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.

Pathophysiology doi: 10.3390/pathophysiology32030040

Authors: Daniel Landínez-Martínez Andres Grisales-Aguirre

Background/Objectives: Post-stroke cognitive impairment significantly impacts long-term functional outcomes, particularly in instrumental activities of daily living (IADLs). Working memory training (WMT) has emerged as a potential cognitive rehabilitation strategy; however, its transfer to real-world functionality remains unclear. This study evaluated whether adaptive computerized WMT enhances IADLs performance compared to a non-adaptive control condition in chronic stroke survivors. Methods: A single-blind, randomized controlled trial was conducted with 50 adults aged 50–79 years, ≥12 months post-ischemic stroke, and diagnosed with a mild neurocognitive disorder. Participants were randomized to adaptive WMT or non-adaptive cognitive training, each completing 25 home-based sessions over 12 weeks via a standardized online platform. Primary outcomes included the Lawton and Brody IADL Scale and the Working Memory Questionnaire (WMQ); secondary outcomes included the Working Memory Index (WMI) from the WAIS-IV. Analyses included frequentist and Bayesian methods. Results: Both groups showed significant pre–post improvements in IADL independence and WMI (p < 0.05; BF10 > 10), with no significant between-group differences on overall IADL outcomes. The adaptive WMT group demonstrated specific gains in WMQ—Storing (p = 0.033; BF10 = 3.83), while the control group improved in WMQ—Attention and IADL—Assistance Required (p = 0.004–0.035; BF10 > 6). Bayesian ANOVA indicated that these effects were primarily driven by the interventions, with minimal influence from depressive symptoms or global cognition. Conclusions: Adaptive WMT yielded domain-specific cognitive benefits but did not enhance IADL performance beyond non-adaptive training. These findings highlight the limited far transfer of WMT and the importance of designing ecologically valid, multimodal rehabilitation strategies post-stroke.

Pathophysiology doi: 10.3390/pathophysiology32030039

Authors: Praise Tatenda Nhau Mlindeli Gamede Andile Khathi Ntethelelo Sibiya

Background: There is growing evidence suggesting that SARS-CoV-2 may contribute to metabolic dysfunction. SARS-CoV-2 infection is associated with systemic inflammation, oxidative stress, and metabolic dysregulation, all of which may impair liver function and promote glucose intolerance. This study investigated the role of SARS-CoV-2, specifically its Main Protease (Mpro), in accelerating insulin resistance and metabolic dysfunction in HepG2 cells in vitro. Methods: HepG2 cells were treated with varying concentrations of Mpro (2.5, 5, 10, 20, 40, 80, and 160 nmol/mL) for 24 h to assess cytotoxicity and glucose uptake. Based on initial findings, subsequent assays focused on higher concentrations (40, 80, and 160 nmol/mL). The effects of Mpro on cell viability, protein kinase B (AKT) expression, matrix metallopeptidase-1 (MMP1), dipeptidyl peptidase 4 (DPP4), interleukin-6 (IL-6) expression, and lipid peroxidation were investigated. Results: Our findings reveal that the SARS-CoV-2 Mpro treatment led to a concentration-dependent reduction in glucose uptake in HepG2 cells. Additionally, the Mpro treatment was associated with reduced insulin-stimulated AKT activation, particularly at higher concentrations. Inflammatory markers such as IL-6 were elevated in the extracellular medium, while DPP4 expression was decreased. However, extracellular soluble DPP4 (sDPP4) levels did not show a significant change. Despite these changes, cell viability remained relatively unaffected, suggesting that the HepG2 cells were able to maintain overall metabolic functions under Mpro exposure. Conclusions: This study demonstrated the concentration-dependent impairment of hepatic glucose metabolism, insulin signaling, and inflammatory pathways in HepG2 cells acutely exposed to the SARS-CoV-2 Mpro. These findings warrant further investigation to explore the long-term metabolic effects of SARS-CoV-2 and its proteases in the liver and to develop potential therapeutic approaches for post-viral metabolic complications.

Pathophysiology doi: 10.3390/pathophysiology32030038

Authors: Lorand-Tibor Reman Ovidiu Malau Daniel Porav-Hodade Calin Chibelean Arpad-Oliver Vida Ciprian Todea Veronica Ghirca Alexandru Laslo Raul-Dumitru Gherasim Rares Vascul Orsolya-Brigitta Katona Raluca-Diana Hagău Orsolya Martha

Secondary lymphoma of the prostate is described as the involvement of the prostate gland by lymphomatous spread from a primary site. This condition is exceedingly rare and often presents diagnostic and therapeutic challenges. The symptoms often mimic those of benign prostatic hyperplasia or prostate cancer, including LUTS (lower urinary tract symptoms) and even complete urinary retention. Here, we present a rare case of a 62-year-old male patient undergoing chemotherapy for stage IV mantle cell stomach lymphoma and subsequently secondary prostatic involvement. The patient presented with complete urinary retention, accompanied by biochemical (PSA = 11.7 ng/mL) and imaging (Magnetic Resonance Imaging-PIRADS V lesion) suspicion for prostate cancer. Histopathologic analysis of the MRI-targeted prostate fusion biopsy revealed secondary prostatic lymphoma. The chosen treatment was transurethral resection of the prostate (TUR-P) for relief of symptoms, which significantly improved urinary function (postoperative IPSS = 5 and Qmax = 17 mL/s). This case underscores the importance of considering prostatic lymphoma in the differential diagnosis of bladder outlet obstruction, especially in patients with a known lymphoma history. This report also provides a focused review of the literature on secondary prostatic lymphoma, highlighting the diagnostic challenges, treatment options, and clinical outcomes.

Pathophysiology doi: 10.3390/pathophysiology32030037

Authors: Michał Mazur Magdalena Szymańska Agnieszka Malik Wojciech Szlasa Joanna Popiołek-Kalisz

Thyroid hormones play a crucial role in regulating metabolism and cardiovascular function, with even mild dysfunction—such as subclinical hypothyroidism—negatively impacting heart health. While previous studies have confirmed the effects of iodine, selenium, and vitamin D on thyroid regulation and inflammation, the combined role of these nutrients in reducing cardiovascular disease (CVD) risk in autoimmune thyroid disorders remains insufficiently understood. This review explores the influence of specific micronutrients—including selenium, iodine, and zinc—and dietary patterns, particularly the Mediterranean diet, on the pathophysiology of hypothyroidism and Hashimoto’s thyroiditis. We introduce a novel framework that integrates emerging data on sex-specific micronutrient interactions and nutritional immunomodulation. Unlike the existing literature, this review introduces original hypotheses related to sex-specific nutritional immunomodulation and proposes a novel framework for micronutrient-driven dietary intervention in Hashimoto’s thyroiditis.

Pathophysiology doi: 10.3390/pathophysiology32030036

Authors: Asya Kuliyeva Natalia Serejnikova Gulnara Eshmotova Yulya Teslya Anastasia Ivina Alexey Zarov Michael Panin Alexey Prizov Vera Lyalina Dmitry Shestakov Alexey Fayzullin Peter Timashev Alexey Volkov

Background/Objectives: Osteonecrosis of the femoral head (ONFH) is a common condition in hip surgery, which is characterized by the death of bone cells due to disruption of the blood supply and ultimately irreversible destruction of the hip joint. As a result of the COVID-19 pandemic, a significant increase in the incidence of ONFH has been identified. To better understand the pathogenesis of ONFH in the context of COVID-19, our research aimed to determine pathomorphological changes in articular tissues specific to post-COVID-19 ONFH. Methods: Using morphological, morphometric, and statistical methods, the femoral heads after hip arthroplasty were retrospectively studied in patients with post-COVID-19 ONFH (n = 41) compared to a non-COVID-19 group of patients (n = 47). Results: Our results revealed that the key morphofunctional biomarkers of post-COVID-19 ONFH were clusters of mast cells, extensive areas of fibrosis, numerous arterial and venous thrombi, and giant cell granulomas. The potential relationship of those morphological features with the action of the SARS-CoV-2 coronavirus was discussed. Conclusions: Mast cells have been proposed as the leading players that may trigger the main molecular and cellular mechanisms in the development of post-COVID-19 ONFH and can be considered a diagnostic sign of the disease.

