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20 pages, 848 KB  
Article
The COX Pathway Alters Hematopoiesis in Hashimoto’s Thyroiditis
by Karolina Wrońska, Maciej Ziętek, Magdalena Marciniak and Małgorzata Szczuko
Cells 2025, 14(22), 1796; https://doi.org/10.3390/cells14221796 (registering DOI) - 15 Nov 2025
Abstract
Introduction: There is limited data in the literature on the effect of prostaglandins (PG) and thromboxanes (TX) on the development and severity of Hashimoto’s Thyroiditis (HT). This article aimed to analyze the association between blood count and the cyclooxygenase (COX) pathway in 39 [...] Read more.
Introduction: There is limited data in the literature on the effect of prostaglandins (PG) and thromboxanes (TX) on the development and severity of Hashimoto’s Thyroiditis (HT). This article aimed to analyze the association between blood count and the cyclooxygenase (COX) pathway in 39 women with HT. Methods: Biochemical analysis of PGE2 and TXB2 was performed using liquid chromatography (HPLC). Results: Morphological abnormalities were found in the women studied, particularly with regard to white blood cell parameters. An increase in thyroid-stimulating hormone (TSH) was associated with significantly higher levels of monocytes (p = 0.041). Correlations were also noted between the neutrophil-to-lymphocyte ratio (NLR) and the platelet-to-lymphocyte ratio (PLR) with TXB2 and PGE2. Furthermore, a very strong correlation was demonstrated for the first time between antibodies against tissue transglutaminase (anti-tTG) and antibodies against thyroglobulin (ATG) (r = 0.781; p = 0.007). Correlations between blood count and eicosanoids were also demonstrated. Conclusions: The results suggest the involvement of COX products in the pathogenesis of HT and hematopoiesis; therefore, this study may contribute not only to advancing knowledge, but also to developing new guidelines for diagnosing and treating autoimmune diseases. Full article
(This article belongs to the Special Issue The Role of T Cells and Cellular Signalling in Immune Diseases)
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13 pages, 1114 KB  
Article
Cord Blood-Based Neonatal Screening for Hemoglobinopathies in Northern Tunisia
by Houyem Ouragini, Nizar Ben Halim, Sana Zitouni, Dorra Chaouachi, Imen Boudrigua, Naima Saidani, Imen Kraiem, Amira Ayachi, Salem Abbes, Mechaal Mourali and Samia Menif
Int. J. Neonatal Screen. 2025, 11(4), 107; https://doi.org/10.3390/ijns11040107 - 14 Nov 2025
Abstract
Hemoglobinopathies represent a major public health concern in Tunisia. Although early diagnosis is essential, systemic neonatal screening has not yet been implemented at the national level. We conducted a screening study in Northern Tunisia (Bizerte region) using cord blood samples. Complete blood counts [...] Read more.
Hemoglobinopathies represent a major public health concern in Tunisia. Although early diagnosis is essential, systemic neonatal screening has not yet been implemented at the national level. We conducted a screening study in Northern Tunisia (Bizerte region) using cord blood samples. Complete blood counts and hemoglobin analysis by capillary electrophoresis were performed. Samples showing abnormal profiles (HbBart’s, HbS, HbC, or HbA < 20%) underwent molecular testing. Correlations between hematological parameters, hemoglobin fractions, and β mutation types were assessed. Among 328 neonatal cord blood samples analyzed, we detected 3 silent α+-thalassemia, 6 β+-thalassemia traits, 3 β0-thalassemia traits, 7 HbS traits, 2 HbC traits, and 1 compound heterozygous for α+-thalassemia/HbC. No homozygous cases were identified. The heterozygous frequency was estimated at 1.2%, 2.7%, and 2.1% for α-thalassemia, β-thalassemia, and sickle cell disease, respectively. HbF levels were significantly associated with the β-thalassemia trait. This study represents the first hemoglobinopathy screening in Northern Tunisia using cord blood, highlighting the feasibility and reliability of this approach. While pilot programs have already been initiated in some regions, our findings reinforce the need for broader implementation to ensure early and accurate diagnosis across the country. Full article
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23 pages, 2863 KB  
Article
Evaluation of Functional Marine Protein Hydrolysates as Fish Meal Replacements in Low-Fish-Meal Diets: Effects on Growth Performance, Feed Utilization, and Health Status of Asian Seabass (Lates calcarifer)
by Dachawat Poonnual, Siriporn Tola and Bundit Yuangsoi
Animals 2025, 15(22), 3285; https://doi.org/10.3390/ani15223285 - 13 Nov 2025
Abstract
An eight-week study was conducted to evaluate the effects of dietary marine protein hydrolysates as fish meal replacements in low-fish-meal diets on the growth performance, feed utilization, and health status of Asian seabass (Lates calcarifer). The high-fish-meal (HFM) diet contained 25% [...] Read more.
