Neutrophilic Leukocytes Research Articles

An In Vitro Alveolar Model Allows for the Rapid Assessment of Particles for Respiratory Sensitization Potential.

Dust, both industrial and household, contains particulates that can reach the most distal aspects of the lung. Silica and nickel compounds are two such particulates and have known profiles of poor health outcomes. While silica is well-characterized, nickel compounds still need to be fully understood for their potential to cause long-term immune responses in the lungs. To assess these hazards and decrease animal numbers used in testing, investigations that lead to verifiable in vitro methods are needed. To understand the implications of these two compounds reaching the distal aspect of the lungs, the alveoli, an architecturally relevant alveolar model consisting of epithelial cells, macrophages, and dendritic cells in a maintained submerged system, was utilized for high throughput testing. Exposures include crystalline silica (SiO2) and nickel oxide (NiO). The endpoints measured included mitochondrial reactive oxygen species and cytostructural changes assessed via confocal laser scanning microscopy; cell morphology evaluated via scanning electron microscopy; biochemical reactions assessed via protein arrays; transcriptome assessed via gene arrays, and cell surface activation markers evaluated via flow cytometry. The results showed that, compared to untreated cultures, NiO increased markers for dendritic cell activation, trafficking, and antigen presentation; oxidative stress and cytoskeletal changes, and gene and cytokine expression of neutrophil and other leukocyte chemoattractants. The chemokines and cytokines CCL3, CCL7, CXCL5, IL-6, and IL-8 were identified as potential biomarkers of respiratory sensitization.

Circulating neutrophils activated by cancer cells and M2 macrophages promote gastric cancer progression during PD-1 antibody-based immunotherapy.

Aims: To analyze the correlation between the neutrophil-to-lymphocyte ratio (NLR) and prognosis of advanced gastric cancer (AGC) patients treated by PD-1 antibody-based therapy and to delineate molecular characteristics of circulating neutrophils by single-cell RNA sequencing (scRNA-seq). Methods: The clinicopathological information of 45 AGC patients receiving PD-1 antibody-based regimens at the Department of Oncology, Ruijin Hospital, was reviewed. Treatment outcomes including objective response rate (ORR), progression-free survival (PFS), and overall survival (OS) were recorded. The correlation between NLR and efficacy of PD-1 antibody-based treatment was analyzed. Single-cell RNA sequencing (scRNA-seq) analysis was performed based on multisite biopsy samples from two AGC patients to explore the molecular characteristics of circulating neutrophils and their pro-tumor mechanisms. Tissue samples from 88 gastric cancer patients who underwent radial gastrectomy were collected for immunochemistry staining. Results: A high posttreatment NLR was associated with poor outcomes of AGC patients receiving PD-1 antibody-based regimens. scRNA-seq analysis showed that an increased number of circulating neutrophils were found in peripheral blood samples after treatment in which neutrophil cluster 1 (NE-1) was the major subcluster. NE-1 was featured with a neutrophil activation phenotype with the high expression of MMP9, S100A8, S100A9, PORK2, and TGF-β1. NE-1 displayed an intermediate state in pseudotime trajectory analysis with gene function enrichment found in neutrophil activation, leukocyte chemotaxis, and negative regulation of MAP kinase activity. Cellular interaction analysis showed that the chemokine signaling pathway is the major interactional pathway of NE-1 between subclusters of malignant epithelial cells (EP-4) and M2 macrophages (M2-1 and M2-2). In turn, the MAPK signaling pathway and Jak-STAT signaling pathway of EP-4, including IL1B/IL1RAP, OSM/OSMR, and TGFB1/TGFBR2 axes, were identified as interacting pathways between EP-4 and NE-1. The high expression of OSMR in tumor cells was closely correlated with lymph node metastasis of gastric cancer. Conclusion: The posttreatment NLR could be a poor prognostic marker of AGC patients treated with immune checkpoint inhibitors (ICIs). Subclusters of circulating neutrophils activated by tumor cells and M2 macrophages could participate in gastric cancer progression through signaling interactions with tumor cells.

The role of the host-Neutrophil biology.

