Search Results (242)

Background/Objectives: Septic arthritis of native joints is an orthopedic emergency in which rapid discrimination from non-infectious arthritis is crucial. Because cartilage damage can occur within hours, urgent irrigation and debridement are often pursued on an emergency basis (ideally within the first 6–8 h) of presentation, underscoring the need for rapidly available biomarkers. The delta neutrophil index (DNI) quantifies circulating immature granulocytes and may complement conventional inflammatory biomarkers such as C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), white blood cell count (WBC), and procalcitonin (PCT). We evaluated the diagnostic performance of DNI for native-joint septic arthritis against both microbiologic and clinical reference standards. Methods: We retrospectively analyzed 85 adults who underwent surgical irrigation and debridement for suspected native joint septic arthritis at a tertiary center. Serum CRP, ESR, WBC, DNI, and PCT (available in 67 patients) were recorded together with synovial leukocyte counts. Infection status was defined using either positive synovial culture (microbiologic reference) or clinical adjudication according to the Guideline for management of septic arthritis in native joints (SANJO). Diagnostic performance was assessed using receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC); exploratory cut-offs were identified by the Youden index, and pairwise AUCs were compared using DeLong’s test. Results: Synovial leukocyte analysis was highly sensitive but poorly specific (sensitivity 92.9%, specificity 10.3%). Against culture, DNI showed the highest discrimination (AUC = 0.914), exceeding CRP (0.687), ESR (0.643), WBC (0.648), and PCT (0.697); DeLong ΔAUC vs. CRP 0.227 (p < 0.001), ESR 0.270 (p < 0.001), WBC 0.266 (p < 0.001), PCT 0.227 (p = 0.001). At pre-specified cut-offs, DNI showed the most balanced sensitivity/specificity (94.3%/84.0%), corresponding to a negative predictive value (NPV) of 95.5% (42/44) and a positive predictive value (PPV) of 80.5% (33/41) against culture in this cohort. Against clinical infection, DNI outperformed others (AUC:0.921; ΔAUC vs. CRP = 0.204, ESR = 0.343, WBC = 0.244, PCT = 0.295; all p < 0.001). As a rule-in threshold, DNI ≥ 0.6 yielded a specificity of 100% with a sensitivity of 73.2%. In culture-negative patients (infected n = 21, uninfected n = 29), DNI remained discriminatory (AUC 0.80, p < 0.001), whereas other biomarkers were not. Conclusions: DNI demonstrated superior diagnostic accuracy compared with conventional inflammatory biomarkers. As a rapid parameter available with the initial complete blood count, DNI may support early risk stratification and rule-in decisions within the first hours of presentation; however, it should be used as a supplementary indicator alongside synovial fluid analysis and clinical assessment rather than as a stand-alone diagnostic tool. Full article

Background: Cytokine oncostatin M (OSM) is implicated in inflammatory conditions. The plant sterol stigmasterol (ST) is found in diverse plant foods and exerts various benefits, such as antitumor, antioxidant, and anti-inflammatory effects. However, the inhibitory mechanism of ST on OSM production in neutrophils needs to be elucidated. Methods: To evaluate the modulatory effects of ST, this investigation employed neutrophil-like differentiated (d)HL-60 cells. ELISA, real-time PCR, Western blotting, and immunofluorescence staining were conducted. dHL-60 cells were pretreated with ST (0.02 to 2 µg/mL) for 1 h, and then stimulated with GM-CSF (5 ng/mL). Results: Our results showed that addition of granulocyte–macrophage colony-stimulating factor (GM-CSF) leads to up-regulation of OSM mRNA and protein in dHL-60 cells, while pretreatment with ST reduces OSM mRNA and protein levels. Mechanistically, the highest dose (2 µg/mL) of ST significantly decreased phosphorylation of phosphatidylinositol 3-kinase, protein kinase B (Akt), and nuclear factor-κB. Conclusions: Our findings suggest that the plant sterol ST shows potential and warrants in vivo validation on OSM regulation via suppressing PI3K/Akt/NF-κB Signaling Processes. Full article

