Neutrophils Research Articles

S100A8/A9hi neutrophils induce mitochondrial dysfunction and PANoptosis in endothelial cells via mitochondrial complex I deficiency during sepsis

S100a8/a9, largely released by polymorphonuclear neutrophils (PMNs), belongs to the S100 family of calcium-binding proteins and plays a role in a variety of inflammatory diseases. Although S100a8/a9 has been reported to trigger endothelial cell apoptosis, the mechanisms of S100a8/a9-induced endothelial dysfunction during sepsis require in-depth research. We demonstrate that high expression levels of S100a8/a9 suppress Ndufa3 expression in mitochondrial complex I via downregulation of Nrf1 expression. Mitochondrial complex I deficiency contributes to NAD+-dependent Sirt1 suppression, which induces mitochondrial disorders, including excessive fission and blocked mitophagy, and mtDNA released from damaged mitochondria ultimately activates ZBP1-mediated PANoptosis in endothelial cells. Moreover, based on comprehensive scRNA-seq and bulk RNA-seq analyses, S100A8/A9hi neutrophils are closely associated with the circulating endothelial cell count (a useful marker of endothelial damage), and S100A8 is an independent risk factor for poor prognosis in sepsis patients.

Correlation analysis of patients with diabetic foot ulcers treated with tibial cortex transverse transport surgery and platelet-to-lymphocyte ratio and monocyte-to-neutrophil ratio.

Diabetic foot ulcers (DFUs) represent one of the most severe late-stage complications of diabetes. Tibial cortex transverse transport (TTT) surgery stands as the prevailing method for addressing DFUs. This surgical intervention holds the promise of expediting DFU wound healing and diminishing the rate of amputations, with the mitigation of inflammatory responses playing a pivotal role. In this study, we aim to explore the correlation between inflammation and TTT surgery, with the overarching goal of facilitating swift prognostic assessments in clinical practice. The correlation between the severity of DFUs and clinical test results remains ambiguous. A clinical prediction model was devised to explore the connection between DFU severity and the efficacy of TTT surgery, utilizing straightforward and efficient clinical indicators. Clinical data and examination results were gathered by tracking hospitalized DFU patients who underwent TTT surgery at the First Affiliated Hospital of Guangxi Medical University (Nanning, China). Indicators associated with DFU severity and wound healing time post-surgery were identified through logistic regression and least absolute shrinkage and selection operator (LASSO) regression analyses. Subsequently, a clinical prediction model was constructed. Finally, the intersection of these 2 sets of indicators revealed factors correlated with wound severity and post-operative healing duration. Our study was comprised of 202 patients who were categorized into 2 groups based on Wagner's grading classifications. Utilizing Student's t-tests, LASSO regression and logistic regression analyses, we identified 3 factors indicative of DFU severity: platelet-to-lymphocyte ratio (PLR), mixed lymphocyte reaction (MLR) and hemoglobin (HGB). Univariate COX regression analysis revealed 12 factors such as: white blood cells (WBC), neutrophils (NEUT), monocytes (MO), PLR, MLR, neutrophil-to-lymphocyte ratio (NLR), erythrocyte sedimentation rate (ESR), age, lymphocytes (LY), monocyte-to-neutrophil ratio (MNR), uric acid (UA), and albumin (ALB) associated with the postoperative healing duration. Ultimately, we identified 2 factors, PLR and MNR, at the intersection of these 2 datasets. Platelet-to-lymphocyte ratio and MNR were identified as factors associated with both the severity of DFUs and the prognosis following TTT surgery.

Short-term exposure to air pollution on peripheral white blood cells and inflammation biomarkers: a cross-sectional study on rural residents