Neutrophilic polymorphonuclear leukocytes (neutrophils) are myeloid cells packed with lysosomal granules (hence also called granulocytes) that contain a formidable antimicrobial arsenal. They are terminally differentiated cells that play a critical role in acute and chronic inflammation, as well as in the resolution of inflammation and wound healing. Neutrophils express a dense array of surface receptors for multiple ligands, ranging from integrins to support their egress from bone marrow into the circulation and from the circulation into tissues, to cytokine/chemokine receptors that drive their navigation to the site of infection or tissue damage and also prime them for a second stimulus, to pattern recognition receptors and immunoglobulin receptors to facilitate the destruction and removal of infective agents or debridement of damaged tissues. When afferent neutrophil signals are proportionate and coordinated they will phagocytose opsonized and unopsonized bacteria, activating the nicotinamide adenine dinucleotide phosphate oxidase (respiratory burst) to generate reactive oxygen species, which augment the proteolytic destruction of microbes secured within the phagosome. A highly orchestrated process of apoptosis follows, forming membrane-bound substructures that are removed by macrophages. Neutrophils are capable of various other forms of programmed cell death, such as NETosis and pyroptotic cell death, as well as nonprogrammed cell death by necrosis. In recent years, research has revealed that neutrophils are capable of far more subtle cell-cell interactions than previously thought possible. This includes synthesis of various inflammatory mediators and also myeloid cell training within bone marrow, where epigenetic and metabolic signals associated with returning neutrophils that undergo reverse egress from tissues into the vasculature and back to bone marrow program a hyperreactive subset of neutrophils during myelopoiesis that are capable of hypersensitive reactions to microbial aggressors. These characteristics are evident in various neutrophil subsets/subpopulations, creating broad heterogeneity in the behavior and biological repertoire of these seemingly schizophrenic immune cells. Moreover, neutrophils are critical effector cells of adaptive and innate immunity, binding to opsonized bacteria and destroying them by extracellular and intracellular methods. The former creates substantial collateral host tissue damage, as they are less specific than T-cytotoxic cell-killing mechanisms, and in conditions such as peri-implantitis, where plasma cells and neutrophils dominate the immune infiltrate, bone and tissue destruction are rapid and appear relentless. Finally, the role of neutrophils as conduits for periodontal-systemic disease connections and for oxidative damage to act as a causal link between the two has only recently been realized. In this chapter, we attempt to expand on these issues, emphasizing the contributions of European scientists throughout a detailed appraisal of the benefits and side effects of neutrophilic inflammation and immune function.

Assessment of correlation between clinical, radiographic, microbiological, and histopathological examinations in identification of pulpal diseases - a single-centre study

AimThis study aimed to analyse the presence of pulpitis using different techniques and compare the findings of the various examination methods. MethodsA total of 108 patients were enrolled and randomly divided into two groups: 56 patients whose pulp samples were sent for histopathological analysis and 52 patients whose samples were sent for microbiological analysis. All participants underwent endodontic procedures, with clinical evaluation and assessment using periapical radiography. Bacteria were isolated and identified using agar culture and VITEK 2 identification cards. ResultsHistopathology confirmed chronic pulpitis in 33 samples (58.9%) and acute pulpitis in 23 samples (41.1 %). For chronic pulpitis, the histopathological diagnosis agreed with the clinical evaluation diagnosis in 65.2% of cases, and a similar percentage of agreement was observed for acute pulpitis. Chronic pulpitis was observed in 34.8% of patients on clinical examination; however, according to histopathology, these cases were acute. Dilated blood vessels were detected in 56.5% of patients with acute pulpitis and 15.2% of patients with chronic pulpitis. Neutrophilic leucocytes were observed in 43.5% of patients with acute pulpitis and 69.7% of patients with chronic pulpitis. Lymphocytes were observed in 17.4% of acute pulpitis samples but zero chronic pulpitis samples. Microbiological analysis identified gram-positive bacilli in 22 samples, gram-positive cocci in 51 samples, and fungi in 2 samples. Acute pulpitis was typically found to be associated with anaerobic Clostridium bifermentans, aerobic Streptococcus mitis, and Granulicatella elegans, whereas chronic pulpitis was more often associated with two facultative anaerobes, Streptococcus oralis and Streptococcus mitis. ConclusionComparison of clinical, radiographic, and histological examination techniquesrevealed several notable discrepancies. Radiographic imaging only suggested the presence of pulpal pathologies; therefore, histopathological analysis of the pulp material was still ultimately required to verify the clinical diagnosis and exclude other pathologies. Although histopathology remains the gold standard for assessing pulpal disease, performing additional examinations may provide the most comprehensive, and perhaps the most effective, approach.