An eight-week study was conducted to evaluate the effects of dietary marine protein hydrolysates as fish meal replacements in low-fish-meal diets on the growth performance, feed utilization, and health status of Asian seabass (Lates calcarifer). The high-fish-meal (HFM) diet contained 25% fish meal, while the low-fish-meal (LFM) diet replaced 60% of the fish meal with soybean meal. Three experimental diets were formulated by supplementing the LFM diet with 5% tuna hydrolysate (TH), 2% shrimp hydrolysate (SH), and 5% salmon silage (SS), each replacing an equivalent amount of fish meal. These diets were designated as LFM + TH, LFM + SH, and LFM + SS, respectively. The results showed that the LFM + TH diet significantly improved the percentage of weight gain, average daily growth, specific growth rate, protein efficiency ratio, and feed conversion ratio compared to the LFM diet (p < 0.05), without negatively affecting feed intake or metabolic markers. Histological analysis revealed improved villus length and goblet cell count in the intestine, indicating better nutrient absorption (p < 0.05). However, no significant differences were observed in hematological and immunological parameters, blood plasma metabolic markers, or carcass proximate composition (p > 0.05). Furthermore, the LFM + TH diet exhibited superior survival rates under ammonia stress, highlighting its potential to enhance stress tolerance. These findings suggest that marine protein hydrolysates, particularly 5%TH, can serve as a sustainable and efficient alternative to fish meal protein in diets with up to 60% in soybean meal compensation, promoting better growth and survival in Asian seabass. Full article
(This article belongs to the Section Aquatic Animals)
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12 pages, 374 KB  
Article
Biomarkers Linked to Malnutrition Identified According to GLIM Criteria Among Older Community-Dwelling Adults: Results from the ilSIRENTE Study
by Hélio José Coelho-Júnior, Riccardo Calvani, Anna Picca, Matteo Tosato, Andrea Russo, Francesco Landi and Emanuele Marzetti
Nutrients 2025, 17(22), 3543; https://doi.org/10.3390/nu17223543 - 13 Nov 2025
Abstract
Objective: This study aimed to examine the associations between malnutrition and circulating blood markers in older adults. Methods: We conducted a prospective cohort study on octogenarians residing in the mountain community of the Sirente geographic area in Central Italy. Data collection [...] Read more.
Objective: This study aimed to examine the associations between malnutrition and circulating blood markers in older adults. Methods: We conducted a prospective cohort study on octogenarians residing in the mountain community of the Sirente geographic area in Central Italy. Data collection was conducted from December 2023 to September 2024. Malnutrition was defined based on the Global Leadership Initiative on Malnutrition (GLIM) criteria. A panel of blood markers was examined, and principal component analysis (PCA) was used to identify clusters of related molecules. Both unadjusted and adjusted binary logistic regression models were applied to investigate the associations between malnutrition and these molecular clusters. Results: Data from 196 older adults (mean age: 86.2 years) were analyzed. Malnutrition was positively associated with PC 2 (i.e., urea, c-terminal agrin fragment, and potassium) (odds ratio [OR] = 1.647, p-value: 0.039) and negatively associated with PC 3 (i.e., hemoglobin, hematocrit, and red blood cell count) (OR = 0.567, p-value: 0.022) and PC 4 (i.e., calcium, albumin, total protein levels, and HDL cholesterol) (OR = 0.607, p-value: 0.035). Conclusions: Findings of the present study suggest that different clusters of blood markers are associated with malnutrition in older adults. Specifically, malnutrition is associated with clusters related to kidney function, anemia, neuromuscular function, and nutrient availability. These associations likely reflect the underlying biological mechanisms contributing to the development of malnutrition in this population. Full article
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18 pages, 1905 KB  
Article
Novel Neutrophilic Parameters of the Sysmex XN-1000V for the Prediction of Inflammation in Dogs
by Leandra C. Schöb, Melanie Ginder, Martina Stirn, Regina Hofmann-Lehmann, Heiner M. Hipp and Barbara Riond
Animals 2025, 15(22), 3275; https://doi.org/10.3390/ani15223275 - 12 Nov 2025
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Abstract
Background: Inflammation is a common reason for dogs to present to veterinary clinics. Early diagnosis of systemic inflammation is important. Acute phase proteins, like C-reactive protein, are useful but not specific to infection. In human medicine, the intensive care infection score (ICIS) offers [...] Read more.
