Search Results (160)

Sarcoptic mange is a skin disease caused by Sarcoptes scabiei infestations, characterized by dermatitis, pruritus, and exudative responses in both humans and animals. Biologically, the life cycle of S. scabiei is confined to the host’s skin (stratum corneum), where mite-derived molecules trigger the influx of innate immune cells, including polymorphonuclear neutrophils (PMN), which play a central role in skin inflammatory responses. The antimicrobial activity of PMNs is regulated by Ca²⁺ fluxes and includes the generation of reactive oxygen species (ROS), degranulation, and the release of neutrophil extracellular traps (NETs). NETs are web-like structures composed of chromatin and enzymes that can trap and eventually kill pathogens; however, their involvement in S. scabiei infestations in bovines remains unclear. Here, we investigated interactions between bovine PMN and S. scabiei mites, as well as PMN responses to S. scabiei antigen (ScAg). Functional parameters included NET release, Ca²⁺ fluxes, ROS production and phagocytic activity, determined by fluorescence microscopy, Fluo-4 staining, luminol-derived luminescence and flow cytometry, respectively. Current data show that ScAg, but not whole mites, induces a weak NET release in exposed bovine PMN. Additionally, ScAg drives rapid and sustained Ca²⁺ fluxes and ROS production over time, without altering the phagocytic capacity of PMN. Full article

Neutrophil extracellular traps (NETs) contribute to innate immunity in sepsis, but their diagnostic value in neonates is unclear. We evaluated whether circulating NET-associated biomarkers discriminate septic from non-infected neonates. In this prospective observational study 96 neonates (≥34 weeks gestational age) with clinical suspicion of infection were enrolled (36 sepsis, 60 controls). Serum cell-free DNA (cfDNA), myeloperoxidase–DNA complexes (MPO-DNA), neutrophil elastase–DNA complexes (NE-DNA), and citrullinated histone H3 (H3cit) were measured alongside CRP and IL-6 at days 1, 3, and 5. Diagnostic performance was assessed by receiver operating characteristic (ROC) analysis with bootstrap confidence intervals. CRP (AUC 0.75, 95% CI 0.66–0.85) and IL-6 (AUC 0.73, 95% CI 0.61–0.83) showed the best diagnostic performance. cfDNA demonstrated moderate discrimination (AUC 0.72, 95% CI 0.60–0.84) but was only transiently elevated at day 1. MPO-DNA (AUC 0.47), NE-DNA (AUC 0.44), and H3cit (AUC 0.47) performed no better than chance. Within the sepsis group, MPO-DNA and NE-DNA at day 3 strongly correlated with the immature-to-total neutrophil ratio (ρ = 0.76 and 0.72), suggesting these markers reflect neutrophil degranulation rather than NET formation. NET-associated biomarkers do not improve diagnostic accuracy for neonatal sepsis beyond CRP and IL-6. These findings support the concept that neonatal innate immune responses differ fundamentally from adult patterns. Full article

Lung cancer is a leading cause of death worldwide and is often accompanied by declines in musculoskeletal health (i.e., cachexia). Despite affecting a majority of lung cancer patients, cachexia remains understudied and currently has no cure. We have previously demonstrated that liver metastases (LMs) exacerbate cachexia in murine models of colorectal cancer, and, while the liver represents a common site of metastases and is associated with poor prognosis in patients with lung cancer, whether LMs heighten musculoskeletal wasting in mice bearing lung cancer is unknown. Here, we aimed to characterize the impact of LMs on musculoskeletal health in a mouse model of lung cancer cachexia. C57BL/6J male mice were injected with LLC tumor cells either subcutaneously or intrasplenically (LMs) to mimic hepatic metastases (n = 6–9/group). Upon sacrifice, skeletal muscle, bone, and plasma were collected for morphological and molecular analyses. Consistently, compared to healthy controls, metastatic tumor hosts displayed greater reductions in muscle weights (~17%), in line with decreased muscle torque (~23%) and reduced muscle cross-sectional area (~10%). On a molecular level, skeletal muscle from mice bearing LMs had elevated levels of pStat3, Murf1, and Atrogin-1, suggesting enhanced protein catabolism. Similar to skeletal muscle, metastatic tumor hosts displayed greater losses in trabecular bone and increased skeletal fragility. Plasma proteomics identified 211 and 131 differentially expressed proteins in metastatic hosts compared to control animals and subcutaneous LLC hosts, respectively. Top regulated pathways in mice bearing LMs included neutrophil degranulation, BAG2 signaling, and cachexia signaling. Overall, our findings demonstrate that LMs are accompanied by accelerated musculoskeletal wasting and weakness in a mouse model of lung cancer cachexia. This work highlights the need for animal models that mimic advanced cancer, thus providing a better understanding of the mechanisms that mediate cachexia. Full article