In this study, we report the development of a recombinant human G-CSF fused with apolipoprotein A-I. The chimeric protein was expressed in Pichia pastoris. Using human bone marrow cells, the fusion protein was shown to retain the granulocyte activity of authentic G-CSF, more effectively inducing the differentiation and maturation of segmented neutrophils and maintaining the viability of progenitor cells. Using human mononuclear cells and THP cells, the resulting protein demonstrated monocytic activity, manifested by an increase in both total and CD14+ cell counts. By maintaining cell viability, the chimeric protein reduced the number of cells expressing caspase 3/7. G-CSF-ApoAI demonstrated accelerated cytokine regulation, promoting a more rapid transition of inflammation phases, accompanied by increased phagocytosis of latex particles, compared with G-CSF, increasing phagocytosis by 1.4-fold in the LPS-induced inflammation model. This suggests that this new pleotropic factor may be useful for pathogen clearance in infected wounds. Full article

Glioblastoma (GBM) is the most common malignant primary brain tumor in adults. Since numerous studies highlight the significance of cholinergic system components in tumor development, acetylcholine (ACh) and the differential activation of its receptors could play a crucial role in GBM progression. The aim of this study was to test this hypothesis by assessing the relevance of the cholinergic system in GBM cells and their microenvironment. We analyzed bulk RNA-seq expression data using the TIMER2.0 web server, focusing on the impact of patient survival in relation to muscarinic receptors (CHRM) and neutrophil infiltration in low-grade glioma (LGG) and GBM. Our analysis revealed a marked decrease in survival associated with all CHRMs, particularly in LGG. Moreover, GBM showed higher neutrophil infiltration and reduced survival, especially in relation to CHRM3. These findings were validated in the U251 cell line and in human GBM tumor biopsies (GBM-b), which also displayed CHRM3 expression. Additionally, we show that GBM cells exposed to cholinergic stimulation exhibited increased vascular endothelial growth factor (VEGF), IL-8 production, and PD-L1 expression, while the VEGF increase was blocked by tiotropium (Tio), a CHRM3 antagonist. Similarly, polymorphonuclear cells from GBM patients (PMN-p) displayed increased PD-L1 expression and IL-8 production upon cholinergic stimulation. Finally, as we previously reported on the relevance of thymic stromal lymphopoietin (TSLP) in GBM pathophysiology, here, we found that TSLP upregulated CHRM3 expression. Our findings highlight the importance of the cholinergic system in the tumor microenvironment, where it may act directly on tumor cells or influence neutrophil physiology, thereby modulating tumor progression. Full article

Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by autoantibody production and the formation of immune complexes (ICs), which lead to widespread inflammation and tissue damage. Neutrophil extracellular traps (NETs), web-like structures composed of DNA, histones, and antimicrobial proteins released by activated neutrophils, play a crucial role in innate immunity by defending against pathogens. However, excessive NET formation and ineffective clearance of these structures contribute to the development of SLE. This review explores the mechanisms behind NET formation in SLE, their relationship with oxidative stress, and the potential role of antioxidants in treatment. Research indicates that SLE patients exhibit two key abnormalities: excessive NET formation and impaired NET clearance. Excessive NET formation is driven by proinflammatory low-density granulocytes (LDGs) and immune complexes (ICs). Impaired NET clearance stems from reduced DNase1/DNase1L3 activity or anti-nuclease autoantibodies. These two abnormalities lead to elevated circulating NETs. These NETs act as autoantigen reservoirs, forming pathogenic NET–ICs that amplify autoimmune responses. Oxidative stress drives NET formation by activating NADPH oxidase. In contrast, various antioxidants, including enzymatic and non-enzymatic types, can inhibit NET formation via scavenging reactive oxygen species (ROS) and blocking NADPH oxidase activation. Preclinical studies show that antioxidants such as curcumin, resveratrol, and mitochondrial-targeted MitoQ reduce NET formation and ameliorate lupus nephritis; clinical trials confirm that curcumin and N-acetylcysteine (NAC) lower SLE disease activity and reduce proteinuria, supporting their role as safe adjuvant therapies. However, high-dose vitamin E may exacerbate autoimmunity, highlighting the need for dose optimization. Future research should aim to clarify the mechanisms underlying NET formation in SLE and to optimize new antioxidant therapies, including assessments of their long-term efficacy and safety. Full article