Effects of short-term exposure to ambient air pollution on systemic immunological and inflammatory biomarkers in rural population have not been adequately characterized. From May to July 2021, 5816 participants in rural villages of northern Henan Province, China, participated in this cross-sectional study. Blood biomarkers of systemic inflammation were determined including peripheral white blood cells (WBC), eosinophils (EOS), basophils (BAS), monocytes (MON), lymphocytes (LYM), neutrophils (NEU), neutrophil-lymphocyte ratio (NLR), and serum high-sensitivity C-reactive protein (hs-CRP). The concentrations of ambient fine particulate matter (PM2.5), PM10, nitrogen dioxide (NO2), carbon monoxide (CO), and ozone (O3) were assessed up to 7 days prior to the blood draw. A generalized linear model was used to analyze the associations between air pollution exposure and the above-mentioned blood biomarkers. Significantly positive associations were revealed between PM2.5, CO and WBC; CO, O3 and LYM; PM2.5, PM10, SO2, CO and NEU; PM2.5, PM10, SO2, CO and NLR; PM2.5, PM10, SO2, NO2, CO, O3 and hs-CRP. Meanwhile, negative associations were found between SO2 and WBC; PM2.5, PM10, NO2, CO, or O3 and EOS; PM2.5, SO2, or CO and BAS; SO2, NO2 or O3 and MON; PM2.5, PM10, SO2, or NO2 and LYM. Moreover, men, individuals with normal body mass index (BMI), current smokers, and those older than 60 years were found vulnerable to air pollution effects. Taken together, short-term exposure to air pollution was associated with systemic inflammatory responses, providing insight into the potential mechanisms for air pollution-induced detrimental systemic effects in rural residents.

Features of the neutrophil granulocyte system in patients with common scalp defects

Objective. To identify the dynamics of changes in the structure of the neutrophil granulocyte system in patients with extensive scalp defects before and after their surgical closure. Materials and methods. A computer morphometric analysis of the quantitative and qualitative characteristics of neutrophilic granulocytes of patients with common scalp defects who were treated at the Department of Microvascular Plastic and Reconstructive Surgery of the Shalimov National Scientific Centre of Surgery and Transplantation from 2017 to 2023. The main group included 15 patients with widespread defects of the soft tissues of the scalp in the I – II stage of the wound process (10) and arteriovenous angiodysplasias in the III stage according to Schobinger (5). The control group consisted of 15 volunteers without pathology. The structure features (optical cytoplasmic density in units, which indicates the presence of bactericidal potential, and cell area in μm², which indicates the level of activation) of peripheral blood neutrophil granulocytes before and after surgical closure of the defect were studied. Results. Before the surgical intervention, a pronounced polymorphism was detected, which meant a functional failure of the neutrophil granulocyte system. After surgery and healing of the defect, excessive polymorphism of the neutrophil granulocyte system was not detected in patients. Conclusions. The assessment of the degree of cellular polymorphism of neutrophil granulocytes determines the diagnostic efficiency of the study and makes it possible to identify and predict the development of local inflammation and infectious complications in patients with scalp defects.

BMAL1 plays a crucial role in immune homeostasis during sepsis-induced acute lung injury

Sepsis is a widespread and life-threatening disease characterised by infection-triggered immune hyperactivation and cytokine storms, culminating in tissue damage and multiple organ dysfunction syndrome. BMAL1 is a pivotal transcription factor in the circadian clock that plays a crucial role in maintaining immune homeostasis. BMAL1 dysregulation has been implicated in inflammatory diseases and immunodeficiency. However, the mechanisms underlying BMAL1 disruption in sepsis-induced acute lung injury (ALI) remain poorly understood. In vitro, we used THP1 and mouse peritoneal macrophages to elucidate the potential mechanism of BMAL1 function in sepsis. In vivo, an endotoxemia model was used to investigate the effect of BMAL1 on sepsis and the therapeutic role of targeting CXCR2. We showed that BMAL1 significantly affected the regulation of innate immunity in sepsis-induced ALI. BMAL1 deficiency in the macrophages exacerbated systemic inflammation and sepsis-induced ALI. Mechanistically, BMAL1 acted as a transcriptional suppressor and regulated the expression of CXCL2. BMAL1 deficiency in macrophages upregulated CXCL2 expression, increasing the recruitment of polymorphonuclear neutrophils and the formation of neutrophil extracellular traps (NETs) by binding to the chemokine receptor CXCR2, thereby intensifying lung injury in a sepsis model. Furthermore, a selective inhibitor of CXCR2, SB225002, exerted promising therapeutic effects by markedly reducing neutrophil infiltration and NETs formation and alleviating lung injury. Importantly, CXCR2 blockade mitigated multiple organ dysfunction. Collectively, these findings suggest that BMAL1 controls the CXCL2/CXCR2 pathway, and the therapeutic efficacy of targeting CXCR2 in sepsis has been validated, presenting BMAL1 as a potential therapeutic target for lethal infections.