Endometrial cancer (EC) is the leading gynecological malignancy worldwide. Obesity is reported to be associated with 50% of EC cases. Significant gaps remain in investigating specific molecular mechanisms behind the obesity-driven EC. Women diagnosed with EC undergoing total hysterectomy were recruited. Patients were divided into two groups: EC-obese with BMI > 29.9 kg/m² (n = 10) and EC-Non-obese with BMI ≤ 29.9 kg/m² (n = 10). Tumor tissues were subjected to label-free quantitative proteomic analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Differentially expressed proteins were identified and subjected to pathway enrichment and network analyses to characterize obesity-associated alterations. Proteomic profiling showed a significant dysregulation of 456 proteins: 171 upregulated and 285 downregulated. Proteins involved in endoplasmic reticulum quality control particularly endoplasmic reticulum lectin 1 (ERLEC1), were reduced. Conversely, EC-obese demonstrated upregulation of hepatocyte growth factor (HGF), integrin-linked kinase (ILK), CTTNBP2 N-terminal-like protein (CTTNBP2NL), and lysyl oxidase homolog 1 (LOXL1), implicating activation of inflammatory pathways. Bioinformatic analysis showed downregulation of immune-related pathways, including neutrophil degranulation, complement activation in the EC-obese group. ROC analysis identified apolipoprotein(a), phospholipase B-like 1, CTTNBP2NL, and ILK as significant proteins that can differentiate between the obese and non-obese states. Our findings provide insights into obesity-associated proteomic changes in EC and highlight candidate proteins that can be used for molecular stratification after further validation. Full article

Background: Dysregulated host response to infection plays vital roles in the high mortality rate of sepsis. Better understanding of the dysregulated host response is required for the improved management of sepsis. Methods: In this work, we conducted side-by-side comparison of several components of host immunity in sepsis with a composite marker gene panel. This gene panel consisted of five sets of marker genes, including the NDrG gene set for neutrophil degranulation-related genes, the ISGa gene set for type I interferon signaling, the GBPs gene set for type II interferon signaling, the HLAd gene set for major histocompatibility (MHC) class II, and the LYMd gene set for lymphocytes and dendritic cells. Seven relevant transcriptome datasets were used for the evaluation. Results: It was found that sepsis was characterized by simultaneous activation of the NDrG gene set and suppression of the HLAd and LYMd gene sets. In contrast, both activation and suppression of the ISGa and GBPs gene sets were observed in sepsis. Compared to sepsis patients with hypo-inflammation, those with hyper-inflammation displayed higher values of NDrG and lower values of HLAd and LYMd. Reversal of gene dysregulation was observed in the NDrG, HLAd and LYMd gene sets for sepsis patients after treatment at intensive care unit (ICU), especially for those who responded better to the treatment. In a cohort of sepsis patients admitted to the ICU, the initial expression values of the NDrG, HLAd and LYMd gene sets were associated with the outcome. Notably, all patients with (NDrG-HLAd) < 1.5 survived, accounting for 75% of the survivors (45 out of 60). A simple combination of the (NDrG-HLAd) value and the sequential organ failure assessment (SOFA) score could identify 90% of the survivors with 3.5% false positive rate. Conclusions: Overall, this composite gene panel is applicable to the serial monitoring of immune dysregulation in sepsis. It also suggests that the NDrG and HLAd gene sets may be promising targets for immune modulation in sepsis. Full article

Epidemiological data link a lack of breastfeeding with an increased risk of breast cancer. Breast tissue remodels after pregnancy through involution. Long-term breastfeeding results in gradual involution (GI), and a lack of breastfeeding leads to abrupt involution (AI). AI causes increased mammary gland estrogen signaling, causing adipocyte redifferentiation through neutrophil infiltration. Adipocyte differences and metabolic implications of involution have not been explored between AI and GI. As breast cancer is characterized as highly metabolic, we explored how adipocyte differences and metabolism during involution may support breast cancer risk. FVB/n was randomized to AI/GI and standardized to 6 pups on day 0/birth. AI mice had pups removed on day 7. GI mice had 3 pups removed on days 28 and 31. Mammary glands were harvested at 28, 56, and 120 days. A subset of AI mice were given tamoxifen for 21 days. Day 28 AI glands had upregulation of estrogen signaling, neutrophil degranulation, and glucose metabolism and downregulation of adipogenesis and glycolysis compared to Day 56 GI. Day 120 AI glands had downregulation of oxidative phosphorylation and upregulation of mitochondrial dysfunction similar to estrogen receptor-negative (ER−) pregnancy-associated breast cancer (PABC). AI with tamoxifen resulted in a similar metabolic phenotype to GI. Early metabolic phenotypes in AI and GI glands may be related to estrogen signaling. AI long-term transcriptional metabolic effects were similar to breast cancer. Full article

