Pathophysiology doi: 10.3390/pathophysiology33030070

Authors: Kawther Zaher Jehan Alrahimi

Background: Chimeric antigen receptor (CAR) T cell therapy has revolutionized the treatment of select hematologic malignancies and is increasingly being explored for non-oncological indications, including autoimmune diseases and persistent infectious diseases. This systematic review synthesized current evidence on CAR T cell and CAR-Treg approaches beyond oncology, with emphasis on therapeutic targets, translational and clinical outcomes, safety, and implementation barriers. Methods: The review followed PRISMA 2020 guidelines. PubMed/MEDLINE, Scopus, Web of Science, Cochrane Library, ClinicalTrials.gov, and major trial registries were searched for studies published from 1 January 2010 to 31 March 2026. Eligible studies included clinical, preclinical, and translational investigations of CAR T cell or CAR-Treg strategies in autoimmune or infectious diseases. Results: Because of heterogeneity in disease indications, CAR constructs, endpoints, and study designs, findings were synthesized descriptively. The strongest early clinical signals were observed with B cell-directed CAR T cell therapy for severe, refractory autoimmune diseases, particularly systemic lupus erythematosus, systemic sclerosis, and inflammatory myopathies. Infectious disease applications, mainly HIV and hepatitis B, showed preliminary safety, persistence, and partial antiviral activity, but durable pathogen eradication remains unproven. Conclusion: Overall, CAR T cell therapy beyond oncology is promising but remains preliminary, requiring standardized reporting, long-term safety monitoring, scalable manufacturing, and carefully defined risk–benefit thresholds. Human clinical evidence was interpreted separately from preclinical and mechanistic evidence, and clinical conclusions were based primarily on human studies.

Pathophysiology doi: 10.3390/pathophysiology33030069

Authors: Zelimkhan G. M. Berikkhanov Zakhar A. Akulov Maria A. Sukhanova Alexey L. Shestakov Aleksey G. Kotelnikov Andrey M. Nikolaev Vadim S. Razumovsky Evgeniy A. Tarabrin Milena Yu. Ivanova Sergey Yu. Muraviev

Modern abdominal wall hernia surgery is centered on the biomechanics of the fascial-aponeurotic framework and on intra-abdominal pressure (IAP) control; neuromuscular regulation of the wall remains outside perioperative planning. This review reframes the abdominal wall as a coordinated neuromuscular system and outlines a framework in which ventral herniation is hypothesized to represent a compensatory response to altered regulation of elevated IAP, while discoordinated contraction of the muscular layers is proposed as a potential predisposing factor. A structured literature search was performed in PubMed/MEDLINE, Embase and the Cochrane Library from inception to March 2026 across four thematic blocks: basal tone and innervation, spinal pathology and coordination, skeletal muscle memory, and electromyographic and clinical models of denervation. Synthesis followed SANRA criteria. This is not a formal systematic review or meta-analysis. Basal tone of the abdominal muscles is individual and depends on spinal innervation, lifestyle, comorbidity, and hernia duration; obstructive pulmonary disease yields a hypertonic phenotype, and persistent meteorism a hypotonic one. In a patient predisposed to herniation, elevated IAP may be partially accommodated through redistribution of abdominal contents; within the proposed framework, herniation is hypothesized to function as a pressure-relief mechanism. Discoordination of the muscular layers may reduce the capacity of the abdominal wall to adapt to rapid or unfamiliar loading, potentially contributing to hernia formation in predisposed patients. Six meta-analyses of preoperative botulinum toxin type A confirm lateral muscle elongation; none assessed basal-tone recovery after toxin washout. Each hernia patient presents with an individual neuromuscular phenotype shaped by basal tone, intermuscular coordination and muscle memory. The proposed framework complements mechano-fascial concepts with a neurophysiological dimension and provides a hypothesis-generating rationale for investigating phenotype-tailored prehabilitation and postoperative rehabilitation. Prospective electromyographic mapping with functional maneuvers is the immediate research priority.

Pathophysiology doi: 10.3390/pathophysiology33030068

Authors: Madison L. Wise Abdel A. Alli

Sodium glucose cotransporter-2 inhibitors (SGLT2is) have rapidly evolved from glucose-lowering agents to multifaceted therapies with significant renoprotective and cardioprotective potential. Although originally developed to inhibit glucose reabsorption within the renal proximal tubule for the treatment of Type 2 diabetes mellitus (T2DM), growing evidence indicates that SGLT2is exert broad systemic actions extending beyond glycemic control. Among this drug class, dapagliflozin has emerged as a clinically important agent with pleiotropic effects involving renal hemodynamics, inflammatory signaling, mitochondrial function, fibrosis regulation, and cellular stress adaptation. This review outlines the historical progression from the discovery of phlorizin to the development of highly selective modern SGLT2 inhibitors while emphasizing mechanistic insights gained from experimental and clinical studies of dapagliflozin. In addition to the established effects on sodium–glucose transport, dapagliflozin modulates multiple epithelial transport proteins including NHE3, NaPi-2a, NCC, and NCX1, highlighting complex regulatory effects on sodium handling and tubular electrolyte transport. Emerging evidence further demonstrates that dapagliflozin suppresses inflammatory and profibrotic pathways involving YAP/TAZ, STAT1, TGF-β, NLRP3, and NF-KB signaling. Restoration of tubuloglomerular feedback, attenuation of oxidative stress, and preservation of mitochondrial function also appear to contribute substantially to the renoprotective actions of SGLT2 inhibition. Beyond the kidney, dapagliflozin and related SGLT2is exert cardioprotective effects through coordinated improvements in cardiac energetics, inflammatory regulation, and hemodynamic function. Emerging studies additionally suggest potential pulmonary benefits, including reductions in inflammatory signaling, pulmonary edema, and respiratory complications. Collectively, these findings support a shift in understanding SGLT2is from targeted metabolic therapies to broader regulators of cellular and organ function. Continued investigation into the glucose-independent mechanisms of dapagliflozin may reveal additional therapeutic applications across chronic metabolic, cardiovascular, and inflammatory diseases.

Pathophysiology doi: 10.3390/pathophysiology33030067

Authors: Eleftherios Panteris Ioanna Kakatsaki Zoi Koukou Charalambos Kolvatzis Styliani Papanikolaou Eleftheria Hatzidaki

Non-communicable diseases remain the dominant cause of adult morbidity and mortality, yet cardiometabolic, vascular and renal risk may originate before birth. This structured narrative review critically evaluates human evidence linking pregnancy-related exposures, placental pathophysiology and later offspring health within a maternal–placental–fetal–life-course framework. Evidence published from 2020 to 2026 was emphasised, while seminal earlier cohorts were retained when they provided uniquely long follow-up or direct placental measurements. The most consistent associations concern maternal obesity, gestational diabetes, excessive gestational weight gain and hypertensive disorders, which are linked with higher offspring blood pressure, greater adiposity and adverse glucose–insulin profiles. Human placental studies implicate vascular malperfusion, altered nutrient transport, inflammatory and oxidative signalling, endocrine function and epigenetic regulation. Few studies, however, have measured the prenatal exposure, a specific placental phenotype and a long-term offspring outcome within the same longitudinal design; most proposed placental pathways are supported by convergent evidence or biological plausibility rather than demonstrated mediation. Preterm birth and fetal growth restriction are clinically observable, etiologically heterogeneous sentinel phenotypes, not obligatory mediators. Placental and offspring epigenetic signatures, telomere biology and mitochondrial function represent distinct domains of biological embedding; their causal and prognostic significance remains uncertain. Longitudinal studies integrating maternal exposures, placental histopathology and molecular phenotypes, fetal organ development, childhood cardiometabolic trajectories and adult clinical outcomes are needed to determine whether, and under what circumstances, the placenta plays an intermediary role rather than merely serving as a marker of an adverse pregnancy environment.

Pathophysiology doi: 10.3390/pathophysiology33030066

Authors: Sayandeep K. Das Prithviraj Karak Afsona Parveen Swastika N. Das Savitri M. Nerune

Background/Objectives: Microplastics (MPs) are increasingly reported in human reproductive tissues and fluids, but detection alone does not establish tissue injury or disease. Recent reviews have synthesised occurrence, reproductive toxicity, placental transfer, fertility, and pregnancy outcomes. The unresolved problem is that occurrence, model-response, lesion-comparison, clinical-association, and causal evidence are not interchangeable. This review asks what minimum evidence is required for an MP finding in the female genital tract (FGT) or fetoplacental continuum to become lesion-relevant and where inference must stop. Methods: We performed a critical narrative search of PubMed, Scopus, Web of Science, ScienceDirect, and Google Scholar from database inception through 10 July 2026. The final synthesis comprised 59 sources: 28 primary studies, 24 reviews or systematic syntheses, and 7 regulatory, consensus, or professional-guidance documents.. Methodological recommendations were graded as claim-essential, context-dependent, or exploratory and as routine, specialised, or research-intensive. Results: The distinct contribution is an FGT-specific mechanistic and claim-to-evidence architecture rather than another environmental or reproductive-toxicity catalogue. We propose an ordered, claim-calibrated evidence ladder linking external exposure, internal dose, reproductive-fluid burden, spatial localisation, lesion response, and clinically annotated phenotype. The framework specifies the minimum evidence for occurrence, localisation, lesion, clinical, and causal claims and provides explicit stopping and downgrading rules rather than treating all desirable measurements as mandatory. The FGT Lesion Atlas is a compartment-by-compartment map of prioritised, non-exhaustive endpoints and sampling requirements. The FGT Lesionome is a cross-compartment synthesis of recurring barrier-receptivity, stromal-fibrotic, vascular-perfusion, immune-microbiome, endocrine-steroidogenic, and particle-cargo axes. An integrated hypothesis-generating pathway links systemic or local exposure, internal and target-compartment burden, physicochemical conditioning in reproductive-fluid microenvironments, particle or particle-associated-constituent interaction with compartment-specific cells and matrices, the six response axes, physiological dysfunction, and clinically annotated outcomes. A reproductive-fluid model, an assisted-reproductive-technology sentinel sequence, and an FGT Microplastic Pathology Reporting Checklist rank study elements by evidentiary necessity, feasibility, and interpretive consequence. Conclusions: The framework is hypothesis-generating, not a validated causal map or a universal core outcome set. It separates essential validity safeguards from context-dependent best practices and exploratory endpoints. Human reproductive and fetoplacental disease causation remains unproven without contamination-controlled sampling, polymer confirmation, spatial co-localisation with lesions, temporality, exposure-response assessment, confounder control, replication, and appropriately powered, temporally informative clinical outcomes.

Pathophysiology doi: 10.3390/pathophysiology33030065

Authors: Thiago Guedes Pinto Barbara dos Anjos Rosario Pedro Everson Alexandre de Aquino Glauce Socorro de Barros Viana Nágila Maria Pontes Silva Ricardo Edilberto Rocha Silveira Débora Hellen Almeida de Brito Dávila Zampieri Milena de Barros Viana Daniel Araki Ribeiro

Background/Objectives: The increasing therapeutic use of cannabinoid-based formulations highlights the need to better characterize their safety profile, particularly in organs involved in xenobiotic elimination, such as the kidneys. This study evaluated the renal effects of cannabidiol (CBD) and Δ9-tetrahydrocannabinol (Δ9-THC) nanoemulsions following subchronic oral administration in male Wistar rats. Methods: Forty animals (n = 8/group) were randomly allocated into five groups: vehicle control (Licuri oil + Pluronic® F127), CBD 2.5 mg/kg, CBD 5 mg/kg, CBD+THC 2.5 mg/kg, and CBD+THC 5 mg/kg. Treatments were administered daily by oral gavage for 21 consecutive days. Kidney samples were analyzed by histopathology and immunohistochemistry (Ki-67, GST-P, and cleaved caspase-3). Results: High-dose groups (5 mg/kg) exhibited marked histopathological alterations, including tubular and glomerular damage, hemorrhagic areas, inflammatory infiltration, and necrosis. These groups also showed increased Ki-67 and cleaved caspase-3 immunoexpression compared to controls. GST-P expression was significantly increased only in the CBD 5 mg/kg group. No significant changes were observed in the low-dose groups. Conclusions: These findings suggest dose-dependent morphological and molecular alterations in renal tissue associated with cannabinoid nanoemulsion exposure under experimental conditions.

Pathophysiology doi: 10.3390/pathophysiology33030064

Authors: Martina Cacciapuoti Lucia Federica Stefanelli Ilaria Caputo Giulia Driussi Monica Ceol Giovanna Priante Lorenzo A. Calò Federico Nalesso

Background/Objectives: Oxidative stress is a major contributor to the pathogenesis of cardiovascular diseases, including hypertension, ischemic cardiomyopathy, and heart failure. MicroRNAs (miRNAs) have been extensively investigated in various contexts, and some of them have been identified to play a role in cardiovascular disease. This narrative review focuses on miR-103 and miR-155, two miRNAs implicated in the modulation of oxidative stress and cardiovascular remodeling. Methods: The following queries were used in PubMed since inception until May 2026: ((“miR-155” OR “microRNA-155” OR miR155) AND (“oxidative stress” OR ROS OR “reactive oxygen species”) AND (“cardiovascular disease” OR cardiovascular OR cardiac OR heart OR vascular)); ((“miR-103” OR “microRNA-103” OR miR103) AND (“oxidative stress” OR ROS OR “reactive oxygen species”) AND (“cardiovascular disease” OR cardiovascular OR cardiac OR heart OR vascular)). Results: A total of seven citations for miR-103 and 79 citations for miR-155 were identified. Reviews and papers about diseases other than those on cardiac/vascular involvement were excluded. miR-155 emerges as a potential regulator of inflammatory-redox signaling, whereas miR-103 appears more closely linked to cell fate and metabolic pathways. In both cases, available evidence supports a context-dependent role that challenges simplistic classification as pro- or antioxidant miRNAs. Conclusions: Available evidence suggests that both miR-103 and miR-155 are important regulators of oxidative stress-related pathways in cardiovascular disease. Nevertheless, the context-dependent effects observed across different cardiovascular disorders raise concerns regarding the safety of systemic miRNA modulation-based therapeutic strategies. Future studies should clarify the determinants of this context-dependent behavior and identify the specific conditions under which these miRNAs exert protective or harmful effects, which might pave the way for the development of miRNA-based therapeutic strategies targeting oxidative stress and cardiovascular remodeling.

Pathophysiology doi: 10.3390/pathophysiology33030063

Authors: Ali Moradi Kurt Ramey Kutiba Tabbaa Abdullah Sahyouni Kevin Sanchez Garcez Shivanshu Kumar Elaine Pan Grant Barton Olugbenga Oyesanmi Robert Subbiondo

Heart failure with preserved ejection fraction (HFpEF) is a type of heart failure in which the ejection fraction remains within the normal range (≥50%); however, patients still experience typical symptoms of heart failure. One of the most common manifestations in this population is exercise intolerance, which in some cases may be the only presenting symptom. Exercise intolerance refers to a reduced capacity to perform physical activity. Several processes, including impaired cGMP–PKG signaling, increased collagen deposition, endothelial dysfunction, increased arterial stiffness, pulmonary hypertension, and vascular remodeling, contribute to the pathophysiology of HFpEF. The disruption of these fundamental physiological processes significantly impairs exercise capacity, leading to attenuated increases in heart rate, stroke volume, and/or contractility, along with abnormal ventricular–vascular coupling during exertion. When EI is suspected, a spectrum of diagnostic modalities—from simple, low-cost tools such as the 6 min walk test to advanced imaging—can be used to evaluate its presence and its association with HFpEF. When EI in HFpEF is diagnosed, management should focus on addressing this limitation to improve quality of life and reduce morbidity and mortality. Exercise training and pharmacological therapies, such as SGLT2 inhibitors, may be beneficial in this population. In conclusion, EI is a common manifestation of HFpEF that significantly impacts patients’ quality of life. Understanding its underlying mechanisms and addressing it appropriately are essential to improving patient outcomes.

Pathophysiology doi: 10.3390/pathophysiology33030062

Authors: Gabriela Chlebowska Krzysztof Michalak Łukasz Mazur Wioletta Szczurek-Wasilewicz

Erythropoiesis-targeted doping remains a major challenge for sports medicine because pharmacological and genetic manipulation of erythropoiesis can improve oxygen transport and endurance performance while increasing the risk of serious cardiovascular complications. Erythropoiesis-targeting strategies extend beyond recombinant erythropoietin (EPO) to include hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs), modified erythropoietin receptor (EPOR) agonists, transforming growth factor beta (TGF-β) signaling inhibitors, cytoprotective EPO derivatives, and gene- or cell-based approaches, and require complementary detection strategies based on direct analytical methods and the Athlete Biological Passport (ABP). Although they may enhance oxygen delivery and endurance performance, excessive stimulation of erythropoiesis may increase blood viscosity, impair vascular function, and elevate the risk of hypertension, thromboembolic complications, and other cardiovascular (CV) events. Erythropoiesis-targeted doping has evolved beyond recombinant EPO into a diverse group of pharmacological and genetic strategies that require increasingly sophisticated detection approaches. A thorough understanding of their molecular mechanisms and cardiovascular consequences is essential for improving anti-doping surveillance and protecting athlete health. This review summarizes current erythropoiesis-targeting agents, their mechanisms of action, detection strategies, cardiovascular risks, and implications for anti-doping practice.

Pathophysiology doi: 10.3390/pathophysiology33030061

Authors: Milica Milinkovic Sorgic Aleksandar Matic Vladimir Jakovljevic Nikola Jovic Sladjana Novakovic Teodora Todorovic Jovan Milosavljevic Bozidar Pindovic Jasmina Sretenovic Petar Canovic Jovana Jakovljevic Uzelac Jovana Joksimovic Jovic

Background/Objectives: Polyendocrine metabolic ovarian syndrome (PMOS) is a complex endocrine–metabolic disorder of multifactorial etiology, driven by interactions among hyperandrogenism, metabolic dysfunction, oxidative stress, and chronic low-grade inflammation that interact to promote reproductive dysfunction and increase risk of cardiometabolic complications. Although the combination of letrozole and high-fat diet (LET + HFD) represents a widely used experimental approach for inducing a PMOS-like phenotype in rats, its multisystem pathophysiological features remain incompletely characterized. The present study aimed to characterize reproductive, cardiometabolic, hormonal, inflammatory, oxidative, and morphometric alterations associated with a LET + HFD-induced PMOS-like phenotype in rats. Methods: PMOS-like model was induced in rats over a 21-day period of orally administered letrozole combined with a high-fat diet. Results: Body weight gain was higher, while uterine weight was lower in the PMOS group compared with controls. Metabolic parameters suggested insulin resistance, while hormonal profiling revealed increased total testosterone and LH levels, accompanied by reduced FSH, estradiol, and progesterone levels. Elevated triglycerides and reduced HDL levels were also observed in the PMOS group. Morphometric analysis revealed numerous atretic and large thin-walled cystic follicles in the ovaries, accompanied by thinning of the uterine luminal and glandular epithelium, stromal layer, and hypoplasia of endometrial glands. Both the longitudinal diameter and cross-sectional area of cardiomyocytes were increased in the PMOS group, together with nuclear hypertrophy and enhanced myocardial collagen deposition. In addition, altered oxidative stress markers, including increased lipid peroxidation and reduced glutathione levels, were observed, whereas IL-6, IL-1β, and IL-23 were not significantly altered. Conclusions: The LET + HFD model reproduces key reproductive and cardiometabolic features of PMOS, extending beyond reproductive dysfunction to involve oxidative and structural alterations in multiple organs. These findings support its use for investigating PMOS pathophysiology and evaluating potential preventive and therapeutic strategies.

Pathophysiology doi: 10.3390/pathophysiology33030060

Authors: Derek Tai Navneet Sandhu Aren Dermarderosian Norayr Mkrtchyan Daniel Park Vinisha Garg Angel Nguyen Mojtaba Akhtari

Introduction: Bispecific antibodies (BsAbs) are a promising and potent therapeutic intervention for relapsed or refractory (R/R) hematologic malignancies, including acute lymphoblastic leukemia (ALL), multiple myeloma (MM), and B-cell lymphomas. These recombinant molecules are designed to target two distinct antigens or epitopes. While BsAbs have shown significant efficacy in managing hematologic malignancies, toxicities must be carefully monitored and their safety profiles require further investigation. This meta-analysis investigates cardiovascular adverse effects associated with seven BsAbs across multiple clinical trials. Methods: A systematic literature review identified seven BsAbs, blinatumomab, elranatamab, epcoritamab, glofitamab, mosunetuzumab, talquetamab, and teclistamab, analyzing 25 clinical trials (Phase I–III) that included monotherapy regimens. The cardiovascular adverse events studied were arrhythmias, heart failure, blood pressure changes, and myocardial infarctions. A meta-analysis using the R meta package calculated proportions and 95% confidence intervals (CIs) and employed a random-effects model to assess heterogeneity. Results: Among 3215 patients, pooled analysis revealed significant findings with 13.6% cardiac arrhythmia (95% CI 10.1–18.2%, p = 0.05), 13.3% hypotension (95% CI 10–17.3%, p < 0.0001), 10.2% tachycardia (95% CI 7.6–13.5%, p = 0.0003), and 3.4% sudden death (95% CI 0–80%, p = 0.046). Less frequent events were hypertension (8%), acute myocardial infarction (1.5%), heart failure (1.4%), and atrial fibrillation (1.3%). Discussion: As BsAbs see increased use in R/R hematologic malignancies, cardiovascular complications must be closely monitored. This analysis highlights hypotension, cardiac arrhythmias, tachycardia, and sudden death as significant adverse effects. Although study heterogeneity and limited patient-level data may have influenced incidence estimates, these findings support routine cardiovascular monitoring and underscore the need for prospective studies to identify high-risk patients and optimize prevention and management strategies. Oncologists, cardiologists, and pharmacists should establish strategies for early detection and management to optimize patients’ outcomes.