Performance evaluation of the digital morphology analyser Sysmex DI-60 for white blood cell differentials in abnormal samples

Sysmex DI-60 enumerates and classifies leukocytes. Limited research has evaluated the performance of Sysmex DI-60 in abnormal samples, and most focused on leukopenic samples. We evaluate the efficacy of DI-60 in determining white blood cell (WBC) differentials in normal and abnormal samples in different WBC count. Peripheral blood smears (n = 166) were categorised into normal control and disease groups, and further divided into moderate and severe leucocytosis, mild leucocytosis, normal, mild leukopenia, and moderate and severe leukopenia groups based on WBC count. DI-60 preclassification and verification and manual counting results were assessed using Bland–Altman and Passing–Bablok regression analyses. The Kappa test compared the concordance in the abnormal cell detection between DI-60 and manual counting. DI-60 exhibited notable overall sensitivity and specificity for all cells, except basophils. The correlation between the DI-60 preclassification and manual counting was high for segmented neutrophils, band neutrophils, lymphocytes, and blasts, and improved for all cell classes after verification. The mean difference between DI-60 and manual counting for all cell classes was significantly high in moderate and severe leucocytosis (WBC > 30.0 × 109/L) and moderate and severe leukopenia (WBC < 1.5 × 109/L) groups. For blast cells, immature granulocytes, and atypical lymphocytes, the DI-60 verification results were similar to the manual counting results. Plasma cells showed poor agreement. In conclusion, DI-60 demonstrates consistent and reliable analysis of WBC differentials within the range of 1.5–30.0 × 109. Manual counting was indispensable in examining moderate and severe leucocytosis samples, moderate and severe leukopenia samples, and in enumerating of monocytes and plasma cells.

Multiplex cytokine analysis for the identification of novel potential synovial fluid biomarkers for periprosthetic joint infections

IntroductionPeriprosthetic joint infections (PJIs) are a devastating complication of total hip (THA) and knee (TKA) arthroplasty. The use of novel techniques like multiplex cytokine analysis could contribute immensely to the identification of potential novel biomarkers. Patients and MethodsThis is a single-centre study of patients that were treated with revision TKA, THA or hemiarthroplasty. Serum's white blood cells (WBCs), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) and synovial fluid's WBCs, percentage of polymorphonuclear neutrophils (%PMNs) and CRP were measured. Proteomic analysis targeting the secreted cytokines in synovial fluid was conducted using a 73-plex assay panel. The results were statistically compared between the septic and aseptic cases and ROC analysis to establish the area under the curve (AUC), sensitivity and specificity of each biomarker. ResultsThe study included 30 patients (18 revision THA cases; 3 conversion of hemiarthroplasty to THA and 9 revision TKA cases); 14 cases were considered infected, 1 likely infected and 15 not infected. The results showed statistically significant differences (p < 0.05) between infected and not infected cases in serum's ESR, CRP and synovial fluid's%PMNs, growth-regulated oncogene alpha (GROA), interleukin-8, interleukin-5, S100-A8/calprotectin and resistin (RETN) with AUCs of 0.75, 0.72, 0.95, 0.75, 0.72, 0.95, 0.83, 0.73, 0.75, 0.81 and 0.76 respectively. ConclusionsIn the present study, serum ESR and CRP as well as synovial %PMNs, GROA, IL-8, IL-5, calprotectin and RETN protein levels were identified as potential biomarkers. Further studies are needed to further investigate their diagnostic utility and optimal cut-off values.

Chronic Heat Exposure Modulates Innate and Adaptive Immune Responses in Firefighters

Global fire activities, which are getting worse due to climate change, cause both environmental and human health hazards. Firefighters, being the first responders, are frequently exposed to heat which may impact their immune system and overall health. However, the nature of the impact of chronic heat exposure on immune function has not been studied in-depth in firefighters. In this study, 22 firefighters exposed to “heavy-smoke fires (structural fires)”, categorized as the “high-exposure group” (&gt;0.15 structural fires/week) and “low-exposure group” (&lt;0.15 structural fires/week), were sampled. Peripheral blood was examined for immune cell profile based on total and differential cell counts, immune function based on the transcriptional expression of drivers of innate and adaptive immunity and key inflammation mediators, and heat stress marker HSP70. The white blood cell (WBC) count, mean corpuscular volume, mean corpuscular hemoglobin, and absolute and segmented neutrophil counts decreased below the normal range in both exposure groups. The gene transcript levels for toll-like receptors (TLR2, TLR4, but not TLR7) and their adaptor protein MYD88 were lower whereas those for T-cell transcription factors (RORC/RORγ, FoxP3) and inflammatory mediators (TNF-α, Granzyme-B) were higher in the “high-exposure group”, indicating mixed response; however, the ratios between pro-inflammatory and anti-inflammatory transcription factors of adaptive immunity, namely T-bet/FoxP3 (Th1/Treg) and RORC/FoxP3 (Th17/Treg), were lower. Collectively, decreased immune cell landscape, downregulated key innate immunity receptors, and Tregs’ dominance suggested that chronic heat exposure in firefighters dysregulated innate and adaptive immunity, skewed towards an overall immunosuppressive condition with inflammation.