Background/Objectives: The delta neutrophil index (DNI)—a hematology analyzer-derived measure of circulating immature granulocytes—may assist pre-biopsy decision-making, yet its behavior across tumor types is incompletely defined. We examined whether pre-biopsy DNI differs by pathology category, tumor class, and definitive histology, and evaluated diagnostic performance. Methods: In this retrospective, single-center cohort, consecutive inpatients with malignancy were screened (n = 2009). Exclusions included positive blood cultures, prior chemotherapy/radiotherapy before index labs, and lack of definitive pathology, yielding 1313 analyzable cases. All laboratories, including DNI, were obtained before diagnostic biopsy. DNI was assessed as a continuous variable and categorized (Zero = 0; High > 0.6). Groupwise differences used Kruskal–Wallis and χ² tests with FDR control; discrimination used ROC analyses (one-versus-rest/pairwise). Results: DNI distributions differed across pathology, tumor class, and definitive diagnoses (all p < 0.001). High DNI (>0.6) and Zero DNI (=0) proportions also varied significantly by grouping. Hematologic malignancies showed the highest DNI (median ~1.0) compared with sarcoma and carcinoma (medians ~0.4). Using DNI alone, one-versus-rest AUCs were 0.735 (hematologic), 0.692 (melanoma), 0.672 (sarcoma), and 0.652 (carcinoma); the strongest pairwise separation was hematologic versus sarcoma (AUC 0.780). For specific solid tumors, including breast and renal cell carcinoma, single-marker discrimination was modest; no clinically actionable RCC cutoff emerged. Sensitivity analyses restricted to culture-negative cases yielded consistent findings. Conclusions: Pre-biopsy DNI exhibits tumor-type-dependent variation and provides adjunct diagnostic signal—the strongest for hematologic malignancy—yet is insufficient alone for solid tumor subtyping. Integration with clinical assessment and routine biomarkers, and multi-center validation with device harmonization are warranted. Full article

Granulocyte-macrophage colony-stimulating factor (GM-CSF) has emerged as a key cytokine in the pathogenesis of rheumatoid arthritis, an autoimmune disease distinguished by synovial inflammation and progressive joint destruction. GM-CSF orchestrates the activation, proliferation, and differentiation of myeloid cells (mainly macrophages and neutrophils) thereby sustaining the pro-inflammatory synovial milieu. Recent advances in monoclonal antibody immunotherapy have enabled selective inhibition of GM-CSF or its receptor. Clinical data on several monoclonal antibodies are presented, focusing on their pharmacodynamic properties and efficacy results documented in phase II and III clinical studies. Cumulative evidence supports GM-CSF inhibition as a compelling strategy for modulating inflammation and improving clinical outcomes in rheumatoid arthritis. Full article

Contact dermatitis (CD) is a common inflammatory skin condition with irritant etiology or a delayed-type hypersensitivity called allergic contact dermatitis (ACD). Contact hypersensitivity (CHS) is a widely used rodent model of ACD and similarly consists of two phases: sensitization and elicitation. To trigger CHS, low-molecular-weight haptens, such as DNFB or TNCB, are commonly applied. However, the characterization of the induced immune response remains incomplete. Our aim was to characterize the immune response after first and repeated exposures to model haptens. First exposure to DNFB or TNCB led to significant ear swelling, with DNFB causing a more pronounced effect. DNFB enhanced neutrophil infiltration, whereas TNCB led to macrophage, dendritic cell, and helper T cell accumulation. Repeated DNFB exposure did not aggravate edema significantly, while TNCB re-exposure enhanced edema formation and induced neutrophil granulocyte, dendritic cell, and helper and cytotoxic T cell accumulation. Our results demonstrate that a hapten-specific immune response is induced during both phases of CHS. A detailed understanding of allergen-specific immune responses is crucial for the appropriate selection of the model and for gaining deeper insight into the mechanisms of inflammatory skin diseases. These findings may contribute to the targeted selection of haptens. Full article

Tamibarotene (AM80) is an agonist of retinoic acid receptor alpha. It is licensed in Japan for the treatment of acute promyelocytic leukemia. Results from preclinical models suggest that tamibarotene might also be effective in the treatment of diverse autoimmune diseases. The effect of tamibarotene on autoimmune diseases of the skin, however, has not been explored. We therefore examined the effect of tamibarotene on disease in the antibody-transfer mouse model of bullous pemphigoid (BP)-like epidermolysis bullosa acquisita (EBA), a prototypical example for pemphigoid diseases. Pemphigoid diseases are a group of autoimmune blistering skin diseases driven by autoantibodies and the recruitment and activity of granulocytes in the dermis. In sharp contrast to its effect in models of other autoimmune diseases, tamibarotene aggravated EBA pronouncedly. At the peak of disease, skin inflammation in tamibarotene-treated mice involved, on average, 1.6-fold more of the total body surface compared to vehicle-treated mice. Tamibarotene markedly reduced the recruitment of regulatory T cells (T_regs) into the dermis. This blunted the counterregulatory mechanisms that normally curb skin inflammation in this model. The effect aligns with previous reports describing tamibarotene-mediated downregulation of skin-homing receptors on T_regs. In addition, tamibarotene prolonged the responsiveness of aging neutrophils to immune complexes in vitro, providing another mechanism that may exacerbate EBA. Collectively, our results suggest that tamibarotene may elicit detrimental effects in patients with EBA by abolishing the recruitment of T_regs into skin. This warrants great caution when using tamibarotene in patients with EBA and possibly other pemphigoid diseases. Full article