Pathophysiology doi: 10.3390/pathophysiology32030035

Authors: Roberto Paparella Carolina Putotto Marco Fiore Fiorenza Colloridi Paolo Versacci Mauro Ceccanti Bruno Marino Luigi Tarani

Fetal alcohol spectrum disorder (FASD) is a preventable cause of developmental disabilities linked to prenatal alcohol exposure (PAE). Congenital heart defects (CHDs) are frequently observed in FASD, with a notable association between PAE and dextro-type transposition of the great arteries (d-TGA). A potential pathogenetic mechanism of d-TGA in FASD, involving retinoic acid (RA) deficiency due to the interference of ethanol with RA biosynthesis, is proposed. Further investigation is required to understand the timing and impact of alcohol exposure on congenital anomalies, particularly in the context of CHDs.

Pathophysiology doi: 10.3390/pathophysiology32030034

Authors: Nikhila Kalapatapu Samantha G. Skinner Emma G. D’Addezio Srija Ponna Enrique Cadenas Daryl L. Davies

Despite the growing morbidity associated with alcohol use disorder (AUD), current FDA-approved therapeutics fail to adequately address the condition. This is in part due to the complex systemic effects of ethanol (EtOH), which have particularly negative consequences on the gut–liver–brain axis. Importantly, two systemic mechanisms underlying the progression of AUD remain underemphasized in therapeutic development: thiamine deficiency and neuroinflammation. Alcohol-induced thiamine deficiency leads to reduced activity of key metabolic enzymes, thereby resulting in energy deficits, oxidative stress, and severe clinical implications. EtOH also activates TLR4 and NLRP3, both of which play critical roles in the regulation of neuroimmune responses. While research directly investigating the relationship between thiamine deficiency and neuroinflammation is still in its early stages, our review highlights the emerging connections between these two seemingly distinct pathomechanisms. Additionally, potential therapeutic approaches and targets for addressing AUD at a systemic level are discussed.

Pathophysiology doi: 10.3390/pathophysiology32030033

Authors: Mario Alamilla-Sanchez Miguel Angel Alcalá Salgado Victor Manuel Ulloa Galván Valeria Yanez Salguero Martín Benjamin Yamá Estrella Enrique Fleuvier Morales López Nicte Alaide Ramos García Martín Omar Carbajal Zárate Jorge David Salazar Hurtado Daniel Alberto Delgado Pineda Leticia López González Julio Manuel Flores Garnica

Renal function refers to the combined actions of the glomerulus and tubular system to achieve homeostasis in bodily fluids. While the glomerulus is essential in the first step of urine formation through a coordinated filtration mechanism, the tubular system carries out active mechanisms of secretion and reabsorption of solutes and proteins using specific transporters in the epithelial cells. The assessment of renal function usually focuses on glomerular function, so the tubular function is often underestimated as a fundamental part of daily clinical practice. Therefore, it is essential to properly understand the tubular physiological mechanisms and their clinical association with prevalent human pathologies. This review discusses the primary solutes handled by the kidneys, including glucose, amino acids, sodium, potassium, calcium, phosphate, citrate, magnesium and uric acid. Additionally, it emphasizes the significance of physicochemical characteristics of urine, such as pH and osmolarity. The use of a concise methodology for the comprehensive assessment of urine should be strengthened in the basic training of nephrologists when dealing with problems such as water and electrolyte balance disorders, acid-base disorders, and harmful effects of commonly used drugs such as chemotherapy, antibiotics, or diuretics to avoid isolated replacement of the solute without carrying out comprehensive approaches, which can lead to potentially severe complications.

Pathophysiology doi: 10.3390/pathophysiology32030032

Authors: Lorrany da Silva Avanci Daniel Vitor de Souza Gabriel Carvalhal de Aguiar Thiago Guedes Pinto Barbara dos Anjos Rosario Milena de Barros Viana Yasmin Alaby Martins Ferreira Viviane Carlin Cordaro Luciana Pellegrini Pisani Daniel Araki Ribeiro

Background: This study investigated the impact of crack cocaine smoke exposure on the submandibular salivary gland of Wistar rats. Methods: The animals were distributed into four groups: control (CTRL); 25 mg exposure (CK25); 50 mg exposure (CK50); and 100 mg exposure (CK100). The animals were exposed to crack cocaine smoke once a day for five consecutive days. Results: Exposure to crack cocaine smoke-induced histopathological changes in submandibular salivary glands in all groups under exposure. The immunohistochemical analysis demonstrates that exposure to crack cocaine smoke led to an increase in BCL-2 and P16 expression in all groups exposed to crack cocaine (p < 0.05). The analysis of Ki-67 expression revealed a significant increase in immunoreactive cells across all exposure groups (p < 0.05). Although MYD88 expression was observed in all crack cocaine-exposed groups, only the group treated with the highest dose (100 mg) exhibited a statistically significant increase compared to the control group (p < 0.05). Conclusions: In summary, this study demonstrates that exposure to crack cocaine smoke-induced tissue degeneration in the submandibular salivary gland, increasing cellular senescence and promoting compensatory cell proliferation in Wistar rats.

Pathophysiology doi: 10.3390/pathophysiology32030031

Authors: Elyaa Saleh Nour Abdelaziz Malaak Ramahi Antonia Loukousia Theodossios Birbilis Dimitrios Kanakis

Amyloidosis, often referred to as “the great imitator”, is a condition characterized by the abnormal deposition of amyloid proteins in various tissues, potentially leading to organ dysfunction. When these deposits localize in the brain, they can disrupt neurological function and present with diverse clinical manifestations, making diagnosis particularly challenging. Cerebral amyloidosis is a rare entity that frequently mimics other neurological disorders, often resulting in significant delays in recognition and management. This case highlights the diagnostic challenge posed by cerebral amyloidosis and underscores its unique presentation. We present the case of a 76-year-old male with a history of rheumatoid arthritis (RA) who developed progressive right-sided weakness over several months. Three years prior, he experienced numbness on the right side of his face and upper limb. Initial imaging identified a small lesion in the left thalamic region, which was originally diagnosed as a glioma. However, due to the worsening of his clinical symptoms, further evaluation was warranted. Subsequent imaging revealed lesion growth, prompting a biopsy that ultimately confirmed the diagnosis of intracerebral amyloidoma. This case underscores the necessity of considering amyloidosis in the differential diagnosis of atypical neurological deficits, particularly in patients with systemic inflammatory conditions such as RA. The initial presentation of hemiparesis resembling a stroke, coupled with non-specific imaging findings and a prior misdiagnosis of glioma, highlights the complexity of cerebral amyloidosis. Only through brain biopsy was the definitive diagnosis established, emphasizing the need for improved diagnostic modalities to facilitate early detection. Further subtyping of amyloidosis, however, requires mass spectrometry-based proteomics or immunohistochemistry to accurately identify the specific amyloid protein involved. Clinicians should maintain a high index of suspicion for cerebral amyloidosis in patients with RA who present with progressive neurological deficits and atypical brain lesions. Early recognition and accurate diagnosis are essential to guiding appropriate management and improving patient outcomes.