Cytotoxic Oxidative Stress Effects of Neutrophil Extracellular Traps' Components on Cattle Spermatozoa.

Bovine spermatozoa are highly susceptible to oxidative stress (OS), and it is known to affect their cellular functions. The main leukocyte producers of reactive oxygen species (ROS) in mammalian semen are polymorphonuclear neutrophils (PMN). PMN activation can result in the formation of neutrophil extracellular traps (NETs), which have been shown to affect the motility and function of spermatozoa. However, OS effects on bull spermatozoa derived from individual NETs components have not been investigated. The hypothesis of this study was that specific NETs components might generate OS on bull spermatozoa. Bovine sperm cells were incubated with five NETs-associated molecules, including 30 μg/mL histone 2A (H2A), neutrophil elastase (NE), 1 μg/mL myeloperoxidase (MPO), cathepsin G (Cat-G), and cathelicidin LL37 (LL-37), for a time course ranging from 15 to 240 min. Fluorescence microscopy was used to evaluate the coincubation of bovine PMN and sperm cells. Within 15 min, H2A, NE, and LL-37 caused membrane disruption, while MPO and Cat-G caused OS on bull spermatozoa after 1 h of coincubation. NET formation was observed within 15 min of coincubation in co-cultures of bovine PMN/sperm cells. This study is the first to report on the role of cytotoxic OS effects caused by NETs-derived components in bovine sperm in vitro.

In-vivo Screening for the Antiasthmatic Potential of Salicin through the Murine Models of Asthma and Airway Remodeling

ABSTRACT Purpose: To determine the antiasthmatic potential of salicin in experimental animals. Materials and Methods: The study was undertaken in two phases: clonidine-induced catalepsy (Phase I) and ovalbumin (OVA) induced lung inflammation (Phase II). In Phase I, 36 Swiss Albino mice were pretreated with clonidine (1 mg/kg) subcutaneously for induction of catalepsy. Different doses of salicin (100, 200, and 300 mg/kg) and pheniramine maleate (10 mg/kg) were administered through the oral route, and the cataleptic score was calculated. In Phase II, 36 Albino Wistar rats were sensitized and challenged with 1 mg OVA absorbed on 20 mg aluminum hydroxide (Al(OH)3) intraperitoneally on days 0, 7, and 14 followed by the treatment with salicin doses (100, 200, and 300 mg/kg) and dexamethasone (0.5 mg/kg). Blood parameters, including total cells (TC), eosinophils (EOS), neutrophils (NEU), and macrophages (MAC), were recorded. Levels of tumor necrosis factor-α (TNFα), interleukin-4 (IL-4), interleukin-6 (IL-6), and interleukin-13 (IL-13) were collected from bronchoalveolar lavage fluid. Levels of OVA-specific IgE were estimated from spleens, Peyer’s patches, and mesenteric lymph nodes of rats. Results: Animals treated with salicin showed a significant reduction (P-value &lt;0.05) in cataleptic scores. Significant reduction (P-value &lt;0.05) in levels of TC, NEU, EOS, and MAC was observed in animals treated with salicin. Levels of TNF-α, IL-4, IL-6, and IL13 also reduced significantly (P-value &lt;0.05) in salicin-treated animals. The concentration of OVA-specific IgE reduced significantly (P-value &lt;0.05) in salicin-treated animals. Conclusion: Salicin ameliorates catalepsy and lung inflammation in asthmatic conditions.

Predictive value of immature granulocyte percentage and neutrophil lymphocyte ratio in terms of prognosis in the course of acute pancreatitis.

