Leukocyte Protein Research Articles

P-2363. Efficacy Of SARS–CoV-2 Specific Antiviral Therapy for Enteroviral Myalgic Encephalomyelitis/ChronicFatigue Syndrome

Abstract Background Etiology remains elusive for Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS), and no treatment exists. Antivirals had no efficacy in randomized clinical trials (RCT) for Epstein-Barr Virus and HHV-6. Enteroviruses (EV) have been implicated, but no antivirals are available. Many patients who received SARS–CoV-2-specific antiviral drugs for acute Covid-19 (COV19) infection experienced significant improvement of prior ME/CFS symptoms. This study summarizes their responses to antivirals for SARS-Cov-2. Methods Neutralizing Antibody (NA) for Coxsackievirus B (CVB)1-6 and Echovirus 6, 7, 9, 11, 30 were done by ARUP lab. Enterovirus Protein (EVP) of Peripheral Blood Leukocytes (PBL) was determined by Western Blot. ME/CFS patients fulfilled Canadian consensus criteria, and had either elevated NA for enteroviruses and/or positive EVP in PBL. ME/CFS patients hospitalized for acute COV19 infection and patients without COV19, were given 5-10 days of IV Remdesivir (Rem) +/- immune modulators. Controls: 20 ME/CFS patients seen concurrently without Remdesivir treatment. Other ME/CFS patients (non-COV19) were given Nirmaltrelvir/Ritonavir (PAX) daily for 10 days +/- one repeat. The energy index (EI) was monitored by the patients before, during and after treatment. Significant improvement was defined as > 30%. Results 15/20 (75%) ME/CFS patients – 10/12 hospitalized, 5/8 non-COV19 patients – responded to IV Rem 2-6 weeks after infusions; remission lasted 6-8 weeks to 6-9 months before relapse. Of Controls: 2/20 had mild improvement (< 0.001, X 2 ). 104/200 (52%) of PAX-treated ME/CFS patients improved, often within 2-3 days; all relapsed within days to weeks after treatment. 66%, 33% and 44% of CVB4+, CVB3+, non-CVB3,4+ patients responded to treatment, respectively. EVP decreased and increased with clinical response and relapse. Conclusion Rem and PAX demonstrated clinical efficacy in ME/CFS patients with chronic enterovirus infections. Placebo-controlled RCT will be needed to clarify the role of antivirals in ME/CFS. Disclosures All Authors: No reported disclosures

Lung immune incompetency after mild peritoneal sepsis and its partial restoration by type 1 interferon: a mouse model study

BackgroundSepsis is commonly associated with acute respiratory distress syndrome (ARDS). Although the exaggerated inflammation may damage intact lung tissues, a percentage of patients with ARDS are reportedly immunocompromised, with worse outcomes. Herein, using a murine sepsis model, time-course immune reprogramming after sepsis was evaluated to explore whether the host is immunocompromised. Leukocyte kinetics in the lung tissue were evaluated in a male C57/BL6 mouse model of mild peritoneal sepsis induced by cecal ligation and puncture, with the survival rate exceeds 90%. Lung immune reactivity was evaluated by intratracheal instillation of lipopolysaccharide (LPS; 30 µg). Furthermore, the effect of interferon (IFN)-β in vivo and ex vivo was evaluated.ResultsFour days after sepsis, the lung water content remained high, even among mice in clinical recovery. While monocytes and neutrophils gradually accumulated in the lung interstitium, the inflammatory cytokine/chemokine expression levels in the lungs continued to decline. Intratracheal LPS instillation induced more leukocyte trafficking and protein leakage into the alveoli in the septic lung, indicating more severe lung injury. However, LPS stimulation-associated mRNA expression of tnf, il6, ccl2, and cxcl1 was suppressed. Intra-alveolar expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1, and keratinocyte-derived cytokine (KC) was also suppressed. Monocytes isolated from the lung tissue showed an impaired response in il6, ccl2, and cxcl1 to LPS. Systemic IFN-β restored the above impaired regulator function of monocytes, as did coculturing these cells from lung tissue with IFN-β.ConclusionsHistologically accelerated inflammation and paradoxically suppressed immunological regulator signaling were observed in the early recovery phase of sepsis. This observation may provide a model for the immunologically irresponsive state that occurs in some patients with sepsis. Systemic IFN-β partly restored the post-septic immunocompromised state, indicating its therapeutic potential for the immunosuppressive state seen in some patients with sepsis/ARDS.

Personalized, disease-stage specific, rapid identification of immunosuppression in sepsis.