Accuracy of NGAL as a Biomarker for Urinary Tract Infection in Young Febrile Children: An Individual Patient Data Meta-Analysis

To compare the accuracy of urine neutrophil gelatinase-associated lipocalin (NGAL) and leukocyte esterase (LE) for the diagnosis of urinary tract infection (UTI) in children. We performed a systematic review and individual patient data meta-analysis of studies that examined urine NGAL as a marker of UTI in children <18years of age. We created a standardized definition of UTI and applied it to all included children. We compared sensitivity, specificity, and the area under the receiver operating characteristic curve (AUC) of NGAL with LE. We included individual patient data from 3 studies for a total of 845 children. Included children had a mean age of 0.9years (SD, 0.6years). Using a cutoff of 32.7ng/mL, NGAL had a sensitivity of 90.3% (95% CI: 83.2%-95.0%) and specificity of 93.7% (95% CI: 91.7%-95.4%) for the diagnosis of UTI. LE, using a cutoff of ≧ trace had a sensitivity of 81.1% (95% CI: 72.5%-87.9%) and specificity of 97.0% (95% CI: 95.4%-98.1%). The AUC for NGAL was 0.95 (95% CI: 0.92-0.98). The AUC for LE was 0.90 (95% CI: 0.86-0.93). In young, febrile children, urinary NGAL is more sensitive for the diagnosis of UTI than LE but is slightly less specific.

The incidence, risk factors, and outcome of new-onset diabetes among post-COVID-19 patients: A single-center study

Background: Coronavirus disease 2019 (COVID-19) infection may elevate the risk of hyperglycemia and other complications in patients with and without prior diabetes history. It is not clear whether the virus induces type 1 or type 2 diabetes or instead causes a novel form of diabetes. Precise mechanism of diabetes onset in COVID-19 patients remains unresolved. Aims and Objectives: The aims of this study were to know the incidence, risk factors, and outcome of new-onset diabetes among post-COVID-19 patients and association of disease severity and occurrence of new-onset diabetes in post-COVID-19 Patients. Materials and Methods: Patients age more than 18 years, not known diabetic, tested positive with rapid antigen test or reverse transcription polymerase chain reaction admitted to a tertiary care hospital were included in the present prospective observational study. The patients who developed new-onset diabetes during the 3 months follow-up and, the risk factors associated with new-onset diabetes are assessed. Patients with hemoglobin (HbA1c) &gt;6.5% were diagnosed with new-onset diabetes. Results: Total 246 patients were non-diabetics at admission, at 1 week 188 were non-diabetics and 49 were diabetics, and nine were prediabetics. Patients were within the age range of 21–95 years with mean age of 49.46±17.02 years and male predominance (59.76%). Out of 188 non-diabetics, 19 (10.10%) developed new-onset diabetes, and 2 (1.06%) developed new-onset prediabetes after 3 months. Out of 49 diabetics, 19 (38.77%) became non-diabetic, 30 (61.22%) remained diabetic, and out of nine prediabetes 2 (22.22%) developed new-onset diabetes, 5 (55.55%) reversed to non-diabetic, and 2 (22.22%) remained prediabetic after 3 months. In total, from HbA1c at admission and 3 months, 51 subjects had new-onset diabetes (20.73%). Most common risk factors found with occurrence of new-onset diabetes were those on high dose of steroid (P=0.0001), family history of diabetes mellitus (DM) (P=0.001), over weight and obesity (P=0.0001), fungal infection (P=0.0001), and need of oxygen and intensive care unit requirement (P=0.0001). The patient with increased laboratory markers of inflammation such as ferritin, neutrophil leukocyte ratio, lactate dehydrogenase, and C-reactive protein D-dimer had strong association with occurrence of new-onset diabetes (P=0.0001). Conclusion: COVID-19 infection confers an increased risk for type 2 diabetes. Patients of all ages and genders had an elevated incidence and risk for occurrence of new-onset diabetes. Moreover, it was strongly associated with overweight and obesity, steroid dosage, and its duration, disease severity, positive family history of DM, and increased laboratory markers of inflammation. Hence, particular attention should be paid during the first 3 months after COVID-19 infection and patients need to be under follow-up for blood glucose monitoring.

May the Systemic Immune-Inflammation Index be an Indicator of Premature Ovarian Insufficiency?

Objective: This study aimed to determine whether there was a correlation between the systemic im-mune-inflammation index and ovarian reserve markers such as follicle stimulant hormone, estradiol, and anti-mullerian hormone Methods: The study comprised 65 people with premature ovarian insufficiency and 71 controls with sim-ilar demographics. The concentrations of hemoglobin, hematocrit, platelets, white blood cells, neutro-phils, and lymphocytes were evaluated. The neutrophil leukocyte ratio, platelet lymphocyte ratio, and systemic immune-inflammation index were calculated. The antral follicle count reserves of all patients were evaluated by transvaginal ultrasonography. An independent t-test was used for the comparison of the study and control groups. Correlations between variables were analyzed using Pearson's correlation test. A p value of 0.05 was considered significant. Results: The results of the neutrophil-to-lymphocyte ratio and the platelet-lymphocyte ratio showed a significant difference between the groups (p = 0.043). The Systemic Immune Inflammation Index value was the statistically significant difference found between the groups. There was a significant positive correlation between the systemic immune-inflammation index, neutrophil-to-lymphocyte ratio, platelet-lymphocyte ratio, and follicle stimulant hormone, while a significant negative correlation was found between the systemic immune-inflammation index, neutrophil-to-lymphocyte ratio, platelet-lymphocyte ratio, antral follicle count, and anti-mullerian hormone. In ROC analysis for SII at a cut-off level of 441.35, the sensitivity was 72.1% and the specificity was 68.9. Conclusion: Our study was the first in this field to reveal the relationship between premature ovarian failure and the systemic immune-inflammation index. According to our study results, the systemic im-mune-inflammation index, neutrophil-to-lymphocyte ratio, and platelet-lymphocyte ratio are significant-ly higher in individuals with ovarian failure.