We aimed to investigate the role and importance of immature granulocyte percentage and neutrophil/lymphocyte ratio in the etiology, diagnosis and follow-up of acute pancreatitis (AP) in patients tentatively diagnosed with AP in the emergency department. We evaluated these factors alongside other established markers proven effective in the diagnosis and follow-up of AP. Atotal of 139 patients with atentative diagnosis of acute pancreatitis who were hospitalized and followed up in the gastroenterology clinic in 2021‒2022 were included in the study. In addition, acontrol group, consisting of 139 individuals admitted to the clinic for various other reasons, was established. The cases were also compared with the control group in terms of NLR, ICG and IG%. There was asignificant difference in the NLR, IGC and IG% measurements between the patients in the AP group and the control group. In all three markers, the average values of the patient group were higher than those of the control group. Furthermore, asignificant difference in IGC and IG% blood measurements was noted between sub-groups of patients categorized based on the severity of acute pancreatitis, particularly the patients with severe pancreatitis exhibited higher mean IGC and IG% blood measurements compared to those with mild or moderate pancreatitis. IGC and IG% values emerged as superior indicators to other acute-phase reactants for detecting inflammation, determining its severity, and establishing prognosis in acute pancreatitis. While the N/L ratio remains an important parameter in acute pancreatitis, our findings indicate that it was not significantly superior to other investigated markers in terms of prognosis (Tab. 5, Ref. 35).

Chronic Septic Granulomatosis Revealed in Adulthood by Respiratory Manifestations

Chronic septic granulomatosis is a rare genetic disease that affects the immune system. It is characterized by a primary immune deficiency that makes it more difficult for phagocytic cells (such as polymorphonuclear neutrophils, eosinophils, monocytes, and macrophages) to eradicate some harmful bacteria and fungi. It is considered an orphan disease because it affects less than 0.05% of a given population.

The band count imprecision - a Croatian multicentric pilot study.

Due to high inter-observer variability the 2015 International Council for Standardization in Haematology (ICSH) recommendations state to count band neutrophils as segmented neutrophils in the white blood cell (WBC) differential. However, the inclusion of bands as a separate cell entity within the WBC differential is still widely used in hematology laboratories in Croatia. The aim of this multicentric study was to assess the degree of inter-observer variability in enumerating band neutrophils within the WBC differential among Croatian laboratories. Seven large Croatian hospital laboratories from different parts of the country participated in the study. In each of 7 participating laboratories, one blood smear, that was flagged by the analyzer as possibly having bands, was evaluated by all personnel participating in the analysis of hematology samples. Between-observer manual smear reproducibility was expressed as coefficient of variation (CV) and calculated using the following formula: CV (%) = (standard deviation (SD)/mean value) x 100%. The CVs (%) and relative band neutrophil counts in participating laboratories were as follows: 15.4% (16-24), 19.2% (16-32), 19.5% (17-40), 21.1% (17-44), 35.0% (8-26), 51.9% (3-29), and remarkably high 62.4% (12-59). For segmented neutrophils CVs were lower, ranging from 7.4% to 32.2%. The CVs did not correlate with the number of staff members in each hospital (P = 0.293). This study revealed very high variability in enumerating band neutrophil count in the blood smear differential among all participants, thus prompting a need for action on a national level.

Predictive value of immature granulocyte percentage and neutrophil lymphocyte ratio in terms of prognosis in the course of acute pancreatitis

Predictive value of immature granulocyte percentage and neutrophil lymphocyte ratio in terms of prognosis in the course of acute pancreatitis

Nomogram established on account of Lasso-logistic regression for predicting hemorrhagic transformation in patients with acute ischemic stroke after endovascular thrombectomy

BackgroundHemorrhagic transformation (HT) is a common and serious complication in patients with acute ischemic stroke (AIS) after endovascular thrombectomy (EVT). This study was performed to determine the predictive factors associated with HT in stroke patients with EVT and to establish and validate a nomogram that combines with independent predictors to predict the probability of HT after EVT in patients with AIS. MethodsAll patients were randomly divided into development and validation cohorts at a ratio of 7:3. The least absolute shrinkage and selection operator (LASSO) regression was used to select the optimal factors, and multivariate logistic regression analysis was used to build a clinical prediction model. Calibration plots, decision curve analysis (DCA) and receiver operating characteristic curve (ROC) were generated to assess predictive performance. ResultsLASSO regression analysis showed that Alberta Stroke Program Early CT Scores (ASPECTS), international normalized ratio (INR), uric acid (UA), neutrophils (NEU) were the influencing factors for AIS with HT after EVT. A novel prognostic nomogram model was established to predict the possibility of HT with AIS after EVT. The calibration curve showed that the model had good consistency. The results of ROC analysis showed that the AUC of the prediction model established in this study for predicting HT was 0.797 in the development cohort and 0.786 in the validation cohort. ConclusionThis study proposes a novel and practical nomogram based on ASPECTS, INR, UA, NEU, which can well predict the probability of HT after EVT in patients with AIS.