Background: Inflammation is a common reason for dogs to present to veterinary clinics. Early diagnosis of systemic inflammation is important. Acute phase proteins, like C-reactive protein, are useful but not specific to infection. In human medicine, the intensive care infection score (ICIS) offers a faster, cost-effective alternative using advanced hematological parameters. While ICIS is not available for veterinary use, some components (e.g., neutrophil side fluorescent light) can be measured using analyzers like the Sysmex XN-1000V. Objectives: This study aimed to establish a control group of healthy dogs for the novel parameters neutrophil side fluorescent light (NE-SFL), neutrophil side scattered light (NE-SSC), and neutrophil forward scattered light (NE-FSC) and assess their utility in detecting inflammation in diseases such as sepsis, pyometra, steroid-responsive meningitis-arteritis (SRMA), and idiopathic epilepsy. Methods & Results: Value ranges were calculated based on 21 healthy dogs. Compared to controls, NE-SFL levels were significantly elevated in sepsis, pyometra, and SRMA, while NE-SSC was only elevated in sepsis and pyometra and NE-FSC only in sepsis. No increases were observed in idiopathic epilepsy. Manual gating of the white blood cell differential scattergram was necessary in samples showing high neutrophil toxicity and the presence of bands. Conclusion: NE-SFL and NE-SSC, obtainable from routine complete blood count, may serve as novel, accessible markers for inflammation in dogs. Further research is needed to validate their broader diagnostic use. Full article
(This article belongs to the Collection Clinical Pathology in Animals)
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11 pages, 737 KB  
Article
Increased Mortality with Intermediate Ascitic Polymorphonuclear Cell Counts Amongst Patients with Cirrhosis: Time to Redefine the Care Approach
by Shahid Habib, Michael Ball, Chris Thomas, Traci Murakami, Nehali Patel, Sandeep Yarlagadda, Sarah Patel, Courtney Walker, Varun Takyar, Krunal Patel, Christian Domingues and Chiu-Hsieh Hsu
Pathophysiology 2025, 32(4), 62; https://doi.org/10.3390/pathophysiology32040062 - 11 Nov 2025
Viewed by 133
Abstract
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 [...] Read more.
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. Full article
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17 pages, 1119 KB  
Article
Cytokine and Metabolomic Signatures of Mepolizumab Response Across Upper and Lower Airway Compartments in Severe Eosinophilic Asthma: An Exploratory Analysis
by Mauro Maniscalco, Pasquale Ambrosino, Claudio Candia, Antonino Di Stefano, Isabella Gnemmi, Martina Zappa, Nicolino Ambrosino, Dina Visca and Andrea Motta
Pharmaceuticals 2025, 18(11), 1704; https://doi.org/10.3390/ph18111704 - 10 Nov 2025
Viewed by 237
Abstract
Background: Mepolizumab improves asthma control in severe eosinophilic asthma (SEA). However, its multidimensional effects on airway and systemic biomarkers are still incompletely understood. Methods: In this prospective study, 15 SEA patients were evaluated at baseline (T0), 6 (T6 [...] Read more.