Granulomatous perioral dermatitis

Parents of patient S., born in 2018 applied with complaints of rashes on the skin of her daughter's face, accompanied by itching for a year. Topically used cream methylprednisolone aceponate and methylene blue. Objectively: the pathological process was widespread, localized on the skin of the face. On the skin of the face, congestive hyperemia, multiple miliary pustules, vesicles, already opened in places, covered with serous crusts, multiple pink papules were visualized periorally. When conducting a histological examination of the affected area of the skin of the face, characteristic signs for granulomatous dermatitis were revealed: in the follicles, foci of necrosis, macrophages with an admixture of neutrophilic leukocytes, single eosinophils and giant multinucleated cells. In the deep layers of the dermis, rough, unevenly thickened and in places multidirectional bundles of collagen fibers with signs of necrosis and the presence of giant cell granulomas, as well as numerous giant cell granulomas without necrosis, exocytosis of lymphocytes, which is characteristic of granulomatous dermatitis, were determined. The diagnosis was made: Perioral dermatitis, granulomatous form. We were prescribed a long-term oral administration of an antibiotic of the macrolide group, externally calcineurin blockers, emollients and ointments containing zinc. On the background of the ongoing treatment, the rash resolved. There was a stable remission.

Treatment of Acute Respiratory Distress Syndrome Caused by COVID-19 with Human Umbilical Cord Mesenchymal Stem Cells

This study aimed to identify the impact of mesenchymal stem cell transplantation on the safety and clinical outcomes of patients with severe COVID-19. This research focused on how lung functional status, miRNA, and cytokine levels changed following mesenchymal stem cell transplantation in patients with severe COVID-19 pneumonia and their correlation with fibrotic changes in the lung. This study involved 15 patients following conventional anti-viral treatment (Control group) and 13 patients after three consecutive doses of combined treatment with MSC transplantation (MCS group). ELISA was used to measure cytokine levels, real-time qPCR for miRNA expression, and lung computed tomography (CT) imaging to grade fibrosis. Data were collected on the day of patient admission (day 0) and on the 7th, 14th, and 28th days of follow-up. A lung CT assay was performed on weeks 2, 8, 24, and 48 after the beginning of hospitalization. The relationship between levels of biomarkers in peripheral blood and lung function parameters was investigated using correlation analysis. We confirmed that triple MSC transplantation in individuals with severe COVID-19 was safe and did not cause severe adverse reactions. The total score of lung CT between patients from the Control and MSC groups did not differ significantly on weeks 2, 8, and 24 after the beginning of hospitalization. However, on week 48, the CT total score was 12 times lower in patients in the MSC group (p ≤ 0.05) compared to the Control group. In the MSC group, this parameter gradually decreased from week 2 to week 48 of observation, whereas in the Control group, a significant drop was observed up to week 24 and remained unchanged afterward. In our study, MSC therapy improved lymphocyte recovery. The percentage of banded neutrophils in the MSC group was significantly lower in comparison with control patients on day 14. Inflammatory markers such as ESR and CRP decreased more rapidly in the MSC group in comparison to the Control group. The plasma levels of surfactant D, a marker of alveocyte type II damage, decreased after MSC transplantation for four weeks in contrast to patients in the Control group, in whom slight elevations were observed. We first showed that MSC transplantation in severe COVID-19 patients led to the elevation of the plasma levels of IP-10, MIP-1α, G-CSF, and IL-10. However, the plasma levels of inflammatory markers such as IL-6, MCP-1, and RAGE did not differ between groups. MSC transplantation had no impact on the relative expression levels of miR-146a, miR-27a, miR-126, miR-221, miR-21, miR-133, miR-92a-3p, miR-124, and miR-424. In vitro, UC-MSC exhibited an immunomodulatory impact on PBMC, increasing neutrophil activation, phagocytosis, and leukocyte movement, activating early T cell markers, and decreasing effector and senescent effector T cell maturation.