Colony-stimulating factor 3 (CSF3), additionally called granulocyte colony-stimulating factor (G-CSF), is the major cytokine regulating neutrophil production and also impacting their function. The actions of this cytokine are mediated through its unique receptor, the colony-stimulating factor 3 receptor (CSF3R). Several classes of pathogenic mutations in the CSF3R gene have been identified that have distinct biological properties and clinical impacts. This review provides an overview of CSF3R, the various pathogenic CSF3R mutations/variants and their biological effects. It also details the diseases to which they contribute, notably including chronic neutrophilic leukemia (CNL) and other myeloproliferative neoplasms (MPNs), myelodysplastic neoplasms (MDS), combined MDS/MPN disorders such as atypical chronic myeloid leukemia (aCML) and chronic myelomonocytic leukemia (CMML), as well as acute myeloid leukemia (AML) and lymphoid malignancies. Full article

Background: The aim of this study was to evaluate the changes in platelet indices (PLT) provided by the ADVIA 2120 hematology analyzer (Siemens Hematology System) in the early stages of onset of infections and acute coronary syndromes (ACSs). Methods: Samples were selected from 40 patients admitted to the intensive care unit with suspected uncomplicated sepsis at presentation, from 40 patients with a biochemical diagnosis of ACS at presentation and from 40 apparently healthy subjects. These samples were tested for PLT and PLT indices [mean platelet volume (MPV); mean platelet mass (MPM); mean platelet component (MPC); immature platelets (RtcPlts)] obtained by automation with the ADVIA 2120 and specific biomarkers for sepsis [white blood cells (WBCs); neutrophil granulocytes (NGs); presepsin (PSP); procalcitonin (Pct); C-reactive protein (CRP)] and for SCA (hs cTnI). Results: Platelet indices (RtcPlts, MPV, MPM) were significantly altered (p > 0.005) in patients with suspected sepsis and patients with ACS compared to control subjects; however, no statistically significant difference was observed between the two groups of patients with disease. Cutoff values (ROC curves) were obtained for platelet indices that best discriminated healthy subjects from subjects with severe infection or ACS. Conclusions: Our data show that, in subjects with suspected sepsis and ACS at disease onset, a state of early platelet activation exists that is not disease-specific. Immature platelets (RtcPlts) and the platelet indices MPM and MPV, provided by the ADVIA 2120 hematology analyzer, showed high sensitivity in subjects with suspected sepsis or ACS at disease onset. Full article

The compatible solute ectoine is known to attenuate inflammatory effects in the airways after exposure to combustion-derived nanoparticles. Pro-inflammatory signaling in epithelial cells, as well as antiapoptotic mechanisms in neutrophilic granulocytes, both triggered by particles, are reduced by this substance. Here we investigated the preventive potential in airway inflammation of additional compounds originating from the ectoine metabolism, Nγ-acetyl-L-2,4-diaminobutyric acid (NADA), and 5-hydroxyectoine in a mouse model and in human neutrophilic granulocytes. Furthermore, effects of these molecules on the reduction in cystic fibrosis transmembrane conductance regulator (CFTR), as an additional pathogenic endpoint of nanoparticle exposure, were investigated. All three solutes exhibited beneficial effects at the level of inflammatory cells in lung lavages from exposed mice. The decrease in CFTR in lung tissue of exposed mice was mitigated by the substances. In primary human neutrophils and in neutrophilic differentiated HL-60 cells, the delay of apoptosis rates after particle exposure was effectively abolished. The decline in CFTR from the cytoplasmic membrane in neutrophilic cells was also counteracted by the compatible solutes. The data identify both NADA and 5-hydroxyectoine as additional substances for molecular prevention of airway effects of environmental particles. Furthermore, the reduction in CFTR might be a relevant finding for patients suffering from impaired function of this ion channel. Full article