Ozone (O₃) is a reactive oxidant increasingly applied in biomedical settings, yet its dose-dependent effects on innate immune cells, particularly those from non-human species, remain insufficiently defined. Within a One Health framework, this study examined how two clinically relevant O₃ exposure regimens (30 µg/mL and 90 µg/mL) affect porcine neutrophils and monocytes isolated from peripheral blood. Cell viability, reactive oxygen and nitrogen species (RONS) production, and the activity of key enzymes (myeloperoxidase, elastase, alkaline phosphatase, arginase) were assessed at 1 h and 24 h post-exposure. The lower dose induced mild functional activation without compromising viability, whereas the higher dose triggered pronounced oxidative stress, enhanced degranulation, and reduced neutrophil viability by more than 60%. Neutrophils exhibited a stronger and more dynamic response than monocytes, which retained viability and differentiation capacity at 30 µg/mL but showed impaired function at 90 µg/mL. These findings highlight the dual nature of O₃, where controlled exposure may support immunomodulation, while excessive dosing disrupts cell function. Defining safe and effective therapeutic windows remains critical for future applications. Full article

Medication-related osteonecrosis of the jaw (MRONJ) is a serious adverse effect of antiresorptive and antiangiogenic therapies, yet its molecular mechanisms remain poorly defined. The present study employed an analysis of microarray data (GSE7116) from peripheral blood mononuclear cells of patients with multiple myeloma, myeloma patients with MRONJ, and healthy controls. Differentially expressed genes were identified using the limma package, followed by functional enrichment analysis, weighted gene co-expression network analysis, and LASSO regression and CytoHubba network ranking. The predictive performance was validated by means of nested cross-validation, Firth logistic regression, and safe stratified 0.632+ bootstrap ridge regression. The profiling revealed distinct gene expression patterns between the groups: the upregulation of ribosomal and translational pathways, as well as the suppression of neutrophil degranulation and antimicrobial defense mechanisms, and identified key candidate genes, including PDE4B, JAK1, ETS1, EIF4A2, FCMR, IGKV4-1, and XPO7. These genes demonstrated substantial discriminatory capability, with an area under the curve ranging from 0.95 to 0.99, and were found to be functionally linked to immune system dysfunction, cytokine signaling, NF-κB activation, and a maladaptive stress response. These findings link MRONJ to systemic immune-inflammatory imbalance and translational stress disruption, offering novel insights and potential biomarkers for diagnosis and risk evaluation. Full article

Neutrophils are the most abundant circulating leukocytes and essential components of innate immunity. Through mechanisms such as phagocytosis, reactive oxygen species (ROS) production, degranulation, and neutrophil extracellular trap (NET) formation, they play a crucial role in host defense. However, dysregulated neutrophil responses are linked to chronic inflammatory conditions, including periodontitis. This review summarizes current evidence on neutrophil biology in periodontal health and disease, focusing on functional mechanisms, recruitment pathways, the influence of dysbiosis, and their potential as biomarkers and therapeutic targets. Neutrophils display a dual role in periodontal tissues: while protecting against microbial invasion, their excessive or impaired activity contributes to tissue destruction. Altered chemotaxis, defective phagocytosis, and uncontrolled NET release perpetuate inflammation and alveolar bone loss. Neutrophil-derived enzymes, including myeloperoxidase, elastase, and matrix metalloproteinases, emerge as promising biomarkers for early diagnosis. In parallel, therapeutic strategies targeting oxidative stress, NET regulation, or neutrophil hyperactivity are being explored to preserve periodontal tissues. Neutrophils are central players in periodontal pathophysiology. Understanding their regulation and interaction with the oral microbiome may enable the development of novel diagnostic and therapeutic approaches, ultimately improving periodontal disease management. Full article