Pathophysiology doi: 10.3390/pathophysiology32030030

Authors: Ye Zheng Haitao Tong Wenjuan Guo Ao Wang Wenxing Hu Min Wu Xiaonan Zhang

Background: Although immune complex formation is widely acknowledged as the etiological agent for the development of systemic lupus erythematosus, polyarteritis nodosa, reactive arthritis, etc., its roles in chronic hepatitis are less understood. This study aims to compare the immunohistochemistry profile of immune complex deposition in patients with chronic hepatitis B (CHB) and autoimmune hepatitis (AIH). Methods: Immunohistochemistry of C4d, a widely used marker for complement deposition was employed on liver biopsies from 72 and 15 patients with CHB and AIH, respectively. Statistical analysis was performed to analyze its prevalence and its association with a range of clinical and histological parameters. Results: Among the 15 AIH biopsies examined, C4d deposition was observed in 11 cases (73.3%), the majority of which showed a periportal staining pattern (10/11). In CHB, 61 (84.7%) of 72 cases tested positive for C4d, which did not differ significantly with that of AIH. While the periportal pattern was predominantly observed in CHB cases, positive staining in central veins, sinusoids, and hepatic parenchyma were also documented. In particular, C4d deposition is significantly associated with elevated serum ALT and liver inflammation in CHB. Of note, in specimens with a patchy parenchymal C4d staining pattern, a spatially correlated HBsAg IHC signal was observed in adjacent sections from the same tissue. Conclusions: These data suggest an involvement of immune complex-mediated immunopathy in autoimmune hepatitis and HBV-induced hepatitis. The positive intrahepatic C4d signal was associated with heightened liver inflammation. The colocalization of the C4d signal on hepatocytes with HBsAg strongly suggests a causal relationship between viral activity and complement deposition. These observations align with our recent evidence implicating the contribution of capsid–antibody complexes in the pathogenesis of CHB.

Pathophysiology doi: 10.3390/pathophysiology32030029

Authors: Ibrahim Aslan Tuğçe Çeker Tayfun Ustabaş Vuslat Zorlu Çağatay Yılmaz Mutay Aslan

Background/Objectives: Diabetic ketoacidosis (DKA) is an acute and severe complication of diabetes mellitus, marked by hyperglycemia, ketosis, and acidosis. It is associated with significant metabolic and inflammatory adjustments that can impact multiple biochemical pathways. This study aimed to determine the serum sphingolipid profile in DKA and investigate its relationship with neutral sphingomyelinase (N-SMase), pro-inflammatory cytokines, β-hydroxybutyrate (β-OHB), and lactate levels. Methods: Thirty-three participants were divided into three groups: control (BMI ≤ 30, no health issues), obese (BMI > 30), and DKA (BMI ≤ 30). Sphingomyelins (16:0–24:0 SMs) and ceramides (C16–C24 CERs) were measured using ultra-fast liquid chromatography combined with tandem mass spectrometry (LC-MS/MS). N-SMase, interleukin 1 beta (IL-1β), and tumor necrosis factor alpha (TNF-α) levels were assessed by enzyme-linked immunosorbent assay. Evaluations were done in the DKA group before and after standard clinical treatment for DKA (post-DKA group), which included intravenous insulin therapy, fluid resuscitation, and electrolyte replacement, as per established clinical guidelines. Results: β-OHB levels were significantly higher in the DKA group than in the control, obese, and post-DKA groups. Although β-OHB levels decreased in the post-DKA group, they remained elevated compared to the control and obese groups. Lactate levels were also higher in the DKA group, with a significant decrease in the post-DKA group. TNF-α and IL-1β were higher in the obese group compared to control and DKA groups, and TNF-α decreased significantly in the post-DKA group compared to DKA. N-SMase, 16:0–18:0 SMs, and C18-C24 CER levels were lower in the DKA and post-DKA groups compared to obese and control groups. Serum β-OHB and lactate levels were significantly correlated with S1P, total CER, total SM, and N-SMase values. Conclusions: The study reveals significant metabolic and inflammatory differences in DKA and post-DKA states, suggesting a relationship between sphingolipids, N-SMase, and these alterations, which could offer insights into DKA pathophysiology and therapeutic targets.

Pathophysiology doi: 10.3390/pathophysiology32020028

Authors: Gianluca Rigatelli Marco Zuin Niva Mileva Dobrin Vassilev Giuseppe Marchese Ervis Hiso Andrea Bertolini Claudio Bilato

Background/Objectives: The similarities and differences from a rheological perspective between significant short focal and mild long coronary lesions warrant investigation to elucidate wall shear stress (WSS) angiographic discrepancies. Methods: Patients who underwent coronary computed tomography angiography (CCTA) between 1 January 2023 and 1 September 2024 were selected for computational fluid dynamics (CFD) analysis. The selection criteria included either a focal (≤20 mm) hemodynamically significant stenosis, defined as ≥75% lumen narrowing, or a long (30–40 mm) non-hemodynamically significant lesion showing ≤50% stenosis of the left anterior descending (LAD) artery. Patient-specific models were reconstructed from ECG-gated CCTA images. Wall shear stress (WSS, measured in Pascals) and residence time (RT) were evaluated for each patient. Results: The LAD arteries of 30 patients (mean age 54 years, 63.3% men) were evaluated: 16 with focal, hemodynamically significant coronary stenosis, while 14 with diffuse, long, non-hemodynamically significant coronary lesions. Both groups exhibited a lower mean WSS compared to the non-stenosed segment, with no significant difference in mean WSS between the two groups (p = 0.84). Conversely, both groups demonstrated a higher mean residence time (RT) compared to the non-stenosed segments (0.2 ± 0.06 vs. 0.60 ± 0.03, p < 0.001 and 0.2 ± 0.006 vs. 0.59 ± 0.02, p < 0.001, respectively), and no significant difference in mean RT (p = 0.82). Conclusions: Long, angiographically mild coronary stenoses show similar WSS and RT characteristics compared to short hemodynamically significant coronary stenosis.

Pathophysiology doi: 10.3390/pathophysiology32020027

Authors: Basak G. Aydemir Andrea Berenyiova Martina Cebova John D. Henderson Andrej Barta Sona Cacanyiova

Background/Objectives: Metabolic syndrome is one of the leading causes of mortality worldwide, with high-fat diet (HFD) intake being a significant driving force. Despite long-term research, new interventions are still being sought to improve cardiovascular disorders associated with metabolic syndrome. Methods: To explore the therapeutic potential of a slow-releasing H2S donor, we evaluated the effects of 3 weeks of treatment with GYY-4137 on systolic blood pressure (sBP), cardiac parameters, adiposity, selected plasma markers, and the vascular function of the thoracic aortas (TAs) and mesenteric arteries (MAs) isolated from male spontaneously hypertensive rats (SHRs) fed an HFD for 8 weeks. Results: HFD administration induced cardiac remodeling, increased adiposity, and decreased adrenergic contractility in both TAs and MAs. Moreover, although high-fat intake improved TAs relaxation, it decreased aortic protein expression of endothelial NO synthase and the involvement of NO in vasoactive responses of both TAs and MAs. In addition, protein expression of inducible NOS and tumor necrosis factor alpha (TNFα) in aortas was increased, as were plasma levels of chemerin, which has been proposed as a possible link among metabolic and vascular disorders and inflammation. Treatment with GYY-4137 reduced sBP, improved relaxation of the MAs, partially restored the contractility of the TAs, generally restored NO signaling, and decreased the protein expression of the inducible NOS and TNFα, as well as plasma chemerin levels. Conclusions: A slow H2S-releasing donor could partially ameliorate the metabolic changes induced by increased fat intake during essential hypertension and trigger beneficial vasoactive effects associated with the NO signaling restoration and suppression of inflammation.