Pathophysiology doi: 10.3390/pathophysiology33030059

Authors: Elena Sanchis-Sanchis José Enrique de la Rubia Ortí David Sancho-Cantus Cristina Cunha-Pérez Jorge Casaña-Mohedo

Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disorder in which systemic pathophysiological alterations significantly contribute to disease progression and non-motor manifestations, such as depression and anxiety. The microbiota–gut–brain axis represents a critical bidirectional pathway in which intestinal dysbiosis and epithelial barrier disruption catalyze central neuroinflammation. This scoping review synthesizes evidence from 43 empirical and analytical studies across 28 countries and maps the findings under the WHO International Classification of Functioning (ICF) framework. Pathophysiological data reveal a profound taxonomic shift in patients with ALS, characterized by severe depletion of neuroprotective, butyrate-producing genera (Akkermansia and Prevotella) and enrichment of pro-inflammatory Enterobacteriaceae. This dysbiotic state leads to structural damage to the intestinal mucosa, alteration of Paneth cells, and downregulation of tight junction proteins (zonulin), triggering a “leaky gut” phenomenon. Subsequent systemic translocation of lipopolysaccharides (LPS) induces TLR4-mediated endotoxemia, microglial hyperactivation, and accelerated motor neuron apoptosis. Conversely, therapeutic modulation via Fecal Microbiota Transplantation (FMT), psychobiotics, and metabolic interventions (ketogenic or Mediterranean diets) has demonstrated significant efficacy in restoring epithelial integrity, mitigating mitochondrial hypermetabolism, and reducing emotional distress. This review identifies a critical research gap in the microstructural characterization of the enteric nervous system in ALS. Incorporating microbiome-targeted biomarkers into clinical protocols is crucial for implementing a stratified, multi-systemic therapeutic strategy aimed at enhancing patient prognosis and psychological well-being.

Pathophysiology doi: 10.3390/pathophysiology33030058

Authors: Berk Rasheed Jacob Freed Ruhi Parikh Aman Singh Krishna K. Singh

The intricate relationship between the heart and the brain has transitioned from historical, cardio-centric theories to a modern, circuit-based, bi-directional framework known as neurocardiology. This review provides a comprehensive anatomical, physiological, and clinical synthesis of the bidirectional heart–brain axis, highlighting how disruptions in one organ systematically impact the other. Structurally, the heart–brain axis operates as a closed-loop feedback network. Vagal parasympathetic control dominates at rest, driven by an asymmetrical fiber distribution where sensory afferents actively supply interoceptive data to the brainstem. Hemodynamically, cerebral blood flow delivery is linked to cardiac output in an age-dependent fashion with the related vascular dysfunction. Clinically, brain-to-heart pathologies like acute brain injury trigger life-threatening catecholamine surges, neuroinflammation, and myocardial stunning, including Takotsubo cardiomyopathy. Conversely, heart-to-brain pathways reveal that atrial fibrillation and heart failure can independently trigger cardioembolic stroke, long-term neuroinflammation, and a profound, progressive burden of cognitive decline. The heart and brain are inexorably linked through complex structural, mechanical, and paracrine pathways. Ameliorating clinical outcomes demands integrated cross-specialty diagnostic screening, digital rhythm monitoring, and holistic neurocardioprotective therapeutic strategies.

Pathophysiology doi: 10.3390/pathophysiology33030057

Authors: Linghong Li Wayne F. Patton

Background/Objectives: Time-varying low-frequency electromagnetic fields (LF-EMFs), including pulsed electromagnetic fields (PEMFs) and sinusoidal low-frequency electromagnetic fields, have been investigated as modulators of tissue repair and vascular remodeling. Although numerous studies report pro-angiogenic effects in endothelial systems, the underlying mechanisms remain incompletely understood. This review synthesizes evidence for LF-EMF-induced endothelial angiogenesis and evaluates emerging mechanistic explanations. Methods: Literature was identified through searches of PubMed, Web of Science, and Google Scholar through March 2026. Peer-reviewed studies emphasizing in vitro endothelial angiogenesis models were prioritized, with selected in vivo studies included when relevant to mechanistic interpretation. The review was conducted as a narrative synthesis rather than a systematic review or meta-analysis. Results: Experimental evidence indicates that LF-EMF exposure can enhance endothelial proliferation, migration, sprouting, tube formation, and survival across diverse model systems. Reported mechanisms include modulation of growth factor signaling, calcium-dependent pathways, nitric oxide production, cytoskeletal remodeling, and mechanotransduction. Reexamination of early ultrastructural studies suggests that membrane ruffling, vacuole formation, and vesicular structures observed following LF-EMF exposure may be interpreted within the framework of macropinocytosis and vesicular trafficking, providing a potential mechanistic link between electromagnetic stimulation and endothelial morphogenesis. Conclusions: Current evidence supports the conclusion that LF-EMFs can modulate key endothelial processes involved in angiogenesis and vascular remodeling. Membrane trafficking and macropinocytosis provide a biologically plausible framework linking historical observations with contemporary endothelial cell biology, although direct experimental validation is needed. Future progress will require standardized exposure paradigms and mechanistic studies using physiologically relevant angiogenesis models.

Pathophysiology doi: 10.3390/pathophysiology33030056

Authors: Jelena Milenkovic Dijana Stojanovic Branka Djordjevic Sanja Velickovic Vladana Stojiljkovic Milica Veljkovic Maja Milojkovic

Background/Objectives: Quiescent leukemia stem cells (LSCs) are self-renewing, pluripotent cells that present a major obstacle to the successful curative treatment of chronic myeloid leukemia (CML). LSCs function independently of BCR::ABL1 signaling and persist following tyrosine kinase inhibitor treatment. The mechanisms enabling LSC survival are a central focus of current CML research. This review details the complex relationship between signaling pathways and discusses recent advancements in energy metabolism research within the pathogenesis of CML. Discussion: Energy metabolism is critical to the biology of CML LSCs. These cells depend on oxidative phosphorylation (OXPHOS) and mitochondrial homeostasis, utilizing fatty acid oxidation as their primary ATP source. Research highlights significant alterations in signaling networks, marked by a dynamic interplay among dominant pathways within the CML clone. While TGF-β-FOXO signaling maintains the self-renewal capacity of quiescent LSCs, proliferating mature CML cells rely heavily on glycolysis and the PI3K/Akt pathway. Furthermore, unique metabolic traits of LSCs underscore the impact of leukemic cell–microenvironment interactions in fostering a permissive niche. Conclusions: Fatty acid oxidation is critical to the survival and self-renewal of CML LSCs. This adaptation of mitochondrial function is closely linked to signaling alterations and entails an adjustment of mitochondrial respiration alongside stimulated OXPHOS. Emerging research unveils many potential targets within metabolic signaling that can be exploited to overcome these survival mechanisms, highlighting the disruption of mitochondrial energy support as a promising strategy to selectively eradicate CML LSCs.

Pathophysiology doi: 10.3390/pathophysiology33030055

Authors: Sara Puddu Stefano Vicentin Filippo Zemin Paola Veronese Elisa Cainelli

Background: During intrauterine life, the fetus is continuously exposed to rhythmic sensory stimuli; among these, the maternal heartbeat is one of the earliest and most biologically salient, contributing to the early organization of physiological regulation. This exploratory longitudinal observational study examined whether exposure to the maternal heartbeat influences neonatal autonomic activity and whether this response is moderated by prenatal maternal psychological distress. Methods: Sixteen dyads of mother–child were included in the analyses. Pregnant women were first assessed between 24 and 28 weeks of gestation: maternal distress was measured using the Depression Anxiety Stress Scales—21 (DASS-21), and maternal heartbeat recordings were obtained. At 3–4 weeks postpartum, neonates were exposed to recordings of their own mother’s heartbeat. Neonatal heart rate variability (HRV) was assessed using electrocardiography at rest and during the hearing of the maternal heartbeat. Results: Exposure to the heartbeat significantly reduced high-frequency (HF) HRV compared to baseline (p = 0.037), providing evidence for modulation of parasympathetic activity. Importantly, an interaction between condition and maternal distress was observed (p = 0.021), suggesting that neonatal autonomic responses differ as a function of maternal psychological state. Conclusions: Overall, these preliminary findings suggest that neonatal autonomic responses to maternal heartbeat exposure may not be uniform but may vary according to maternal psychological distress, highlighting the potential role of the maternal psychophysiological context in shaping early autonomic regulation.

Pathophysiology doi: 10.3390/pathophysiology33030054

Authors: Afonso Brito Bruno Daniel Carneiro Daniel Humberto Pozza

Background/Objectives: Chronic orofacial pain represents a heterogeneous group of disorders affecting the trigeminal system and remains difficult to treat due to its complex pathophysiology, which involves glial cells in the development and maintenance of persistent pain states. This systematic review aimed to synthesize evidence from animal studies investigating the pathophysiological role of glial cells in chronic orofacial pain. Methods: After registering on PROSPERO, a systematic search of the literature, based on PRISMA guidelines, was conducted to identify experimental animal studies evaluating glial involvement in orofacial pain models. After screening and eligibility assessment, 10 studies met the inclusion criteria. Data regarding experimental models, glial populations investigated, and pain-related behavioral outcomes were extracted and qualitatively synthesized. Results: Activation of satellite glial cells, microglia, and astrocytes was consistently associated with increased neuroinflammatory signaling and enhanced neuronal excitability within trigeminal pathways, demonstrating that both peripheral and central nervous tissues were involved. Several studies reported that pharmacological modulation of glial activity may reduce pain-related behaviors. Conclusions: Glial cells are key modulators of chronic orofacial pain through neuroimmune interactions that contribute to peripheral and central sensitization. Although these findings highlight promising therapeutic targets, further translational research is required to clarify their relevance for human pain conditions.

Pathophysiology doi: 10.3390/pathophysiology33030053

Authors: Sante Di Gioia Annalucia Carbone Pamela Vitullo Domenico Tierno Domenico Larobina Gabriele Grassi Mario Grassi Massimo Conese

Background/Objectives: Cystic fibrosis (CF) is a genetic disease whose hallmarks include chronic inflammation of the airways causing tissue damage with increased levels of alarmins in the respiratory secretions, as well as activation of the coagulation/fibrinolytic systems that are linked to inflammation. However, no information is available about the fibrinolytic system and how fibrinolysis is involved in airway injury. On the road to understanding this issue, we determined levels of two key markers, High Mobility Group Box 1 (HMGB1), an alarmin, and D-dimers, associated with fibrin breakdown and airway inflammation, in the sputum from patients with cystic fibrosis. Methods: Sputum samples were collected from 13 individuals with CF and subjected to two different treatment protocols. In the first protocol, the sample was treated with dithiothreitol (DTT) and then centrifuged in order to collect the supernatant (SED). In the second protocol, the sample was centrifuged and the supernatant was obtained (SE). The pellet obtained was treated with DTT and then centrifuged in order to collect the supernatant (SPE). ELISA assays were performed on all samples. Results: HMGB1 and D-dimer levels were significantly lower in supernatants from sputum centrifuged before DTT treatment (SE) compared to those processed after DTT (SPE and SED). However, no significant difference was observed between SPE and SED samples for both markers. D-dimer levels in SED correlated positively with FEV1 and blood monocyte counts. Conclusions: Direct sputum processing with DTT may be a useful procedure for assessing biomarkers of inflammation and fibrinolysis in CF.

Pathophysiology doi: 10.3390/pathophysiology33030052

Authors: Konstantinos Stamopoulos Georgios Pilichos Eleni Karatrasoglou Penelope Korkolopoulou Stratigoula Sakellariou

Background/Objectives: Lung cancer tumors are the most frequent malignant tumors and the primary cause of cancer-related deaths. Despite the improvement in survival related to the implementation of immunotherapy, a significant proportion of patients fail to demonstrate satisfactory response. As a result, the need for new biomarkers, which will predict patient response, as well as for novel therapeutic targets, is urgent. SHP-2 is an intracellular signal transduction molecule, modifying signal transduction of numerous intracellular cascades, acting primarily as an oncogene. In this study we analyzed the expression pattern of SHP-2 in a cohort of non-small-cell lung cancer cases and attempted to correlate it with available clinicopathological parameters. Methods: We performed immunohistochemistry for SHP-2 detection in a cohort of 258 NSCLC cases. H-score was estimated and statistical analyses correlated it with survival and other clinicopathological and molecular parameters. Results: SHP-2 expression was observed in 60 cases (23.3%), with an H-score ranging from 150 to 300. Positive samples were distributed among the three major histological subtypes, without statistically significant differences. Statistical analysis revealed significant positive correlation of SHP-2 expression with smoking, PD-L1 levels, and KRAS mutations. Moreover, positive SHP-2 immunohistochemistry was associated with better clinical outcome in both the total cohort and in the group of patients who received immunotherapy. Finally, the simultaneous presence of SHP-2 expression and KRAS mutation was correlated with prolonged survival. Conclusions: Our findings indicate that SHP-2 could be a useful prognostic factor and a very reliable biomarker in predicting response to immune checkpoint inhibitors.

Pathophysiology doi: 10.3390/pathophysiology33030051

Authors: Priscilla Vogt Fernandes de Souza Maria Clara Salgado-Ramos Jhenifer de Paiva Quadros Juliana Cardoso Moraes Daniel Luiz Santos da Silva Alex Oliveira da Camara Rafaella de Carvalho Cardoso Hércules Rezende Freitas

Background/Objectives: Hypertensive disorders of pregnancy, particularly preeclampsia and eclampsia, are associated with high maternal and fetal morbidity and mortality, with maternal obesity representing a major, modifiable risk factor. This study aimed to estimate the association between obesity and preeclampsia/eclampsia (PEC) in obstetric hospitalizations in the state of São Paulo, Brazil, and to examine risk trends across maternal age groups and between two distinct temporal cohorts. Methods: This population-based observational study analyzed 2,338,934 obstetric hospitalizations using data from the Hospital Information System of the Unified Health System (SIH/SUS). PEC and obesity diagnoses were coded according to the International Classification of Diseases, 10th Revision (ICD-10). Logistic regression models were applied to estimate associations adjusted for maternal age, complemented by adjusted marginal predictions. Results: PEC occurred more than twice as often among women with obesity than without (5.4% vs. 2.2%; OR 2.13, 95% CI 1.84–2.47) and was associated with longer hospital stays (3.52 vs. 2.70 days) and higher costs (R$797.91 vs. R$602.69). The absolute risk difference between obese and non-obese women declined from 24.7% to 13.6% at age 20 between the 2017–2019 and 2023–2025 cohorts, while the relative risk gradient remained stable across all ages. Conclusions: Obesity is strongly associated with increased PEC risk, longer hospitalization, and higher costs. The observed attenuation in absolute risk differences in the most recent cohort reinforces the need for preventive interventions, nutritional surveillance, and public health policies aimed at reducing hypertensive complications during pregnancy for both obese and non-obese women.

Pathophysiology doi: 10.3390/pathophysiology33030050

Authors: Meri Hristamyan Stoilka Mandadzhieva Plamena Stoimenova Nikolay Mandadzhiev Blagoi Marinov

Background/Objectives: There is increasing evidence on the effect of obesity on lung function in children with and without respiratory diseases. The aim of was to evaluate the prevalence of overweight and obesity and their impact on lung function parameters among healthy children. Methods: Six hundred and seventy-one healthy Bulgarian school children (339 males) aged 7–18 years were included in the study. All participants completed anthropometric measurements, including standing height, weight, and BMI. The studied group underwent comprehensive pulmonary function assessment. Results: The overweight and obese children group consisted of 131—overweight, 97, and obese, 34, children, accounting for 19.5% or every fifth child. The highest prevalence of overweight was at the ages of 10, 11, 12 and 17 years, and that of obesity was 7, 10 and 14 years, respectively. We compared normal weight, overweight and obese children in every age group and found that an increase in weight was associated with increased height and increased lung function parameters, such as FVC, FEV1, PEF, and FEF50. To overcome the effect of height, we compared normal weight and overweight obese children in height groups and found nonsignificant differences in the spirometric indices. Only in the 130–139 cm group the obese children had lower values of FEV1(L): 1.78 ± 0.13 vs. 1.91 ± 0.19 (NS) in children with normal weight. Conclusions: The increase in weight is associated with height growth, and the same pattern is observed for the mean spirometry indices, with lowest values found only in obese 7-year-old children.

Pathophysiology doi: 10.3390/pathophysiology33030049

Authors: Ali Sadek Yulia Khramtsova Boris Yushkov

Background: The microenvironment of spermatogonial stem cells plays a crucial role in determining their developmental trajectory. Mast cells (MCs) are an important component of this microenvironment and are present along the pathway of sperm production and maturation. Elevated temperature is associated with various disorders of male fertility and its effect on MCs has not been previously studied. Methods: The experiment was conducted on male Wistar rats, which were divided into four groups: intact, control, single, and repeated heat exposure. Morphofunctional and morphometric parameters of MCs were investigated in testes, epididymides, and seminal vesicles. Correlation analysis was performed between MC parameters, ejaculated and epididymal spermatozoa, and testosterone levels. Results: MCs become activated even after a single heat exposure, manifested by increased degranulation. Repeated exposures lead to an increase in MC numbers in the reproductive organs and an enhancement of their functional activity. Testicular MCs exhibit the highest sensitivity to elevated temperature, followed by epididymal MCs. These findings reveal organ-specific differences in MCs responses, which may be explained by the anatomical location of these organs. Correlation analysis revealed a negative relationship between MC morphofunctional parameters and sperm functional characteristics in epididymis and ejaculate, as well as testosterone levels. Conclusions: Heat exposure activates MCs in male reproductive organs, and the changes in their morphofunctional parameters support a potential role in heat-related male reproductive impairment that requires further evidence.

Pathophysiology doi: 10.3390/pathophysiology33030048

Authors: Nokwanda N. Ngcobo Ntethelelo H. Sibiya

Type 1 diabetes mellitus (T1DM) is regarded as an autoimmune disorder characterized by a progressive loss of β-cells, culminating in insulin deficiency and hyperglycemia. Currently, T1DM is managed through exogenous insulin administration. Continued efforts to decipher T1DM pathogenesis have yielded significant progress, which can be harnessed to develop disease-modifying therapeutic modalities. Immunological and genetic studies have demonstrated pathogenetic mechanisms that underpin the sensitivity and risk of developing autoimmunity, ultimately leading to the destruction of β-cells. Genetic and immunological studies, therefore, suggest that certain individuals are at risk of developing T1DM; and that autoantibodies against β-cells develop and circulate years before the onset of symptomatic T1DM. Furthermore, the onset of autoimmunity has been associated with specific triggers in genetically susceptible individuals. Recent developments have revealed how viral infections, gut dysbiosis, dietary factors, and obesity trigger autoimmunity and β-cell damage. In this review, we present the current, consolidated understanding of T1DM pathogenesis, informed by recent research. We further identify opportunities for early-detection strategies and drug development targeting the asymptomatic phase of T1DM to slow disease progression. Currently, immunomodulatory strategies have yielded promising outcomes in clinical trials. These strategies seek to target immune cells and inflammatory mediators implicated in the pathogenesis of T1DM. Other strategies include tolerogenic strategies against autoimmune cells, whilst others employ β-cell protection. The emergence of regenerative therapies also offers promising avenues toward T1DM.

Pathophysiology doi: 10.3390/pathophysiology33030047

Authors: Anastasia P. Sklifasovskaya Mikhail L. Blagonravov Madina M. Azova Sergey V. Kurevlev Vyacheslav A. Goryachev Sergey P. Syatkin Tatyana Yu. Zotova Daniil Yu. Prokofiev

We thank the commentators for their thoughtful and constructive review of our work [...]

Pathophysiology doi: 10.3390/pathophysiology33030046

Authors: Bişar Amaç

We read with great interest the article by Sklifasovskaya et al [...]

Pathophysiology doi: 10.3390/pathophysiology33030045

Authors: Muralidharan Mani Thomas F. J. Martin

Pancreatic β-cells maintain glucose homeostasis through tightly regulated insulin biosynthesis, storage, and secretion. To prevent pathological accumulation of excess or aging secretory granules (SGs), β-cells use crinophagy, a selective lysosomal degradation pathway in which mature insulin-containing granules fuse directly with lysosomes to form hybrid organelles termed crinosomes. Crinophagy was historically considered a simple mechanism for discarding obsolete, aged SGs. The acidic, protease-rich environment of crinosomes is proposed to generate unconventional insulin-derived epitopes through cathepsin-mediated proteolysis and transpeptidation reactions. These cryptic epitopes, which include hybrid insulin peptides (HIPs) resulting from the covalent fusion of insulin fragments with peptides from co-resident granule proteins, are largely absent from the thymic epitope repertoire. This creates a “peripheral–thymic mismatch” that allows autoreactive CD4+ T cells to escape central tolerance, ultimately driving β-cell destruction in type 1 diabetes (T1D). Recent studies demonstrate that pharmacological or genetic inhibition of crinophagy reduces crinosome abundance, narrows the pathogenic epitope repertoire, and delays the onset of diabetes in preclinical models. In type 2 diabetes (T2D), a related pathway termed stress-induced nascent granule degradation (SINGD) diverts newly synthesized insulin granules to lysosomes under glucolipotoxic conditions, contributing to insulin depletion and progressive β-cell failure. This review summarizes the current understanding of the molecular mechanisms behind crinophagy. It discusses its two main functions: maintaining physiological quality control and generating pathological antigens. Additionally, the review explores how crinophagy interacts with other cellular stress pathways and highlights new therapeutic strategies aimed at targeting this process to protect pancreatic β-cell function and potentially prevent or delay diabetes.