Background: Mepolizumab improves asthma control in severe eosinophilic asthma (SEA). However, its multidimensional effects on airway and systemic biomarkers are still incompletely understood. Methods: In this prospective study, 15 SEA patients were evaluated at baseline (T0), 6 (T6), and 12 months (T12) after starting mepolizumab. Lung function, FeNO values, asthma control, blood eosinophil count (BEC), cytokines, and metabolomic profiles (1H-NMR) were evaluated in serum, nasal secretions, and exhaled breath condensate (EBC). Univariate and multivariate (PCA, OPLS-DA) analyses were performed. Results: Mepolizumab reduced exacerbations, from a median of 2 at T0 to 0 at both T6 (p = 0.001) and T12 (p = 0.003). ACT improved from 18.7 ± 4.7 at baseline to 23.0 ± 2.8 at T6 (p = 0.026) and 23.4 ± 3.3 at T12 (p = 0.032), while FEV1 increased by 270 mL at T6 (p = 0.032) and remained stable at T12. Median BEC decreased from 450.0 (350.0–560.0) to 65.0 (50.0–87.5) cells/μL at T6 and to 50.0 (35.0–160.0) at T12 (p < 0.001), while FeNO showed a non-significant downward trend. IL-13 significantly decreased in serum and nasal secretions at T6 and T12, while IL-5 increased in nasal secretions at both timepoints and remained unchanged in serum. IL-2 showed opposite trends in serum and nasal samples, whereas GM-CSF and IFN-γ increased in nasal secretions at T12. Metabolomic profiling suggested compartment-specific changes, with decreased short-chain alcohols in EBC, increased amino acids in nasal secretions and serum at T6, and elevated pyruvate in serum at T12, although none reached statistical significance in univariate analysis. Conclusions: Mepolizumab induced consistent clinical, immunologic, and metabolic changes across compartments, supporting the use of integrated cytokine and 1H-NMR metabolomic profiling as a complementary approach for response assessment in SEA. Full article
(This article belongs to the Special Issue Advances in Pharmacotherapy for Nasal Disorders in Rhinology)
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14 pages, 1430 KB  
Article
Ensemble-Based Refinement of Landmark Annotations for DNA Ploidy Analysis in Digital Pathology
by Viktor Zoltán Jónás, Dániel Küttel, Béla Molnár and Miklós Kozlovszky
Appl. Sci. 2025, 15(22), 11892; https://doi.org/10.3390/app152211892 - 8 Nov 2025
Viewed by 172
Abstract
Reliable evaluation of image segmentation algorithms in digital pathology depends on high-quality annotation datasets. Landmark-type annotations, essential for cell-counting analyses, are often limited in quality or quantity for segmentation benchmarking, particularly in rare assays where annotation is scarce and costly. In this study, [...] Read more.
Reliable evaluation of image segmentation algorithms in digital pathology depends on high-quality annotation datasets. Landmark-type annotations, essential for cell-counting analyses, are often limited in quality or quantity for segmentation benchmarking, particularly in rare assays where annotation is scarce and costly. In this study, we investigate whether ensemble-inspired refinement of landmark annotations can improve the robustness of segmentation evaluation. Using 15 fluorescently imaged blood samples with more than 20,000 manually placed annotations, we compared three segmentation algorithms—a threshold-based method with clump splitting, a difference-of-Gaussians (DoG) approach, and a convolutional neural network (StarDist)—and used their combined outputs to generate an ensemble-derived ground truth. Confusion matrices and standard metrics (F1 score, precision, and sensitivity) were computed against both manual and ensemble-derived ground truths. Statistical comparisons showed that ensemble-refined annotations reduced noise and decreased mean offsets between annotations and detected objects, yielding more stable evaluation metrics. Our results demonstrate that ensemble-based ground truth generation can guide targeted revision of manual markers, provide a quality measure for annotation reliability, and generate new annotations where no human-generated landmarks exist. This methodology offers a generalizable strategy to strengthen annotation datasets in image cytometry, enabling robust algorithm evaluation in DNA ploidy analysis and potentially in other low-frequency assays. Full article
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15 pages, 1122 KB  
Case Report
Training, Immunity, and Health in Elite Kayaking: A Longitudinal Study Monitoring a World-Class Marathon Paddler with Exercise-Induced Bronchoconstriction
by José Augusto Rodrigues dos Santos, Tiago Rama, Liliana Carina Baptista, Ana Isabel Padrão and Rodrigo Zacca
Sports 2025, 13(11), 401; https://doi.org/10.3390/sports13110401 - 7 Nov 2025
Viewed by 285
Abstract
Background: Exercise-induced bronchoconstriction (EIB) is common in athletes, being more frequent in outdoor endurance-based/long-distance sports. We followed a World-Class marathon paddler’s season with recurrent episodes of EIB, which intensified during cold exposure workouts. This unique immunophenotype profile during the season and its variations [...] Read more.