Pathophysiology doi: 10.3390/pathophysiology32020026

Authors: Stanislav Kotlyarov

Lipids play important roles in maintaining pulmonary structure, performing physiological functions and controlling the immune status of the lung. There is increasing evidence that lipid metabolism and immune activity are closely linked and that dysfunction in lipid metabolism contributes to the development and progression of chronic respiratory diseases such as COPD and asthma. These diseases are characterized by metabolic and immune dysregulation, with lipid mediators playing a key role in both the development and resolution of inflammation. In this regard, lipid metabolic pathways are attracting increasing attention as promising targets for biomarker detection and therapeutic intervention.

Pathophysiology doi: 10.3390/pathophysiology32020025

Authors: Grigol Keshelava Zurab Robakidze Igor Mikadze

A four-year-old female patient was admitted for evaluation after a mass on the right side of her neck was noticed during straining (Valsalva maneuver). The family first observed the mass when the patient was one year old, and noted that it gradually increased in size over time. A family history assessment revealed no known genetic disorders. The patient underwent neck ultrasonography and computed tomography angiography (CTA), which revealed two aneurysms in a right-sided communicating vein. One aneurysm was located above the jugular notch, and the other was located in the retro-parotid region. The presence of two venous aneurysms in a right-sided communicating vein—one above the jugular notch and the other in the retro-parotid region—suggests a rare and apparently benign congenital anomaly. The progressive enlargement of these malformations warrants close monitoring and surgical intervention, and long-term follow-up may be necessary to prevent complications such as thrombosis, rupture, or compression of adjacent structures.

Pathophysiology doi: 10.3390/pathophysiology32020024

Authors: Nazerke Oshakhtiyeva Dmitriy Klyuyev Zhanna Amirbekova Rinat Gatin Anar Turmukhambetova Kamilya Makhambetova Irina Kadyrova Yevgeniy Kamyshanskiy

Background/Objectives: Normal remodeling of the extracellular matrix of the endometrium is a necessary condition for the implantation of a blastocyst. We evaluated whether the use of histochemical reticulin–collagen staining can improve the assessment of the extracellular matrix of the mid-secretory endometrium in recrudescent clinical/biochemical pregnancy losses in comparison with ultrasound and routine histological examination. Methods: We compared the histochemical pattern of reticulin–collagen endometrial biopsy (21st day of the cycle) with ultrasonography and standard histological examination of the endometrium in the following groups: (1) fertile women with gravidity ≥ 2, (gravidity = parity) and (2) women with two or more clinical/biochemical pregnancy losses. Results: A normal pattern (NP) with ordered reticulin fibers forming cellular structures was determined in 92% of biopsies with physiological reproductive status and 44% of biopsies with recrudescent reproductive failure (p < 0.05), despite the fact that there were no differences in ultrasonography and standard histological examination between the groups (p > 0.05). A histochemical pattern of insufficient secretory endometrial transformation with abnormal noodle-like pattern (aNP) collagen fibers was more common in recrudescent reproductive failure (56%) than in women with physiological reproductive status (8%) (p < 0.001), despite the fact that insufficient secretory endometrial transformation with an abnormal noodle-like pattern with collagenization (aNPC) was detected only in recrudescent reproductive failure, and it was not detected in women with physiological reproductive status (p < 0.00001). Conclusions: We determined the histochemical pattern of the extracellular matrix of the endometrium in terms of the type of reticulin–collagen, associated in our study with recrudescent clinical/biochemical pregnancy loss, with improved predictability compared to ultrasonography and standard histological examination. We propose to use the method of histochemical evaluation of the reticulin–collagen pattern in order to stratify groups of women of fertile age at risk of reproductive failure.

Pathophysiology doi: 10.3390/pathophysiology32020023

Authors: Maciej Jaromin Marcin Cichocki Tomasz Konecki Piotr Kutwin Waldemar Maniukiewicz Piotr Wysocki Magdalena Gajek Małgorzata Iwona Szynkowska-Jóźwik Dariusz Moczulski

Background/Objectives: Urolithiasis is a common disease in Western societies, affecting approximately 10% of the population, and more often men than women. The formation of renal calculi is a complex process, including various compounds and proteins. The aim of this study is to compare differences between the trace element concentrations in male and female renal calculi as well as differences between the trace element concentrations in different stone types. Material and Methods: Renal calculi specimens were obtained during elective nephrolithotripsy procedures. Crystallography of renal calculi was performed using X-ray diffraction; an elemental analysis was performed using Inductively Coupled Plasma–Optical Emission Spectrometry. Statistical analysis was performed to assess the differences in the metal element concentration between men and women. The second part of the analysis measured the differences in the metal element concentration between stones containing calcium phosphate (CaP) and pure calcium oxalate (CaOx) stones. Results: The renal calculi (n = 20) obtained from the male patients had a lower potassium concentration than the calculi (n = 24) from the female patients: 393.4 vs. 792.3 mg/kg, p = 0.007. A comparison of the CaP calculi and CaOx calculi showed a higher zinc concentration (p < 0.001) and potassium concentration (p < 0.001) in the stones containing calcium phosphate. Conclusions: The renal calculi from females had a significantly higher potassium content than those from males. This difference was not correlated with hyperkalemia or the blood potassium levels, suggesting a sex-dependent role of uromodulin in stone formation. The stones containing calcium phosphate exhibited higher zinc and potassium concentrations compared to the pure calcium oxalate stones. The increased presence of zinc and potassium in urine may accelerate the formation of calcium phosphate calculi.

Pathophysiology doi: 10.3390/pathophysiology32020022

Authors: José C. De La Flor Marco Dominguez Davalos Tania Linares Grávalos Marina Alonso-Riaño Francisco Díaz Celia Rodríguez Tudero Rocío Zamora González-Mariño Michael Cieza Terrones Jesús Hernández Vaquero

Background: Fibrillary glomerulonephritis (FGN) is a rare and poorly understood kidney disease characterized by the deposition of non-amyloid fibrils in the glomeruli. Its clinical heterogeneity and high rate of progression to end-stage renal disease (ESRD) pose significant diagnostic and therapeutic challenges. This case series aims to enhance awareness of FGN and emphasizes the need for further research to improve patient outcomes. Case Reports: We reviewed the clinical, histopathological, and therapeutic data of three patients with FGN diagnosed by kidney biopsy. The cases included variations in clinical presentation from nephrotic syndrome to rapidly progressive glomerulonephritis (RPGN). Diagnostic methods incorporated light microscopy, immunofluorescence, and electron microscopy, with the integration of DnaJ homolog subfamily B member 9 (DNAJB9) staining for confirmation. Patient 1 showed a more favorable response to rituximab, achieving complete remission (CR) at 6 months and maintaining CR after 3 years. Patient 2 showed only partial remission after 2 years following treatment with rituximab. Patient 3 presented with RPGN and rapidly progressed to ESRD despite aggressive immunosuppressive therapy. Discussion: DNAJB9 has emerged as both a specific and sensitive biomarker in patients with FGN and has facilitated accurate differentiation from other glomerulopathies. This series underscores the variability in clinical outcomes and responses to therapy as well as the importance of early and accurate diagnosis. Conclusions: FGN remains a diagnostic and therapeutic challenge due to its rarity and heterogeneity. Advances in biomarkers like DNAJB9 have improved diagnostic accuracy, distinguishing FGN from similar conditions such as immunotactoid glomerulopathy. Further research into pathophysiological mechanisms and targeted therapies is essential to optimize management and outcomes for affected patients.