Pathophysiology doi: 10.3390/pathophysiology33030044

Authors: Salvatore Crucillà Asia Berlato Antonietta Gerarda Gravina Anneline Cremer Piotr Eder Massimo Claudio Fantini Stefano Festa Daniela Pugliese Andreas Blesl Anna Viola Chiara Viganò Sophie Vieujean Ioannis Koutroubakis Edoardo Vincenzo Savarino Lieven Pouillon Mathieu Uzzan Pierre Ellul Marie Truyens Federico Caldart Rachele Ciccocioppo Luca Frulloni Maria Cristina Conti Bellocchi

Background and Aims: Pancreatic hyperenzymemia in inflammatory bowel disease (IBD) is an under-recognized and challenging condition, as elevated pancreatic enzymes may arise from heterogeneous mechanisms and do not necessarily reflect true pancreatic disorder. This multicenter study aimed to characterize the clinical spectrum, diagnostic work-up, and outcomes of hyperenzymemia in IBD. Methods: This retrospective multicenter study was conducted within the PANDORA network, including 34 international IBD centers. For the present analysis, only centers providing patient-level data on pancreatic hyperenzymemia were considered. Collected variables included demographic, clinical, biochemical, imaging, and therapeutic data. Cases were categorized into three predefined phenotypes: chronic asymptomatic pancreatic hyperenzymemia (CAPH), reclassified acute pancreatitis not meeting Atlanta criteria (recAP), and autoimmune pancreatitis (AIP). Results: A total of 148 IBD patients with elevated pancreatic enzymes were included (CAPH 54.7%, RecAP 35.1%, AIP 10.1%). Ulcerative colitis (UC) accounted for 56.8% of cases and Crohn’s disease (CD) for 43.2%. Overall, 73.6% of patients were asymptomatic at the time of enzyme elevation. Marked hyperenzymemia (≥3× ULN) occurred in 22.3% of patients and was more frequent in RecAP than in CAPH or AIP (50.0%, 6.2%, and 13.3%, respectively). IBD was clinically active in 48.6% of patients, with higher rates in CD. Notably, the clinical meaning of hyperenzymemia differed across IBD phenotypes: in CD it was more often associated with active inflammation, whereas in UC it more frequently prompted advanced imaging and led to the identification of AIP. Imaging strategies differed significantly across phenotypes, and drug withdrawal was common in CAPH and RecAP but unnecessary in AIP. Only two rechallenges confirmed a drug-related mechanism. Conclusions: Pancreatic hyperenzymemia in IBD encompasses a spectrum of conditions with different implications. A phenotype-oriented diagnostic approach is essential to avoid misclassification and unnecessary treatment changes.

Pathophysiology doi: 10.3390/pathophysiology33030043

Authors: Maryam Adenike Salaudeen Stuart M. Allan Emmanuel Pinteaux

Background: Hypoxic–ischaemic brain injury (HI) is a major contributor to neurological deficits following stroke. Understanding what happens to the smallest functional and structural unit of the central nervous system in the face of oxygen and nutrient deprivation is essential to fully comprehend the pathogenesis of diseases and disorders associated with HI, such as ischaemic stroke. Aim: The aim of this study was to develop a robust in vitro tool for initial screening of potential therapeutics and identification of diagnostic markers of brain hypoxic injury. Methods: This study details and validates a comprehensive protocol for modelling HI using differentiated SH-SY5Y neuroblastoma cells (Neuron-like Cells, NLCs). First, we optimized the differentiation process and confirmed the maturity and purity of NLCs via standard molecular markers. The NLCs exhibited functional excitotoxicity, demonstrating a graded cell death response to N-methyl-D-aspartate (NMDA), thus validating their functional application. To simulate HI, we initially optimized the oxygen-glucose deprivation (OGD) treatment using graded concentrations of CoCl2 (0.125 mM to 2 mM) in glucose-free media. The validated NLCs were then subjected to the refined OGD protocol (1 mM CoCl2 in glucose-free media) for 3 h, followed by various periods of reoxygenation (1 h, 3 h, 6 h, 12 h, 18 h, and 24 h). Result: Bulk RNA-sequencing revealed a distinct temporal transcriptional response to HI. Injury-associated genes, including heat shock proteins and stress markers, were significantly (p < 0.05) upregulated at 3 h of reoxygenation, peaked at 6 h, and declined thereafter, remaining above baseline at 24 h. Upstream regulator analysis identified IL-1β, TNF-α, and HIF-1α as key drivers during OGD, with additional regulators emerging during reoxygenation. TNF-α and β-oestradiol were consistently identified across time points, while TGF-β1 and NTRK1 became prominent during peak injury and later phases. Analysis of secreted factors showed increased release of inflammatory (TNF-α) and neurotrophic (β-NGF, BDNF, VEGF) mediators with reoxygenation, while maximal cell death occurred at 24 h. Conclusions: This study identifies a transient, time-dependent transcriptional cascade following hypoxic–ischaemic injury, highlighting a critical window for early neuronal response. The model provides a reproducible platform for studying neuronal injury and recovery, and identifies known (TNF-α, IL-β, and HIF-1α), context-specific (NTRK1 and TGF-β) and novel (β-oestradiol) regulators of the injury response with potential relevance for therapeutic targeting.

Pathophysiology doi: 10.3390/pathophysiology33030042

Authors: Igor Manzhulo Yuliya Kipryushina Ekaterina Gromova Olga Manzhulo Elena Milkina Darya Ivashkevich

Background/Objectives. Parkinson’s disease (PD) is a multifactorial neurodegenerative disorder characterized by dopaminergic neuron loss, α-synuclein pathology, neuroinflammation, and cognitive decline. Synaptamide (N-Docosahexaenoylethanolamine (DHEA)) is an endogenous lipid mediator with documented anti-inflammatory and neurogenic properties, but its effects in PD models remain unexplored. This study aimed to evaluate the neuroprotective potential of synaptamide in a subchronic MPTP-induced mouse model of PD. Methods. Male C57BL/6 mice received MPTP (30 mg/kg/day, i.p., 5 days) with or without synaptamide (10 mg/kg/day, s.c., 13 days). Behavioral tests (open field, Y-maze, elevated plus maze, novel object recognition (NOR)) were performed, followed by immunohistochemical analysis of tyrosine hydroxylase (TH)-positive neurons in the substantia nigra, and Western blotting for α-synuclein, p-α-synuclein, TH, and IL1β in brain homogenates and serum. In vitro Neuro-2a cells were co-treated with MPP+ (100 µM) and synaptamide (0.1–10 µM) for cytotoxicity assessment (MTS assay). Results. Synaptamide (10 µM) significantly attenuated MPP+-induced cytotoxicity in Neuro-2a cells. In vivo, MPTP caused a marked loss of TH+-neurons in the substantia nigra, which was prevented by synaptamide treatment. Importantly, this subchronic MPTP model recapitulates early biochemical alterations (e.g., α-synuclein phosphorylation at Ser129) rather than mature Lewy body pathology, a limitation that should be considered when interpreting these findings. Although no motor deficits or anxiety-like behavior were observed, the NOR test revealed MPTP-induced long-term memory impairment, which was fully restored by synaptamide. Conclusions. These findings suggest that synaptamide may exert effects on pathological processes associated with PD, warranting further investigation into its potential role in combination or supportive therapy for this disease.

Pathophysiology doi: 10.3390/pathophysiology33020041

Authors: Ryoji Tanei Yasuko Hasegawa

Atopic dermatitis (AD) is increasingly interpreted through frameworks emphasizing barrier dysfunction, type 2 cytokine signaling, pruritus pathways, and microbial dysbiosis, often relegating IgE-mediated mechanisms to secondary roles. In this narrative review, we synthesize historical, clinical, immunologic, and histopathologic evidence to propose a conceptual model in which IgE-bearing antigen-presenting cells (APCs)—including Langerhans cells, inflammatory dermal dendritic cells, and inflammatory dendritic epidermal cells (IDECs)—participate in an IgE-dependent amplification loop that may contribute to the chronicity of extrinsic (IgE-associated) AD. Evidence from human studies indicates that FcεRI-expressing APCs can acquire environmental allergens through IgE, enhancing antigen uptake and T-cell activation, while mast cells and basophils further reinforce type 2 inflammation through IgE-dependent and IgE-augmented pathways. Although these mechanisms have been described across distinct experimental and clinical contexts, their integration into a unified pathogenic circuit remains hypothesis-driven. We therefore present an interpretive framework that organizes these partially validated mechanisms into a coherent model linking cutaneous sensitization, allergen capture, APC activation, Th2 polarization, and spongiosis formation. This conceptual synthesis aims to reposition IgE-mediated processes within the broader pathophysiology of extrinsic AD and to highlight potential therapeutic implications for targeting IgE–FcεRI signaling and IgE-dependent APC biology.

Pathophysiology doi: 10.3390/pathophysiology33020040

Authors: Bianca Herrmann Horia Sirbu Hayk Kikoyan Mostafa Higaze Abbas Agaimy Arndt Hartmann Ralf Rieker Mohamed Anwar Haj Khalaf

Background: Spread through air spaces (STAS) is a recognized histologic pattern of invasion associated with poor prognosis in non-small-cell lung cancer (NSCLC), particularly adenocarcinoma. However, its presence in pulmonary squamous cell carcinoma (SCC), whether primary or metastatic, remains largely unexplored. Given the limited available evidence, this study was designed as an exploratory analysis to evaluate the prevalence and potential prognostic significance of STAS in pulmonary SCC. Material and Methods: In this exploratory retrospective study, we analyzed 57 patients who underwent surgical resection for pulmonary squamous cell carcinoma (P-SCC) at the Department of Thoracic Surgery at the University Hospital Erlangen between 2008 and 2020. The cohort included both primary lung SCC and metastatic SCC to the lung from extrapulmonary sites, primarily from ear, nose, and throat (ENT) tumors. Histological slides were reviewed to assess the presence of STAS, as defined by established morphological criteria. The Chi-square test was used to investigate the presence of STAS. Disease-free survival (DFS) and overall survival (OS) was evaluated using Kaplan–Meier analysis, and the prognostic impact of STAS along other variables were assessed using Cox proportional hazards regression. Results: A total of 57 patients with squamous cell carcinoma (SCC), 22 (39%) had primary lung SCC and 35 (61%) had metastatic SCC from head and neck tumours (ENT). Spread through air spaces (STAS) was detected in 20 patients (35%). Disease-free survival (DFS) differed according to primary tumour location (p-value of 0.009), with higher 1-, 3-, and 5-year DFS in patients with primary lung SCC (86.4%, 77.3%, 63.3%) than in those with head and neck SCC (54.3%, 31.4%, 22.2%). DFS was also significantly higher in patients undergoing solitary resections compared with multiple resections (78.6%, 64.3%, 49.5% vs. 33.3%, 6.7%, not estimable; p-value < 0.001). DFS was slightly longer in STAS-negative patients compared with STAS-positive patients (1-, 3-, 5-year DFS: 64.9%, 51.4%, 40.5% vs. 70%, 45%, not estimable), (median DFS 36 vs. 25 months; p-value of 0.776). Overall survival (OS) was significantly longer in patients with primary lung SCC (median OS 125 months) than in those with head and neck SCC (27 months; p-value of 0.039). STAS-negative patients had also a longer OS than STAS-positive patients (median OS 46 vs. 38 months; HR = 1.11, 95% CI 0.56–2.20; p-value of 0.771). Conclusions: STAS was identified in metastatic pulmonary SCC lesions as well as in primary lung SCC, occurring in approximately one-third of cases. However, due to the limited cohort size and the exploratory univariate design of the study, the prognostic significance of STAS could not be definitively established and requires further investigation in larger, adequately powered studies.

Pathophysiology doi: 10.3390/pathophysiology33020039

Authors: Dominika Jabłonka Stefan Modzelewski Napoleon Waszkiewicz

Background: Atopic dermatitis is a chronic inflammatory skin disease characterized by epidermal barrier dysfunction and immune dysregulation, whereas major depressive disorder is a common psychiatric condition with a substantial impact on quality of life; increasing attention has been given to oxidative and nitrosative stress as a potential biological link between these disorders. Methods: This narrative review synthesizes current evidence on molecular biomarkers of oxidative and nitrosative stress in AD and MDD and examines shared mechanisms within the skin–brain axis. Results: Across both conditions, studies consistently report increased markers of lipid peroxidation (e.g., malondialdehyde, 4-hydroxynonenal), oxidative DNA damage (8-hydroxy-2′-deoxyguanosine), and nitrosative stress, alongside impaired antioxidant defenses, particularly involving glutathione; these alterations are closely associated with chronic inflammation, cytokine signaling, mitochondrial dysfunction, and dysregulation of neuroimmune and hypothalamic–pituitary–adrenal axis pathways. Conclusions: Although the available evidence is heterogeneous and largely based on cross-sectional studies, limiting causal inference, the findings support a biologically plausible link between AD and depression mediated by shared redox pathways and highlight the need for further longitudinal and mechanistic research.

Pathophysiology doi: 10.3390/pathophysiology33020038

Authors: Ilyas Kabdesh Aizilya Bilalova Yana Mukhamedshina Yuri Chelyshev

NG2 glia, also known as oligodendrocyte progenitor cells, represent a unique population of glial cells characterized by dynamic morphology and the ability to extend branched processes that actively contact neurons and other cellular elements. These structural and functional interactions enable NG2 glia to contribute to the regulation of axonal excitability, electrical activity, and axonal architecture. Unlike most other glial cells, NG2 glia receive direct synaptic input from neurons and can generate action potentials, defining their distinctive physiological status. A particularly important feature of this cell population is the expression of the chondroitin sulfate proteoglycan NG2/CSPG4, which serves as a key molecular marker and plays an essential role in intercellular interactions. Following spinal cord injury (SCI), NG2 glia rapidly become activated, undergo phenotypic changes, and engage in extensive interactions with neurons, astrocytes, microglia, and endothelial cells. These interactions form a complex regulatory network that influences both the severity of secondary injury and the effectiveness of remodeling and repair processes. Mechanisms of particular importance include the secretion of chondroitin sulfate proteoglycans and alterations in extracellular matrix properties. Finally, this review highlights potential therapeutic approaches aimed at modulating NG2 glial activity and their intercellular interactions. The focus is on strategies designed to reduce the inhibitory effects of proteoglycans while enhancing the remyelinating and neuroprotective potential of these cells, thereby opening new perspectives for regenerative therapies after SCI.

Pathophysiology doi: 10.3390/pathophysiology33020037

Authors: Ghassan Bkaily Alexandre Normand Ashley Jazzar Houssein Najibeddine Danielle Jacques

Background/objectives: The development of hereditary cardiomyopathy associated with early death was known to take place during the patients’ lifetime. Previous work showed that remodeling of heart cells in hereditary cardiomyopathy of hamsters (HCMH) took place as early as during postnatal development. The present work tested the hypothesis that cardiomyocyte remodeling in HCMH occurred early in fetal development. Methods and results: Using quantitative 3D confocal microscopy associated with ionic and immunofluorescence, our results showed that, as in the adult heart, cardiomyocytes isolated from 15-day-old fetal HCMH hearts showed an increase in intracellular calcium, sodium, ROS, and sodium-hydrogen exchanger 1 (NHE-1) when compared to age-matched cardiomyocytes of a normal hamster (NH). Conclusions: These results demonstrated that hereditary cardiomyopathy occurred during fetal heart development, and that early treatment with an NHE-1 blocker might have prevented the development of hereditary cardiomyopathy and heart failure that occur during the hamsters’ lifetime.

Pathophysiology doi: 10.3390/pathophysiology33020036

Authors: Jhan S. Saavedra-Torres Lady Viviana Acosta Castillo Alexandra Montoya Rendon Daniel Esteban Castro Valencia Diego A. Lucero Guanga Manuela Garzon Ovalle Fabián Darío Arias Rodríguez Andrés López-Cortés Juan S. Izquierdo-Condoy

Sepsis is a systemic disorder in which infection-induced inflammation progressively disrupts vascular homeostasis and drives organ dysfunction. This review reframes septic pathophysiology as a sequential and self-amplifying process centered on endothelial failure. Early activation of innate immune pathways by pathogen- and damage-associated molecular patterns promotes cytokine release, oxidative stress, and enzymatic degradation of the endothelial glycocalyx. Loss of this protective surface layer exposes endothelial cells to unbuffered inflammatory and mechanical injury, impairing mechanotransduction, increasing leukocyte and platelet adhesion, and destabilizing vascular barrier function. Subsequent disruption of intercellular junctions promotes capillary leakage, tissue edema, and impaired oxygen diffusion, while mitochondrial dysfunction and redox imbalance reduce endothelial repair capacity. In parallel, complement activation, neutrophil extracellular trap formation, platelet–leukocyte interactions, and loss of anticoagulant signaling shift the microvasculature toward a prothrombotic and proinflammatory state. These interconnected mechanisms culminate in microvascular incoherence, characterized by heterogeneous capillary flow, regional hypoxia, impaired oxygen extraction, and progressive organ failure despite apparent restoration of systemic hemodynamics. Within this framework, biomarkers such as syndecan-1, soluble thrombomodulin, angiopoietin-2, von Willebrand factor, and plasminogen activator inhibitor-1 are best interpreted as mechanistic readouts of glycocalyx shedding, endothelial injury, permeability imbalance, and thromboinflammatory activation. Understanding sepsis as an evolving endothelial pathophysiological process provides a coherent framework for integrating inflammation, vascular leakage, hypoxia, coagulation, and organ dysfunction while identifying mechanistic biomarkers that reflect distinct stages of microvascular collapse.

Pathophysiology doi: 10.3390/pathophysiology33020035

Authors: Alireza Minagar Mohammadali Sahraian

Multiple sclerosis (MS) is a chronic, immune-mediated demyelinating disease of the central nervous system whose heterogeneous clinical, radiological, and biological course has long resisted precise individual-level prediction. The recent convergence of large longitudinal datasets, advanced computational methods, and increasingly informative biomarkers has created conditions in which artificial intelligence (AI) and machine learning (ML) can begin to address that problem substantively. This review surveys the current evidence for AI/ML applications across the MS care continuum, with particular focus on the literature from 2022 through early 2026. Nine domains are examined: automated MRI lesion segmentation and quantification, fluid biomarker interpretation, unsupervised disease subtyping, disability progression prediction, treatment response stratification, drug repurposing and molecular discovery, digital biomarker monitoring, mechanistic interpretability, and integrated clinical management protocols. Notable recent contributions include the SuStaIn-based identification of two biologically distinct MS trajectories distinguished by early versus late serum neurofilament light chain elevation, the MindGlide deep learning platform enabling longitudinal analysis of archived routine clinical MRI data, the T-cell morphological classifier predicting natalizumab treatment response before drug initiation, and the fenebrutinib Phase III program that produced the first Bruton’s tyrosine kinase inhibitor results meeting primary endpoints in both relapsing and primary progressive MS. A proposed AI-Enhanced Management Protocol (AMP-26) reflecting 2026 clinical standards is included as an appendix. Throughout, emphasis is placed on mechanistic interpretability: the distinction between models that correlate features with outcomes and models whose decision logic reflects established MS pathobiology is considered a prerequisite for clinical credibility and regulatory readiness.

Pathophysiology doi: 10.3390/pathophysiology33020034

Authors: Matteo Gelardi

Biomarker-driven approaches have markedly improved the stratification and management of airway inflammatory diseases. However, in everyday clinical practice, these strategies still rely mainly on systemic indicators, which often provide only an indirect view of the inflammatory processes occurring within the airway mucosa. This limitation becomes particularly evident in chronic conditions such as chronic rhinosinusitis with nasal polyps (CRSwNP), where local inflammatory patterns may not relate to circulating biomarkers. Nasal cytology represents a simple, non-invasive, and reproducible technique that allows direct evaluation of the cellular components of the nasal mucosa. By identifying distinct inflammatory patterns, it offers a real-time snapshot of the local inflammatory microenvironment, bringing the clinician closer to the site of disease. In this hypothesis, we propose that airway inflammation is primarily driven by local cytological patterns. In particular, we suggest that the interaction between eosinophils and mast cells constitutes a key pathogenic axis underlying disease activity, severity, and progression. From a pathophysiological perspective, eosinophils may reflect a more chronic component of inflammation, whereas mast cells are more closely associated with active and dynamic phases of the disease. Their coexistence may therefore identify a state of amplified inflammatory activity, often associated with more severe clinical phenotypes. We further propose that integrating cytological findings into clinical–cytological grading (CCG) systems could improve patient stratification and support more personalized therapeutic strategies. This model is readily testable in current clinical and research settings and may contribute to a progressive shift toward the use of local biomarkers in precision medicine.

Pathophysiology doi: 10.3390/pathophysiology33020033

Authors: Reese Leonhard Zachary Beau Phillips Jamie Wilson Zaid Abu-Mowis John DiGiorgi Epiphany N. Wilson Zane Borenstein Laura Wilson Richard Tang Elizabeth H. Stephens Adrian Crucean Michael S. Shillingford Giles J. Peek Mark Steven Bleiweis J. Steven Alexander Jeffrey Phillip Jacobs

Background: Hypoplastic left heart syndrome (HLHS) is defined as “a spectrum of congenital cardiovascular malformations with normally aligned great arteries without a common atrioventricular junction, characterized by underdevelopment of the left heart with significant hypoplasia of the left ventricle including atresia, stenosis, or hypoplasia of the aortic or mitral valve, or both valves, and hypoplasia of the ascending aorta and aortic arch”. Without treatment, HLHS is usually lethal in the neonate. Many hypotheses have been advanced to explain the etiology of HLHS; however, no single theory appears to fully explain the phenotypic variability seen in HLHS. Furthermore, many of these theories offer no explanations regarding the precipitating events which lead to the development of HLHS. Objective: This review considers and critically evaluates the strengths and weaknesses of the leading theories proposed to explain the pathogenesis of HLHS—including hemodynamic disturbances, primary myocardial structural defects, valvar malformations, and genetic or epigenetic alterations that may provoke developmental and anatomic abnormalities. After presenting each model, we propose a novel, comprehensive, and data-driven framework which may assist researchers in developing models for the pathogenesis of the various subtypes of HLHS. Methods: Key findings from human fetal imaging, histopathology, genetic studies, and animal models were considered, as well as the hypothetical contribution of each in observed HLHS phenotypes. The rationales for these findings as causal factors initiating individual HLHS patterns, as well as how they might contribute to HLHS in general, were critically analyzed. Results: The flow theory is strongly supported by animal models and in utero interventions that demonstrate the impact of altered hemodynamics on cardiac morphogenesis. However, the flow theory fails to identify initial causes of disturbed flow or related histological features of HLHS like endocardial fibroelastosis. The myocardial and valve-first models suggest an important role in developmental defects, but do not necessarily have a strong experimental basis that provides explanations for how they mediate HLHS. Genetic studies in patients with HLHS have identified several candidate causal mutations. However, such genetic causes of HLHS exhibit incomplete phenotypic penetrance and clinical impact. A multifactorial framework attempts to integrate these diverse mechanisms and may provide the most coherent explanation that can accommodate the heterogeneity and variable presentation of HLHS. Such a framework may identify multiple forces that drive disease but does not provide useful pathways for future research about HLHS. Conclusions: No single hypothesis has fully explained how HLHS is initiated, progresses, and presents with the clinical conditions that are encountered by cardiac surgeons and cardiologists. The most current models suggest that the spectrum of HLHS reflects acomplex interaction between genetic susceptibility, flow-dependent cardiac remodeling, and environmental factors in utero. A multifactorial model integrates these diverse mechanisms and may provide the most coherent explanation for the various phenotypic variations in HLHS. Based on our analysis of the most current data and the strengths and weaknesses of the current theoretical frameworks, we propose a novel research strategy aimed at identifying specific cardiac progenitor cell populations whose dysregulation may represent a unifying explanation for the etiology of the various phenotypes of HLHS. Based on the arguments made throughout this manuscript that evaluate the various genetic, structural, and hemodynamic models of initiation of disease, we believe that the significant phenotypic variability across the spectrum of HLHS (i.e., the different anatomic subtypes for “classic” HLHS) most likely reflects different underlying etiologies and mechanisms. At the very least, it is very likely that the timing of the insult is critical in determining anatomic subtype. Based on the published data and the arguments within this manuscript, it seems naive to think that there is a single unifying mechanism explain all forms of HLHLS.

