT07406386; 6 February 2026.

Life doi: 10.3390/life16081230

Authors: Deborah A. Jehu Mitchell Hanson Ying Huang Andre Soares Colleen Hergott Jennifer L. Waller Lufei Young William Hall Dawnchelle Robinson-Johnson Crystal Allen Richard Sams Ryan M. Carrick Sadanand Fulzele Mark Hamrick Haidong Zhu Yanbin Dong

The objective of this randomized controlled trial was to evaluate the impact of a multifactorial exercise program on circulating inflammation in people living with dementia (PWD) in residential care. This parallel-group, 6-month assessor-blinded trial (NCT05488951) allocated (1:1) 42 PWD to a multifactorial exercise intervention or usual care alone in residential care settings between July 2022 and January 2023. The exercise group engaged in 30 min of physical therapist-led strength and balance training, followed by 30 min of walking 3 x/week over 6 months, and received usual care. The usual care group only received care from healthcare providers, ongoing medical treatment, and opportunities to participate in social activities. Fasted blood was drawn at baseline and 6 months. Intention-to-treat (ITT) and per-protocol (PP; ≥2 x/week exercise vs. usual care) analyses were conducted. The ITT analysis revealed no differences between groups over time (p > 0.05). In adjusted PP analyses, there was a group-by-time interaction trend resulting in decreases in IL-1β (−3.6; +5.0 pg/mL) and IL-8 (−13.4; +14.0 pg/mL) in the exercise group compared with usual care (IL-1β: p = 0.089; IL-8: p = 0.087). Findings suggest adherence-dependent reductions in pro-inflammatory cytokines, indicating potential benefits of exercise. However, this trial was not powered for this secondary analysis, necessitating the need for larger, adequately powered trials.

Life doi: 10.3390/life16081232

Authors: João Pereira Soares Andreas E. Kremer Jérôme Bonzon

Vanishing bile duct syndrome (VBDS) is a rare cholestatic liver disease often associated with drug-induced liver injury, yet systematic data on pharmaceutical triggers remain limited. Using WHO VigiBase, we applied Bayesian disproportionality analysis (IC0.25) to identify drug-event associations that may not be readily apparent in clinical trials or pre-marketing studies. Product labels approved by Swissmedic or the FDA, as well as LiverTox, were reviewed to determine whether VBDS was already acknowledged as an adverse event. Signal detection was deliberately restricted to reports naming a single suspect drug. Among these single-agent reports, 22 drugs demonstrated a positive IC0.25 signal, of which nevirapine, dapsone and azithromycin showed the strongest disproportionality signal. Only one of these agents (carbamazepine) explicitly labelled VBDS as an adverse event. These findings are based on spontaneous reporting data: disproportionality analysis is a hypothesis-generating signal-detection method that does not establish causality and requires further validation. This study expands the current understanding of drug-induced VBDS by reinforcing the associations with known drugs and generating pharmacovigilance signals for new potential VBDS triggers across several drug categories.

Life doi: 10.3390/life16081231

Authors: Liancheng Liu Huanhuan Li Gonghua Lin Tianjuan Su Fang Zhao Bo He Zuhao Huang

This study sequenced and comparatively analyzed the mitochondrial genomes of three agriculturally important Aulacophora pests: the polyphagous A. indica, the oligophagous A. lewisii, and the early-diverging A. nigripennis. All three mitogenomes contained the typical 37 genes with conserved gene order. However, total length varied substantially (15,258–18,766 bp), driven primarily by control region length variation. All species exhibited pronounced AT bias, and relative synonymous codon usage analysis revealed marked interspecific divergence. Phylogenetic analysis based on 13 protein-coding genes resolved A. nigripennis as the basal lineage and A. lewisii and A. indica as closely related sister species. Both widespread species displayed maximum haplotype diversity (Hd = 1.000); however, A. indica exhibited approximately 2.4-fold higher nucleotide diversity (Pi = 0.00478) than A. lewisii (Pi = 0.00198), indicating differential population genetic architectures between the two species. These findings established a comparative mitogenomic framework for Aulacophora and provide baseline data for future phylogenomic and phylogeographic investigations.