Pathophysiology doi: 10.3390/pathophysiology32020021

Authors: Jelena Gulišija Vesna Čapkun Stefan Golic Sanda Stojanović Stipić

Background/Objectives: The pathogenesis of intensive care unit-acquired weakness (ICU-AW) is multi-factorial, with some of the main risk factors being sepsis, multiorgan failure, and the inflammatory response related to critical illness. Vitamin D is crucial for muscle function, the immune response, and inflammation, and has been identified as a predictor of negative outcomes in intensive care unit (ICU) patients with COVID-19. The objective of this preliminary study was to examine the relationship between vitamin D serum levels and the incidence of ICU-AW in a cohort from the University Hospital of Split. Methods: A prospective observational cohort study was conducted in the University Hospital of Split in ICU from December 2021 to March 2022. The inclusion criteria were as follows: patients over 18 years old who had a confirmed severe acute respiratory coronavirus disease 2 (SARS-CoV-2) infection, patients who were mechanically ventilated for more than 48 h, and patients who were weaned from a ventilator over at least 24 h. The exclusion criteria were a history of neurological or musculoskeletal disorders and a pre-existing poor functional status. Vitamin D was detected in the first routine blood sample. Results: A total of 77 patients were observed, with 36 patients who were successfully weaned from a ventilator over at least 24 h and 1 patient who could not be examined because of impaired consciousness (this patient was excluded from further analysis), and thus a total of 35 patients were analyzed. Of these 35 patients, 12 (34%) developed ICU-AW. The median vitamin D serum level in the ICU-AW group was 17 (7.5–73.3), while that in the non-ICU-AW group was 25.2 (12.3–121). The difference in vitamin D serum levels between the groups was not significantly different from zero (p = 0.567). All patients, except for one, were vitamin D insufficient. Conclusions: Vitamin D serum levels in the ICU-AW group were not statistically different from the non-ICU-AW group, possibly due to the small sample size. Given the known roles of vitamin D in muscle function, immune modulation, and inflammation, a potential etiopathogenetic role in ICU-AW cannot be excluded without additional studies. Therefore, further studies with larger sample sizes than ours are necessary to determine whether vitamin D deficiency contributes to the development of ICU-AW and whether supplementation could have preventive or therapeutic value.

Pathophysiology doi: 10.3390/pathophysiology32020020

Authors: In-sop Kim Jaejin Hwang Chorong Oh Richard J. Morris

Background/Objectives: This study explores variations in brain activity between individuals with dementia of the Alzheimer’s type (DAT) and healthy older adults during a resting state using functional near-infrared spectroscopy (fNIRS). Methods: FNIRS measured brain activity in ten AD patients and six healthy individuals. A device with 16 channels was placed on each participant’s forehead to measure oxygenation levels while they kept their eyes closed. The data were analyzed using a support vector machine (SVM) model. Results: The results indicated differences in oxygenated hemoglobin (HbO) levels between the two groups. Specifically, HbO levels were generally higher in the dementia group in the left hemisphere, with a sharp increase after 26 s. Conversely, HbO levels were consistently lower in the right hemisphere of the dementia group. The SVM analysis demonstrated high accuracy in differentiating between the AD and healthy groups based on HbO levels. Conclusions: The study indicates that differences in brain activity during resting state can potentially distinguish people with DAT from healthy individuals. We found relatively reduced hemoglobin activity in the prefrontal areas of those with DAT. Furthermore, the concentration changes in the HbO in the left lateral prefrontal and right medial brain regions emerged as the most informative in distinguishing individuals with DAT from healthy individuals. The results of the current study show that this method could improve current DAT diagnostic practices due to its efficiency.

Pathophysiology doi: 10.3390/pathophysiology32020019

Authors: Yusuke Iizuka Masanori Sasaki Kojiro Terada Takuro Sakai Yoshinobu Nagaoka Shinobu Fukumura Jeffery D. Kocsis Takeshi Tsugawa Osamu Honmou

Background: Podocytes are essential for kidney function, and their dysfunction can result in nephrotic syndrome, such as minimal change disease (MCD). Oxidative stress contributes to podocyte damage. We investigated the therapeutic potential of intravenously infused mesenchymal stem cells (MSCs) in a puromycin aminonucleoside (PAN)-induced rodent MCD model, focusing on oxidative stress modulation. Methods: Sprague-Dawley rats were divided into three groups: intact, PAN-Vehicle, and PAN-MSC. MCD was induced through subcutaneous PAN injection. MSCs were infused intravenously in the PAN-MSC group on day 7. Urinary albumin, serum albumin, and creatinine levels were assessed. Histological analysis of the renal cortex was performed. Podocyte protein (NPHS1, NPHS2, and PODXL) and antioxidant enzyme (SOD1, SOD2, and GPX1) levels were measured using quantitative real-time reverse-transcription PCR (qRT-PCR). Results: MSC infusion significantly reduced proteinuria and restored podocyte structure in the PAN-MSC group. Electron microscopy revealed that infused MSCs could inhibit the fusion of the foot process induced by PAN injection. qRT-PCR showed that intravenous infusion of MSCs rescued the inhibition of GPX1 expression. GFP-labeled MSCs accumulated at the podocyte injury sites. Conclusion: Systemic MSC infusion mitigates PAN-induced MCD by reducing proteinuria, preserving podocyte structure, and modulating oxidative stress via the GPX1 pathway, offering a potential therapeutic approach for nephrotic syndrome.

Pathophysiology doi: 10.3390/pathophysiology32020018

Authors: Charles Bitamazire Businge Benjamin Longo-Mbenza

Background: Pregnancy simulates a metabolic syndrome-like state and predisposes to iodine deficiency and hypothyroidism through increased iodine renal loss and transplacental transfer to the fetus. Iodine deficiency is thought to predispose to dyslipidemia through elevation of serum TSH. Obesity, dyslipidemia, and hypothyroidism are established risk factors of preeclampsia. Hence, pregnant women with iodine deficiency are likely to be at increased risk of dyslipidemia and preeclampsia. We investigated the pattern of dyslipidemia among preeclamptic and normotensive pregnant women with and without iodine deficiency. Methods: The pathophysiological mechanisms linking iodine deficiency and dyslipidemia were delineated using bivariate correlations, logistic regression, and exploratory factor analysis of anthropometric, lipid profile, urine iodine concentration (UIC), and thyroid function data from 240 women with preeclampsia and 120 normotensive pregnant controls at term who attended Lomo Medical Centre, Democratic Republic of Congo (DRC). Results: Preeclamptic women with iodine deficiency had significantly lower HDL-C but higher triglyceride levels than those with sufficient iodine intake. Both normotensive and preeclamptic participants with elevated TSH had high serum oxidized LDL-C but low NO, p < 0.001. Conclusions: SCH, secondary to iodine deficiency, is associated with elevated serum oxidized LDL and decreased Nitric Oxide (NO) among both normotensive and preeclamptic women, while insufficient iodine nutrition among preeclamptic women predisposes to reduced HDL-C and increased serum Triglycerides, which are risk factors of atherosclerosis and cardiovascular disease.