Investigación con PCR en Tiempo Real e Histopatología sobre la presencia de H. felis, H. heilmannii y H. pylori en Perros

Helicobacter species such as H. heilmannii, H. pylori, H. felis, H. bizzozeronii and H. salomonis have been identified in cats and dogs, and research suggesting that these species may be zoonotic agents and has been studied intensified in recent years. The aim of this study was to reveal the presence, comparing the histopathological findings and Real-time PCR results of H. felis, H. heilmannii, and H. pylori in the stomach and liver tissues taken during the necropsies of owned, stray or shelter dogs. The material of the study consisted of stomach and liver tissues taken from 35 dogs that died for different reasons and were brought for necropsy. DNA copies of H. heilmannii were detected by Real-time PCR in the liver samples of 30 dogs using H. heilmannii-specific primers. In the case of gastric samples, Real-time PCR detected H. heilmannii in 13 cases, H. pylori in 3 cases, both H. heilmannii and H. pylori in 13 cases, and H. felis, H. heilmannii and H. pylori in 3 cases. Microscopically, neutrophil leukocyte infiltration, epithelial degeneration, fibrosis and oedema in the lamina propia, and lymphoplasmacytic cell infiltration were determined in the stomachs. In the Hemotoxylin Eosin staining of the sections, 5 cases and 14 cases in the Warthin–Starry staining were found positive for Helicobacter-like microorganisms. Microscopically, dissociation of the remark cords and hydropic degeneration in hepatocytes, and focal mononuclear cell infiltrations in some sections were determined in the livers. In conclusion, with this study, it was understood that Real-time PCR analyzes are very useful in the diagnosis of H. felis, H. heilmannii, and H. pylori. However, histopathological examinations are necessary to associate the presence of bacteria with the development of the disease.

Neutrophilic Leukocytes and Neutrophil Extracellular Traps in Native Aortic Valve Endocarditis

Abstract Infective endocarditis is a disease that affects the endocardium and often alters heart valves, notably the aortic valve. Bacteraemia and valvular endothelial damage play an essential role in the pathogenesis of infective endocarditis. The pertinent literature suggests that neutrophil extracellular traps are important contributors to the development of the disease. However, features of the valvular damage and contribution of neutrophils to the alteration of cardiac tissue are not explored sufficiently. The purpose of this study was to investigate the occurrence and distribution of neutrophilic leukocytes and neutrophil extracellular traps in native aortic valves affected by infective endocarditis, using histopathology and immunohistochemistry assays. In addition, the presence of vegetations on the heart valve was determined. Infiltration of neutrophils into the valvular leaflet was significantly more severe at the free margin (mean 5.89 ± 3.00, p &lt; 0.001) and the middle portion (mean 4.58 ± 3.64, p = 0.032) when compared to the base portion (2.05 ± 1.90). No significant differences in neutrophilic leukocyte infiltrating inflammatory lesions were found between cusp layers. The presence of myeloperoxidase and citrullinated histone expression characteristic of neutrophil extracellular traps was demonstrated by the use of immunohistochemistry in IE-affected valvular leaflets and vegetations. Collectively, the study results suggest that the free cusp margin and its middle portion of the aortic valve are exposed to enforced blood flow; endothelial damage and vegetation formation are likely to occur along with the presence of infective endocarditis-related bacteraemia.

Flow Cytometric Characterization of Hematopoietic Stem and Progenitor Cell Subpopulations in Autologous Peripheral Blood Stem Cell Preparations after Cryopreservation

Introduction: Autologous stem cell transplantation is a successful routine procedure with only a small number of non-engraftment cases, although the time to hematopoietic recovery may vary considerably across patients. While CD34 has been the decisive marker for enumerating hematopoietic stem and progenitor cells (HSPCs) for more than 30 years, the impact of CD34-positive cellular subpopulations in autologous HSPC grafts on hematopoietic reconstitution remains unclear. Methods: The two-color ISHAGE protocol represents the current gold standard for CD34+ cell enumeration but includes only the number of viable CD45+/CD34+ cells relative to the body weight of the recipient. We adapted a multicolor flow cytometry marker panel for advanced characterization of CD34 subpopulations in retained samples of autologous peripheral blood stem cell products (n = 49), which had been cryostored for a wide range from 4 to 15 years. The flow cytometric analysis included CD10, CD34, CD38, CD45, CD45RA, CD133, and viability staining with 7AAD. The findings were correlated with clinical engraftment data, including reconstitution of leukocytes, neutrophils, and platelets after transplantation (TPL). Results: We demonstrated that the identification of autologous HSPC subpopulations by flow cytometry after cryopreservation is feasible. Regarding the distribution of HSPC subpopulations, a markedly different pattern was observed in comparison to previously published data obtained using fresh autologous material. Our data revealed the largest ratio of lympho-myeloid progenitors (LMPPs) after freezing and thawing, followed by multipotent progenitors and erythroid-myeloid progenitors. A high ratio of LMPPs, representing an immature stage of differentiation, correlated significantly with early neutrophilic granulocyte and leukocyte engraftment (p = 0.025 and p = 0.003). Conversely, a large ratio of differentiated cells correlated with late engraftment of neutrophilic granulocytes (p = 0.024). Overall, successful engraftment was documented for all patients. Conclusion: We established an advanced flow cytometry panel to assess the differentiation ability of cryostored autologous peripheral blood stem cell grafts and correlated it with timely hematopoietic reconstitution. This approach represents a novel and comprehensive way to identify hematopoietic stem and progenitor subpopulations. It is a feasible way to indicate the engraftment capacity of stem cell products.