Data overlapping of different biological conditions prevents personalized medical decision-making. For example, when the neutrophil percentages of surviving septic patients overlap with those of non-survivors, no individualized assessment is possible. To ameliorate this problem, an immunological method was explored in the context of sepsis. Blood leukocyte counts and relative percentages as well as the serum concentration of several proteins were investigated with 4072 longitudinal samples collected from 331 hospitalized patients classified as septic (n=286), non-septic (n=43), or not assigned (n=2). Two methodological approaches were evaluated: (i) a reductionist alternative, which analyzed variables in isolation; and (ii) a non-reductionist version, which examined interactions among six (leukocyte-, bacterial-, temporal-, personalized-, population-, and outcome-related) dimensions. The reductionist approach did not distinguish outcomes: the leukocyte and serum protein data of survivors and non-survivors overlapped. In contrast, the non-reductionist alternative differentiated several data groups, of which at least one was only composed of survivors (a finding observable since hospitalization day 1). Hence, the non-reductionist approach promoted personalized medical practices: every patient classified within a subset associated with 100% survival subset was likely to survive. The non-reductionist method also revealed five inflammatory or disease-related stages (provisionally named 'early inflammation, early immunocompetence, intermediary immuno-suppression, late immuno-suppression, or other'). Mortality data validated these labels: both 'suppression' subsets revealed 100% mortality, the 'immunocompetence' group exhibited 100% survival, while the remaining sets reported two-digit mortality percentages. While the 'intermediary' suppression expressed an impaired monocyte-related function, the 'late' suppression displayed renal-related dysfunctions, as indicated by high concentrations of urea and creatinine. The data-driven differentiation of five data groups may foster early and non-overlapping biomedical decision-making, both upon admission and throughout their hospitalization. This approach could evaluate therapies, at personalized level, earlier. To ascertain repeatability and investigate the dynamics of the 'other' group, additional studies are recommended.

Validation of a blood biomarker panel for machine learning-based radiation biodosimetry in juvenile and adult C57BL/6 mice

Following a large-scale radiological event, timely collection of samples from all potentially exposed individuals may be precluded, and high-throughput bioassays capable of rapid and individualized dose assessment several days post-exposure will be essential for population triage and efficient implementation of medical treatment. The objective of this work was to validate the performance of a biomarker panel of radiosensitive intracellular leukocyte proteins (ACTN1, DDB2, and FDXR) and blood cell counts (CD19+ B-cells and CD3+ T-cells) for retrospective classification of exposure and dose estimation up to 7 days post-exposure in an in-vivo C57BL/6 mouse model. Juvenile and adult C57BL/6 mice of both sexes were total body irradiated with 0, 1, 2, 3, or 4 Gy, peripheral blood was collected 1, 4, and 7-days post-exposure, and individual blood biomarkers were quantified by imaging flow cytometry. An ensemble machine learning platform was used to identify the strongest predictor variables and combine them for biodosimetry outputs. This approach generated successful exposure classification (ROC AUC = 0.94, 95% CI: 0.90–0.97) and quantitative dose reconstruction (R2 = 0.79, RMSE = 0.68 Gy, MAE = 0.53 Gy), supporting the potential utility of the proposed biomarker assay for determining exposure and received dose in an individual.

Evaluating The Factors Affecting Mortality and Survival of the Patients Who Underwent Percutaneous Endoscopic Gastrostomy; The Short-term and Long-term Results

Summary Aim Percutaneous endoscopic gastrostomy (PEG) is a common minimally invasive technique performed in patients with oral feeding problems. The aim of the current study is to evaluate the associated factors that have an impact on the short- and long-term survival and mortality in these patients who received PEG tube placement. Materials and Methods The patients who underwent PEG placement between June 2020 and June 2023 are enrolled in the study. Mortality data was extracted from the National Death Registry database. Data regarding the demographics, indications of PEG, co-morbidities, length of hospital stay prior to PEG procedure, the need for mechanical ventilator support, body-mass index, serum levels of hemoglobin, leukocyte, albumin, and c-reactive protein are extracted retrospectively. Kaplan Meier and Cox regression analyses were used to evaluate the factors affecting survival. Results A total of 137 patients are enrolled in the study (71 female and 66 male). One-month mortality was 16.7% and overall mortality during a follow-up of 38 months was 51%. Survival was significantly higher in patients with motor neuron diseases than the patients with Alzheimer’s (p=0.036). Length of hospital stay before PEG placement and hemoglobin levels were found to have a significant impact on survival in Cox regression analysis (p=0.000, p=0,009). Conclusion Length of hospital stay before PEGplacement, need for mechanical ventilator support and hemoglobin levels were found significantly associated with survival. Higher mortality in Alzheimer's patients may indicate that indications of PEG should be re-evaluated in these patients.