Life doi: 10.3390/life16081229

Authors: Adelina-Mihaela Sorescu Cristina Isabel Viorica Ghiță Gabriela Duică Alin Nicolescu Eliza Elena Cinteză Rachele Adorisio Antonio Amodeo Paola Francalanci Gianluca Brancaccio Gessica Ingrasciotta Erica Mencarelli Oana Andreia Coman

Introduction: Dilated cardiomyopathy (DCM) is a relatively rare manifestation of pediatric heart failure. Despite recent diagnostic advancements and expanded screening modalities, the underlying cause of DCM remains elusive in over 50% of pediatric cases. We report the case of a 1-year-old girl presenting with newly diagnosed, rapidly progressive dilated cardiomyopathy, caused by a novel, rare homozygous pathogenic variant in the PPP1R13L gene. Case presentation: The patient, with no significant family history of cardiac disease, previously asymptomatic and in good health, presented with a 2-day history of nausea, vomiting, difficulty breathing, and low urine output, following a recent respiratory infection. The echocardiography identified a dilated left ventricle (Z score > +2 for age) and severe systolic left ventricular dysfunction, and a positive diagnosis of dilated cardiomyopathy was confirmed by the cardiac magnetic resonance (CMR). Genetic testing identified a homozygous pathogenic variant in the PPP1R13L gene, classified as a novel pathogenic variant that encodes the inhibitor of apoptosis-stimulating protein of p53 (iASPP). Despite targeted treatment, the severe systolic dysfunction persisted, and the general state progressively deteriorated and warranted the need for a cardiac transplant, which she underwent one year following the initial diagnosis. Conclusions: Patients diagnosed with underlying genetic DCM, particularly those harboring PPP1R13L mutations, carry an exceedingly poor prognosis in the absence of orthotopic heart transplantation. Existent case reports confirm that disruptions in this gene predictably yield highly aggressive, life-threatening cardiomyopathies.

Life doi: 10.3390/life16081228

Authors: Sondos Eladawi Prut Koonalintip Amelia Ngomuo Emily Leech Mark Thaller Benjamin R. Wakerley

Background and objectives: A migraine is a highly prevalent and disabling neurological disorder that mainly affects women of childbearing age. Treatment options for migraine prevention are limited during pregnancy and breastfeeding. Although OnabotulinumtoxinA (OnabotA) has proven efficacy and safety in chronic migraine prophylaxis in adults, its safety in pregnancy and breastfeeding remains understudied. Methods: Using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses methodology, this review systematically appraised the evidence on the use of OnabotA for chronic migraines and other neurological conditions during pregnancy and breastfeeding across four electronic databases between January 2000 and December 2025. Results: In summary, of the 555 OnabotA-exposed pregnancies, 80.2% resulted in full-term births, with a 17.5% loss rate (73 spontaneous, 21 elective, three unspecified) and 1.6% reporting foetal anomalies. These findings align with general population baselines for miscarriages (12.5–18.7%) and foetal anomalies (3–5%), respectively. Among 94 cases of reported breastfeeding, no adverse infant events or developmental delays were observed at 1 year. Maternal outcomes showed favourable symptom control without adverse effects. Conclusions: Although there is limited data on the use of OnabotA in pregnancy and breastfeeding, no significant adverse effects have been identified in pregnant mothers or neonates. OnabotA should therefore be considered on a case-by-case basis for treatment of various neurological conditions, including migraines, when there are no better safe alternatives.

Life doi: 10.3390/life16081227

Authors: Marcella Chagas-Sena Wei Ling Lau Thomas Edward Lane Paola Cristina Resende Ane Claudia Fernandes Nunes