Neutrophilic asthma (NA) is an inflammatory phenotype of asthma, characterized by predominantly neutrophilic infiltrations in bronchial mucosa. It is usually diagnosed on the basis of high neutrophil count in induced sputum (from >40% to >76%) with low eosinophils (<2%). The prevalence of NA ranges from 16% to 28% of the adult asthma population depending on the definitions and study methods applied. A clinical picture of NA is characterized by late onset of symptoms, higher exacerbation rate, lower level of symptoms control, and poorer response to steroids compared to eosinophilic phenotype. Comorbidities such as obesity and GERD as well as the influence of environmental factors (air pollution, smoking, bacterial infections) contribute to the development and severe course of the disease. NA is T2-low disease with predominantly Th1/Th17-type inflammation. Neutrophils are key cells responsible for initiating and sustaining inflammation. In addition to their primary functions like phagocytosis, degranulation, and NETosis, neutrophils release several pro-inflammatory cytokines (IL-1α, IL-1β, IL-6, TNF) and chemokines (CXCL-1, -2, -8, -9, -10) responsible for the recruitment of other neutrophils or T cells. Increasing knowledge about the biology of neutrophiles and their role in asthma results in new potential therapies that could improve control of NA, particularly new biologicals targeting Th1/Th17-related cytokines. In this review, we discuss the prevalence, mechanisms, and clinical features of neutrophilic asthma. Furthermore, current therapeutic options and some promising perspectives for the near future are presented. Full article

Clinical studies have linked glyphosate exposure to substantial morbidity, with acute kidney injury occurring in some cases. Although the toxic effects of glyphosate-based herbicides (GBHs) have been reported in several studies, their molecular impact on renal function remains poorly understood. Given the kidney’s critical role in excretion, it is particularly susceptible to damage from xenobiotic exposure. In this study, we aim to identify N-glycomics and proteomics change in the kidney following chronic GBH exposure, to better understand the mechanisms behind glyphosate-induced kidney damage. Kidney tissues from female and male rats were analyzed using liquid chromatography–tandem mass spectrometry. The results revealed notable changes in the N-glycan composition, particularly in the fucosylated and sialofucosylated N-glycan types. The proteomic analysis revealed the activation of immune signaling and inflammatory pathways, including neutrophil degranulation, integrin signaling, and MHC class I antigen presentation. Transcription regulators, such as IL-6, STAT3, and NFE2L2, were upregulated, indicating a coordinated inflammatory and oxidative stress response. Sex-specific differences were apparent, with female rats exhibiting more pronounced alterations in both the N-glycan and protein expression profiles, suggesting a higher susceptibility to GBH-induced nephrotoxicity. These findings provide new evidence that chronic GBH exposure may trigger immune activation, inflammation, and potentially carcinogenic processes in the kidney. Full article

Community-acquired pneumonia (CAP) is a leading cause of death, with mortality linked to an unbalanced host response. Sirtuin (Sirt)1, a histone deacetylase, regulating metabolism and epigenetics, may be fundamental in activating the innate immune response. Sirt1 mRNA expression was significantly reduced in monocytes from CAP patients (n = 76) upon admission compared to healthy controls (n = 42), with levels returning to normal after 30 days. Pharmacological activation of Sirt1 with SRT1720 decreased LPS- and K. pneumoniae-induced IL-6 release in primary human monocytes and decreased NF-κB activation in THP1 cells. In a mouse K. pneumoniae pneumosepsis model, SRT1720 strongly reduced neutrophil influx and degranulation markers in bronchoalveolar lavage fluid, lowered pulmonary concentrations of IL-6 and TNF-α, and reduced lung pathology scores. Simultaneously, it reduced neutrophil content in liver tissue and plasma transaminase levels, alongside a trend toward reduced liver necrosis. Plasma IL-6 and TNF-α were significantly lower in SRT1720-treated mice at 42 h. Finally, while SRT1720 did not impact bacterial loads in the lungs, it reduced bacterial burden in blood, with a similar trend observed in liver homogenates. In conclusion, the Sirt1 activator SRT1720 exerts anti-inflammatory effects on human monocytes, reduces local and systemic inflammation and organ injury, and diminishes bacterial dissemination in murine pneumosepsis. Full article