Background: Exercise-induced bronchoconstriction (EIB) is common in athletes, being more frequent in outdoor endurance-based/long-distance sports. We followed a World-Class marathon paddler’s season with recurrent episodes of EIB, which intensified during cold exposure workouts. This unique immunophenotype profile during the season and its variations were reflected in acute and chronic inflammatory markers. Methods: A longitudinal case study was conducted with blood sampling obtained from a single paddler after overnight fasting at three timepoints: T1 (beginning of season, after 15-day rest period), T2 (post-Winter National Championship), and T3 (post-Summer National Championship). Complete blood counts and lymphocyte immunophenotyping were performed using automated hematology analysis and multiparametric flow cytometry. Results: The total numbers of leukocytes (T1: 6.3; T2: 5.0; T3: 5.5 × 109/L), neutrophils (3.1; 2.5; 2.8 × 109/L), and lymphocytes (2.4; 1.8; 2.2 × 109/L) declined between T1 and T2, followed by a partial recovery at T3. In contrast, monocyte counts exhibited the reverse pattern (0.41; 0.62; 0.31 × 109/L). The two T cell subsets (αβ and γδ) remained relatively stable, showing only minor seasonal fluctuations. CD19+ B cells, initially at very low levels, increased steadily as the season progressed (0.05; 0.07; 0.16 × 109/L). During T2, the proportion of memory lymphocytes (CD45RO+) rose, while naive cells (CD45RA+) declined; this trend was subsequently inverted at M3. Although the CD4+/CD8+ ratio varied over time, it consistently stayed below the normal reference range established for healthy controls (0.50; 0.83; 0.60 for T1, T2, and T3, respectively). Conclusions: The immune assessment of the World-Class marathon paddler revealed transient immunosuppression early in the season, marked by reduced neutrophils, a low CD4+/CD8+ ratio, and diminished CD19+ lymphocytes. Over time, immune parameters showed signs of recovery, indicating a temporary imbalance that did not impair the athlete’s physical performance. Conclusions: This case study of an elite marathon kayaker revealed transient immune fluctuations across a competitive season, including early immunosuppression (low neutrophils, CD4+/CD8+ ratio 0.50, and minimal CD19+ B cells) followed by partial recovery mid- and late-season. Despite persistently inverted CD4+/CD8+ ratios suggesting chronic immune dysregulation, the athlete maintained competitive performance, highlighting the temporary nature of these changes and emphasizing that regular immune monitoring can help optimize health and performance in elite athletes. Full article
(This article belongs to the Collection Human Physiology in Exercise, Health and Sports Performance)
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10 pages, 467 KB  
Article
Type I Interferon-Related Gene Expression and Laboratory Abnormalities in Acute Infection Are Associated with Long COVID Symptom Burden
by Mary Emmanouil, Vasiliki E. Georgakopoulou, Konstantinos Drougkas, Panagiotis Lembessis, Charalampos Skarlis, Aikaterini Gkoufa, Nikolaos V. Sipsas and Clio P. Mavragani
J. Clin. Med. 2025, 14(21), 7875; https://doi.org/10.3390/jcm14217875 - 6 Nov 2025
Viewed by 199
Abstract
Background: Long COVID—defined as the persistence of symptoms or the development of new symptoms beyond four weeks after acute SARS-CoV-2 infection—affects an estimated 10–30% of individuals recovering from COVID-19, posing a significant public health burden. Emerging evidence suggests that type I interferons (IFNs) [...] Read more.