Pathophysiology doi: 10.3390/pathophysiology33020032

Authors: Dmitry S. Semenovich Polina A. Abramicheva Ljubava D. Zorova Andrey V. Elchaninov Maria A. Kozlova David A. Areshidze Nadezda V. Andrianova Nina P. Kanunnikova Andrey G. Moiseenok Irina B. Pevzner Egor Y. Plotnikov Dmitry B. Zorov

Background/Objectives: Inflammation and oxidative stress are key factors contributing to the initiation and progression of liver fibrosis in chronic obstructive cholestasis. Pantothenic acid (PA) and some of its derivatives have been reported to exhibit moderate anti-inflammatory, antioxidant, and regenerative effects. This study aimed to evaluate the redox-modulating effects of PA derivatives—panthenol (PL), pantethine (PT), and hopantenic acid (HPA) in a rat model of chronic obstructive cholestasis induced by common bile duct ligation (BDL). Methods: Macroscopic, histological, and ultrastructural alterations in the liver were assessed, along with molecular markers of oxidative stress, inflammation, and parameters of the glutathione (GSH) system. Results: BDL-induced liver injury was associated with enhanced lipid peroxidation, mitochondrial structural alterations, depletion of GSH, increased levels of protein S-glutathionylation (PSSG), and elevated thiobarbituric acid-reactive substances in mitochondria. Treatment with PL and, to a lesser extent, PT was associated with attenuation of hepatocellular ultrastructural damage, reduced bile duct hyperplasia, decreased inflammatory and necrotic changes, and moderate improvement in fibrosis-related parameters. In contrast, HPA (a PA antagonist) did not demonstrate hepatoprotective effects and it was associated with more pronounced liver injury. Conclusions: Chronic BDL is accompanied by suppression of glutathione redox capacity and enhanced oxidative stress. PL and PT, but not HPA, were associated with reduced levels of protein S-glutathionylation and partial restoration of redox balance. The protective effects of PL and PT may contribute to their antifibrotic activity, potentially through direct antioxidant capacity or redox-modulating mechanisms associated with the GSH system.

Pathophysiology doi: 10.3390/pathophysiology33020031

Authors: Pedro Céspedes Cristina Buigues María Dolores Torregrosa Francisco M. Martínez-Arnau Omar Cauli Isabel Trapero

Background: Fibroblast growth factor 21 (FGF21) is a peptide hormone that is synthesized by several organs and regulates energy homeostasis, including reducing fat mass and lowering hyperglycemia, insulin resistance and dyslipidemia. It also increased metabolic syndrome (MS) and cardiovascular risk in breast cancer (BC) survivors treated with aromatase inhibitors (AIs) aimed at reducing cancer recurrence. We evaluated whether blood FGF21 concentration is associated with MS and its five criteria in postmenopausal women treated with AIs, and whether this persists after multimodal interventions that reduce MS. Methods: A quasi-experimental longitudinal study in 31 postmenopausal women with localized BC on Ais, assessed via a 12-week multimodal program. Their MS was evaluated per the NCEP-ATP III guidelines (waist circumference, blood pressure, fasting glucose, triglycerides, HDL-cholesterol). Plasma FGF21 was measured pre/post-intervention via fasting blood samples, centrifugation, and ELISA assay. Results: Pre-intervention FGF21 median: 377.62 pg/mL (38.40–1616.42). Plasma FGF21 concentrations positively correlated with MS criteria number pre- and post-intervention (all p < 0.05). Linear regression confirmed pre-intervention MS criteria (β = 127.262, p = 0.006) and antihypertensive drugs as predictors of post-FGF21 levels. Analysis of individual MS criteria revealed significant associations with blood pressure (p = 0.028 and p = 0.022 for systolic and diastolic pressure, respectively) and fasting glucose changes (p = 0.008). Conclusions: Plasma FGF21 may act as a biomarker for monitoring exercise-based interventions which reduce MSs, particularly hypertension and hyperglycemia, in AI-treated BC survivors.

Pathophysiology doi: 10.3390/pathophysiology33020030

Authors: Felix Y. Narvaez Irizarry Tyrel R. Porter Neisha Ramirez Serrano Lilia Y. Kucheryavykh

Background: Amyloid-related pathways are well studied in neurodegenerative diseases but remain poorly characterized in gliomas. Amyloid-related transcriptional programs in low-grade gliomas (astrocytoma grade II-III) and oligodendrogliomas, and their association with patient survival, were analyzed in this study. Methods: Transcriptomic data from 193 grade II-III astrocytomas and 191 oligodendrogliomas were analyzed to evaluate histology-specific expression patterns and prognostic significance. Differential and single-sample gene set enrichment analyses (ssGSEA) were used to calculate per-sample enrichment scores for 30 amyloid-related Gene Ontology biological process gene sets across the combined cohort. These scores were used to compare pathway activity between grade II-III astrocytoma and oligodendroglioma samples. Pathway-level survival analyses were performed for each tumor type using ssGSEA enrichment scores to evaluate associations with overall survival. Results: Distinct amyloid-related transcriptional programs were identified between glioma subtypes. Grade II-III astrocytomas showed enrichment of pathways related to amyloid precursor protein (APP) processing and amyloid-β clearance, whereas oligodendrogliomas were enriched in lipid transport and negative regulation of amyloid formation. Survival analyses revealed that higher activity of the positive regulation of APP biosynthetic process and amyloid-β clearance by transcytosis was significantly associated with worse overall survival in grade II-III astrocytoma, but not in oligodendroglioma. Gene-level analyses in astrocytoma demonstrated consistent survival associations across multiple genes within these pathways, supporting coordinated pathway-level effects rather than isolated single-gene prognostic markers. Conclusions: Amyloid-related transcriptional programs differ substantially between diffuse glioma subtypes. Increased APP biosynthesis and amyloid-β transcytosis pathways are associated with poorer survival specifically in grade II-III astrocytoma, suggesting a potential role for amyloid metabolism in tumor progression. These findings identify APP-related pathways as candidates for further mechanistic investigation and potential therapeutic targeting in grade II-III astrocytoma.

Pathophysiology doi: 10.3390/pathophysiology33020029

Authors: Weiwei Gong Yueyang Liu Natalie Falkenberg Marion Kiechle Holger Bronger Julia Dorn Viktor Magdolen Tobias Dreyer

Background: Triple-negative breast cancer (TNBC) is associated with poor prognosis and limited targeted treatment options. The urokinase plasminogen activator receptor (uPAR) contributes to tumor aggressiveness and may be regulated by microRNAs such as miR-221. This study aimed to evaluate the prognostic relevance of uPAR mRNA and miR-221 expression in TNBC. Methods: uPAR mRNA and miR-221 expression levels were quantified by real-time PCR in tumor tissues from 101 patients with TNBC. Associations with clinicopathological parameters and disease-free survival (DFS) were analyzed using univariate and multivariable Cox regression models. In silico analyses of publicly available datasets were performed for validation and, in addition, for further miR-221 target prediction. Results: In both univariate and multivariable analyses, high uPAR mRNA expression was associated with shorter DFS, whereas, in contrast, elevated miR-221 expression correlated with improved DFS. No inverse correlation between uPAR and miR-221 expression was observed, making a direct regulatory miR-221/uPAR axis in TNBC unlikely. Still, combined analysis revealed a pronounced additive prognostic effect, with high uPAR and low miR-221 expression identifying patients with the poorest DFS. These findings were supported by in silico analysis with publicly available patient data. Finally, other potential miR-221 targets were identified by applying in silico target prediction. Conclusions: uPAR and miR-221 represent independent prognostic markers in TNBC. Their combined expression provides additional prognostic value for disease-free survival and supports their potential relevance as biomarkers and therapeutic targets in TNBC.

Pathophysiology doi: 10.3390/pathophysiology33020028

Authors: Yuchun Chen Megan Chiem Nandini Joshi Paul C. H. Li

Background/Objectives: Multidrug resistance (MDR) remains a major pathophysiological barrier to effective chemotherapy based on anthracyclines, including daunorubicin (DNR), in the treatment of leukemia. However, conventional population-level measurements of drug uptake do not resolve variability in uptake kinetics among individual leukemia cells, which may influence intracellular drug accumulation and therapeutic response. Methods: In this study, real-time DNR uptake was quantified at the single-cell level using a microfluidic biochip that enabled long-term cellular retention and continuous monitoring. Both wild-type drug-sensitive leukemia cells and a multidrug-resistant mutant overexpressing the P-glycoprotein (P-gp) efflux pump were examined. Results: Kinetic analysis revealed that DNR uptake in drug-sensitive cells was well described by a single dominant uptake process, whereas uptake in MDR cells required a model incorporating two kinetically distinct processes. In both cell populations, pronounced cell-to-cell variation was observed in uptake rates and intracellular drug retention, indicating substantial functional heterogeneity within phenotypically similar cells. This variability persisted following the treatment with an MDR inhibitor and obscured the differences between inhibitor-treated and untreated cells when the uptake was compared across different single cells. To overcome this limitation, a same-single-cell analysis (SASCA) approach was employed, enabling direct comparison of DNR uptake in the same individual cell before and after inhibitor exposure, thereby revealing enhanced intracellular DNR retention and accelerated uptake kinetics following inhibition. Conclusions: Together, these results demonstrate that real-time single-cell kinetic analysis reveals functionally relevant heterogeneity in multidrug-resistant leukemia cells and provides insight into the pathophysiology of MDR that cannot be obtained from population-averaged measurements.

Pathophysiology doi: 10.3390/pathophysiology33020027

Authors: Akram M. Eraky Yasser Mokhtar Guy Grabau Adnan Khan Mark Jarosz Alisha Wright Matthew Grounds Kyle Kennedy

Patients with preload-dependent conditions are at high risk of hemodynamic instability from both hypovolemia and hypervolemia. In hypovolemic states, the presence of third spacing may be misleading and obscure true intravascular volume status. Therefore, management of critically ill patients should be guided by a thorough understanding of physiology and pathophysiology to appropriately address hemodynamic derangements. Overreliance on rigid protocols and protocol-driven care without adequate clinical judgment may, in some cases, adversely affect patient outcomes. Herein, we present a case of hypovolemia-induced hypotension in the setting of third spacing and concentric left ventricular hypertrophy.

Pathophysiology doi: 10.3390/pathophysiology33020026

Authors: Andreas Pfützner Tobias Gantner Harald Burgard Tilman Steinmeier Eduard Stappler Julia Jantz Petra Wiechel

Atherosclerosis remains the leading cause of death worldwide and imposes a major healthcare burden. Physiologically, elimination of cholesterol from the arterial wall depends on reverse cholesterol transport (RCT). RCT requires access to HDL and apolipoprotein A-I (ApoA-I) to lesional macrophages/foam cells. The endothelial glycocalyx is a dynamic and injury-sensitive layer of proteoglycans and glycosaminoglycans (including hyaluronan). It contributes to vascular barrier properties, leukocyte adhesion, mechanotransduction, and macromolecular transport. In atherosclerosis, glycocalyx structure and function are altered; this may facilitate entry/retention of atherogenic lipoproteins and may also alter transport conditions relevant to cholesterol efflux pathways. This article presents a mechanistic hypothesis: short, transient, systemic hyaluronidase exposure could temporarily remodel glycocalyx/extracellular matrix components and thereby facilitate conditions permissive for regulated transport processes relevant to RCT. However, the proposed link between glycocalyx remodeling and improved lesional cholesterol efflux remains theoretical. Direct in vivo evidence that the endothelial glycocalyx is a dominant barrier limiting HDL- or ApoA-I-mediated cholesterol efflux from plaque macrophages is currently limited. Moreover, glycocalyx degradation is widely associated with endothelial dysfunction, increased permeability, inflammation, and thrombosis, all of which could aggravate rather than ameliorate atherosclerosis. Human pharmacokinetic data indicate a very short plasma half-life of circulating hyaluronidase activity, suggesting that any systemic enzymatic effect is brief. Nevertheless, the biological consequences of repeated degradation–regeneration cycles, especially in high-risk states such as diabetes, inflammation, oxidative stress, or chronic kidney disease, remain incompletely understood. Evidence supporting clinical benefit in atherosclerosis is currently limited to heterogeneous animal experiments, historical uncontrolled reports, and a small number of anecdotal case observations, whereas randomized trials have only been performed in other settings such as acute myocardial infarction and do not establish efficacy for plaque regression. We therefore provide a balanced evaluation of knowns, uncertainties, alternative interpretations, potential risks, dosing unknowns, and a translational research agenda including mechanistic preclinical studies, biomarker development, imaging, and carefully designed early-phase clinical investigation.

Pathophysiology doi: 10.3390/pathophysiology33020025

Authors: Reona Shiro Ikuo Tsunoda

Human papillomavirus (HPV) can cause cervical cancer. Global viral eradication relies on specific criteria, including a single host species and effective vaccines, a feat successfully achieved with smallpox and rinderpest. Although measles is also a candidate for elimination, its progress has been hindered by vaccine hesitancy based on misinformation about vaccine safety. Similarly, HPV is an ideal candidate for eradication due to its strict human infectivity and the proven vaccine efficacy in reducing cancer rates and establishing herd immunity. We highlighted the growing global consensus on single-dose HPV vaccination to improve feasibility and compliance with comparable effectiveness and safety to three-dose vaccination. Supporting this, we demonstrated that mice receiving a single HPV vaccine produced anti-HPV antibodies without a prolonged pro-inflammatory cytokine profile. On the other hand, in Japan, a nine-year suspension of proactive government recommendations occurred due to alleged adverse events termed “HPV vaccination-associated neuro-immunopathic syndrome (HANS),” drastically reducing vaccination rates, despite rigorous international studies have confirmed the vaccine’s safety. Critical scientific evaluation demonstrated that HANS failed to meet the criteria for autoimmune diseases (Witebsky’s postulates); no evidence has been presented that HANS is a novel autoimmune disease. The claim of molecular mimicry between HPV L1 and human proteins was based solely on flawed computational analyses. Furthermore, the hypothesis implicating a pathogenic role for aluminum adjuvants was unsupported by experimental evidence; HANS animal models were flawed methodologically and unreproducible experimentally. In summary, we believe that implementing worldwide HPV vaccination strategies, including gender-neutral and single-dose programs, as well as denouncing pseudoscientific claims hold the potential to eliminate high-risk HPV types globally.

Pathophysiology doi: 10.3390/pathophysiology33020024

Authors: Harjinder Singh Nida Qadir Malti Bhamrah William Rosales-Gonzalez Paul Bhamrah Naomi Ghildiyal Brittany Monceaux Cesar Liendo Sheila Asghar Jonathan Steven Alexander Oleg Y. Chernyshev

Background: Obstructive sleep apnea (OSA) is a complex and diverse disorder affecting almost one billion individuals worldwide. Severity of untreated OSA, measured by the apnea–hypopnea index (AHI), is noted to be associated with an increased all-cause and cardiovascular mortality. Although widely used, AHI insufficiently captures disease variability as there is a poor correlation of symptoms with the AHI. There lies individual susceptibility to the effects of OSA and that parameter alone poorly predicts cardiovascular outcomes without considering intermittent hypoxia and the hemodynamic effects of OSA. Recognition of clinical, polysomnographic, and neurophysiological phenotypes offers an opportunity to refine diagnosis, prognosis, and management strategies. Methods: We conducted a narrative synthesis of the literature involving 70 articles, focusing on quantitative and qualitative (Q2) clinical traits, polysomnographic parameters, and mechanistic insights that enable subclassification of OSA beyond AHI. Evidence from large cohorts, animal models, and pathophysiological studies were reviewed. Results: Phenotyping based on a Q2 analysis of polysomnographic respiratory event predominance, event duration, positional and REM dependence, hypoxic burden, and arousal characteristics reveals significant heterogeneity in risk profiles and therapeutic response. Apnea-predominant OSA correlates with a higher oxygen desaturation index and Epworth sleepiness scale. Hypopnea-predominant OSA correlates with a cardiometabolic disease burden and may show a more favorable response to surgical therapies. The duration of respiratory events is related to cardiovascular risk, and REM-predominant OSA independently predicts hypertension and adverse cardiovascular outcomes. Supine-predominant OSA demonstrates treatment responsiveness to auto-positive airway pressure and positional therapy. Respiratory effort–related arousals (RERAs), RERA-predominant OSA and the broader respiratory disturbance index (RDI) provide neurophysiological insight often missed by AHI-based classifications. Hypoxic burden, rather than AHI, emerged as a superior predictor of cardiovascular events and mortality. Finally, arousal frequency and periodic limb movements independently predict cardiovascular morbidity. Conclusions: Employing Q2-based phenotyping that incorporates clinical, polysomnographic, and neurophysiological markers improves risk stratification, prognosis, and individualized management of OSA. Future investigations should prioritize integrating phenotypic subclassification into diagnostic criteria and treatment planning to advance precision medicine in sleep apnea care.

Pathophysiology doi: 10.3390/pathophysiology33010023

Authors: Lorena Oreščanin Zrinka Biloglav Ivana Škrlec

Introduction: Apert syndrome is a rare genetic disorder characterized by craniofacial anomalies and limb malformations, often accompanied by neurodevelopmental abnormalities that can considerably affect motor development. Aim: The aim of this study was to document the progress in motor development of a girl with Apert syndrome, with an emphasis on assessing functional needs and evaluating the effects of a multidisciplinary rehabilitation approach. Materials and Methods: Motor functions were evaluated using the Gross Motor Function Measure (GMFM-88) at 16 and 24 months of age. Rehabilitation consisted of an intensive physiotherapy program, Dynamic Movement Intervention (DMI), delivered in monthly cycles over eight months. The therapeutic approach focused on developing postural control, transitional positions, and functional mobility while stimulating sensorimotor integration and neuroplasticity. Results: The initial GMFM score was 29.00%, and the final assessment score reached 68.68%, representing a relative improvement of 136.83%. The most considerable progress was observed in sitting, crawling, and kneeling, with initial improvements in standing. Despite the limitations of this study, the results suggest a positive effect of early, intensive, and individualized rehabilitation combined with active family involvement. Conclusions: The outcomes highlight the importance of early assessment, continuous monitoring of motor development, and a multidisciplinary rehabilitation approach in children with Apert syndrome, with the GMFM serving as a valuable tool for evaluating gross motor function.

Pathophysiology doi: 10.3390/pathophysiology33010022

Authors: Said José Serrano Guzmán Carlos Leyber Vargas Juárez Marcos Hernández Gómez José Roberto Luis Vásquez Sergio Roberto Aguilar Ruiz Juan Carlos Ramos Martínez Joscelin Amaranta Macías Ríos Edgar Gustavo Ramos Martínez José Luis Cano Pérez Jesús David Guzmán Ortiz Martha Silvia Martínez Luna Leticia Lorena Hernández González

Background/Objectives: Sex and age influence inflammatory responses, but researchers have not fully characterized their combined association with complicated acute appendicitis (CAA). This study assessed the independent and interactive associations of sex, age, and inflammatory cell counts with CAA. Methods: We conducted a retrospective observational study of 708 patients with histopathologically confirmed uncomplicated appendicitis (UAA) or CAA. We analyzed demographic and clinical data, including preoperative complete blood counts, stratified by sex. We used multivariable logistic regression models with interaction terms to evaluate associations and possible effect modification by sex and age. We explored the direction and magnitude of these interactions by estimating marginal predicted probabilities. Results: The incidence of CAA was significantly higher in men than in women. In men with CAA, complete blood count analysis showed elevated neutrophil and monocyte counts and reduced lymphocyte counts. Male sex (odds ratio (OR) 2.197, 95% confidence interval (CI) 1.610–2.999), continuous age (1.017, 1.002–1.033), lymphocyte count (0.656, 0.526–0.820), monocyte count (1.551, 1.036–2.321), and platelet count (1.004, 1.001–1.006) were independently associated with CAA. Interaction analysis revealed significant interactions between neutrophils and both sex and age (p < 0.05), while lymphocyte counts showed significant interaction with age but not with sex. Conclusions: This study provides new insight into complex sex- and age-related immune cell patterns in CAA and may inform future diagnostic and management strategies by highlighting immune profile variability.