Caused by the SARS-CoV-2 virus, the COVID-19 pandemic is still considered a complex challenge, with manifestations not only of respiratory issues, but also conditions related to chronic cerebrovascular damage. Endothelial biomarkers, neuropathological and neuroimaging findings indicate endothelial dysfunction, microthrombosis, and disruption of the blood–brain barrier (BBB) are central mechanisms for acute and chronic ischemic and hemorrhagic cerebral events. Understanding these mechanisms is vital to reducing their impact on population health, whether through treatment or prevention of adverse outcomes. The objective of this study is to perform a review of the scientific literature on the post-infection effects of SARS-CoV-2 affecting the cerebral endothelium and the BBB, correlating them with potential clinical outcomes. Material and Methods: Analysis of studies extracted from the PubMed database using the following terms: Long-COVID “AND” SARS-CoV-2 “AND” blood–brain barrier. Inclusion criteria: keywords, publications related to the topic, and primary studies published after peer review. Exclusion criteria: preprint studies, publication outside of the timeframe 2020–2025, study design not compatible with this research, and full text not available. Results: An initial 121 studies were identified, of which 105 were excluded due to not meeting all inclusion criteria and 6 studies were inaccessible due to not being in the English language and full-text access limitations. Fifteen articles were included in the analysis, for topics as expression of viral receptors in the endothelium, markers of their activation, cerebral microvascular injury and coagulopathies. To clarify the pathogenic cascade, the evidence was stratified by biological model where in vitro evidence demonstrates that the Spike protein induces direct endothelial toxicity and platelet aggregation, establishing the primary molecular insult. Animal models confirm the translation of this insult into structural degradation of the BBB and pericyte loss. Clinically, infection-phase findings, characterized by multifocal microthrombosis and permeability spikes, act as the determining event that predisposes to the persistent neuroinflammatory environment. Biomarkers of BBB disruption and neuronal damage were consistently reported, with persistence of BBB dysfunction modifying risk stratification and rehabilitation efforts. Reported cases of Long-COVID demonstrated normalization of BBB markers without correlation with long-term symptoms, suggesting that other mechanisms are involved in Long-COVID. Conclusion: Cerebral endotheliopathies and BBB dysfunction in patients with COVID-19 continue to impact the health of the population. The scientific literature indicates that SARS-CoV-2 induces cerebral endothelial injury, BBB disruption, and an increased risk of vascular events related to endotheliopathy, inflammation, and hypercoagulability. Understanding the impact of COVID-19 pathology on the population and developing prospective studies is essential to quantify the prevalence and mechanisms of brain injury. The identification of molecular targets and infection pathways are promising toward defining both preventive and therapeutic strategies to improve outcomes in the Long-COVID population.

Life doi: 10.3390/life16081226

Authors: Longzhen Qiu Jiling Liang Ling Yu Siyao Gao Yineng Tan Xueliang Li Jianyu Gan

Time-efficient strategies are needed to enhance physical fitness in sedentary university students. This pilot study examined the preliminary effects of maximal-effort stair-climbing exercise snacks performed alone (ESA) or combined with practical blood flow restriction (ES + BFR). Twenty-three sedentary male university students were included in the per-protocol analysis (ESA, n = 7; ES + BFR, n = 8; control, n = 8). Over 4 weeks, both intervention groups completed 48 supervised sessions, each consisting of three 20 s maximal stair climbs separated by 2 min rest. Assessed outcomes were forced vital capacity (FVC), 50 m sprint time, 1000 m run time, standing long jump (SLJ), sit-and-reach flexibility, and body mass index (BMI). Group × Time interactions were tested using mixed-model ANOVA. Significant interactions emerged for all metrics (p ≤ 0.01, partial η2 = 0.35–0.70), except BMI. Both interventions significantly improved FVC, flexibility, 1000 m run and 50 m sprint performance (p < 0.05; absolute Cohen’s d range = 1.01–2.68). Although no significant between-group differences were observed, ES + BFR produced larger within-group effect sizes than ESA for 1000 m run time (d = −2.14 vs. −1.66) and 50 m sprint time (d = −2.21 vs. −1.89). Only ES + BFR significantly increased SLJ (p < 0.05), whereas the control group exhibited a significant decline (p < 0.05). BMI remained unchanged. The 4-week stair-climbing exercise snack protocol is feasible and may improve pulmonary function, endurance, speed, and flexibility in sedentary male students. Adding practical BFR shows promising numerical benefits for sprint, power, and endurance performance, but these exploratory findings require confirmation in larger trials.