Upon activation, neutrophils perform three distinct functions: phagocytosis, degranulation of antimicrobial substances into the extracellular medium, and release of neutrophil extracellular traps. Determination of the nuclear area expansion of neutrophils activated to release neutrophil extracellular traps has become critical in demonstrating early neutrophil activation and has become standard. Here, we demonstrate an automated method for measuring nuclear area expansion in two different mammalian species: canine and bovine. For both species, neutrophils were isolated from peripheral blood and co-incubated with fresh spermatozoa for up to 120 min for canine neutrophil–spermatozoa and recently thawed cryopreserved spermatozoa up to 240 min for bovine neutrophil–spermatozoa. Fluorescence images were acquired using a TissueFAXS microscope and then analyzed using StrataQuest v.7.0 software. The images show the release of neutrophil extracellular traps upon activation with spermatozoa for both species, as evidenced by the co-localization of neutrophil elastase and DNA staining. Neutrophil nuclei were expanded as early as 15 min and were detected at up to 120 min in both species. Analysis by nuclei segmentation showed that the data sets generated for both species were reliable and consistent with previously published methods. The method was developed as an automated alternative for measuring the area expansion of neutrophil nuclei in different species. Full article

Background and objective: Periodontitis, a chronic inflammatory disease driven by host immune dysregulation, leads to progressive destruction of periodontal tissues. This study employed an integrative approach combining single-cell transcriptomics, hierarchical weighted gene co-expression network analysis (hdWGCNA), and deep learning algorithms to identify key biomarkers associated with neutrophil degranulation in periodontitis, aiming to establish diagnostic models for early detection and precision interventions. Methods: We integrated single-cell RNA sequencing (scRNA-seq) data from human gingival tissues with bulk transcriptomic datasets. Pathogenic neutrophil subsets were characterized via pseudotime trajectory and cell–cell communication analyses. Hierarchical weighted gene co-expression network analysis (hdWGCNA) identified functional modules linked to degranulation. Machine learning and a convolutional neural network (CNN) model combining gene expression and immune cell profiles were developed for diagnosis. Results: scRNA-seq revealed a neutrophil subpopulation significantly increased infiltration in periodontitis, with cell–cell communication and pseudotime trajectory analyses demonstrating amplified inflammatory crosstalk. hdWGCNA identified the turquoise module enriched in PD-KEY-Neutrophils, containing hub genes linked to neutrophil degranulation and complement activation. Immune infiltration and non-negative matrix factorization linked high-degranulation neutrophil signatures to the periodontal immunity microenvironment. Machine learning demonstrated that the neutrophil degranulation-associated genes effectively distinguish diseased gingival tissue, suggesting their potential to predict periodontitis. Finally, integrating transcriptomic and immunological data, we developed a gene-immune CNN deep learning model accurately diagnosed periodontitis in diverse cohorts (AUC = 0.922). Conclusions: Our study identified a pathogenic neutrophil subpopulation driving periodontitis through degranulation and inflammation. The neutrophil degranulation genes serve as critical biomarkers, offering new insights for clinical diagnosis and treatment of periodontitis. Full article

Ankylosing spondylitis (AS) displays wide inter-patient variability that is not accounted for by HLA-B27 alone, suggesting that additional immune and metabolic modifiers contribute to disease severity. Using a genetically matched design, we profiled peripheral blood mononuclear cells from two brother pairs discordant for AS severity and one healthy brother pair. Strand-specific RNA-seq was analyzed with a family-blocked DESeq2 model, while untargeted metabolites were quantified using gas chromatography–mass spectrometry (GC-MS) and liquid chromatography–mass spectrometry (LC-MS). Differential features were defined as follows: differentially expressed genes (DEGs) (|log₂FC| ≥ 1 and FDR < 0.05) and metabolites (VIP > 1, FC ≥ 1.2, and BH-adjusted p < 0.05). Pathway enrichment was performed with KEGG and Gene Ontology (GO). A total of 325 genes were differentially expressed. Type I interferon and neutrophil granule transcripts (e.g., IFI44L, ISG15, S100A8/A9) were markedly up-regulated, whereas mitochondrial β-oxidation genes (ACADM, CPT1A, ACOT12) were repressed. Metabolomics revealed 110 discriminant features, including 25 MS/MS-annotated metabolites. Primary bile acid intermediates were depleted, whereas oxidized fatty acid derivatives such as 12-Z-octadecadienal and palmitic amide accumulated. Spearman correlation identified two antagonistic modules (i) interferon/neutrophil genes linked to pro-oxidative lipids and (ii) lipid catabolism genes linked to bile acid species that persisted when severe and mild siblings were compared directly. Enrichment mapping associated these modules with viral defense, neutrophil degranulation, fatty acid β-oxidation, and bile acid biosynthesis pathways. This sibling-paired peripheral blood mononuclear cell (PBMC) dual-omics study delineates an interferon-driven lipid–bile acid axis that tracks AS severity, supporting composite PBMC-based biomarkers for future prospective validation and highlighting mitochondrial lipid clearance and bile acid homeostasis as potential therapeutic targets. Full article