Pathophysiology doi: 10.3390/pathophysiology33010021

Authors: Kristof Babarczy Bence L. Radics Lili Kiss Alexandra Graczer Bence Nagy Sandor Dosa Gyongyi Kelemen Marton Balazsfi Pal Barzo Andras Voros Peter Klivenyi Levente Szalardy

Background/Objectives: Diffuse midline glioma (DMG), H3 K27M-altered, represents a rare group of gliomas arising in midline structures of the central nervous system. Historically regarded as a pediatric entity, it is now increasingly recognized in adults. Although its relative prevalence among all midline diffuse gliomas and its clinical-radiological characteristics are well defined in children, these tumors remain less characterized in adults, and comparative evaluations with H3 K27 wildtype midline diffuse gliomas are limited. Methods: Consecutive adult patients with histopathologically confirmed diffuse glioma (WHO grade ≥ 2) diagnosed between 2016 and 2025 were retrospectively screened for midline tumor location, with systematic revision of imaging and pathology. For identified midline diffuse gliomas, comprehensive clinical, imaging, and immunohistochemical data were collected, and a detailed morphometric analysis was performed. H3 K27 alteration status was established immunohistochemically, with supplementary immunostaining when necessary. Descriptive and comparative analyses were conducted. Results: A total of 5% of the 541 adult diffuse gliomas were midline, and 23% of IDH wildtype midline gliomas were consistent with DMG, H3 K27-altered (all H3 K27M-mutant). The affected patients were significantly younger, and these tumors predominantly involved the thalamus and mesencephalon. Morphometric analyses revealed trends toward fewer high-grade features in H3 K27-altered tumors, with composite scores demonstrating significant discriminatory ability. The overall survival was not significantly different between groups but showed associations with ring-like enhancement as well as adjuvant and salvage therapies in the overall midline cohort. Conclusions: This study provides population-based prevalence estimates for DMG, H3 K27M-altered, and complements the limited literature with comparative clinical-radiological and morphometric data of potential prognostic relevance.

Pathophysiology doi: 10.3390/pathophysiology33010020

Authors: Jose Redondo Kori B. Ascher Alexandre R. Abreu

Background: Obstructive sleep apnea (OSA) is a highly prevalent and heterogeneous disorder associated with substantial cardiometabolic morbidity. Although continuous positive airway pressure (CPAP) remains first-line therapy, long-term effectiveness is frequently limited by suboptimal adherence. Advances in airway devices, surgical techniques, neuromodulation, and pharmacologic therapies have expanded the therapeutic landscape and created opportunities for individualized, mechanism-based treatment. Methods: We conducted a selective, narrative review with structured quantitative synthesis of randomized controlled trials, comparative cohorts, long-term follow-up studies, registries, and mechanistic investigations addressing OSA therapies beyond CPAP. Evidence spanning oral appliances, upper-airway and skeletal surgery, hypoglossal nerve stimulation, neuromuscular electrical stimulation, positional therapy, and pharmacologic interventions targeting metabolic and non-anatomical endotypes was integrated. Outcomes of interest included apnea–hypopnea index (AHI), oxygenation, blood pressure, patient-reported symptoms, durability, safety, and real-world adherence. Results: Mandibular advancement devices (MADs) consistently reduced AHI relative to placebo and produced symptom relief comparable to CPAP in mild-to-moderate OSA, largely due to superior adherence. Palatal surgery yielded meaningful short-term improvement in selected patients but demonstrated limited long-term durability. In contrast, maxillomandibular advancement (MMA) achieved the largest and most durable reductions in OSA severity, with efficacy comparable to CPAP and superior to other surgical modalities in appropriate skeletal phenotypes. Hypoglossal nerve stimulation (HNS) produced substantial, durable improvements in AHI and symptoms with high adherence, supported by randomized trials, long-term follow-up, and real-world registry data; newer bilateral and proximal stimulation systems may further broaden candidacy. Neuromuscular electrical stimulation and positional therapy provided modest, phenotype-dependent benefits, primarily as adjunctive or early-stage interventions. A major advance is the emergence of metabolic and endotype-targeted pharmacotherapy: longitudinal data demonstrate a dose-dependent relationship between weight change and OSA progression or regression, while randomized trials show that GLP-1-based therapies—particularly dual GLP-1/GIP agonism with tirzepatide—produce large, clinically meaningful reductions in AHI and cardiometabolic risk in obesity-associated OSA. Additional pharmacologic strategies targeting ventilatory loop gain and arousal threshold further support an endotype-driven treatment paradigm. Conclusions: Contemporary OSA management is shifting from a CPAP-centric model toward a precision-guided, multimodal framework that aligns therapy with dominant anatomic and physiological contributors to airway collapse. Integrating metabolic, neuromodulatory, and structural interventions—often in combination—offers the potential for durable disease control and improved patient-centered outcomes. Future priorities include head-to-head and combination trials, long-term cardiovascular outcomes, cost-effectiveness analyses, and pragmatic tools to operationalize personalized OSA therapy in routine clinical practice.

Pathophysiology doi: 10.3390/pathophysiology33010019

Authors: Anastasia P. Sklifasovskaya Mikhail L. Blagonravov Madina M. Azova Sergey V. Kurevlev Vyacheslav A. Goryachev Sergey P. Syatkin Tatyana Yu. Zotova Daniil Yu. Prokofiev

Background: Arterial hypertension (AH) and insulin-dependent diabetes mellitus (DM) are major comorbid risk factors for accelerated myocardial damage, yet the behavior of key stress-adaptive heat shock proteins HSP70 and HSP90 under combined stress remains unclear. This study aimed to characterize the expression profiles of HSP70 and HSP90 in left ventricular cardiomyocytes during isolated and comorbid AH and DM, and to evaluate their association with structural remodeling and expansion of interstitial elements. Methods: The study was conducted in accordance with the European Convention for the Protection of Vertebrate Animals (ethical approval No. 26, RUDN Institute of Medicine, 18 February 2021) on 25 male rats divided into five groups (n = 5 each): control—38-week-old Wistar–Kyoto (WKY) rats; AH—38-week-old spontaneously hypertensive rats (SHR); long-term AH—57-week-old SHR; DM—38-week-old WKY rats with streptozotocin-induced insulin-dependent DM (65 mg/kg, i.p.); AH+DM—38-week-old SHR with STZ-induced DM. After 30 days of DM, left ventricular (LV) tissue was analyzed by immunohistochemistry (IHC) for HSP70/HSP90 protein expression and by RT-qPCR for mRNA levels. Increased stromal elements in myocardium were quantified morphometrically as interstitial stromal volume fraction (%) on hematoxylin and eosin-stained sections. Results: HSP90 was significantly upregulated in all pathological groups. The most pronounced increase occurred in isolated DM, with a 4.0-fold rise in HSP90-positive area (21.80% vs. 5.45% in control) and a 1.82-fold increase in mRNA. In the AH+DM group, HSP90 mRNA expression was extremely elevated (25.93-fold), accompanied by a 3.7-fold increase in protein. In contrast, HSP70 protein was elevated only in the 38-week AH group (27.68% vs. 19.70% control, p ≤ 0.05), remained unchanged in isolated DM (19.50%), and was significantly reduced in AH+DM (14.71%, p ≤ 0.05), despite a modest 1.64-fold mRNA upregulation in DM. Morphometric analysis revealed progressive expansion of interstitial elements, most severe in AH+DM (9.43% stromal volume vs. 4.81% in control, p ≤ 0.05). Conclusions: Comorbid AH and DM provoke synergistic HSP90 upregulation, while HSP70 expression is markedly suppressed, indicating a shift from an adaptive to a maladaptive cellular-stress response. The imbalance between HSP90 and HSP70 may represent a key molecular mechanism underlying accelerated structural and functional deterioration of the myocardium in cardiometabolic comorbidity.

Pathophysiology doi: 10.3390/pathophysiology33010018

Authors: Bozidar Pindovic Vladimir Zivkovic Radisa Pavlovic Djurdjina Petrovic Maja Muric Ivan Srejovic Dmitry Kolesov Marina Kolotilova Sergey Bolevich Zarko Finderle Vladimir Jakovljevic Aleksandra Stojanovic

Myocarditis is still a major global health issue that frequently manifests due to oxidative stress, immune-mediated myocardial damage, and unpredictable clinical progression. Experiments with autoimmune myocarditis (EAM) models have shown different ways that T-cell subsets, proinflammatory cytokines, macrophage polarization, and mitochondrial dysfunction are all connected and play a part in both acute inflammation and chronic remodeling of the heart. As a possible multimodal intervention that could affect several of these disease-causing pathways, hyperbaric oxygen therapy (HBOT) has become popular. This therapy delivers 100% oxygen to different tissues at higher atmospheric pressures. Early research shows that HBOT improves the delivery of oxygen to the inflamed myocardium, suppress the activation of NF-κB and NLRP3 inflammasomes, lowers oxidative stress, protects mitochondrial function, and boosts immune-regulatory T-cell responses. Despite these potentially promising findings, there are still a number of important translational obstacles to overcome, such as inconsistent protocols, a lack of long-term outcome data, insufficient mechanistic profiling, and doubts about the best protocol length and patient selection. To assess safety and effectiveness in human myocarditis, future studies should aim to integrate multi-omics analyses, HBOT regimens that are already standardized, sophisticated imaging, and carefully planned early-phase clinical trials. Overall, the currently available evidence supports HBOT as a biologically plausible and potentially valuable adjunct therapy for autoimmune myocarditis, expressing the need for further mechanistic and clinical investigation.

Pathophysiology doi: 10.3390/pathophysiology33010017

Authors: Jonathan Steven Alexander

As the Year of the Snake draws to a close and we ‘gallop’ into the Year of the Horse, we find a fitting metaphor for the work of science: continued effort, coordination, and progress that depends as much on balance in the saddle as on speed [...]

Pathophysiology doi: 10.3390/pathophysiology33010016

Authors: Talib Hussain Monika Verma Sagarika Biswas

Background: Rheumatoid arthritis (RA) is a systemic, pro-inflammatory, autoimmune disease that mainly affects the joints in a symmetrical manner. Differential proteomic profiling through Sequential Window Acquisition of all Theoretical Fragment Ion Mass Spectra (SWATH-MS/MS) helps in a better understanding of the RA pathogenesis. In this study, we compared the differentially upregulated proteins with those associated with fibrosis to gain a deeper understanding of the fibrotic aspect of RA. Methods: We analyzed plasma proteomics data, previously obtained by SWATH-MS/MS. Our focus was on proteins associated with Leucine Rich Alpha2glycoprotein1 (LRG1) and we employed an in silico method. Results: We identified common proteins between RA and fibrosis. Among them, LRG1 and Serine Protease Inhibitor Clade A, Member 1 (SERPINA1) showed a high co-expression score in the gene clusters. LRG1 is both pro-inflammatory and pro-fibrotic, while SERPINA1 is an anti-inflammatory protein that inhibits pro-inflammatory and pro-fibrotic molecules (Elastase). Further, docking studies and a simulation study of the docked complexes with the analysis of Hydrogen bonds, Solvent Accessible Surface Area (SASA), Root Mean Square Deviation (RMSD), Root Mean Square Fluctuation (RMSF) and Radius of gyration (Rg), suggested a strong interaction between the two partners, LRG1 and SERPINA1. Conclusions: Our study suggests that LRG1 may inhibit SERPINA1 and promote inflammation and fibrotic processes by disrupting SERPINA1’s primary function.

Pathophysiology doi: 10.3390/pathophysiology33010015

Authors: Julieta M Ramírez-Mejía Geysson Javier Fernandez Silvio Urcuqui-Inchima

Background: Zika virus (ZIKV), a mosquito-borne flavivirus, is associated with congenital malformations and neuroinflammatory disorders, highlighting the need to identify host factors that shape infection outcomes. Macrophages, key targets and reservoirs of ZIKV, orchestrate both antiviral and inflammatory responses. Methods: Vitamin D (VitD) has emerged as a potent immunomodulator that enhances macrophage antimicrobial activity and regulates inflammation. To investigate how VitD shapes macrophage responses to ZIKV, we reanalyzed publicly available RNA-seq and miRNA-seq datasets from monocyte-derived macrophages (MDMs) of four donors, differentiated with or without VitD and subsequently infected with ZIKV. Results: Differential expression analysis identified long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and mRNAs integrated into competing endogenous RNA (ceRNA) networks. In VitD-conditioned and ZIKV-infected MDMs, 65 lncRNAs and 23 miRNAs were significantly modulated. Notably, lncRNAs such as HSD11B1-AS1, Lnc-FOSL2, SPIRE-AS1, and PCAT7 were predicted to regulate immune and metabolic genes, including G0S2, FOSL2, PRELID3A, and FBP1. Among the miRNAs, let-7a and miR-494 were downregulated, while miR-146a, miR-708, and miR-378 were upregulated, all of which have been previously implicated in antiviral immunity. Functional enrichment analysis revealed pathways linked to metabolism, stress responses, and cell migration. ceRNA network analysis suggested that SOX2-OT and SLC9A3-AS1 may act as molecular sponges, modulating regulatory axes relevant to immune control and viral response. Conclusions: Despite limitations in sample size and experimental validation, this study provides an exploratory map of ncRNA–mRNA networks shaped by VitD during ZIKV infection, highlighting candidate molecules and pathways for further studies on host–virus interactions and VitD-mediated immune regulation.

Pathophysiology doi: 10.3390/pathophysiology33010014

Authors: Yadira Gasca-Martínez Miguel Angel Ontiveros-Torres Isaías López-Gallegos José Jaime Jarero-Basulto

Alzheimer’s disease (AD) is characterized by progressive cognitive decline, with amyloid beta oligomers (AβOs) emerging as the most neurotoxic species and acting as early triggers of cellular alterations. Before the appearance of other protein aggregates, AβOs disrupt the dynamics and stability of the neuronal cytoskeleton, a structure essential for maintaining neuronal morphology, axonal transport, and synaptic plasticity. Experimental evidence demonstrates that AβOs promote microtubule disassembly, Tau hyperphosphorylation, reduced kinesin levels, impaired axonal transport, and alterations in actin dynamics through the LIMK–cofilin signaling pathway. In addition, increased levels of neurofilament light chain have been identified as an early biomarker of axonal damage. Notably, these cytoskeletal disturbances arise in the absence of extensive neuronal death, underscoring the cytoskeleton as a critical early target in AD pathogenesis. In this review, we analyze cytoskeletal alterations induced by AβOs in neurons and discuss how these changes may contribute to disrupted neuronal communication, a defining early hallmark of AD pathology.

Pathophysiology doi: 10.3390/pathophysiology33010013

Authors: Lorena Cayetano-Salazar Jhactcidi Jackeline García-López Dania A. Nava-Tapia Eymard Hernández-López Caroline Weinstein-Oppenheimer Julio Ortiz-Ortiz Marco Antonio Leyva-Vázquez Miguel Ángel Mendoza-Catalán Adán Arizmendi-Izazaga Napoleón Navarro-Tito

Background/Objectives: Although tissue inhibitors of metalloproteinases (TIMPs) are key regulators in breast cancer, their differential expression, clinical relevance, and molecular roles remain unclear. This study aimed to compare the expression patterns of the four TIMPs in breast cancer and evaluate their molecular interactions and associated pathways through an integrated bioinformatic analysis. Methods: The expression of TIMPs and their correlations with MMPs were analyzed using the TCGA PanCancer, cBioPortal, and GEO datasets. Associations between TIMP expression and overall survival were assessed in the TCGA Breast Invasive Carcinoma PanCancer cohort. Pathway enrichment analysis was performed using GO, KEGG, and DAVID. The relationships between immune cell infiltration, stromal cells, and TIMP expression were assessed using the EPIC algorithm. Statistical analyses were performed using R. Results:TIMP1 was the only inhibitor overexpressed in breast tumors and showed significant associations with the Luminal B, HER2, TNBC, and normal-like subtypes, along with a modest increase across stages. TIMP2, TIMP3, and TIMP4 were downregulated in tumors. High expression of TIMP1 and TIMP4 correlated with better overall survival. TIMP1-associated genes were enriched in NF-kappa and PI3K–Akt signaling and actin cytoskeleton components. TIMP2 was linked to Hedgehog and MAPK pathways and actin-related elements. TIMP3 correlated with Hedgehog and PI3K–Akt signaling, DNA damage response, and membrane components. TIMP4 was associated with VEGF, MAPK, PI3K–Akt, DNA damage pathways, and actin organization. TIMP2 showed strong positive correlations with MMP2 and MMP14, while TIMP4 showed negative correlations with MMP1 and MMP9. Interestingly, we found a strong positive correlation between TIMP2 and TIMP3 with ADAM12, as well as between TIMP2 and TIMP3 with ADAM10, and negative correlations with ADAM15. The differential expression of TIMPs favors greater infiltration of immune cells related to tumor progression and poor prognosis in breast cancer patients. Conclusions: TIMPs display contrasting expression profiles and distinct pathway associations in breast cancer. TIMP1 emerges as the only consistently overexpressed inhibitor, while TIMP4 appears as a promising prognostic marker with unique MMP correlations that may influence tumor behaviors.

Pathophysiology doi: 10.3390/pathophysiology33010012

Authors: Taishi Imoto Junpei Ishizaka Yukinori Tamura

Background: AMP-activated protein kinase (AMPK) acts as a key energy sensor that negatively regulates skeletal muscle mass. Zinc is an essential trace element that is required for myogenic differentiation and protein synthesis, while zinc deficiency has been associated with muscle atrophy in vivo. However, how zinc status modulates AMPK activation itself or alters downstream responses to AMPK signaling in muscle cells remains unclear. Methods: C2C12 myotubes were cultured under zinc-depleted (ZnD), zinc-sufficient (20 μM; Zn20), or zinc-supplemented (40 μM; Zn40) conditions. AMPK was activated by AICAR, and zinc status–dependent responses were evaluated using molecular and morphological analyses. Results: AICAR increased intracellular zinc levels in Zn20 and Zn40 but not in ZnD. Zinc transporter expression exhibited gene-specific regulation: Zip3 was upregulated across all zinc conditions, Zip14 was significantly induced in ZnD and Zn40, and Zip10 was selectively upregulated in Zn40. AICAR induced myotube atrophy in all groups; however, the reduction in myotube diameter was significantly greater under zinc-depleted conditions. Zinc depletion was associated with transcriptional upregulation of FoxO1, FoxO3, Atrogin-1, and MuRF1 in response to AICAR, while AMPK activation and suppression of S6K1 phosphorylation occurred to a similar extent regardless of zinc status. Conclusions: These findings indicate that zinc availability does not alter AMPK activation itself but modulates downstream atrophic responses to AMPK signaling. Under conditions of AMPK activation, adequate zinc availability is accompanied by increased intracellular zinc levels and stress-responsive ZIP regulation, which may limit excessive atrophic gene induction, whereas zinc depletion increases susceptibility to AMPK-induced atrophic responses in skeletal muscle cells.

Pathophysiology doi: 10.3390/pathophysiology33010011

Authors: Fiorela Gorea Martina Pelle Federico Fiori Nastro Carmine Gelormini Fatime Elezi Michele Ribolsi Giorgio Di Lorenzo

Background/Objectives: Cannabis use has a particularly high prevalence in individuals with psychotic disorders. Although cannabis use is generally associated with cognitive impairments in the general population, its impact on cognition in psychosis remains controversial. This study aimed to investigate the association between cannabis use and cognitive performance in a cohort of individuals affected by psychotic disorders. Methods: A total of 105 inpatients with psychotic disorders (mean age: 40.3 years; 34 females) were recruited from the University Hospital Center “Mother Teresa” in Tirana. Data collection included socio-demographic and clinical variables. Cognitive functioning was evaluated using the Montreal Cognitive Assessment (MoCA), while psychopathology was assessed with the Brief Negative Symptom Scale (BNSS), the Calgary Depression Scale for Schizophrenia (CDSS), the Psychotic Symptom Rating Scales (PSYRATS), and the Scale for the Assessment of Thought, Language, and Communication (TLC). Results: Cannabis users (CU) were more frequently male, younger, and exhibited an earlier onset of psychosis compared to non-users (No-CU). Importantly, CU demonstrated higher MoCA scores, with the most favorable outcomes observed among daily users. Conclusions: Contrary to the prevailing assumption that cannabis use exacerbates cognitive decline, our findings indicate an unexpected association between cannabis use and preserved cognitive functioning in psychosis. These results underscore the need to consider dosage, frequency, and cannabinoid composition (THC/CBD ratio) when interpreting cannabis-related cognitive outcomes in psychotic disorders.

Pathophysiology doi: 10.3390/pathophysiology33010010

Authors: Zheyi Wang Keiji Naruse Ken Takahashi

For more than a century, pathology has served as a cornerstone of modern medicine, relying primarily on static microscopic assessment of tissue morphology—such as H&E staining—which remains the “gold standard” for disease diagnosis. However, this conventional paradigm provides only a snapshot of disease states and often fails to capture their dynamic evolution and complex functional mechanisms. Moreover, animal models are constrained by marked interspecies differences, creating a persistent gap in translational research. To overcome these limitations, we propose the concept of New Pathophysiology, a research framework that transcends purely morphological descriptions and aims to resolve functional dynamics in real time. This approach integrates Organ-on-a-Chip (OOC) technology, multi-omics analyses, and artificial intelligence to reconstruct the entire course of disease initiation and to enable personalized medicine. In this review, we first outline the foundations and limitations of traditional pathology and animal models. We then systematically summarize more than one hundred existing OOC disease models across multiple organs—including the kidney, liver, and brain. Finally, we elaborate on how OOC technologies are reshaping the study of key pathological processes such as inflammation, metabolic dysregulation, and fibrosis by converting them into dynamic, mechanistic disease models, and we propose future perspectives in the field. This review adopts a relatively uncommon classification strategy based on pathological mechanisms (mechanism-based), rather than organ-based categorization, allowing readers to recognize shared principles underlying different diseases. Moreover, the focus of this work is not on emphasizing iteration or replacement of existing approaches, but on preserving past achievements from a historical perspective, with an emphasis on overcoming current limitations and enabling new advances.