Life doi: 10.3390/life16081224

Authors: Mikołaj Basza Wojciech Bojanowicz Fryderyk Zawadzki Dagmara Galle Mateusz Soliński Weronika Kowalczyk Łukasz Kołtowski Marek Ochman

Background: Lung transplantation (LTx) is the final therapeutic option for patients with end-stage irreversible respiratory failure. Chronic lung allograft dysfunction (CLAD) remains a major determinant of long-term outcomes, but early detection of functional decline may enable timely intervention and partial reversibility. Therefore, systematic monitoring of graft function is essential, and telemedicine-based spirometry may facilitate early identification of clinically relevant decreases in lung function. Objective: The aim of our study was to evaluate the feasibility of spirometry telemonitoring (TM) and its association with healthcare utilisation in lung transplant patients. Methods: This retrospective study compared lung transplant recipients: the first group underwent spirometry TM (n = 21), the second group was monitored with standard home spirometry (HS) (n = 23), and the control group underwent routine follow-ups only at the transplant centre (n = 32). Results: In the TM spirometry group, mean adherence was 62.7%, with 4957 examinations performed over a mean monitoring period of 252 days; 59.7% of FEV1 and 58.9% of FVC measurements were technically acceptable. Compared with routine care (RC), TM spirometry was associated with a shorter mean duration of both combined planned and urgent admissions (5.8 vs. 10.0 days, p < 0.001) and urgent admissions analysed separately (14.7 vs. 26.2 days, p < 0.001). Conclusions: TM spirometry was feasible and associated with lower healthcare utilisation in lung transplant recipients, supporting its potential value for post-transplant surveillance. Larger prospective randomised studies are needed to confirm these preliminary findings.

Life doi: 10.3390/life16081225

Authors: Noman Ahmed Muhammad Yousaf Muhammad Muneeb Chaudhry Ahsan Akram Irfan Ahmed Muhammad Qumar Abdul Ghayas Muhammad Asif Ali R. Al Sulaiman Ala E. Abudabos

Guanidinoacetic acid (GAA), a natural precursor of creatine, plays a pivotal role in cellular energy metabolism. This study investigated the effects of dietary GAA supplementation at varying levels (0, 0.6, and 1.2 g/kg) combined with standard or reduced (−100 kcal/kg) dietary energy on growth performance, carcass characteristics, meat quality, and heat stress tolerance (during the finisher phase) in broiler chickens. A total of 360 one-day-old male Ross-308 broiler chicks were randomly allocated to 36 floor pens in a 3 × 2 factorial arrangement under a completely randomized design. Six treatments with six replicates, each having 10 birds, were evaluated over 35 days. Growth performance parameters were recorded weekly. On day 35, samples were collected for carcass characteristics (dressing percentage, breast and thigh yield, and organ weights), meat quality analysis (drip loss, cooking loss, and color parameters), and heat stress response. The heat stress response (panting frequency) was recorded during the finisher phase of the trial. GAA supplementation at 1.2 g/kg significantly (p < 0.001) improved final body weight, total weight gain, feed conversion ratio, dressing percentage, breast yield, and thigh yield. Meat quality parameters showed significant improvement (p < 0.001) with GAA in a dose-dependent manner for drip loss, cooking loss, and all color parameters (L*; a*; b*). Significant GAA × energy interactions (p < 0.001) were observed for final body weight, total weight gain, feed intake, FCR, cooking loss, live body weight at slaughter, breast yield, liver weight, gizzard weight, intestine weight, and panting frequency. The effect of GAA on heat stress tolerance was context-dependent, with a significant GAA × energy interaction (p < 0.001) for panting frequency. Under reduced-energy conditions, 1.2 g/kg GAA decreased panting frequency, suggesting improved thermoregulatory capacity, while under standard-energy conditions, GAA increased panting frequency. The main effect of GAA on panting frequency was not significant (p = 0.38), while the main effect of energy was highly significant (p < 0.001). In conclusion, GAA supplementation at 1.2 g/kg significantly improves growth performance, feed efficiency, meat quality (reduced drip and cooking losses and improved color parameters), and carcass yield in broiler chickens. Importantly, GAA supplementation largely compensated for a 100 kcal/kg reduction in dietary energy while maintaining performance. These findings support GAA as a promising nutritional strategy for sustainable broiler production, particularly in hot-climate regions.