Assessment and correction of local antimicrobial protection factors in women with chronic recurrent vulvovaginal candidiasis and type 2 diabetes mellitus

Aim. Evaluation and correction of local antimicrobial protection factors in women with type 2 diabetes mellitus (DM 2) and chronic recurrent vulvovaginal candidiasis (rVVC). Materials and methods. The study involved 100 women undergoing outpatient follow-up at the age of 40.9±5.8 years, with a body mass index of 29.8±3.5. The period was 2022–2023. The examination included anamnesis, collection of anthropometric data, calculation of body weight, microscopic examination of smears from the cervical canal and vagina, Gram-stained before treatment, in the 1st and 3rd months after its completion, identification pathogenic microorganisms using PCR, colposcopy, ultrasound of the pelvic organs. Indicators of glycosylated hemoglobin in the subjects at the control points of the study: at 1 month the average values were 5.9±2.9%, at 3 months – 5.9±3.0%, the average value was 5.9±3.1%. Candida species were identified by the bacteriological method using Sabouraud dextrose agar (growth of colonies of fungi of the genus Candida in an amount of more than 103 CFU/ml). Immunological methods for studying antimicrobial protection factors included studying the number of neutrophil granulocytes (NG) on the surface of the mucous membranes of the vulva and vagina, their phagocytic and NBT-reducing activity in a latex test. Randomization of patients into groups: group 1 included 50 (50%) women with DM 2 who, as part of complex therapy for rVVC received the drug Genferon® 1 intravaginal suppository of 500 thousand units 2 times a day for 10 days and fluconazole 150 mg orally three times with an interval of 72 hours at the first stage of treatment; group 2 included 50 (50%) women who received therapy with fluconazole 150 mg three times with an interval of 72 hours. The maintenance anti-relapse course therapy in both groups included the use of fluconazole 150 mg once a week for 6 months. In group 1st anti-relapse therapy was supplemented by the administration of the drug Genferon® 1 intravaginal suppository of 500 thousand units at night 3 times a week for 3 months, after which the vaginal microbiota was corrected for 2 weeks using vaginal suppositories, containing Lactobacillus acidophilus in an amount of at least 108 CFU. In group 2nd, basic anti-relapse antimycotic therapy was not accompanied by the prescription of any immunomodulatory or probiotic drugs. Results. The etiological agents of rVVC in women of late reproductive age in 79% (n=79) и 28% (n=28), respectively, were C. albicans and C. glabrata, which during the period of acute of the disease were detected by culture in the vaginal discharge at more than 104 CFU/ml. Chronic recurrent course of vulvovaginal candidiasis in women with DM 2 was characterized by exacerbations 4 or more times a year, accompanied by the corresponding clinical picture: white or yellowish-white cheesy discharge from the genital tract, itching or burning in the anogenital area, discomfort in the external genital area, dyspareunia, dysuria, decreased phagocytic activity of NG in vaginal secretions by 25.8%, impairment of their spontaneous and latex-induced NBT-reducing activity by 35.2%, functional reserve by 23.0% relative to reference values. The use of the drug Genferon® as part of complex therapy for rVVC contributed to a decrease in the number of Candida spp. in 1st and 3d months of observation after completion of anti-relapse therapy, normalization of cellular factors of innate immunity of the mucous membranes, faster resolution of the clinical manifestations of an episode of the disease. The decrease in the number of relapses over a 12-month observation period compared to the control group was also facilitated by an increase in the protective properties of the vaginal mucosa due to the restoration of the vaginal microbiota with the help of lactobacilli acidophilus. Conclusion. The etiological agents of rVVC in women of late reproductive age are C. albicans and C. glabrata. Subcompensated DM 2 is a risk factor for rVVC, which requires additional monitoring of microbiological parameters of the vaginal microbiota. rVVC in women with DM 2 is associated with an increase in the number of NG in the vaginal secretion, a decrease in their phagocytic activity and functional reserve compared to the reference values. Combination therapy of rVVC with topical recombinant interferon α2b, benzocaine and taurine in the formulation of Genferon® (suppositories) is an effective method to improve the clinical and immunological efficacy of therapy.