Pathophysiology doi: 10.3390/pathophysiology33010009

Authors: Guan-Tian Lang Xiao-Ling Weng Yun Liu Xin Hu Zhi-Ming Shao Zhen Hu

Objectives: The molecular characterization of male breast cancer (MaBC) has long been understudied, primarily due to its rare occurrence. Clinical management of MaBC remains profoundly challenging, with current therapeutic strategies largely extrapolated from female breast cancer protocols. Methods: Through panel-based sequencing targeting BRCA1, BRCA2, and PALB2 variants, we delineated the genomic landscape of 96 MaBC cases. Subsequent whole-exome sequencing (WES) of 84 BRCA1/2- and PALB2-mutation-negative MaBC patients, compared against 4480 healthy controls, revealed compelling findings. Results: Pathogenic variants in BRCA1/2 and PALB2 were identified in 14.6% (14/96) of MaBC cases, with BRCA2 mutations predominating at 12.5% (n = 12). Notably, one patient harbored the BRCA1 c.4015G > T stop-gained mutation, while another exhibited the PALB2 c.481_482dupGA alteration. Our analysis further uncovered 170 pathogenic/likely pathogenic mutations, with RAD50, DMD, ARSA, and ABCC6 demonstrating recurrent mutations in MaBC. Conclusions: As the inaugural germline genomic investigation of MaBC in a Han Chinese population, this work reveals clinically actionable alterations with diagnostic and therapeutic implications. These discoveries not only advance our understanding of MaBC’s molecular architecture but also underscore the critical need for dedicated research into this malignancy.

Pathophysiology doi: 10.3390/pathophysiology33010008

Authors: María de Guadalupe Chávez-López Arturo Avalos-Fuentes Estrella del C. Cruz-Manzo Pedro A. Aguirre-Arriaga Benjamín Florán Julio Isael Pérez-Carreón Cecilia Bañuelos Javier Camacho

Hepatocellular carcinoma (HCC) is the main type of liver cancer and one of the malignancies with the highest mortality rates worldwide. HCC is associated with diverse etiological factors including alcohol use, viral infections, fatty liver disease, and liver cirrhosis (a major risk factor for HCC). Unfortunately, many patients are diagnosed at advanced stages of the disease and receive palliative treatment only. Therefore, early markers of HCC and novel therapeutic approaches are urgently needed. The endocannabinoid system is involved in various physiological processes such as motor coordination, emotional control, learning and memory, neuronal development, antinociception, and immunological processes. Interestingly, endocannabinoids modulate signaling pathways involved in cell survival, proliferation, apoptosis, autophagy, and immune response. Consistently, several cannabinoids have demonstrated potential antitumor properties in experimental models. The participation of metabotropic and ionotropic cannabinoid receptors in the biological effects of cannabinoids has been extensively described. In addition, cannabinoids interact with other targets, including several ion channels. Notably, several ion channels targeted by cannabinoids are involved in inflammation, proliferation, and apoptosis in liver diseases, including HCC. In this literature review, we describe and discuss both the endocannabinoid system and exogenous phytocannabinoids, such as cannabidiol and Δ9-tetrahydrocannabinol, along with their canonical receptors, as well as the cannabidiol-targeted ion channels and their role in liver cancer and its preceding liver diseases. The cannabidiol-ion channel association is an extraordinary opportunity in liver cancer prevention and therapy, with potential implications for several environments that are for the benefit of cancer patients, including sociocultural, public health, and economic systems.

Pathophysiology doi: 10.3390/pathophysiology33010007

Authors: Alicia A. Brunet Annie L. Miller Xin Ru Lim Alan R. Harvey Livia S. Carvalho

Background/Objectives: The RhoP23H/WT mouse line is a commonly used model to study rhodopsin P23H-associated autosomal dominant retinitis pigmentosa. Previous studies in RhoP23H/WT mice have largely focused on retinal changes occurring at one month of age and later, and have indicated a compensatory thickening of inner retinal layers in response to rod degeneration. However, the effect of disease processes during early postnatal retinal development remains understudied. Methods: In this study, we investigated the retinal response to rod dysfunction during early postnatal developmental ages P8–P24 in our novel RhoP23H/WT reporter line, RhoP23H.GFP, which expresses green fluorescent protein (GFP) exclusively in cone photoreceptors. Results: Histological analysis revealed no significant difference in retinal thickness in RhoP23H.GFP mice compared to healthy controls at the ages investigated. RhoP23H.GFP retinas initially exhibited a greater mislocalization of rhodopsin to the rod cell bodies at P12, though this mislocalization normalized to wildtype by P24. Most notably, flow cytometry revealed significantly increased cone photoreceptor numbers in P12 (61%), P16 (48%), and P24 (40%) RhoP23H.GFP mice compared to wildtype controls, indicating a possible compensatory response of cone photoreceptors to rod dysfunction. Additionally, cone morphology appeared altered in diseased cones. Conclusions: Our results suggest that cones may undergo a developmental compensatory adaptation in response to rod dysfunction, providing new insights into early disease mechanisms of retinitis pigmentosa.

Pathophysiology doi: 10.3390/pathophysiology33010006

Authors: Matthias Welsner Sarah Dietz-Terjung Svenja Strassburg Dirk Westhölter Sivagurunathan Sutharsan Christoph Schöbel Christian Taube Florian Stehling Cornelius Kürten Cornelius Deuschl Michael Forsting Sebastian Zensen Johannes Haubold Benedikt M. Schaarschmidt Marcel Opitz

Objective: To assess whether chronic rhinosinusitis (CRS) severity is associated with obstructive sleep apnea (OSA) in adult people with cystic fibrosis (pwCF). Methods: We conducted a retrospective single-center study of 44 adults with CF who underwent overnight polysomnography (PSG), Epworth Sleepiness Scale (ESS) assessment, and sinus computed tomography (CT). CRS severity was quantified using the Lund–Mackay score (LMS) and the main nasal cavity score (MNCS). OSA was defined by Apnea–Hypopnea Index (AHI) thresholds per American Academy of Sleep Medicine criteria. Results: Participants had a mean age of 31.1 ± 8.4 years and a mean percent predicted FEV1 of 51.8 ± 15.7. Sinus CT showed radiological evidence of CRS in all participants. Mean AHI was 5.3 ± 4.4/h; 48% had AHI ≥ 5/h. There were no significant differences between pwCF with and without OSA in age, sex, BMI, lung function, total sleep time, sleep efficiency, or ESS score (all p > 0.05). Mean LMS and MNCS did not differ between OSA and non-OSA groups (both p > 0.05), and neither score correlated with PSG parameters or ESS (all p > 0.05). Receiver operating characteristic (ROC) analysis demonstrated low discriminative ability of LMS and MNCS for predicting OSA (AUCs < 0.70, p < 0.05). Conclusions: In this cohort of adults with CF, CT-based CRS severity was not associated with OSA. Given the substantial prevalence of OSA observed, PSG screening should be considered irrespective of CRS severity.

Pathophysiology doi: 10.3390/pathophysiology33010005

Authors: Fani-Niki Varra Olga Pagonopoulou Michail Varras Viktoria-Konstantina Varra Panagiotis Theodosis-Nobelos

Introduction: Multiple sclerosis (MS) is a chronic autoimmune inflammatory disorder of the central nervous system (CNS) that leads to demyelination of CNS neurons and is influenced by genetic, environmental, and lifestyle factors, including diet and obesity. Methods: This review aims to analyze at the molecular level the relationship between obesity, as a chronic inflammatory condition, and the pathophysiology of MS, as a chronic autoimmune inflammatory disease, in order to understand the complex links between obesity and MS through a search of the PubMed and Google Scholar databases. Discussion: Chronic inflammation and OS are interconnected processes, causing a toxic state, which contributes to the development of CNS neuroinflammation and neuronal damage, resulting in neuronal demyelination and the onset of MS. Adipose tissue is a complex endocrine organ; in addition to being a lipid storage organ, it secretes cytokines and adipokines, which are involved in the regulation of hormones, metabolism, inflammation, and whole-body homeostasis. Obesity triggers chronic low-grade inflammation, disruption of the blood–brain barrier (BBB) and brain metabolism, infiltration of the CNS by immune cells, production of ROS, and generation of oxidative stress (OS). Anti-inflammatory and pro-inflammatory adipokines are also implicated in MS and obesity. Conclusions: Obesity affects MS through common underlying mechanisms and seems to be a modifiable risk factor. Antioxidant and anti-inflammatory compounds with multi-functional characteristics could be additional tools to slow the progression of MS and its promotion through obesity while also offering potential treatment options for both conditions via their multi-targeting characteristics.

Pathophysiology doi: 10.3390/pathophysiology33010004

Authors: Ewa Jankowska-Steifer Anna Ratajska Aleksandra Flaht-Zabost Dorota Magdalena Radomska-Leśniewska Iwona Badurek Ewelina Kiernozek Aneta Moskalik Barbara Majchrzak Mateusz Bartkowiak Krzysztof Bartkowiak Bogdan Ciszek Marek Kujawa Justyna Niderla-Bielinska

Background/Objectives: Metabolic syndrome (MetS) conditions lead to structural and functional alterations in cardiomyocytes, microvasculature, and extracellular matrix (ECM), leading to myocardial fibrosis and impaired diastolic function. Cardiac lymphatic vessels (LVs) are increasingly recognized as key regulators of myocardial homeostasis, yet their response to MetS remains poorly understood. Therefore, we aimed to investigate transcriptional changes in cardiac lymphatic endothelial cells (LECs) in db/db mice, a well-established model of MetS. Methods: Using flow cytometry-sorted LECs and RT-PCR, we analyzed mRNA expression of genes involved in lymphangiogenesis, metabolism, mechanotransduction, immune cell trafficking, and ECM interactions. Results: Our findings show the transcriptional plasticity of cardiac LECs in response to MetS. Conclusions: Although our study is limited by the lack of protein-level validation and functional assays, our approach provides a broader interpretative framework and identifies potential directions for future research, including functional studies and pathway-specific investigations of the identified genes to assess their impact on lymphatic flow and cardiac function. Understanding LEC responses to metabolic stress may uncover novel therapeutic targets for heart failure associated with MetS.

Pathophysiology doi: 10.3390/pathophysiology33010003

Authors: Natalya Semenova Nadezhda Garashchenko Olga Nikitina Sergey Kolesnikov Natalia Belkova Elizaveta Klimenko Nadezhda Smurova Elizaveta Novikova Irina Madaeva Liubov Kolesnikova

Background: Menopause, a critical period during a woman’s life, is characterized by various changes, including disturbances in their oxidative balance and circadian rhythm. Currently, the gut microbiome is suggested as an important participant in these processes. Methods: This study involved 96 menopausal women. Their sleep quality was assessed using three questionnaires: the Insomnia Severity Index (ISI), the Pittsburgh Sleep Quality Index (PSQI), and the Epworth Sleepiness Scale (ESS). The GSH and GSTP1 contents in the serum were measured by means of immunoassay methods, while the composition of the gut microbiome was determined via molecular genetic methods. Results: E. coli, K. oxytoca, S. aureus, Enterobacter spp., Shigella spp., Streptococcus spp., Prevotella spp., and M. stadmanae were found to correlate with the GSH content in different sleep groups, while the presence of K. oxytoca, S. aureus, Enterococcus spp., K. pneumoniae, and M. stadmanae is also important for the GSH level in several of these groups. F. prausnitzii, S. aureus, P. micra, Acinetobacter spp., and E. rectale are associated with GSTP1 concentration in various sleep groups, while the presence of F. nucleatum and P. micra is also relevant for the GSTP1 content in some of these groups. Conclusions: Thus, in menopausal women, the composition and structure of the gut microbiota are associated with sleep disorders. GSH and GSTP1 are associated with some gut microbiome markers in menopausal women, but these relationships differ in different sleep disorders.

Pathophysiology doi: 10.3390/pathophysiology33010002

Authors: William Rosales Srija Chowdary Vanka Harjinder Singh Paul Bhamrah Malti Bhamrah Naomi Ghildiyal Cesar Liendo Sheila Asghar J. Steven Alexander Oleg Y. Chernyshev

Background: Obstructive sleep apnea (OSA) is a heterogeneous disorder traditionally classified and stratified by the apnea–hypopnea index (AHI), which fails to capture variability in symptom burden, comorbid associations, and treatment responses. Clinical phenotyping has emerged as a promising strategy to improve disease characterization and management over the last decade. Methods: We conducted a narrative literature review of studies published between January 2014 and December 2022 that used cluster analysis to define OSA phenotypes in adults with moderate-to-severe disease (AHI ≥ 15 events/h). Eligible studies employed validated questionnaires, symptom reporting, and comorbidity profiling to identify subgroups. Findings were summarized across diverse populations, with emphasis on phenotype reproducibility, comorbidity associations, and treatment implications. Results: Across international cohorts, three reproducible symptom-based phenotypes were consistently identified: excessively sleepy (ES), disturbed sleep (DS), and minimally symptomatic (MS). Additional subtypes, such as upper airway dominant (UA) and moderately sleepy (MoS), were described in larger cohorts. Phenotypes differed in demographic profiles, comorbidity burden, and treatment adherence. ES patients exhibited the greatest symptom burden, higher cardiovascular risk, and better adherence to positive airway pressure (PAP) therapy, with significant symptomatic improvement. DS patients frequently reported insomnia symptoms, showed modest PAP-related gains, and may benefit from adjunctive insomnia-targeted interventions. MS patients, despite low symptom burden, often carried substantial comorbidity risk, specifically buildup of OSA-related cardiovascular risk. Conclusions: Symptom-based OSA phenotypes are reproducible across diverse populations and provide clinically meaningful insights beyond AHI. They allow for improved risk stratification, highlight gaps in detection of minimally symptomatic patients, and inform personalized treatment strategies. Integrating phenotyping into clinical practice has the potential to enhance diagnostic accuracy, optimize therapeutic outcomes, and refine cardiovascular risk prediction in OSA.

Pathophysiology doi: 10.3390/pathophysiology33010001

Authors: Michelle Tucci Robert C. O′Brien Joseph D. Lichtenhan Hamed Benghuzzi Drew Hildebrandt

Background/Objectives: No effective intervention currently exists for non-compressible pulmonary injury, especially in a prehospital setting. Visco-liquids like trisilanol i-octyl POSS could remedy this. POSS resists hemorrhage and activates clotting; this can be augmented with kaolin (22.5%; PK) or chitin (10%; PC). Methods: We tested the efficacy of POSS, PK, and PC in treating incisional lung wounds in swine (39 ± 1 kg; n = 10). An incisional wound was made in the lung via a left thoracotomy, allowed to bleed freely for 30 s, and then no treatment (UNT), gauze with compression (GC), or POSS, PK, or PC was applied (1.5 mL). Each treatment was applied once per animal for a total of 5 wounds. Wounds were observed for 10 min for hemostasis and pneumostasis; GC treatments were assessed at 3 min intervals. Results: POSS and PC produced hemostasis in 8 of 10 wounds; GC: 7 (all significant from UNT); PK: 5 and UNT: 1. PK was not different from any group. POSS (2 ± 0.3 min) and PC (1.4 ± 0.4 min) clotted more quickly than GC (8 ± 3 min); PK was intermediate (3.8 ± 2 min) and not different from any other group. Pneumostasis was achieved in all POSS, PC, and PK, and only after hemostasis in the GC group. Conclusions: Because both POSS and PC provided quick and lasting hemorrhage and pneumatic control in this model, without need for compression, these results support the concept that these types of liquid POSS compounds could prove to be efficacious in prehospital treatment of non-compressible trauma wounds.

Pathophysiology doi: 10.3390/pathophysiology32040070

Authors: Gianpiero Greco Alessandro Petrelli Francesco Fischetti Stefania Cataldi

Cancer remains a leading global cause of morbidity and mortality. Modifiable lifestyle factors, including avoidance of tobacco use and excessive ultraviolet radiation, healthy dietary patterns, regular physical activity, and weight management, play key roles in prevention and care. This narrative review synthesizes evidence on lifestyle-based interventions influencing cancer risk, treatment tolerance, and survivorship. A literature search was conducted in PubMed and Scopus, supplemented by manual screening via Google Scholar. The time frame (2001–2025) was selected to reflect evidence produced within the modern era of molecular oncology and contemporary lifestyle medicine research. Eligible publications addressed carcinogen exposure (tobacco, alcohol, ultraviolet radiation), diet and nutritional strategies, physical activity, sedentary behavior, obesity, metabolic health, complementary therapies, and cancer outcomes. Evidence indicates that reducing exposure to tobacco and ultraviolet radiation remains central to cancer prevention. Adherence to predominantly plant-based diets, regular physical activity, and maintenance of healthy body weight are consistently associated with lower incidence of several cancers, including breast, colorectal, and liver cancer. Nutritional strategies such as caloric restriction, ketogenic diets, and fasting-mimicking diets show promise in improving treatment efficacy and quality of life. Complementary and mind–body therapies may alleviate treatment-related symptoms, although high-quality evidence on long-term safety and effectiveness is limited. Integrating lifestyle medicine into oncology offers a cost-effective, sustainable strategy to reduce cancer burden and enhance survivorship. Comprehensive programs combining carcinogen avoidance, dietary regulation, structured exercise, and effective radiation risk mitigation may extend healthspan, improve treatment tolerance, and help prevent recurrence.

Pathophysiology doi: 10.3390/pathophysiology32040069

Authors: Tara Al-Barazenji Asma Allouch Nedhal Al Husaini Sondos Yousef Wisam Nabeel Ibrahim Amal Al-Haidose Hatem Zayed Atiyeh M. Abdallah

In the original publication [...]

Pathophysiology doi: 10.3390/pathophysiology32040068

Authors: Tatyanne L. N. Gomes Thaís C. Borges Jessica F. M. Ivo Lara G. Mainardi Renata G. C. Abadio Benjamin T. Wall Gustavo D. Pimentel

Background and aims: This study aimed to determine whether neuromuscular electrical stimulation (NMES) combined with nutritional counseling promotes an increase in thigh muscle thickness (MT), as well as to assess changes in the relationship between MT and intracellular water (ICW). Body composition methods such as ultrasound may overestimate muscle mass, depending on the context, because they cannot distinguish the contractile protein component from body fluids, including intra- and extracellular water. Methods: A pilot randomized parallel trial was conducted with 25 hospitalized patients with unselected cancer, who were divided into two groups: NMES + Diet and Diet. Both groups received nutritional counseling, but only one group received NMES. NMES was applied bilaterally to the origin and insertion points of the quadriceps twice daily, with a 3 h interval between sessions, for 7 consecutive days. MT and ICW were measured before and after the intervention. Food consumption was assessed using a 24 h dietary recall at baseline and at the end of the study to quantify and adjust macronutrient intake during the intervention. Results: Both treatment groups (Diet × NMES + Diet) showed similar dropout rates which means participants in the more intensive treatment did not quit more frequently, once intervention with NMES was feasible and well tolerated. In addition, both groups showed a reduction in carbohydrate intake (p = 0.012) and an increase in leucine intake (p < 0.001) post-intervention. The increase in leucine intake was significantly greater in the NMES + Diet group (p < 0.001), and the reduction in carbohydrate intake was also greater in this group (p = 0.012). In the delta analysis, the NMES + Diet group showed an increase in thigh MT, whereas the Diet group experienced a decrease (Diet group: ∆ = −2.53 ± 3.73 mm vs. NMES + Diet group: ∆ = 2.09 ± 2.27 mm, p = 0.001). Moreover, the MT/ICW ratio was higher in the NMES + Diet group post-intervention (Diet group: ∆ = −0.15 ± 0.19 mm/L vs. NMES + Diet group: ∆ = 0.11 ± 0.09 mm/L, p < 0.001), while no significant difference in ICW was observed between groups. Conclusions: short-term intervention combining nutritional counseling with NMES increased thigh MT and the MT/ICW ratio, possibly due to NMES-induced extracellular water expansion.

Pathophysiology doi: 10.3390/pathophysiology32040067

Authors: Roman Ryabushko Heorhii Kostenko Oleh Akimov Vitalii Kostenko

Objectives: This study investigates the effects of resveratrol on systemic inflammatory, oxidative, and metabolic responses in a rat model that combines surgical trauma with prior exposure to Single Prolonged Stress (SPS), an established experimental protocol for modeling post-traumatic stress disorder (PTSD). Methods: Male Wistar rats (n = 21) were randomly assigned to three groups: (I) control (polyvinylpyrrolidone, PVP), (II) SPS + laparotomy + PVP), and (III) SPS + laparotomy + resveratrol. Resveratrol (5 mg/kg of body weight/day) or vehicle was administered intragastrically for seven days. Serum concentrations of cortisol, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), glucose, insulin, lipid fractions, and thiobarbituric acid–reactive substances (TBA-RS) were determined by enzyme-linked immunosorbent assay and spectrophotometric methods. Insulin resistance was assessed using the homeostatic model assessment of insulin resistance (HOMA-IR) index. Results: Combined SPS and surgical trauma induced a pronounced systemic inflammatory response characterized by elevated cortisol (+138%), TNF-α (+83%), IL-6 (+465%), and ceruloplasmin (+71%), as well as hyperglycemia, hyperinsulinemia, increased HOMA-IR, and atherogenic dyslipidemia with reduced high-density lipoprotein cholesterol (HDL-CH; −64%), elevated triglycerides (TGs; +216%), and very low-density lipoprotein cholesterol (VLDL-CH; +218%). Marked activation of lipid peroxidation was observed, as indicated by increased TBA-RS levels before and after incubation. Resveratrol administration significantly decreased cortisol (−45%), TNF-α (−47%), and IL-6 (−85%), normalized the IL-10/IL-6 ratio, and reduced ceruloplasmin levels (−13%). The compound improved insulin sensitivity (HOMA-IR −50%), elevated HDL-CH (+115%), and lowered TGs and VLDL-CH (−44%). It also attenuated both basal and inducible lipid peroxidation (TBA-RS −11% and −13%), indicating restoration of antioxidant capacity. Conclusions: Thus, resveratrol effectively counteracts the neuroendocrine, inflammatory, and metabolic disturbances induced by combined PTSD-like stress and surgical trauma.