Pathophysiology doi: 10.3390/pathophysiology32020017

Authors: Anna A. Starshinova Andrey An. Savchenko Alexander Borisov Igor Kudryavtsev Artem Rubinstein Irina Dovgalyuk Anastasia Kulpina Leonid P. Churilov Polina Sobolevskaia Tamara Fedotkina Dmitry Kudlay Evgeny V. Shlyakhto

Currently, understanding the immune response, its abnormalities, and its diagnostic possibilities is a key point in the management of patients with various diseases, from infectious to oncological ones. The aim of this review was to analyze the data presented in the current literature on immune disorders and the possibility of their laboratory diagnostics in combination with clinical manifestations. We have performed a systematic analysis of the literature presented in international databases over the last ten years. We have presented data on the possibility of diagnosing immunopathological processes due to changes in immune cells and soluble molecules involved in the pathogenesis of a wide range of diseases, as well as the determination of antibodies to detect autoimmune processes. By applying laboratory techniques such as hematology, flow cytometry, ELISA, etc., available to most clinical laboratories worldwide, clinical data on immune system dysfunction in a wide range of diseases are being collected. This process is unfortunately still very far from being completed. However, with all the diversity of accumulated knowledge, we can currently state that the pathogenesis of the vast majority of immune-mediated diseases is not yet known. At the same time, the current success in dividing immune-mediated diseases into distinct clusters based on different types of inflammatory responses that are based on the involvement of different populations of T helper cells and cytokine molecules represents significant progress. Further research in this direction seems very promising, as it allows the identification of new target cells and target molecules for both improved diagnostics and targeted therapies.

Pathophysiology doi: 10.3390/pathophysiology32020016

Authors: Darya Ivashkevich Arina Ponomarenko Igor Manzhulo Anastasia Egoraeva Inessa Dyuizen

Background: Metabolic effects of oleoylethanolamide-based dietary supplement (OEA-DS) were studied in a model of dietary-induced obesity in mice. Obesity was induced by a 2-month high-fat, high-cholesterol diet, resulting in significant morphological changes in liver tissues and elevated cholesterol levels in the animals’ blood serum. Elevated levels of proinflammatory cytokines, oxidative stress, and hepatocyte apoptosis were also observed in the liver tissue. The aim of this study was to examine the mechanisms through which an OEA-based dietary supplement (OEA-DS) exerts a comprehensive influence on multiple aspects of the pathogenesis of MASLD, thereby demonstrating a robust hepatoprotective effect. Methods: mice were fed a high-fat, high-cholesterol diet with or without OEA-DS supplementation. Liver tissues and blood serum were analyzed for cholesterol levels, inflammatory markers (CD68, Iba-1, CD163, IL-1β, IL-6, TNFα), apoptotic markers (Bad, Bax, Bcl-2), nuclear receptors (PPAR-α, PPAR-γ, AdipoR1), and enzymes involved in lipolysis (Acox1, Cpt1a) and cholesterol metabolism (Ldlr, Furin, Pcsk9). Immunohistochemistry, Western blotting, and RT-PCR were used to assess protein expression and gene transcription. Results: administration of OEA-DS normalized cholesterol levels, decreased expression of inflammatory markers (CD68 and Iba-1), pro-apoptotic markers (Bad, Bax) and levels of pro-inflammatory cytokines (IL-1β, IL-6, TNFα). In parallel, the expression of nuclear receptors PPAR-α and PPAR-γ, adiponectin receptor 1 (AdipoR1), and anti-inflammatory (CD163) and anti-apoptotic (Bcl-2) markers have risen. OEA-DS administration induced the expression of liver lipolysis enzymes (Acox1, Cpt1a) and cholesterol metabolism factors (Ldlr, Furin), while simultaneously reducing the transcription of the proatherogenic factor Pcsk9. Conclusions: The results of this study suggest a complex action of OEA-DS in obesity-associated liver damage, which includes reduction of systemic inflammation.

Pathophysiology doi: 10.3390/pathophysiology32020015

Authors: Michael F. Hammer Martin E. Weinand

Background/Objectives: For patients with medically refractory temporal lobe epilepsy (TLE), surgery is an effective strategy. However, post-operative seizure recurrence occurs in 20–30% of patients, and it remains challenging to predict outcomes solely based on clinical variables. Here, we ask to what extent differences in gene expression in epileptic tissue can predict the outcome after resective epilepsy surgery. Methods: We performed RNAseq on hippocampal tissue resected from eight patients who underwent anterior temporal lobectomy with amygalohippocampectomy (ATL/AH), half of whom became seizure free (SF) or non-seizure free (NSF). Results: Bioinformatic analyses revealed 1548 differentially expressed genes and statistical enrichment analyses identified a distinct set of pathways in NSF and SF cohorts that were associated with neuroinflammation, neurotransmission, synaptic plasticity, and extracellular matrix (ECM) reorganization. Resected tissue exhibiting strong pro-inflammatory processes are associated with better post-surgery seizure outcomes than patients exhibiting cellular signaling processes related to ECM reorganization, autoantibody production, and neural circuit formation. Conclusions: The results suggest that post-operative targeting of both inhibitory aspects of the ECM remodeling and the autoimmune/inflammatory components may be helpful in promoting repair and preventing the recurrence of seizures.

Pathophysiology doi: 10.3390/pathophysiology32020014

Authors: Patricia Elvira Sánchez-Valencia Juan Daniel Díaz-García Margarita Leyva-Leyva Fabiola Sánchez-Aguillón Nelly Raquel González-Arenas Jesús Guillermo Mendoza-García Erika Karina Tenorio-Aguirre Mercedes Piedad de León-Bautista Aurora Ibarra-Arce Pablo Maravilla Angélica Olivo-Díaz

Background/Objectives: Two of the microvascular complications in type 2 diabetes (T2D) are diabetic retinopathy (DR), which is the most common cause of non-traumatic blindness, and diabetic kidney disease (DKD); the latter generally requires renal replacement therapy. The aim of the present study was to determine the frequency of polymorphisms of Tumor Necrosis Factor-α, interleukin-6, and interleukin-10 (TNF-α, IL-10, and IL-6), as well as to describe the clinical and laboratory characteristics of T2D association with these microvascular complications. Methods: This study included 203 patients with T2D, of which 102 had microvascular complications: 95 with DR, 50 with DKD, and 15 with diabetic neuropathy (the latter were not included in the statistical analysis); those with T2D without confirmed microvascular complications were considered as controls. Clinical and laboratory data were collected from the patient’s medical records. Polymorphism typing of TNF-α rs361525 and rs1800629 and IL-10 rs1800872 and rs1800871 were obtained using MALDI-TOF MS. IL-10 rs1800896 and IL-6 rs1800795 were typed using a quantitative real-time polymerase chain reaction. Results: The results of age, HbA1c, fasting glucose, and arterial hypertension are significantly associated in every group. The TNF-α rs1800629A allele and TNF-α rs1800629G/A genotype were associated with microvascular complications and DR. For IL-10-rs1800896, all the models were associated in DKD. The TNF-α rs361525-rs1800629GA haplotype was associated with microvascular complications and DR, while the IL-10 haplotype, rs1800872-rs1800871-rs1800896 GGC, showed susceptibility in every group. Conclusions: Our results show the contributions of the variants of these cytokines to these microvascular complications, but more studies are required to reach relevant conclusions.