Serum Uric Acid to HDL-Cholesterol Ratio Could Be Promising Predictor of Atrioventricular Nodal Reentrant Tachycardia

Objective: Paroxysmal supraventricular tachycardia (PSVT), particularly atrioventricular nodal reentrant tachycardia (AVNRT), is a common arrhythmia with no associated structural heart disease. Inflammation has been implicated in the pathogenesis of arrhythmias. Uric acid to HDL-cholesterol ratio (UHR) is a novel inflammation marker that has been investigated in various conditions. This study aimed to explore the potential association between UHR and AVNRT. Material and Methods: A total of 136 patients were included in the study, including 86 patients with AVNRT and 50 controls with normal electrophysiologic study. All patients' medical records were reviewed, and data were obtained retrospectively. We recorded baseline features, hematologic and biochemical markers, and determined the UHR value. Results: UHR (p=0.031) and uric acid (p&lt;0.001) levels were significantly higher, while HDL cholesterol (p=0.031) levels were significantly lower in the AVNRT group. Neutrophil leukocyte ratio (NLR) was also higher in the AVNRT group (p=0.034). However, in multivariate analysis, only UHR emerged as an independent predictor for AVNRT (OR: 1.088; 95%CI: 1.022 – 1.159; p=0.008). ROC curve analysis suggested a UHR cut-off &gt; 14.05 for predicting AVNRT with 56% sensitivity and 76% specificity. Conclusion: This study identifies UHR as a promising predictor for AVNRT, shedding light on the potential role of inflammation in the arrhythmia's development. Utilizing UHR as a readily accessible marker in the evaluation of AVNRT patients may have clinical implications.

Association between morphological manifestations of inflammatory bowel disease and biochemical markers of inflammation

Aim. To determine morphological and morphometric changes in the structure of the mucous membrane of the large intestine and their relationship with the content of short-chain volatile fatty acids and fecal calprotectin in patients with inflammatory bowel disease. Materials and methods. We examined 68 patients with inflammatory bowel disease, including 30 women and 38 men aged 20–66 years, 55 patients were with ulcerative colitis and 13 patients – with Crohn’s disease. Histological examination focused on changes in the mucous membrane of the large intestine. Determination of short-chain volatile fatty acids in feces was performed using gas chromatography. Calprotectin content was measured in fecal samples by enzyme-linked immunosorbent assay. Results. Significant correlations were found between epithelial disruption and changes in crypt architecture, infiltration density, and histological activity. The higher degree of inflammation in patients with severe course was combined with a lower level of tissue nonspecific protection, which was detected by the number of goblet cells, eosinophilic and neutrophilic leukocytes. There was a decrease in the fecal concentration of butyric acid in patients with nonspecific ulcerative colitis and Crohn’s disease compared with controls, but a more significant decrease was observed in patients with severe nonspecific ulcerative colitis. The data have confirmed the association between inflammatory bowel disease and the content of short-chain volatile fatty acids, fecal calprotectin in coprofiltrate, which was evidenced by the presence of correlations between low content of short-chain volatile fatty acids and eosinophilic infiltration (r = -0.412; fecal calprotectin levels and disease severity (r = 0.589; P = 0.001), atrophy (r = 0.458; Р &lt; 0.05), infiltration density (r = 0.434; Р &lt; 0.05). Conclusions. All patients with inflammatory bowel disease are characterized by a specific histological picture, which reflected the different degrees of inflammation. Morphometric parameters more accurately show significant atrophic changes in the mucous membrane of the large intestine. The data have confirmed the association between inflammatory bowel disease and the content of short-chain volatile fatty acids and calprotectin in coprofiltrate.