Pathophysiology doi: 10.3390/pathophysiology32040066

Authors: Luisa F. Delgadillo Nabil A. Rashdan Hunter Hamilton Jack H. Pattillo Shuai Yuan Randa S. Eshaq Norman R. Harris Jonathan S. Alexander Christopher B. Pattillo

Background: Vascular calcification is a strong predictor of cardiovascular morbidity and mortality. Oxidative stress plays a key role in promoting vascular calcification. Glutathione (GSH), as a major cellular antioxidant, is produced in response to oxidative stress and is regulated by the enzyme glutamate-cysteine ligase (GCL). In this study, we examined the role of the GCL modifier subunit (GCLm) in regulating vascular smooth muscle cell (VSMC) calcification. Methods: Human coronary artery VSMCs were exposed to phosphate-rich media to induce calcification. Results: Calcification led to a decrease in the GSH:GSSG ratio (reduced glutathione to oxidized glutathione), and elevated GCLm expression, coincident with mobilization of osteogenic genes and loss of contractile phenotype. KEGG pathway analysis of human unstable atherosclerotic plaques similarly showed increased GCLm expression and activation of reactive oxygen species (ROS)-related pathways. Notably, forced overexpression of GCLm in murine VSMCs (MOVAS cells) significantly accelerated calcification. These findings implicate GCLm upregulation in promoting VSMC calcification, potentially by disrupting redox homeostasis and driving phenotypic switching. Further mechanistic studies are warranted to evaluate GCLm as a potential therapeutic target in vascular calcification.

Pathophysiology doi: 10.3390/pathophysiology32040065

Authors: Anne Petzold Jan Dreßler Anne Schrimpf André Gries

Introduction: The prognosis for patients admitted to emergency departments (ED) after drowning or diving accidents is often uncertain. In this study, we evaluated a range of clinical and laboratory parameters as potential indicators of survival. Many of these markers have previously been investigated in the context of survival prediction in both trauma-related and non-trauma-related clinical scenarios. Methods: We conducted a retrospective analysis of 25 patients aged >17 years who were admitted to the ED of the University Hospital Leipzig after drowning or diving accidents between 2012 and 2024. Clinical and laboratory parameters were compared between survivors and non-survivors, with survival defined as discharge from the hospital. Results: Of all cases analyzed—comprising 19 drowning and six diving incidents—10 patients (40%) survived, while 15 (60%) did not. Age, sex, or etiology of the accident were not statistically associated with survival. Compared to survivors, non-survivors were significantly more likely to have received prehospital cardiopulmonary resuscitation (CPR; 20% vs. 86.7%) and to have exhibited lower Glasgow Coma Scale scores and lower pH values (7.4 vs. 6.7). They were also more likely to have shown increased levels of lactate (4.3 mmol/L vs. 14.8 mmol/L), CK-MB quotient (9.7% vs. 51.8%), myoglobin (188.9 µg/L vs. 1930.9 µg/L), and blood glucose (6.6 mmol/L vs. 14.3 mmol/L). Conclusions: The need for CPR appears to be the most significant risk factor for not surviving a drowning or diving accident. Furthermore, certain laboratory parameters, such as pH and lactate, may provide supportive information regarding the severity of hypoxia and could be cautiously considered as indicators of survival likelihood in these patients. Our findings offer a rationale for future prospective studies, aiming to incorporate additional clinical and biochemical markers and potentially develop new prognostic scoring systems for patients following drowning or diving accidents. This study examines the association between clinical and laboratory parameters and survival in patients following drowning and diving accidents. A total of 25 cases from 2012 to 2024 were retrospectively analyzed. The results showed that patients who required CPR had significantly poorer outcomes. Certain laboratory markers; such as pH and lactate levels; were closely related to survival status in this patient group.

Pathophysiology doi: 10.3390/pathophysiology32040064

Authors: Késia Zanuzo Márcia Fernandes Nishiyama Eloá Angélica Koehnlein Sabrina Grassiolli

Objectives: Serum ferritin (SF) reflects iron homeostasis, in addition to being an acute phase reactant protein. Since its levels are altered in the obesity state, we compared body composition, metabolic profile, liver alterations, and dietary patterns in adults stratified by SF levels (normal vs. high). Methods: A cross-sectional study was conducted using secondary data from 113 adults (≥18 years) of both sexes, attended at an outpatient nutrition clinic in southern Brazil and categorized for normal or high SF. Socioeconomic, anthropometric, blood pressure, dietary, biochemical, and liver parameters were assessed and statistical analyses performed. Results: Participants with high SF were more frequently male (p < 0.0001), married or in a civil union (p = 0.012), and had lower educational levels (p = 0.009). Moreover, higher rates of obesity (p = 0.003), cardiovascular risk (p = 0.004), increased body fat percentage (BF%; p = 0.002) and metabolic disturbances such as elevated glucose (p = 0.023), triglycerides (p = 0.003), insulin resistance (p = 0.027), hypertension (p = 0.001), and metabolic syndrome (MS) (p = 0.001) were noted in this group. Liver-related findings comprised increased ALT (p = 0.008), uric acid (p = 0.016), and indicators of steatosis (p = 0.022). Logistic regression demonstrated a higher likelihood of elevated SF among men (OR = 16.82) and individuals with increased BF% (OR = 7.5), without significant influence of diet. Conclusions: Adults with elevated SF were predominantly obese men with excess adiposity, insulin resistance, and metabolic and hepatic dysfunctions, conditions that increase the risk of MS and liver injury. These findings suggest that SF and other iron biomarkers may serve as valuable tools for diagnosing metabolic dysfunctions and obesity-related liver diseases, particularly Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD).

Pathophysiology doi: 10.3390/pathophysiology32040063

Authors: Alexandra Nikolaeva Maria Pospelova Mark Voynov Varvara Krasnikova Albina Makhanova Samvel Tonyan Aleksandr Efimtsev Fionik Olga Anatoliy Levchuk Gennadiy Trufanov Konstantin Samochernykh Tatyana Alekseeva Stephanie E. Combs Maxim Shevtsov

Objectives: Breast cancer survivors often experience long-term neurological complications, including balance impairments, following treatment. This study aimed to investigate microstructural changes in white matter tracts in breast cancer survivors with balance impairment using diffusion tensor tractography. Methods: An open, single-center, prospective study was conducted including two groups—healthy age-matched volunteers (n = 28) and breast cancer survivors (n = 35) with balance impairment. All participants underwent diffusion tensor tractography at baseline and at the end of the follow-up period of six months. Quantitative anisotropy was analyzed using DSI Studio to assess white matter integrity. Results: At baseline, patients with balance impairment exhibited significantly reduced quantitative anisotropy values in the middle cerebellar peduncles (p = 0.046) and cerebellar hemispheres (p = 0.024, 0.055) compared to healthy controls. At the end of the follow-up, quantitative anisotropy values were increased across most tracts, though some differences persisted between groups (p < 0.001). Conclusions: Breast cancer survivors with balance impairment demonstrate sustained microstructural white matter changes, particularly in cerebellar and vestibular pathways. These findings suggest that diffusion tensor tractography can provide valuable insights into central nervous system alterations contributing to post-treatment balance dysfunction and may serve as a potential tool for early diagnosis and rehabilitation planning.

Pathophysiology doi: 10.3390/pathophysiology32040062

Authors: Shahid Habib Michael Ball Chris Thomas Traci Murakami Nehali Patel Sandeep Yarlagadda Sarah Patel Courtney Walker Varun Takyar Krunal Patel Christian Domingues Chiu-Hsieh Hsu

Background: Spontaneous bacterial peritonitis (SBP) is a serious complication in patients with decompensated cirrhosis and ascites. Diagnosis typically relies on an ascitic polymorphonuclear (A-PMN) cell count ≥ 250 cells/high-power field (HPF). Methods: In this retrospective cohort study, 117 hospitalized patients with acute decompensation of chronic liver disease and a diagnostic paracentesis were evaluated. Clinical, laboratory, and imaging data were collected. Patients were stratified by A-PMN counts of ≤50, 51–249, or ≥250 cells/HPF. Additional analysis was performed with patients stratified by ascitic white blood cell (WBC) count and albumin. Mortality risk was assessed at 28, 90, and 365 days. Results: Patients with A-PMN ≤ 50 cells/HPF had the lowest 28-day mortality (8%). At 90 and 365 days, mortality risk was significantly higher for the A-PMN 51–249 cells/HPF group (90-day hazard ratio (HR) 3.55, p = 0.01; 365-day HR 2.43, p = 0.02), but not A-PMN ≥ 250 cells/HPF group (90-day HR 2.95, p = 0.1; 365-day HR 2.95, p = 0.2). Ascitic WBC count did not significantly predict mortality, though higher counts were associated with extraperitoneal infections. Ascitic fluid albumin ≤ 1.0 g/dL was independently associated with increased 365-day mortality (HR 3.53, p = 0.03). Conclusions: Binary SBP A-PMN thresholds may not adequately capture mortality risk in cirrhotic patients with ascites. Low ascitic albumin and intermediate A-PMN counts are associated with increased long-term mortality, suggesting the need for more nuanced diagnostic and prognostic criteria in SBP evaluation.

Pathophysiology doi: 10.3390/pathophysiology32040061

Authors: Arslan Ahmed Siru Li Jane J. Yu Wen-Hai Shao

Systemic lupus erythematosus is a multifactorial autoimmune disease characterized by the dysregulation of both innate and adaptive immunity, resulting in chronic inflammation, autoantibody production, and multi-organ damage. Innate immune dysfunction involves macrophages, neutrophils, plasmacytoid dendritic cells, natural killer cells, and the complement system, which collectively amplify autoimmunity through defective clearance of apoptotic cells, overproduction of pro-inflammatory cytokines, and abnormal type I interferon signaling. Adaptive immune abnormalities, including skewed T-cell subsets, impaired regulatory T and B cells, and autoreactive B-cell hyperactivity, further perpetuate pathogenic autoantibody generation. Gut microbiota dysbiosis contributes to SLE pathogenesis via Th17 activation, loss of mucosal tolerance, and molecular mimicry mechanisms. This review synthesizes current knowledge on the immunopathogenesis of SLE, emphasizing the interplay between innate and adaptive immunity and integrating evidence from both human and experimental murine models to provide a comprehensive understanding of disease mechanisms.

Pathophysiology doi: 10.3390/pathophysiology32040060

Authors: Josip Katic Ante Anic Toni Breskovic Josip Andelo Borovac Branka Kresic Daniela Supe-Domic Marko Kumric Josko Bozic Zrinka Jurisic

Background: Thromboembolic events, though infrequent, remain a significant complication of atrial fibrillation (AF) ablation, largely related to endothelial damage. Cryoballoon (CB) and radiofrequency ablation can induce pro-coagulant responses, whereas pulsed-field ablation (PFA), a novel non-thermal electroporation-based technique, has shown tissue selectivity with potential endothelial-sparing effects. Methods: We aimed to compare PFA and second-generation CB ablation regarding endothelial injury in patients with paroxysmal AF. In this single-center prospective observational study, 25 patients with paroxysmal drug-refractory AF underwent pulmonary vein isolation using either a pentaspline PFA catheter (n = 14) or a second-generation CB catheter (n = 11). Circulating von Willebrand factor antigen (vWF) levels were assessed before and after ablation as a biomarker of endothelial damage, alongside routine laboratory and echocardiographic parameters. Procedural characteristics were also analyzed. Results: Baseline demographic, clinical, and echocardiographic data were comparable between groups. PFA was associated with significantly shorter skin-to-skin procedure time (59 vs. 94 min, p = 0.005) and left atrial dwell time (44 vs. 79 min, p < 0.001) compared with CB ablation. Importantly, vWF levels decreased significantly after PFA (−7.6%, p = 0.007), while CB ablation showed a non-significant increase (+9.5%, p = 0.155). The between-group difference in percent change of vWF was statistically significant (−5.6% vs. +8.3%, p = 0.006). Conclusions: PFA was associated with reduced endothelial injury and shorter procedural times compared with CB ablation, suggesting a potential advantage in lowering thromboembolic risk. These findings support the concept of PFA as an “endothelial sparing” ablation modality. However, the PFA procedure was associated with a significantly greater extent of myocardial injury, as reflected in circulating high-sensitivity cardiac troponin T values, compared to CB ablation (p = 0.007). Larger, randomized studies are warranted to confirm these results and evaluate long-term clinical outcomes.

Pathophysiology doi: 10.3390/pathophysiology32040059

Authors: Catharina C. Gaeth Travis R. Madaris Jamila M. Duarte Amber M. Powers Christina M. Sandoval Stefanie M. Shiels Randolph Stone

Background/Objectives: Extremity trauma represents a significant proportion of battlefield injuries and is prevalent in polytraumatized patients from accidents. Delayed antibiotic treatment and surgical intervention can lead to wound infections, contributing to preventable mortality. This preliminary study aimed to develop a conscious swine model of complex extremity trauma that induces systemic inflammatory response syndrome (SIRS). Methods: All surgical procedures were conducted under anesthesia with sufficient analgesia. All swine were instrumented with a telemetry device and catheters at least 3 days prior to any injury. In phase 1 of model development, a complex extremity injury was performed that consisted of skin and muscle loss, bone defect, severe hemorrhage, and 2 h tourniquet application. In phase 2, multi-drug resistant Gram-positive and Gram-negative bacteria were inoculated topically at the injury site to exacerbate pathophysiological changes towards SIRS. Post-injury, conscious animals were assessed a minimum of twice daily, including pain assessment, neurological response, and vital signs. Blood samples were collected for microbiological testing, complete blood cell counts, and biochemical analysis. Results: After establishing SIRS criteria for Sinclair swine, we developed a model of severe extremity trauma leading to SIRS. During phase 1, resuscitative fluids were reduced and discontinued, with animals surviving 24 h and maintaining SIRS for up to 4 h post-recovery. Phase 2 showed that Gram-negative and Gram-positive pathogens can exacerbate and prolong SIRS. After 72 h, localized infection at the injury site was observed in all animals. Conclusions: We established a new swine model of complex extremity trauma with SIRS. Our model is consistent, reproducible, and relevant to prolonged care scenarios, providing a platform for future research into the evaluation of preventative and therapeutic strategies.

Pathophysiology doi: 10.3390/pathophysiology32040058

Authors: Alexander Ponomarev Daniil Kuznetsov Elena Mukhlynina

Background: To address limitations in static cold storage (SCS) of donor hearts, we developed the High-Pressure Gaseous Perfusion without Fluidic Preservation Media (HIPPER) method, along with the necessary equipment for its application. Methods: 33 Wistar rat hearts were split into five groups: (Control) static cold storage (SCS) in HTK solution, (Exp) HIPPER using oxygen–xenon gas mixtures of varying ratios (“Gas-A”: 1/9, “Gas-B”: 9/1, and “Gas-C”: 1/1), and (Air) HIPPER using air. Hearts were preserved for six hours, followed by a one-hour Langendorff assessment. Results: Beating was restored in 4/10 Control hearts, 15/15 Exp hearts across all gas mixtures (p = 0.001 Control vs. Exp), and 6/8 Air hearts. Among resuscitated hearts, the mean heart rates (in bpm) were 131 ± 10 (Control), 164 ± 21 (Air), and 226 ± 13 (Exp) (p = 0.001 Control vs. Exp; p = 0.015 Exp vs. Air). The mean left ventricular pressures (in mmHg) were 31 ± 5 (Control), 45 ± 9 (Air), and 73 ± 7 (Exp) (p = 0.002 Control vs. Exp; p = 0.014 Exp vs. Air), with dP/dT max/min showing consistent trends (p < 0.006 Control vs. Exp and Air vs. Exp). Infarct size in Exp group was also significantly reduced, averaging 39.6 ± 6.6% (Control), 12.6 ± 3.3% (Air), and 6.3 ± 0.7% (Exp) of total myocardium area (p < 0.014 for Control vs. all). Conclusions: as evidenced by both quantitative and qualitative data, HIPPER consistently outperformed SCS following six hours of storage of rat heart regardless of the gas mixture, highlighting its potential as a more robust preservation method.

Pathophysiology doi: 10.3390/pathophysiology32040057

Authors: Lina Zaripova Abai Baigenzhin Alyona Boltanova Zhanna Zhabakova Maxim Solomadin Larissa Kozina

Background/Objectives: Systemic sclerosis (SSc) is a heterogeneous connective tissue disease characterized by immune dysregulation, vasculopathy, and fibrosis. Objectives: To evaluate the genetic architecture and autoantibody profile in a Kazakh cohort of patients with SSc. Methods: A total of 26 Kazakh patients with diffuse SSc were examined for disease activity and organ impairment using EScSG and the modified Rodnan skin score (mRSS). Eighteen healthy volunteers were enrolled in the control group. Antinuclear factor (ANF) was estimated on HEp-2 cells, while antibodies to Scl-70, CENP-B, U1-snRNP, SS-A/Ro52, SS-A/Ro60, Sm/RNP, Sm, SS-B, Rib-P0, and nucleosomes were determined by immunoblotting. The level of IL-6 cytokine was detected using ELISA. To investigate the genetic basis of SSc in Kazakh patients, a custom AmpliSeq panel including targeting immune/fibrosis pathways and 120 genes was used on the Ion Proton sequencer. The statistical analysis of categorical variables was conducted using Fisher’s exact test and Chi-square (χ2) test. Results: The examination of SSc patients (mRSS 16 ± 7.2; EScSG 3.54 ± 2.18) revealed a broad range of antibodies to Scl-70, CENP-B, SS-A/Ro60, SS-A/Ro52, U1-snRNP, and RNP/Sm, which were undetectable in the control group. Genetic analysis identified multiple variants across immune regulatory genes, including likely pathogenic changes in SAMD9L, REL, IL6ST, TNFAIP3, ITGA2, ABCC2, AIRE, IL6R, AFF3, and TREX1. Variants of uncertain clinical significance were detected in LY96, IRAK1, RBPJ, IL6ST, ITGA2, AIRE, IL6R, JAZF1, IKZF3, IL18, IL12B, PRKCQ, PXK, and DNASE1L3. Novel variants at the following genomic coordinates were identified and have not been previously reported in association with SSc: LY96 (chr8:74922341 CT/C), PTPN22 (chr1:114381166 CT/C), IRAK1 (indels at chrX:153278833), and SAMD9L (chr7:92761606 GT/G; chr7:92764981 T/TT). Conclusions: The first immunogenetic investigation of SSc in Kazakhstan revealed a polygenic architecture involving immune signalling pathways that partially overlap with international cohorts while exhibiting region-specific variation. Although the limited sample size and lack of functional validation constrain the interpretability of the findings, the results provide a framework for larger research to confirm the pathogenic mechanisms and establish clinical relevance.

Pathophysiology doi: 10.3390/pathophysiology32040056

Authors: Ion Prisneac Abigail I. Wald Chelsea Bragg John A. Ozolek

Medullary thyroid carcinoma (MTC) is a thyroid tumor with neuroendocrine properties purportedly derived from C-cells. The biochemical activity of medullary thyroid carcinoma includes the production of calcitonin and carcinoembryonic antigen, which are sensitive tumor markers, facilitating diagnosis, follow-up, and prognostication. Calcitonin-negative medullary thyroid carcinoma is a rare, poorly understood primary neuroendocrine carcinoma of the thyroid characterized by classic medullary thyroid carcinoma morphology without raised serum calcitonin and with or without the expression of calcitonin detected by immunohistochemistry. Previous studies reported that C-cells were derived from the neural crest; however, more recently, C-cells have been indisputably shown to be derived from the pharyngeal endoderm and ultimobranchial bodies. Ultimobranchial body (UBB) remnants can persist in the thyroid and express p63, but their function is poorly understood. Some have postulated that ultimobranchial bodies may be the “stem” cell of the thyroid and may be precursors for thyroid tumors, particularly mixed tumors with follicular and medullary components. We present a unique case of calcitonin-negative MTC in a 58-year-old male arising in an inflamed and fibrotic thyroid with numerous scattered ultimobranchial body remnants and concomitant C-cell hyperplasia/medullary microcarcinoma (CCH/MMC). The ultimobranchial body remnants, C-cell hyperplasia, and medullary thyroid carcinoma were MTC classifier positive according to ThyroSeq®. The areas representing CCH/MMC expressed calcitonin by IHC while the main MTC tumor was negative. An additional unique feature was an area demonstrating a “mixed” C-cell/thyroid follicular epithelial phenotype. In this review we review the possible etiologies of calcitonin-negative MTC, the possibility of a neoplastic sequential progression from ultimobranchial bodies to CCH/MMC to medullary thyroid carcinoma with the individual elements (UBB, CCH/MMC, MTC) demonstrated in this thyroid, and previous postulations that ultimobranchial bodies may be the source of some follicular thyroid cancers, medullary thyroid cancers, and mixed tumors of medullary and follicular epithelial types.

Pathophysiology doi: 10.3390/pathophysiology32040055

Authors: Leila Stabayeva Madina Mergazina Yevgeniy Kamyshanskiy Gulchekhra Ikhtiyarova Zhanna Amirbekova Gulnazira Imanbayeva Olga Kostyleva

Objectives: To evaluate the diagnostic and prognostic value of histophenotyping of the extracellular matrix of the cervical stroma at cervical intraepithelial neoplasia (CIN). Methods: Retrospective analysis of 160 biopsies and surgical preparations of the cervix in women of reproductive age included cases of CIN 1–3 and the group with confirmed persistence or lesion progression (CIN P) at repeated biopsy. The control group (n = 40) consisted of morphologically intact cervical tissue. Histophenotypes were evaluated by staining with hematoxylin, eosin, and Masson trichrome, and classified as follows: normal (dense parallel bundles of type I collagen), intermediate (disorganized and fragmented type I collagen fibers), and myxoid (amorphous weakly fibrillar matrix). The clinical, viral, and inflammatory characteristics between histophenotypes were statistically compared. Results: The distribution of histophenotypes of the extracellular matrix of the cervix varied significantly depending on the CIN degree (p < 0.001). In the control group, the normal pattern was detected in 97.5% of cases; its frequency decreased from CIN 1 (27.5%) to CIN 2 (12.5%) and was absent at CIN 3. The frequency of the myxoid pattern increased significantly in severe and persistent forms: 55% at CIN 3 and 62.5% at CIN P. Human papillomavirus 16/18 was most frequently detected in groups with intermediate (69.1%) and myxoid (27.2%) patterns. Inflammatory changes were more often accompanied by disorganized extracellular matrix; however, intermediate and myxoid types also occurred in the absence of inflammation. Conclusions: The myxoid histophenotype of the extracellular matrix is significantly associated with the high degree of dysplasia and CIN persistence. It can reflect the morphological equivalent of tumor-associated stroma remodeling. Histophenotyping of the extracellular matrix of the cervix appears to be a promising method of risk stratification and may complement existing diagnostic algorithms for CIN.