Pathophysiology doi: 10.3390/pathophysiology32020013

Authors: Angelika Hettenbach Tanja Elger Muriel Huss Gerhard Liebisch Marcus Höring Johanna Loibl Arne Kandulski Martina Müller Hauke Christian Tews Christa Buechler

Background/Objectives: Proprotein convertase subtilisin/kexin type 9 (PCSK9) regulates serum cholesterol levels and inflammation, both of which are dysregulated in inflammatory bowel disease (IBD). Free cholesterol (FC) and the various types of cholesteryl ester (CE) have different functions in the body. However, it is not yet known whether these lipids undergo parallel changes in male and female patients with active IBD, nor whether PCSK9 correlates with these lipids and disease severity in either sex. The present study measured the serum levels of PCSK9, FC, and 15 CE species in IBD patients, focusing on the associations of these molecules with sex, each other, and with disease severity. Methods: The serum PCSK9 levels of 80 IBD patients (42 males and 38 females) and 24 controls (12 males and 12 females) were measured by enzyme-linked immunosorbent assay. In addition, FC and 15 CE species levels of 53 randomly selected IBD patients and 16 controls were determined by direct flow injection analysis (FIA) using a high-resolution hybrid quadrupole-orbitrap mass spectrometer (FIA-FTMS). Results: Serum PCSK9 levels in controls and IBD patients were comparable and did not correlate with disease severity in IBD patients. There was no discernible difference in serum PCSK9, FC, and CE levels between patients with Crohn’s disease (CD) and those with ulcerative colitis (UC). FC and almost all CE species decreased in male patients with active IBD but were not related to disease severity in the female patients. The decrease in different CE species in male IBD patients with diarrhea compared to those with normal stool consistency appears to be related to IBD severity. Bile acids regulate serum cholesterol levels, and FC and CE levels were positively correlated with fecal levels of secondary bile acids in the patients with UC but not CD. This association also existed in male UC patients and could not be evaluated in women due to the small sample size. Conclusions: In active IBD, a reduction in FC and almost all CE species was observed only in males, while serum PCSK9 levels remained within normal ranges in both sexes. It can be hypothesized that blocking PCSK9 may further reduce serum cholesterol levels, which may have adverse effects in male patients with active IBD.

Pathophysiology doi: 10.3390/pathophysiology32010012

Authors: Ashley Jazzar Danielle Jacques Amira Abou-Aichi Ghassan Bkaily

Elevated circulating insulin levels between 80 and 100 µU/mL characterize hyperinsulinemia, which often leads to metabolic disorders such as obesity, insulin resistance, and type 2 diabetes (T2D). Elevated circulating insulin levels can directly affect vascular function and contribute to the pathophysiology of the cardiovascular system, including secondary arterial hypertension (SAH) and atherosclerosis. It is well known that hyperinsulinemia induced remodeling of the heart. However, there is no information on whether intrinsic differences exist between human vascular smooth muscle cells (VSMCs) and if in vitro mimicking hyperinsulinemia induces human VSMCs morphological and intracellular homeostasis remodeling in a sex- and sex hormones-dependent manner. Our in vitro cultured human VSMCs, coupled with quantitative 3D confocal imaging results, show that intrinsic differences exist between VSMCs from young men and women. Chronic hyperinsulinemia (80 µU/mL, 48 h treatment) increases cell and nuclear volumes associated with increased intracellular calcium (Ca2+) and ROS and decreased glutathione. In the absence of hyperinsulinemia, pretreatment with testosterone in VSMCs from men and oestradiol in VSMCs from women had no effect. Both sex hormones partially but not completely prevented hyperinsulinemia-induced remodeling of VSMCs from young men and women. The increase in VSMC volume may increase the thickness of the tunica media, leading to a decrease in the lumen of the blood vessel, which promotes the development of SAH and atherosclerosis in a sex-dependent manner.

Pathophysiology doi: 10.3390/pathophysiology32010011

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

Chemotherapy-related cognitive impairment termed «chemobrain» is a prevalent complication in breast cancer survivors that requires early detection for the development of novel therapeutic approaches. Magnetic resonance voxel morphometry (MR morphometry), due to its high sensitivity, might be employed for the evaluation of the early changes in the volumes of brain structures in order to explore the «chemobrain» condition. Methods: The open, prospective, single-center study enrolled 86 breast cancer survivors (43.3 ± 4.4 years) and age-matched 28 healthy female volunteers (44.0 ± 5.68). Conventional MR sequences (T1- and T2-weighted, TIRM, DWI, MPRAGE) were obtained in three mutually perpendicular planes to exclude an organ pathology of the brain. Additionally, the MPRAGE sequence was performed for subsequent MR morphometry of the volume of brain structures using the open VolBrain program. The evaluation was performed at two follow-up visits 6 months and 3 years after the completion of BC treatment. Results: According to the MR morphometry, breast cancer survivors presented with significantly decreased volumes of brain structures (including total brain volume, cerebellum volume, subcortical gray matter, etc.) as compared to healthy volunteers. Evaluation over the follow-up period of 3 years did not show the restoration of brain volume structures. Conclusions: The data obtained employing MR morphometry revealed significant reductions (that were not detected on the conventional MR sequences) in both gray and white matter in breast cancer survivors following chemotherapy. This comprehensive analysis indicated the utility of MR morphometry in detecting subtle yet statistically significant neuroanatomical changes associated with cognitive and motor impairments in patients, which can in turn provide valuable insights into the extent of structural brain alterations, helping to identify specific regions that are most affected by treatment.

Pathophysiology doi: 10.3390/pathophysiology32010010

Authors: Xiaodan Chu Jie Xu Xinggui Shen Wenji Sheng Jingxia Sun Yang Gu David F. Lewis Danielle Cooper Dani Zoorob Yuping Wang

Background/Objectives: Hydrogen sulfide (H2S) is a vasorelaxant gas and exerts anti-oxidative, anti-inflammatory, and cytoprotective effects. H2S has been implicated in regulating placental vaso-activity and angiogenesis. It is believed that abnormal trophoblast production of vasodilators and angiogenic factors contributes to pre-eclampsia development. However, little is known about whether aberrant H2S production is present in placental trophoblasts from pre-eclamptic pregnancies. Methods: Trophoblasts were isolated from normal and pre-eclamptic placentas. After incubation, cell production of H2S in the culture medium and the cellular levels of H2S were analyzed by reversed phase high-performance liquid chromatography (RP-HPLC). Expression levels of the three key H2S converting enzymes, cystathionine-β-synthase (CBS), cystathionine-γ-lyase (CSE), and 3-mercaptopyruvate sulfurtransferase (3-MST), were determined by immunohistochemistry. The protein expression of CBS and CSE was assessed by Western blot analysis. Results: (1) Trophoblast production and cellular levels of H2S were significantly reduced in cells from pre-eclamptic vs. normal placentas; (2) free H2S production was increased in a time-dependent manner in cultured trophoblasts from normal, but not from pre-eclamptic, placentas; and (3) strong CBS and CSE expression was seen in trophoblasts from normal, as opposed to pre-eclamptic, placentas. Reduced CBS and CSE expression in trophoblasts from pre-eclamptic vs. normal placentas were confirmed by Western blot analysis; and (4) 3-MST expression was undetachable in both normal and pre-eclamptic placentas, but 3-MST expression was strongly expressed in the first and second trimester placentas. Conclusions: These data provide plausible evidence that downregulation of CBS and CSE, but not 3-MST, expression may be responsible for reduced free H2S production and decreased cellular H2S levels in pre-eclamptic placentas. Our data provide further evidence that expression of 3-MST in placental trophoblasts is likely gestational age (developmental)-dependent.