The role of gamma globulin, complement component 1q, factor B, properdin, body temperature, C-reactive protein and serum amyloid alpha to the activity and the function of the human complement system and its pathways

The complement system plays a crucial role in orchestrating the activation and regulation of inflammation within the human immune system. Three distinct activation pathways—classical, lectin, and alternative—converge to form the common lytic pathway, culminating in the formation of the membrane-attacking complex that disrupts the structure of pathogens. Dysregulated complement system activity can lead to tissue damage, autoimmune diseases, or immune deficiencies.In this study, the antimicrobial activity of human serum was investigated by using a bioluminescent microbe probe, Escherichia coli (pEGFPluxABCDEamp). This probe has previously been used to determine the antimicrobial activity of complement system and the polymorphonuclear neutrophils. In this study, blocking antibodies against key serum activators and components, including IgG, complement component 1q, factor B, and properdin, were utilized. The influence of body temperature and acute phase proteins, such as C reactive protein (CRP) and serum amyloid alpha (SAA), on the complement system was also examined.The study reveals the critical factors influencing complement system activity and pathway function. Alongside crucial factors like C1q and IgG, alternative pathway components factor B and properdin played pivotal roles. Results indicated that the alternative pathway accounted for approximately one third of the overall serum antimicrobial activity, and blocking this pathway disrupted the entire complement system. Contrary to expectations, elevated body temperature during inflammation did not enhance the antimicrobial activity of human serum.CRP demonstrated complement activation properties, but at higher physiological concentrations, it exhibited antagonistic tendencies, dampening the response. On the other hand, SAA enhanced the serum's activity.Overall, this study sheds a light on the critical factors affecting both complement system activity and pathway functionality, emphasizing the importance of a balanced immune response.

Unsupervised clustering analysis based on multidimensional features reveals distinct clinical characteristics and associated factors of different phenotypes in patients with chronic rhinosinusitis with nasal polyp

Objective: To utilize routinely available clinical parameters to uncover the clinical features of different clusters in patients with chronic rhinosinusitis with nasal polyp (CRSwNP) through unsupervised clustering analysis. Methods: The clinical data from 155 CRSwNP patients undergoing nasal endoscopic surgery at Renmin Hospital of Wuhan University from 2021 to 2023 were prospectively collected, including 112 males and 43 females, aged from 7 to 87 years. Unsupervised clustering analysis was conducted using various clinical parameters, including age, gender, smoking and drinking history, local eosinophil (EOS) and neutrophil (NEU) counts, comorbid allergic rhinitis (AR), comorbid asthma, recurrence status, serum-specific IgE, total IgE, cytokine levels, peripheral blood EOS count and percentage, Lund-Mackay CT score, the ratio of CT scores for the maxillary sinus and ethmoid sinus (E/M ratio), visual analogue scale (VAS) score, Lund-Kennedy endoscopic score, and other common clinical indicators to elucidate the clinical characteristics of each cluster. Statistical analysis was conducted using GraphPad Prism 9.5 software. Results: Hierarchical clustering analysis identified four main clusters (Cluster A1-A4), showcasing distinct characteristics such as mild nasal polyps with higher peripheral blood cytokines levels, nasal polyps accompanied by allergies and asthma, a subtype of nasal polyps with high recurrence rates dominated by neutrophils, and nasal polyps with high eosinophil levels. Further subset clustering revealed two clusters of mild polyps (Cluster B1-B2) featuring high cytokine expression and comorbid AR; and two clusters of severe polyps (Cluster B3-B4) presented with severe symptoms, high Lund-Mackay CT score, and high Lund-Kennedy endoscopic score. Variations between Cluster B3 and B4 included symptom complexity, the degree of eosinophil infiltration, and the probability of comorbid asthma. Further clustering analysis for eosinophilic nasal polyps revealed a cluster characterized by highly neutrophilic infiltration and recurrent nasal polyps. The comprehensive analysis of multi-index correlations demonstrated valuable insights into the relationships between common clinical parameters of nasal polyps, providing valuable information for a deeper understanding of the pathogenesis of CRSwNP. Conclusion: The clustering analysis in this study categorizes CRSwNP patients into different clusters based on clinical features and disease outcomes, providing a new perspective for more precise clinical treatment strategies.