Background: Long COVID—defined as the persistence of symptoms or the development of new symptoms beyond four weeks after acute SARS-CoV-2 infection—affects an estimated 10–30% of individuals recovering from COVID-19, posing a significant public health burden. Emerging evidence suggests that type I interferons (IFNs) (a critical group of cytokines in the antiviral defense) and hematologic alterations, such as lymphopenia and elevated inflammatory markers, are linked to both the severity of acute COVID-19 and the likelihood of developing long-term symptoms. The aim of this study is to explore the association between type I IFN signatures and long COVID. A second aim is to examine the relationship between laboratory findings during acute infection and long COVID. Methods: The study included 61 patients investigated for the presence of long COVID symptoms 16.5 ±1.5 months after acute infection. Patients were divided into two groups of higher symptom burden of long COVID and those with milder symptoms based on demographic, laboratory, and clinical data as well as type I IFN-inducible gene expression (MX-1, IFIT-1, and IFI-44) measured in peripheral blood by real-time PCR. Data collected during acute infection were recorded. Peripheral blood samples were collected during the acute phase of infection, within the first 48 h of hospital admission. IFN-inducible gene expression was measured prospectively at that time, and RNA was extracted immediately for subsequent analysis. Results: History of intubation emerged as a significant associated factor of severe long COVID, with 75% of intubated patients reporting >8 persistent symptoms approximately 16 months post-infection. Higher white blood cell (WBC) and neutrophil counts but lower eosinophil and monocyte counts in acute infection were found to be associated with a high burden of long COVID symptoms. Interestingly, absolute monocyte count was found to independently correlate with higher long COVID symptom burden. Lactate dehydrogenase (LDH) and serum glutamic-oxaloacetic transaminase (SGOT) also differed significantly between groups, with higher levels correlating with a high burden of long COVID symptoms. Notably, MX-1 transcript levels in peripheral blood at the time of acute infection were reduced in patients with a high burden of long COVID symptoms, suggesting that dysregulated immune responses during the acute phase may contribute to persistent symptoms. Conclusions: These findings suggest the potential association of hematological and immune markers with long COVID severity, as well as the importance of monitoring these parameters to identify at-risk patients for early interventions. Full article
(This article belongs to the Section Clinical Laboratory Medicine)
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16 pages, 1040 KB  
Article
Cell-Free DNA Hypermethylation in Patients with Acute Pancreatitis
by Hassan Al-Mashat, Daniel Roger Baddoo, Søren Lundbye-Christensen, Poul Henning Madsen, Inge Søkilde Pedersen, Henrik B. Krarup, Benjamin Emil Stubbe, Ole Thorlacius-Ussing and Stine Dam Henriksen
Int. J. Mol. Sci. 2025, 26(21), 10792; https://doi.org/10.3390/ijms262110792 - 6 Nov 2025
Viewed by 236
Abstract
Cell-free DNA (cfDNA) promoter hypermethylation shows promise as a blood-based biomarker for pancreatic cancer, and similar alterations may occur in acute pancreatitis (AP). This study investigated the cfDNA hypermethylation profile of AP patients over time, compared with healthy controls, and its association with [...] Read more.
Cell-free DNA (cfDNA) promoter hypermethylation shows promise as a blood-based biomarker for pancreatic cancer, and similar alterations may occur in acute pancreatitis (AP). This study investigated the cfDNA hypermethylation profile of AP patients over time, compared with healthy controls, and its association with AP severity markers. A prospective longitudinal study including hospitalized AP patients and healthy controls was conducted. Methylation-specific PCR of a 23-gene panel was performed on plasma collected at inclusion (T0), 6 weeks (T6W), 6 months (T6M), and 7–8 years (T8Y). Associations between gene hypermethylation and clinical markers of AP severity—CRP, leukocyte count, creatinine, hospital stay, and complications—were evaluated. AP patients had a significantly higher mean number of hypermethylated genes at T0 (7.4, 95% CI: 6.8–8.0) compared with the controls (3.3, 95% CI: 2.8–3.8; p < 0.01). The mean number decreased over time to 3.2 (95% CI: 2.4–4.1) at T8Y. Total hypermethylation was positively associated with CRP (ρ = 0.39; p = 0.0018), leukocytes (ρ = 0.35; p = 0.0052), and hospital stay (ρ = 0.27; p = 0.0375). AP patients exhibited significantly higher cfDNA hypermethylation at disease onset, which normalized over time. Total hypermethylation showed positive associations with several markers of AP severity. Full article
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13 pages, 1157 KB  
Article
Evaluation of Total Eosinophil Counts, Serum Allergen-Specific IgE and Related Cytokines in Dogs with Atopic Dermatitis
by Min-Joo Chae, Min-Hee Kang and Hee-Myung Park
Animals 2025, 15(21), 3219; https://doi.org/10.3390/ani15213219 - 5 Nov 2025
Viewed by 269
Abstract
Canine atopic dermatitis (AD) is a chronic allergic skin disease in which various immunological markers have been investigated. While peripheral eosinophil counts, serum allergen-specific immunoglobulin E (IgE), and cytokines have each been evaluated in allergic disorders, their simultaneous assessment in dogs with AD [...] Read more.