Prognostic Value of Histological and Immunohistochemical Data in Diabetic Foot Ulcers

A clinical prospective observational study of the treatment outcomes of fifty-three patients with diabetic foot ulcers hospitalized twice in one specialized centre over the course of the year was conducted. The analysis of histological and immunohistochemical data of the tissues of the edges and the centre of the ulcer taken during the primary surgical treatment was performed. While performing histological analyses of wound tissues, special attention was given to the determination of cellular characteristics of leukocyte-necrotic masses, granulation tissue, and loose and dense connective tissue. Immunohistochemistry was performed using a set of monoclonal antibodies, allowing verification of neutrophilic leukocytes, fibroblasts, and endothelial cells. Unfavourable outcomes (amputation, reamputation, death from cardiovascular diseases, nonhealing ulcer within a year) were registered in 52.8% of cases. Uniform distribution of neutrophils and endothelial cell fibroblasts in all parts of the wound was recorded in patients with a favourable outcome. An unfavourable outcome was predetermined by the uneven content of these cells with a significant increase in neutrophilic leukocytosis in the bottom of the wounds, as well as a significant decrease in the number of fibroblasts and endotheliocytes in the centre of the wounds. The datasets obtained during primary surgical treatment are extremely informative to predict the outcome of the treatment of diabetic foot ulcers and indicate more active surgical strategies with the potential to reduce the treatment time, increase its effectiveness, and eventually make the treatment cost-effective.

Inflammation promotes resistance to immune checkpoint inhibitors in high microsatellite instability colorectal cancer

Inflammation is a common medical complication in colorectal cancer (CRC) patients, which plays significant roles in tumor progression and immunosuppression. However, the influence of inflammatory conditions on the tumor response to immune checkpoint inhibitors (ICI) is incompletely understood. Here we show that in a patient with high microsatellite instability (MSI-H) CRC and a local inflammatory condition, the primary tumor progresses but its liver metastasis regresses upon Pembrolizumab treatment. In silico investigation prompted by this observation confirms correlation between inflammatory conditions and poor tumor response to PD-1 blockade in MSI-H CRCs, which is further validated in a cohort of 62 patients retrospectively enrolled to our study. Inhibition of local but not systemic immune response is verified in cultures of paired T cells and organoid cells from patients. Single-cell RNA sequencing suggests involvement of neutrophil leukocytes via CD80/CD86-CTLA4 signaling in the suppressive immune microenvironment. In concordance with this finding, elevated neutrophil-to-lymphocyte ratio indicates inhibited immune status and poor tumor response to ICIs. Receiver operating characteristic curve further demonstrates that both inflammatory conditions and a high NLR could predict a poor response to ICIs in MSI- CRCs, and the predictive value could be further increased when these two predictors are combined. Our study thus suggests that inflammatory conditions in MSI-H CRCs correlate with resistance to ICIs through neutrophil leukocyte associated immunosuppression and proposes both inflammatory conditions and high neutrophil-to-lymphocyte ratio as clinical features for poor ICI response.

Clinico-radiological Profile and Outcome of Children with Viral Pneumonia admitted to Paediatric Intensive Care Unit in the pre COVID 19 period

Introduction: Viruses are common etiological agents of severe acute respiratory illness in under five children. Very few studies are available considering the profile of children with viral pneumonia admitted to paediatric intensive care unit (PICU) in our setting. Hence this study was done to describe the clinico- radiological profile and outcome of children diagnosed with viral pneumonia admitted to PICU. Methods: This was a retrospective descriptive study done in the PICU of a tertiary care hospital in South India. The presenting clinical features, blood parameters, chest radiography findings, course during the hospital stay and outcome of children with viral pneumonia (RT- PCR Positive) admitted to PICU were studied. Results: The aetiological profile of 28 children included - Influenza virus - 14 cases, Respiratory Syncytial Virus - 6 cases, Adeno virus - 4 cases, Human Boca virus - 2 cases, Human Rhino Virus - 1 case and Human metapneumo Virus - 1 case. Majority of children (50%) presented with severe respiratory distress. Predominant radiological picture included bilateral interstitial infiltrates followed by patchy alveolar consolidation. Eight children required mechanical ventilation. Complications included septic shock and MODS (n = 5), pneumothorax (n = 2), myocarditis (n = 2), pleural effusion (n = 1), ventilator associated pneumonia (n = 2) and pulmonary artery hypertension (n = 2). Mortality was observed in seven (25%) children. SpO2 / FiO2 ratio &lt; 300, shock at admission, neutrophil leucocyte ratio &gt; 2 and hypoalbuminemia were found to be significant predictors of mortality. Conclusions: Malnutrition and iron deficiency analmia were the common risk factors. Patchy alveolar consolidation is also a common radiological finding along with interstitial infiltrates. Hypoalbuminemia was a common finding among non-survivors.

Oral Epithelial Cell Culture Model for Studying the Pathogenesis of Chronic Inflammatory Disease.