Extrusion of Neutrophil Extracellular Traps (NETs) Negatively Impacts Canine Sperm Functions: Implications in Reproductive Failure.

Reproductive failure in dogs is often due to unknown causes, and correct diagnosis and treatment are not always achieved. This condition is associated with various congenital and acquired etiologies that develop inflammatory processes, causing an increase in the number of leukocytes within the female reproductive tract (FRT). An encounter between polymorphonuclear neutrophils (PMNs) and infectious agents or inflammation in the FRT could trigger neutrophil extracellular traps (NETs), which are associated with significantly decreased motility and damage to sperm functional parameters in other species, including humans. This study describes the interaction between canine PMNs and spermatozoa and characterizes the release of NETs, in addition to evaluating the consequences of these structures on canine sperm function. To identify and visualize NETs, May-Grünwald Giemsa staining and immunofluorescence for neutrophil elastase (NE) were performed on canine semen samples and sperm/PMN co-cultures. Sperm viability was assessed using SYBR/PI and acrosome integrity was assessed using PNA-FITC/PI by flow cytometry. The results demonstrate NETs release in native semen samples and PMN/sperm co-cultures. In addition, NETs negatively affect canine sperm function parameters. This is the first report on the ability of NETs to efficiently entrap canine spermatozoa, and to provide additional data on the adverse effects of NETs on male gametes. Therefore, NETs formation should be considered in future studies of canine reproductive failure, as these extracellular fibers and NET-derived pro-inflammatory capacities will impede proper oocyte fertilization and embryo implantation. These data will serve as a basis to explain certain reproductive failures of dogs and provide new information about triggers and molecules involved in adverse effects of NETosis for domestic pet animals.

GPX4 mRNA levels in the polymorphonuclear neutrophils are negatively correlated with autoantibody production, disease activity, and lupus alopecia in systemic lupus erythematosus

AbstractBackgroundFerroptosis is a type of regulated necrosis and glutathione peroxidase 4 (GPX4) has been recognized as a key enzyme that protects against ferroptosis. However, the significance of GPX4 in polymorphonuclear neutrophils (PMNs) of systemic lupus erythematosus (SLE) has not been explored. So we examined GPX4 mRNA in PMNs and analyzed its association with serological and clinical features.MethodsA single center research from the Department of Rheumatology and Immunology of the Third Affiliated Hospital, Southern Medical University was conducted between December 2020 and September 2022. Real‐time transcription‐polymerase chain reaction analysis was used to determine the expression of GPX4 mRNA in PMNs from patients suffering from different rheumatoid immune diseases and healthy controls. The associations of GPX4 RNA levels in SLE patients with serological and clinical indicators were assessed by Spearman's correlation analysis. Receiver operating characteristic curve analysis was performed to evaluate the diagnostic value of GXP4 mRNA for SLE.ResultsGPX4 mRNA levels were lower in SLE patients than those in healthy individuals, patients with rheumatoid arthritis and axial spondyloarthritis. GPX4 mRNA levels were negatively correlated with the number of positive autoantibodies (r = −0.3072, p = 0.0300), anti‐dsDNA antibody (r = −0.3654, p = 0.0336), antinucleosome antibody (r = −0.4052, p = 0.0263), erythrocyte sedimentation rate (r = −0.3773, p = 0.0069), C‐reactive protein (r = −0.4037, p = 0.0036), and SLE disease activity index (r = −0.3072, p = 0.0300). GPX4 mRNA levels were downregulated in patients with alopecia compared with patients without alopecia (p &lt; 0.05). The diagnostic capacity of GPX4 mRNA achieved high diagnostic accuracy (area under the curve = 0.848) with sensitivity (78.00%) and specificity (80.95%).ConclusionDownregulated GPX4 mRNA in PMNs of SLE patients is negatively associated with the production of antinuclear antibodies, disease activity, and lupus alopecia, suggesting an important role of ferroptosis in SLE, high diagnostic value of GPX4 mRNA in PMNs, and potential therapies targeting GPX4 for SLE patients, especially lupus alopecia.