Canine atopic dermatitis (AD) is a chronic allergic skin disease in which various immunological markers have been investigated. While peripheral eosinophil counts, serum allergen-specific immunoglobulin E (IgE), and cytokines have each been evaluated in allergic disorders, their simultaneous assessment in dogs with AD has rarely been reported in Korea. This study aimed to evaluate the diagnostic and clinical utility of these parameters in affected dogs. A total of 93 dogs were included between August 2019 and February 2020, comprising 65 dogs diagnosed with AD and 28 healthy controls. Clinical information, peripheral blood eosinophil counts and ratios, serum allergen-specific IgE using a multiple allergen panel (60 allergens), and cytokines related to T helper 2 (Th2) and T regulatory (Treg) cells (IL-4, IL-13, IL-31, TGF-β1) were analyzed. The mean age of AD dogs was 6.34 ± 3.99 years, with a predominance of small breeds and males. Eosinophil counts and ratios showed no significant difference between groups. In contrast, allergen-specific IgE levels were significantly elevated for several allergens, including Dermatophagoides pteronyssinus, Acarus siro, Tyrophagus putrescentiae, alder/birch, hazel, oak, cladosporium, and selected dietary antigens (pea, soybean, pumpkin, apple) (p < 0.05). Sensitization rates were also higher for Acarus siro, Tyrophagus putrescentiae, oak, and sheep sorrel (p < 0.05). Th2-related cytokines tended to increase and TGF-β1 tended to decrease in AD dogs, though without statistical significance. These findings indicate that peripheral eosinophil counts have limited diagnostic value, whereas allergen-specific IgE testing provides clinically useful information for the diagnosis and management of canine AD. Further research stratifying disease stages and assessing local tissue cytokine expression is warranted. Full article
(This article belongs to the Section Veterinary Clinical Studies)
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18 pages, 3680 KB  
Article
Polymicrobial Infection (Gram-Positive and Gram-Negative) Exacerbates Systemic Inflammatory Response Syndrome in a Conscious Swine Extremity Trauma Model
by Catharina C. Gaeth, Travis R. Madaris, Jamila M. Duarte, Amber M. Powers, Christina M. Sandoval, Stefanie M. Shiels and Randolph Stone
Pathophysiology 2025, 32(4), 59; https://doi.org/10.3390/pathophysiology32040059 - 4 Nov 2025
Viewed by 262
Abstract
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 [...] Read more.
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. Full article
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23 pages, 9119 KB  
Article
Protective Effects of Velvet Antler Polypeptides on Cyclophosphamide-Induced Myelosuppression in Mouse and Bone Marrow Mesenchymal Stem Cells
by Fusheng Gao, Yusu Wang, Jinze Liu, Yichen Xie, Ying Geng, Zhongmei He, Jianan Geng, Jianming Li, Weijia Chen and Rui Du
Nutrients 2025, 17(21), 3428; https://doi.org/10.3390/nu17213428 - 31 Oct 2025
Viewed by 368
Abstract
Background: Myelosuppression is one of the most common chemotherapy side effects, seriously threatening the quality of life of cancer patients. Studies have shown that velvet antler polypeptides (VAPs) could enhance immunity and anti-aging and also have a hematopoietic-promoting effect. However, there are relatively [...] Read more.