Neutrophils, a first-line defender, has a multifaceted presence in chronic inflammation, autoimmune pathology, and tumor progression. The microenvironmental cues facilitate functional plasticity and phenotypic heterogeneity to neutrophils that enable both their protective and pathogenic roles. Autoimmune diseases including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and juvenile idiopathic arthritis (JIA) display the presence of dysregulated subsets of neutrophil, such as low-density granulocytes (LDGs) that promote proinflammation and contribute to tissue damage via NETosis and type I interferon-mediated signaling. In cancer, particularly tumors, they exhibit tumor-associated neutrophils (TANs) which may polarize either towards anti-tumorigenic ‘N1’ or pro-tumorigenic ‘N2’ phenotypes based on available modulators such as TGF-β and leucine-driven epigenetic modifications. The development in neutrophil biology has introduced several novel therapeutic strategies that allow NET targeting, inhibition of chemokine receptors like CXCR2, and exploration of neutrophil-derived biomarkers for diagnosis and disease monitoring. Such findings encourage the importance of neutrophils as both effectors and therapeutic targets in inflammatory and neoplastic conditions. Full article

Objectives: Gla-rich protein (GRP), a vitamin K-dependent protein, has been increasingly recognized for its dual role in modulating inflammation and inhibiting pathological calcification. Despite its emerging importance in chronic conditions, limited evidence exists regarding its behavior during acute critical illness. This study aimed to investigate the association between GRP, systemic inflammatory markers, oxidative stress (via total thiol oxidation-reduction ratio, TORR), and calcium metabolism in critically ill patients. Materials and Methods: This cross-sectional observational study included 93 critically ill patients admitted to the intensive care unit (ICU) and 60 age- and sex-matched non-critically ill volunteers. Serum GRP levels were measured using ELISA. Other biomarkers including TORR, C-reactive protein (CRP), procalcitonin (PCT), white blood cell count (WBC), immature granulocytes (IGs), and serum calcium were also analyzed. Pearson’s correlation, multivariate linear regression, and ROC analysis were performed to assess the relationships among GRP and biochemical markers, as well as their capacity to differentiate ICU patients from controls. Results: GRP, TORR, CRP, PCT, WBC, IGs, and ferritin levels were significantly elevated in ICU patients compared to the control group, whereas serum calcium levels were markedly reduced (all p < 0.05). GRP levels demonstrated moderate positive correlations with WBC (r = 0.47), neutrophils (r = 0.51), TORR (r = 0.42), CRP (r = 0.30), and IGs (r = 0.46), and a strong negative correlation with calcium (r = −0.63). In multivariate regression, TORR, CRP, WBC, IGs, PCT, and calcium levels showed significant correlations with GRP levels in univariate analysis. ROC analysis revealed that CRP had the highest discriminatory power (AUC = 0.88; 95% CI: 0.82–0.94), followed by TORR (AUC = 0.79; 95% CI: 0.71–0.86), GRP (AUC = 0.76; 95% CI: 0.68–0.84), and IGs (AUC = 0.77; 95% CI: 0.69–0.85), for distinguishing ICU patients from non-critically ill individuals. Conclusions: Our findings demonstrated that GRP is significantly associated with systemic inflammation, oxidative stress, and calcium metabolism disturbances in critically ill patients. The combined evaluation of GRP and TORR may enhance the understanding of inflammatory and oxidative mechanisms in acute critical illness. Although this study did not assess patient outcomes, these biomarkers could serve as promising candidates for future prognostic research in ICU settings. Full article

The purpose of this study was to evaluate normal values of healthy human bone marrow (n = 56) and identify gender- and age-related variations using cell lineage markers and maturational curves. Using 10-color quantitative flow cytometry, various cell types were identified, including B cells, T cells, NK cells, granulocytes, monocytes, erythroblasts, plasma cells, basophils, mast cells, and dendritic cells. Results revealed significant age-related declines in the absolute counts of nucleated cells (p = 0.001), including CD34+ immature B cells (p = 0.006) and CD34- immature B cells (p = 0.004). Declines were also observed for T cells (p = 0.002), cytotoxic T cells (p < 0.001), double-negative T cells (p = 0.0001), NK cells (p = 0.007), CD16- NK cells (p < 0.001), metamyelocytes (p = 0.002), neutrophils (p = 0.001), basophils (p = 0.009), promonocytes (p = 0.001), mature monocytes (p = 0.007), and plasmacytoid dendritic cells (p = 0.001). Gender differences showed males had more intermediate monocytes (p = 0.009) compared to females. In summary, this study provides normal values for hematopoietic cells, highlighting age- and gender-related disparities critical for understanding hematopoietic dynamics. Full article