Pathophysiology doi: 10.3390/pathophysiology32010009

Authors: Franklyn Nonso Iheagwam Amarachi Joy Joseph Eniola Deborah Adedoyin Olawumi Toyin Iheagwam Samuel Akpoyowvare Ejoh

Diabetes mellitus represents a complicated metabolic condition marked by ongoing hyperglycemia arising from impaired insulin secretion, inadequate insulin action, or a combination of both. Mitochondrial dysfunction has emerged as a significant contributor to the aetiology of diabetes, affecting various metabolic processes critical for glucose homeostasis. This review aims to elucidate the complex link between mitochondrial dysfunction and diabetes, covering the spectrum of diabetes types, the role of mitochondria in insulin resistance, highlighting pathophysiological mechanisms, mitochondrial DNA damage, and altered mitochondrial biogenesis and dynamics. Additionally, it discusses the clinical implications and complications of mitochondrial dysfunction in diabetes and its complications, diagnostic approaches for assessing mitochondrial function in diabetics, therapeutic strategies, future directions, and research opportunities.

Pathophysiology doi: 10.3390/pathophysiology32010008

Authors: Aleksandra Damasiewicz-Bodzek Agnieszka Nowak Maciej Maciejczyk Sławomir Waligóra Brygida Przywara-Chowaniec

Psoriasis is a multifactorial inflammatory disease. Methylglyoxal (MG) is a highly reactive dicarbonyl compound responsible for dicarbonyl stress in some inflammatory conditions, and it may play a role in the etiopathogenesis of psoriasis. Methods: A total of 50 patients with psoriasis and 35 healthy individuals participated in this study. The following indices were assessed in patients: Body Surface Area (BSA), Psoriasis Area and Severity Index (PASI), and Dermatology Life Quality Index (DLQI). MG concentration was evaluated in blood samples. The following inflammatory response indices were calculated: Systemic Inflammation Response Index (SIRI), Systemic Immuno-inflammation Index (SII), and Aggregate Index of Systemic Inflammation (AISI). Results: An analysis of the obtained data showed a statistically significant decrease in the mean serum MG concentration in patients with psoriasis when compared to the healthy individuals (1.19 ± 0.4 μg/mL vs. 1.75 ± 0.6 μg/mL; p = 0.000002). In the patients, MG concentration correlated negatively with psoriasis disease severity indicators (BSA and PASI), C-reactive protein (CRP) concentration, and inflammatory response indicators (SII and AISI). Conclusions: The decreased concentration of MG may be attributed to an increased accumulation of its derivatives (advanced glycation end-products) in the inflamed skin and/or scavenging by polyamines.

Pathophysiology doi: 10.3390/pathophysiology32010007

Authors: Shalan Alaamri Abdulhalim S. Serafi Zahir Hussain Shouq K. Bafail Mohammed A. Bafail Lusine Demirkhanyan Christopher S. Gondi Sumera Sohail

Background/Objectives: One of the major causes of hypertension (HT) is the transition of normal weight (NW) status to overweight (OW) status and obesity in a population, which leads to cardiovascular disease (CVD) and other disorders. A variety of factors/variables are involved in the development of HT and OW-related hypertension (OHT). However, we planned to investigate the pathophysiological role of serum leptin (Lep), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), total cholesterol (TC) and serum testosterone (ST) in OHT in middle-aged men. Methods: We consulted three groups of middle-aged men (age: 51–60 years)—an HT group (n: 97, high normal weight (HNW), body mass index (BMI): 23–24.9 kg/m2); an OHT group (n: 97, high overweight (HOW), BMI: 28–29.9 kg/m2) and a normal control group (NC, n: 98, HNW)—to investigate the variations in and correlations of Lep, IL-6, TNF-α, ST, TC and other variables. Results: Significant variations were obtained for the comparisons of TNF-α, Lep, ST and TC for the patient groups. OHT vs. NC showed a significant difference for ST. OHT vs. NC and OHT vs. HT had significant variations for IL-6. Significant changes were obtained for the serum levels of TNF-α, Lep, IL-6, ST and TC among groups. Significant and positive linear associations were obtained for TNF-α, Lep, TC and IL-6. Significant and negative linear associations were found for ST plotted against Lep, TNF-α and IL-6. Conclusions: The current report provides pathophysiological evidence of the interactive role of serum Lep, TNF-α, ST, TC and IL-6 in middle-aged men with HT and OHT. We suggest that the changes we noted in the present study would be helpful for further BMI-based studies in various subcategories of NW, OW and obese subjects with/without HT.

Pathophysiology doi: 10.3390/pathophysiology32010006

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

Background/Objectives: Low bone mineral density increases the risk of bone fractures, and this condition is especially common in postmenopausal women. Genetic factors significantly influence bone mineral density. This meta-analysis examined the relationship between vitamin D receptor (VDR) gene polymorphisms (BsmI, ApaI, and TaqI) and bone mineral density in postmenopausal women in the Middle East and North Africa (MENA) region. Methods: The PubMed, Embase, Scopus, and Web of Science databases were searched from inception to March 2024 for case–control studies on VDR BsmI, ApaI, and TaqI polymorphisms and their relationship with low bone density. Associations with low bone mineral density were tested with respect to different genetic models (dominant, recessive, allelic) using RevMan v5.3. Results: The meta-analysis included seven studies for BsmI, six for ApaI, and seven for TaqI, representing 704/689 cases/controls for BsmI, 914/711 for ApaI, and 974/895 for TaqI. No significant association was found between VDR polymorphisms and low bone mineral density in postmenopausal women, except in the dominant model (CC + CG vs. GG) for the BsmI variant (OR = 1.27, 95% CI: 1.01–1.59, p = 0.04). Conclusions: We found a modest association between the BsmI polymorphism and increased risk of low bone mineral density (BMD) in postmenopausal women from the MENA region, suggesting its potential as a biomarker. No associations were observed for ApaI or TaqI. These findings highlight the complex genetic–environmental interactions influencing BMD.

Pathophysiology doi: 10.3390/pathophysiology32010005

Authors: Martina Cebova Radoslava Bulkova Olga Pechanova

Background/Objectives: Evolocumab inhibits proprotein convertase subtilisin/kexin type 9 (PCSK9) binding to low-density lipoprotein (LDL) receptors, thus allowing more LDL receptors to remove LDL cholesterol from the blood. We aimed to determine the effects of evolocumab on the plasma lipid profile, reactive oxygen species (ROS), and nitric oxide (NO) generation in the heart of adult male obese Zucker rats. Methods: The rats were divided into lean and obese controls and obese rats treated with evolocumab subcutaneously at a dose of 10 mg/kg every two weeks. After 6 weeks, the lipid profile was determined in the plasma, and NO synthase (NOS) activity, thiobarbituric acid reactive substance (TBARS), conjugated diene (CD) concentration, and protein expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, nuclear factor kappaB (NF-κB), endothelial NOS (eNOS), and phosphorylated eNOS (peNOS) were measured in the heart. Results: Evolocumab treatment did not reduce body weight, relative heart weight, or systolic blood pressure in obese Zucker rats. Evolocumab treatment, however, reduced plasma LDL levels, TBARS, and CD concentrations along with decreasing expression of NADPH oxidase and NF-kappaB proteins in the heart. On the other hand, evolocumab had no effect on NOS activity or eNOS and peNOS protein expression. Conclusions: Besides its lipid-lowering effect, evolocumab may exert antioxidant properties and protect cardiomyocytes from lipid peroxidation while not affecting NO production.

Pathophysiology doi: 10.3390/pathophysiology32010004

Authors: Prathyusha Yamarthi Rama Satyasri Kotipalli Samatasai Patnaik Kv Veena Muralidharan Kathirvel Rajkumar Vutukuri Manjula Bhanoori

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