Background: Myelosuppression is one of the most common chemotherapy side effects, seriously threatening the quality of life of cancer patients. Studies have shown that velvet antler polypeptides (VAPs) could enhance immunity and anti-aging and also have a hematopoietic-promoting effect. However, there are relatively few studies on the treatment of myelosuppression with VAPs, and the therapeutic mechanism remains unclear. Methods: This study employed both in vitro and in vivo models to explore the mechanism of VAPs against myelosuppression. In this study, the cyclophosphamide (CTX)-induced bone marrow mesenchymal stem cell (BMSC) injury model was used to evaluate the effects of VAPs on cell viability, apoptosis, reactive oxygen species activity, and protein expression. Furthermore, a CTX-induced myelosuppression mouse model was employed to evaluate peripheral blood counts, organ indices, femoral tissue histopathology, immunohistochemical expression of CD34, VEGF, and Notch1, and key proteins in the Notch1/PI3K/AKT pathway in vivo. Results: Our results showed that VAPs protected BMSCs from CTX-induced apoptosis, inhibited ROS production, and promoted the secretion of VEGF, TPO, and VCAM-1, thereby improving the bone marrow microenvironment. Furthermore, the results showed that VAPs improved the peripheral blood counts and bone marrow nucleated cell (BMNC) count in CTX-induced myelosuppression mice and ameliorated pathological injury of the spleen, thymus, and liver. VAPs inhibited the apoptosis of bone marrow cells, manifested by regulating the expression levels of proteins like PI3K/p-PI3K, AKT/p-AKT, Bcl-2, Bax, and Caspase-3. Simultaneously, it upregulated the expression of Notch1 and Hes1 proteins. The application of the PI3K inhibitor LY294002 and the Notch1 inhibitor DAPT demonstrated that the ameliorative effect of VAPs on myelosuppression was dependent on the activation of both the Notch1 and PI3K/AKT pathways. Conclusions: Our study indicates that VAPs may achieve treatment of myelosuppression by improving the hematopoietic microenvironment, inhibiting apoptosis of mouse bone marrow cells, and regulating the Notch1 and PI3K/AKT signaling pathways. Full article
(This article belongs to the Section Proteins and Amino Acids)
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12 pages, 251 KB  
Article
Evaluation of Sleep Disturbances and Depression in Children with Juvenile Idiopathic Arthritis Using the Beck Depression Inventory and Child Sleep Disorder Scale: Relationship with Leukocyte and Neutrophil Counts
by Seyda Dogantan, Evin İlter Bahadur, Burcu Bozkaya Yücel, Adem Keskin and Esma Bekece
Children 2025, 12(11), 1470; https://doi.org/10.3390/children12111470 - 31 Oct 2025
Viewed by 225
Abstract
Background/Objectives: The pathophysiology of juvenile idiopathic arthritis (JIA), the most widespread rheumatologically illness in juvenile period, is shaped by complex interactions between leukocytes and the cytokines they secrete. The aim of this research was to evaluate the severity of sleep disturbances and [...] Read more.
Background/Objectives: The pathophysiology of juvenile idiopathic arthritis (JIA), the most widespread rheumatologically illness in juvenile period, is shaped by complex interactions between leukocytes and the cytokines they secrete. The aim of this research was to evaluate the severity of sleep disturbances and depression, which are closely associated with many diseases and can negatively impact the course of the illness, in patients with JIA using Beck Depression Inventory (BDI) and Sleep Disturbance Scale for Children (SDSC) scores and to investigate the relationship between these scores and laboratory findings in patients with JIA. Methods: The research involved 58 children with JIA and 71 healthy children as controls. BDI and SDSC scores of these groups were compared with laboratory findings and correlation analysis were performed. Results: In the JIA group, BDI and SDSC scores, C-reactive protein (CRP), red blood cell distribution width (RDW), erythrocyte sedimentation rate, neutrophil, and leukocyte counts, were higher than in the control group, while vitamin D values were lower. A positive relation was determined between BDI and SDSC scores in the JIA group, but no correlation was found in the control group. In the JIA group, both BDI and SDSC scores were found to be negatively related with leukocyte and neutrophil counts. In the control group, the BDI score was determined to be negatively correlated with CRP, vitamin D and RDW levels. Conclusions: Depression and sleep disorders may interact in patients with JIA, and their causal relationship with leukocyte and neutrophil levels should be investigated. Full article
(This article belongs to the Section Pediatric Allergy and Immunology)
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