Pathophysiology doi: 10.3390/pathophysiology32040054

Authors: Margherita Palermo Carolina D’Elia Giovanni Mazzucato Christine Mian Christine Schwienbacher Esther Hanspeter Silvia Clauser Salvatore Mario Palermo Armin Pycha Isabel Heidegger Igor Tsaur Emanuela Trenti

Background/Objectives: Testicular germ cell tumours (GCT) have high cure rates, especially in early stages. MicroRNA-371a-3p (M371) has recently emerged as a highly sensitive biomarker for malignant GCTs, except teratoma. This study aimed to evaluate the diagnostic performance of M371-test in a real-life clinical setting, compared to conventional markers alpha-fetoprotein (AFP), lactate-dehydrogenase (LDH), and beta-human chorionic gonadotropin (β-HCG) in patients with suspected GCT. Methods: The study, approved by the Ethic-Committee of the Provincial Hospital of Bolzano (N.97-2021), included 91 M371-tests, performed from March 2021 to May 2025. A total of 75 patients had suspected GCT; 19 healthy males served as control. Serum levels of M371, AFP, LDH, and β-HCG were compared with final histopathological diagnosis. M371 was also assessed in controls to evaluate test performance. Secondary analyses investigated correlations between preoperative M371 levels and tumour size in non-metastatic patients, and between M371-levels and clinical stage in the entire GCT cohort. A cut-off of RQ > 5 (relative quantification) was used to calculate sensitivity, specificity, and predictive values. Results: M371 showed a sensitivity of 90.9% and specificity of 89.3%, outperforming in terms of sensitivity AFP (20.4%/96.4%), LDH (40.9%/96.4%), and β-HCG (43.1%/100%). Positive predictive value (PPV) and negative predictive value (NPV) were 93.0% and 86.2%, respectively. Sensitivity was 95% for non-seminomas and 87.5% for seminomas. In non-metastatic patients, M371 levels correlated with tumour size and were significantly higher in advanced stages (median RQ 1128.35 vs. 98.36; p = 0.015). Conclusions: M371 showed excellent diagnostic performance, even for small tumours, supporting its clinical use. Further studies are needed to define its role in treatment planning and follow-up.

Pathophysiology doi: 10.3390/pathophysiology32040053

Authors: Nikola Šutulović Neriman Ezgin Nela Puškaš Emilija Đurić Željko Grubač Daniel Škrijelj Milena Vesković Dušan Mladenović Isidora Savić Djuro Macut Yavuz Dodurga Aleksandra Rašić-Marković Olivera Stanojlović Dragan Hrnčić

Current standard treatments for chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), a urological disorder with anxiety as a major comorbidity, are limited in success rates. Recent findings revealed the anti-inflammatory and neuroprotective effects of CO-releasing molecules (CO-RMs), but there is a gap in the knowledge on its effects in CP/CPPS. Therefore, the objective of our study was to investigate the potential therapeutic effects of CORM-A1 on the scrotal pain threshold and anxiety-related behaviors in experimental model of CP/CPPS. Adult Wistar albino male rats were randomized to Sham (intraprostatic saline) or CP/CPPS (intraprostatic λ-carrageenan) groups (n = 12). Half received CORM-A1 (2 mg/kg/day, i.p., days 1–7), others PBS, forming four subgroups (n = 6). The pain threshold (by an electronic von Frey esthesiometer) and anxiety-like behavior (by an open field, elevated plus maze and light/dark test) were assessed; prostates were histologically examined. Carrageenan-induced CP/CPPS caused significant mechanical pain hypersensitivity (p < 0.001), anxiety-like behaviors (p < 0.001–0.05), and histological prostate damage when compared to corresponding Sham groups. CORM-A1 treatment increased pain thresholds (p < 0.001) and improved behavioral outcomes (p < 0.001–0.01) in all ethological tests. These findings indicate that CORM-A1 exerts analgesic and anxiolytic effects in an experimental model of CP/CPPS in rats.

Pathophysiology doi: 10.3390/pathophysiology32040052

Authors: Kitty P. Toews Finn Morgan Auld Terence N. Moyana

The morphogenesis of the primordial gut relies on signaling pathways such as Wnt, FGF, Notch, Hedgehog, and Hippo. Reciprocal crosstalk between the endoderm and mesoderm is integrated into the signaling pathways, resulting in craniocaudal patterning. These pathways are also involved in adult intestinal homeostasis including cell proliferation and specification of cell fate. Perturbations in this process can cause growth disturbances manifesting as adenomas, serrated lesions, and cancer. Significant differences have been observed between right and left colon cancers in the hindgut, and between the jejunoileum, appendix, and right colon in the midgut. The question is to what extent the embryology of the mid- and hindgut contributes to differences in the underlying tumor biology. This review examines the precursor lesions and consensus molecular subtypes (CMS) of colorectal cancer (CRC) to highlight the significance of embryology and tumor microenvironment (TME) in CRC. The three main precursor lesions, i.e., adenomas, serrated lesions, and inflammatory bowel disease-associated dysplasia, are linked to the CMS classification, which is based on transcriptomic profiling and clinical features. Both embryologic and micro-environmental underpinnings of the mid- and hindgut contribute to the differences in the tumors arising from them, and they may do so by recapitulating embryonic signaling cascades. This manifests in the range of CRC CMS and histologic cancer subtypes and in tumors that show multidirectional differentiation, the so-called stem cell carcinomas. Emerging evidence shows the limitations of CMS particularly in patients on systemic therapy who develop drug resistance. The focus is thus transitioning from CMS to specific components of the TME.

Pathophysiology doi: 10.3390/pathophysiology32040051

Authors: Domenico De Falco Dario Di Stasio Alessandra Caggiula Carlo Lajolo Alberta Lucchese Massimo Petruzzi

Background/Objectives: Lichen Planus Pemphigoides (LPP) represents a rare variant of Oral Lichen Planus in which the typical pemphigoid-associated antibodies, BP180 and BP230, are present. The objectives of this Systematic Review are to analyze the data currently available in the literature on this rare condition, with the aim of laying the groundwork for future investigations and research. Methods: This Systematic Review was registered in the International Prospective Register of Systematic Reviews (PROSPERO) under the registration number CRD420251133018. Subsequently, a search was conducted on PubMed/Medline, Scopus, and Ovid using specific keywords combined with Boolean operators. Articles published up to 2025 were included. The following types of studies were considered eligible: case reports, clinical conferences, clinical studies, clinical trials, controlled clinical trials, letters, multicenter studies, observational studies, randomized controlled trials, and human-based studies. Book chapters, systematic reviews, narrative reviews, in vitro studies, and animal models were excluded. Results: A total of 67 articles were initially identified; following thorough review and exclusion, 20 articles were retained. The patient data extracted from these selected studies were used to construct a table in which patients were categorized according to both qualitative and quantitative variables. The results highlight that LPP is a condition requiring a complex diagnostic process involving both histological examination and serological testing (Immunofluorescence and Enzyme-Linked Immunosorbent Assay—ELISA). Conclusions: Furthermore, with the advent of immunotherapy, an increasingly well-documented new category of drug-induced LPP has emerged, associated with PD-1 and PD-L1 inhibitors.

Pathophysiology doi: 10.3390/pathophysiology32040050

Authors: Oscar Arturo Benítez-Cárdenas Néstor Oliver Herrera-Salguero Elhi Manuel Torres-Hernández Miguel Angel Noyola-Frías Ricardo Martínez-Rider Marlen Vitales-Noyola

Background: Obstructive Sleep Apnea Syndrome (OSAS) is a prevalent and underdiagnosed condition with significant systemic and quality-of-life impacts. While polysomnography remains the gold standard for diagnosis, cone-beam computed tomography (CBCT) presents a potential adjunctive imaging tool for anatomical airway evaluation. Objective: We aimed to assess the effectiveness of three-dimensional airway evaluation via CBCT as a complementary diagnostic tool for OSAS. Methods: A diagnostic test study (experimental pilot study) was conducted using CBCT scans of 30 patients, divided into two groups: 15 scans from patients with a confirmed OSAS diagnosis through polysomnography and 15 scans from healthy controls. Five tomographic variables were analyzed: anteroposterior distance, lateral distance, minimum cross-sectional area, airway volume, and airway shape. Statistical analysis was performed comparing both groups. Results: The minimum cross-sectional area and airway volume showed statistically significant differences between the OSAS and control groups (p = 0.038 and p = 0.0055, respectively). Anteroposterior and lateral distances showed trends toward significance but were not statistically significant. Conclusions: CBCT-based airway analysis, particularly focusing on volumetric and cross-sectional area parameters, demonstrates strong potential as a complementary tool in the diagnosis of peripheral-type OSAS. However, it cannot replace polysomnography, especially for central OSAS diagnosis.

Pathophysiology doi: 10.3390/pathophysiology32040049

Authors: Santiago Ruvira Pilar Rodríguez-Rodríguez Metee Iampanichakul Lucía G. Cuquerella David Ramiro-Cortijo Silvia M. Arribas

Background and objectives: Hypertension is a worldwide burden, for which fetal malnutrition is a risk factor. Another societal challenge is environmental waste. Our research focusses on cocoa shell extract (CSE), a cocoa by-product with antioxidant bioactive components. Male rats exposed to fetal malnutrition develop hypertension and endothelial dysfunction, which are improved by CSE supplementation. We hypothesized that effects of CSE are related to an antioxidant action. Methods: Adult male and female offspring of dams exposed to 50% food restriction during gestation (MUN) and controls were supplemented for 3 weeks with CSE (250 mg/kg/day) or a vehicle. We assessed plasma SOD activity, GSH and carbonyls (via spectrophotometry) and aortic expression of enzymes related to ROS degradation or production (via Western blotting). Results: MUN males showed lower Nrf2 expression and increased carbonyls, SOD activity and mitochondrial SOD2 expression, without alterations in GSH or the related enzyme CGLM. No changes in xanthine oxidase or NADPH subunits (p22phox and p47phox) were detected, suggesting a different origin of superoxide anion. Phosphorylated-eNOS/eNOS and 3-nitrotyrosine expression were increased without changes in plasma nitrates. MUN females only showed plasma SOD and aortic 3-nitrotyrosine elevation. CSE supplementation reduced SOD2 and p-eNOS/eNOS expression and SOD activity and increased Nrf2 expression. Conclusions: MUN arteries exhibit oxidative damage, with a higher impact on males. SOD2 and p-eNOS/e-NOS overexpression may be a counteracting mechanism that compensates for superoxide anion overproduction, likely involving mitochondria. The reversal of these alterations by CSE supplementation is probably related to a reduction in vascular superoxide anion through a direct scavenging action of its bioactive components. A longer supplementation period may be needed to increase endogenous antioxidants through Nrf2 and to reduce oxidative–nitrosative damage.

Pathophysiology doi: 10.3390/pathophysiology32030048

Authors: Mark M. Melamud Evgeny A. Ermakov Anna S. Tolmacheva Georgy A. Nevinsky Valentina N. Buneva

Background/Objectives: The pathognomonic feature of systemic lupus erythematosus (SLE) is the formation of antibodies to double-stranded DNA (anti-dsDNA Abs). Cell-free DNA (cfDNA) has been suggested as one of the antigens for the generation of anti-dsDNA Abs, but the temporal changes in these biomarkers are not clear. In this study, the association of dynamic changes in total cfDNA and anti-dsDNA Abs levels in blood plasma during disease progression in a murine model of pristane-induced SLE was examined. Methods: The experimental group consisted of 12 BALB/c pristane-immunized mice; the control group included 8 PBS-treated mice. Blood samples were collected six times during the 38-week study (2 weeks before and 8, 14, 22, 28, and 36 weeks after immunization). Total cfDNA and anti-dsDNA Abs levels were determined at each time point. Results: Pristane-immunized mice showed a significant increase in the concentration of anti-dsDNA Abs. A 14-week delay in the formation of anti-dsDNA Abs was observed after an increase in the concentration of cfDNA in the experimental and control groups. Anti-dsDNA Abs and total cfDNA levels did not correlate at specific time points, but the change in cfDNA concentration from week 14 to week 28 was inversely correlated with the change in the anti-dsDNA Abs level over the same time period (R = −0.71, p = 0.009), i.e., the more the anti-dsDNA Abs level increased, the more the cfDNA concentration decreased. A direct correlation was shown between the increase in body weight of pristane-immunized mice and the increase in total cfDNA concentration in the blood from week 0 to week 14 (R = 0.6, p = 0.04). Conclusions: These findings demonstrate the dynamic nature of cfDNA and anti-dsDNA Abs levels and reciprocal dynamics of these markers in a pristane-induced mouse model of SLE.

Pathophysiology doi: 10.3390/pathophysiology32030047

Authors: Toru Arai Masaki Hirose Eiji Sugimoto Takayuki Takimoto Yoshikazu Inoue Hiromitsu Sumikawa Tamiko Takemura Shigeki Shimizu

Autoimmune pulmonary alveolar proteinosis (aPAP) is characterized by the accumulation of phospholipids and surfactant proteins in the peripheral air spaces due to alveolar macrophage dysfunction caused by anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibodies (GMAb). Hypersensitivity pneumonitis (HP) is a granulomatous lung disease associated with GM-CSF. In this report, we evaluated serial changes in serum GMAb levels in a 67-year-old male current smoker with HP and aPAP and examined their correlation with HP disease activity. GMAb levels increased at HP onset and decreased after HP remission with oral prednisolone therapy. After the first remission, the patient experienced three relapses and remissions. Although GMAb levels were not evaluated for all HP relapses and remissions, GMAb levels increased at one relapse but decreased at two remissions induced by the oral prednisolone therapy. Pulmonary fibrosis progressed, and the patient died of pneumonia. GMAb was at its almost normal levels at 8 months before the onset of pneumonia. We hypothesized that GMAbs may have been induced to improve HP through neutralizing GM-CSF. Although the hypothesis needs to be confirmed in additional patients, serial measurement of GMAb may be useful for a better understanding of the pathophysiology and deciding the appropriate treatment for HP with aPAP.

Pathophysiology doi: 10.3390/pathophysiology32030046

Authors: Sophie Maria Penfold James Cunningham Pauline Whelan Martin G. McCabe John Ainsworth

Background: Fatigue has been associated with poorer quality of life and increased morbidity in multiple clinical fields. Patients with autonomic dysfunction have been found to experience poorer physiological health, as well as having an increased risk of comorbidity and all-cause mortality. Heart rate variability (HRV) has been documented as a validated tool to assess autonomic function in clinical practice. The aim of this systematic review was to understand the relationship between fatigue and HRV in different medical populations. Methods: A systematic search was conducted in MEDLINE via Web of Science and Scopus. Results: A total of seventeen articles were identified for inclusion. Patients with Chronic Fatigue Syndrome were the most investigated population (n = 7), followed by cancer (n = 4) and Multiple Sclerosis (n = 4). The most implemented fatigue measure was the Multidimension Fatigue Inventory Scale used in four studies and HRV was monitored by electrocardiogram in nine studies. The most recorded and analysed domain for HRV was the frequency parameters. A significant association between increased subjective fatigue and imbalanced metrics of HRV (p < 0.05) was identified in fourteen articles. However, results from this review were heterogenous partly owing to the inconsistency with the instruments implemented to monitor HRV and measure fatigue. Additionally, only a small number of medical conditions were investigated, and the patients were predominately older adults (mean age 43.2) and women (64%). Conclusions: Despite these discrepancies, the reviewed evidence suggests that a rise in sympathetic activity and reduced parasympathetic tone are associated with an increased perception of fatigue in medical populations.

Pathophysiology doi: 10.3390/pathophysiology32030045

Authors: Magdalena Zawadzka Ewelina Ejchman-Pac Amelia Kowalska Paweł Szymański Justyna Marszałkowska-Jakubik

Background: Cardiovascular disease (CVD) remains the leading cause of death in Europe. Despite medical advancements, modifiable risk factors—such as obesity, smoking, and physical inactivity—continue to rise, especially in high-demand professional groups like military personnel. The updated SCORE2 model offers broader assessment capabilities compared to the traditional Pol-SCORE system used in Poland. This study aimed to assess and compare cardiovascular risk using both models and evaluate self-awareness of cardiovascular risk factors among military and civilian employees. Methods: The study included military personnel and civilian defense employees who completed a health-related questionnaire and underwent clinical evaluation, including blood pressure measurement and lipid profiling. Cardiovascular risk was assessed using both Pol-SCORE (fatal events only) and SCORE2 (fatal and non-fatal events). Statistical analysis was conducted using standard parametric and nonparametric methods. Results: SCORE2 classified significantly more individuals into high or very high cardiovascular risk categories than Pol-SCORE. Differences were especially pronounced among women and civilians. Elevated blood pressure, overweight, obesity, tobacco use, and stress were commonly observed. Despite a high level of awareness about prevention, regular participation in screening was low, and many respondents underestimated their health risk, indicating the presence of unrecognized or underestimated risk. Conclusions: SCORE2 proves to be a more sensitive and comprehensive tool for cardiovascular risk evaluation. The findings emphasize the urgent need for targeted prevention strategies and health education, especially in high-risk occupational groups such as military personnel.

Pathophysiology doi: 10.3390/pathophysiology32030044

Authors: Juan Manuel Velázquez-Enríquez Jovito Cesar Santos-Álvarez Karina González-García Itayetzi Reyes-Avendaño Víctor Acevedo-Sánchez Ariadna Jalife Gómez Antonio Arcos-Román Jaime Arellanes-Robledo Verónica Rocío Vásquez-Garzón Rafael Baltiérrez-Hoyos

Tenascin-C (TNC) is an extracellular matrix (ECM) protein with key roles in various biological processes, such as embryonic development and tissue regeneration. However, its deregulated expression can contribute to pathological responses, promoting chronic inflammation, fibrosis, or tumor progression. It belongs to the tenascin family, a class of extracellular proteins that interfere with cellular events in both physiological and pathological contexts, interacting specifically with cells and other components of the ECM. TNC has emerged as a key player in the pathogenesis of chronic respiratory diseases (CRDs), including asthma, chronic obstructive pulmonary disease (COPD), lung cancer (LC), pulmonary hypertension (PH), and idiopathic pulmonary fibrosis (IPF). The influence of TNC on cellular responses, which is mediated by precise interactions with cellular receptors and ligands, triggers complex intracellular signaling cascades associated with the inflammatory response, fibrosis, and tumorigenesis in these CRDs. This review synthesizes recent evidence highlighting the multifaceted roles and underlying mechanisms of TNC in the context of these CRDs.

Pathophysiology doi: 10.3390/pathophysiology32030043

Authors: Judyta K. Juranek Bernard Kordas Piotr Podlasz Agnieszka Bossowska Marta Banach

Neurodegenerative diseases are a group of disorders characterized by the progressive deterioration of the structure and function of central nervous system neurons and include, among others, amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), Parkinson’s (PD), Alzheimer’s (AD), and Huntington’s (HD) diseases. And while all these diseases seem to have different genetic and environmental components, growing evidence shows that they share common underlying pathological features such as increased neuroinflammation and excessive oxidative stress. RAGE, the receptor for advanced glycation end-products, is a signal transduction receptor, and its activation triggers an increase in proinflammatory molecules, oxidative stressors, and cytokines. Diaph1, protein diaphanous homolog 1, is an actin modulator and an intracellular ligand of RAGE. Studies demonstrated that RAGE and Diaph1 act together, and their downstream signaling pathways play a role in neurodegeneration. Here, based on current evidence and our own research, we provide an overview of the RAGE–Diaph1 signaling and discuss the therapeutic potential of targeted therapy aimed at RAGE–Diaph1 signaling inhibition in the prevention and treatment of neurodegenerative diseases.

Pathophysiology doi: 10.3390/pathophysiology32030042

Authors: Ruth Solomon Jan Pieter Hommen Francesco Travascio

In the original publication [...]

Pathophysiology doi: 10.3390/pathophysiology32030041

Authors: Minsup Lee Taekyung Ha Ivan A. Alvarez Wendy Leskova Changwon Park Norman R. Harris

Background/Objectives: Methamphetamine (METH), a potent psychostimulant, exerts harmful effects on the vascular system by promoting oxidative stress, inflammation, and endothelial injury. While its impact on the blood–brain barrier is well documented, its influence on the retinal microvasculature remains less understood. This study investigated the effects of METH on syndecan-1 expression and endothelial function in primary rat retinal microvascular endothelial cells (RRMECs) and isolated ophthalmic arteries. Methods: We assessed METH-induced changes in mRNA and protein expression levels of syndecan-1, matrix metalloproteinase (MMP)-2, and MMP-9. Endothelial function was evaluated using scratch migration assays and trans-endothelial electrical resistance (TEER) measurements. The mechanistic involvement of MMP-9 and trace amine-associated receptor 1 (TAAR-1), a known receptor for METH, was examined using selective pharmacological inhibitors. Results: METH exposure significantly decreased syndecan-1 expression and increased MMP-9 levels. These changes were accompanied by impaired endothelial migration and reduced TEER in RRMECs. Similar findings were confirmed in cultured ophthalmic arteries, reinforcing the translational relevance of our in vitro results. Inhibition of MMPs restored syndecan-1 expression and rescued endothelial function. Furthermore, TAAR-1 antagonism protected against syndecan-1 degradation, reduced MMP-9 upregulation, and improved endothelial migration and barrier resistance. Conclusions: Our findings suggest that METH induces loss of syndecan-1 and retinal vascular integrity by promoting TAAR-1–mediated MMP-9 upregulation. Targeting the TAAR-1/MMP-9 axis may offer a promising therapeutic strategy for preventing METH-induced microvascular damage in the retina.