The interactions between bacteria, epithelium, and neutrophilic polymorphonuclear leukocytes (neutrophils) are the key to the initiation and progression of many chronic inflammatory-immune diseases. In addition, all can be influenced by external factors, such as micronutrients, thereby providing potentially novel approaches to therapy. This chapter will therefore provide detailed methods for core techniques involved in studying cellular and molecular epithelial responses to a bacterial challenge in relation to chronic inflammatory disease pathogenesis and therapy.

Ferritin triggers neutrophil extracellular trap-mediated cytokine storm through Msr1 contributing to adult-onset Still’s disease pathogenesis

Hyperferritinemic syndrome, an overwhelming inflammatory condition, is characterized by high ferritin levels, systemic inflammation and multi-organ dysfunction, but the pathogenic role of ferritin remains largely unknown. Here we show in an animal model that ferritin administration leads to systemic and hepatic inflammation characterized by excessive neutrophil leukocyte infiltration and neutrophil extracellular trap (NET) formation in the liver tissue. Ferritin-induced NET formation depends on the expression of peptidylarginine deiminase 4 and neutrophil elastase and on reactive oxygen species production. Mechanistically, ferritin exposure increases both overall and cell surface expression of Msr1 on neutrophil leukocytes, and also acts as ligand to Msr1 to trigger the NET formation pathway. Depletion of neutrophil leukocytes or ablation of Msr1 protect mice from tissue damage and the hyperinflammatory response, which further confirms the role of Msr1 as ferritin receptor. The relevance of the animal model is underscored by the observation that enhanced NET formation, increased Msr1 expression and signalling on neutrophil leukocytes are also characteristic to adult-onset Still’s disease (AOSD), a typical hyperferritinemic syndrome. Collectively, our findings demonstrate an essential role of ferritin in NET-mediated cytokine storm, and suggest that targeting NETs or Msr1 may benefit AOSD patients.

Evaluation of treatment efficiency of diabetic foot syndrome using immunohistochemical studies

Objective. To evaluate the results of complex conservative treatment of patients with diabetic foot syndrome using immunohistochemical methods of studying tissue bioptates of the affected foot regions.&#x0D; Materials and methods. Examination and treatment of 129 patients (aged 16 to 80 years) with 1 and 2 type diabetes mellitus, whose course of the disease was complicate by the development of diabetic foot syndrome (DFS) was carried out. The main group (group 1) included 89 patients, who underwent complex conservative treatment added by the preparations mexidol and roncoleukin. Mexidol was used intravenously 10 ml for 10 days, and then tableted drugs in the dose of 125 ml. Roncoleukin was administered intravenously in the dose of 1.00.500 units: the 1st day after admission to hospital as a preoperative preparation; after the surgery on the 3rd day. The comparison group (group 2) enclosed 40 persons. They received a standard conservative treatment of DFS including sugar-containing, hypotensive and hypolipidemic drugs as well as a local therapy using sorption bandages. Before treatment and two weeks after, the tissue bioptates of the affected regions were taken from patients. For morphological study of bioptates, histochemical methods were applied: Mallorys toluidine blue staining in Heidenhain modification, PAS reaction. The number of leukocytes, lymphocytes, Ki-67 protein proliferation index, P53 protein expression were calculated.&#x0D; Results. The morphological study before the onset of treatment showed that the inflammation had a marked character in all the bioptates. The margins of the ulcerous defect were infiltrated with polymorphonuclear leukocytes. The number of neutrophils was 75 %, lymphocytes 25 %. The results of immunohistochemical analysis were the following: Ki-67 protein proliferation index was 2 %, P53 protein expression 2.9 %. Treatment of the control group (group 2) with the standard method resulted in a complete purification of the trophic ulcer from necrotic mass on the day 21st. In the infiltrate, there prevailed lymphocytes (70 %), the number of neutrophils decreased to 30 %. The result of immunohistochemical study was an increase in Ki-67 biomarker expression up to 7 %, P53 biomarker expression 5 %. Combined treatment of DFS applying mexidol and roncoleukin (group 1) demonstrated a maximum restorative effect already on the 614 day. In the infiltrate, the number of lymphocytes was 85 %, neutrophile leukocytes fell to 15 %, Ki-67 biomarker proliferation index grew to 15 %, P53 biomarker expression to 5.9 %.&#x0D; Conclusions. The use of immunohistochemical methods of research of ulcer bioptates in treatment of DFS makes it possible to reliably evaluate the dynamics of wound process. The application of original preparations roncoleukin and mexidol in complex treatment contributes to a faster reduction of inflammatory process and purification of wounds that is confirmed by an increase in Ki proliferation index and P53 apoptosis.