Human placental mesenchymal stem cells transplantation repairs the alveolar epithelial barrier to alleviate lipopolysaccharides-induced acute lung injury

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are accompanied by high mortality rates and few effective treatments. Transplantation of human placental mesenchymal stem cells (hPMSCs) may attenuate ALI and the mechanism is still unclear. Our study aimed to elucidate the potential protective effect and therapeutic mechanism of hPMSCs against lipopolysaccharide (LPS)-induced ALI, An ALI model was induced by tracheal instillation of LPS into wild-type (WT) and angiotensin-converting enzyme 2 (ACE2) knockout (KO) male mice, followed by injection of hPMSCs by tail vein. Treatment with hPMSCs improved pulmonary histopathological injury, reduced pulmonary injury scores, decreased leukocyte count and protein levels in bronchoalveolar lavage fluid(BALF), protected the damaged alveolar epithelial barrier, and reversed LPS-induced upregulation of pro-inflammatory factors Interleukin-6 (IL-6) and Tumor necrosis factor-α(TNF-α) and downregulation of anti-inflammatory factor Interleukin-6(IL-10) in BALF. Moreover, administration of hPMSCs inhibited Angiotensin (Ang)II activation and promoted the expression levels of ACE2 and Ang (1–7) in ALI mice. Pathological damage, inflammation levels, and disruption of alveolar epithelial barrier in ALI mice were elevated after the deletion of ACE2 gene, and the Renin angiotensin system (RAS) imbalance was exacerbated. The therapeutic effect of hPMSCs was significantly reduced in ACE2 KO mice. Our findings suggest that ACE2 plays a key role in hPMSCs repairing the alveolar epithelial barrier to protect against ALI, laying a new foundation for the clinical treatment of ALI.

Difficulties in diagnosing non-IgE-dependent gastrointestinal forms of food allergy: How informative is fecal calprotectin?

The prevalence of allergic diseases is increasing at rates corresponding to epidemic’s spread rates. Cow’s milk protein allergy (CMPA) is one of the most common food allergies in infancy. Its clinical variants remain hard-to-diagnose diseases due to their multiple clinical faces. Food allergy can develop along the pathway associated with both IgE and non-IgE, or mixed one. Non-IgEassociated variants of food allergies, due to the lack of accurate and specific laboratory markers, can cause significant difficulties in making a diagnosis. The search for new diagnostic markers continues. Faecal calprotectin, a calcium-binding leukocyte protein consisting of a complex of two monomers S100A8 and S100A9, may be one of them. Faecal calprotectin concentration is directly proportional to the number of leukocytes migrating into the intestinal wall, due to which faecal calprotectin becomes an accurate, non-invasive and sensitive indicator reflecting the level of inflammation in the intestines. Today, serum and faecal calprotectin are used as laboratory markers, and the latter, being more accessible and not critical for collection material is widely known as a diagnostic marker of inflammatory bowel diseases. Some studies suggest that faecal calprotectin may be used to diagnose cow’s milk protein allergy in young infants when other diagnostic tests are not available. Data from various studies remain contradictory. Further studies to investigate the possibilities of using faecal calprotectin as a marker of the gastrointestinal food allergy in young children is needed.

Rheological and Biochemical Properties of Blood in Runners: A Preliminary Report

Purpose: Physical activity induces numerous modifications in the morphological, rheological, and biochemical properties of blood. The purpose of this study was to evaluate changes in blood rheological and biochemical indicators among runners. Also, we assessed how the rheological and biochemical properties of blood in people who practised running characterised the range and direction of exercise modifications and allowed for the diagnosis of transient adaptive effects. Methods: This study included 12 athletes who regularly trained in middle- and long-distance running (6–8 times a week) and presented a high sports level (national and international class). The athletes performed a 30 min warm-up consisting of 15 min of jogging and exercises. After a 10 min rest, they completed a 3 km run with submaximal effort. Blood samples were collected at baseline and after the effort. Results: No statistically significant changes were revealed in erythrocyte, leukocyte, platelet, iron, ferritin, transferrin, erythropoietin, or C-reactive protein concentrations in the examined runners. The same applied to the elongation index at a shear stress within the range of 0.30–60.00 Pa, amplitude and total extent of aggregation, aggregation half-life, and aggregation index. A significant increase (within standard limits) was only observed in fibrinogen concentration after running. Conclusions: The lack of post-exercise changes in blood rheological and biochemical indicators in the investigated runners points at an efficient haemorheological system. This, in turn, reflects well-executed training and remarkably well-trained adaptive systems responsible for regeneration.

IMMUNOLOGICAL CRITERIA OF THE EFFECTIVENESS OF IMMUNOPREVENTIVE OF THE NON-SPECIFIC IMMUNOMODULATOR BIVEL AMONG CONTACTED CHILDREN

The aim of the study - to study the feasibility of using the natural immunomodulator BIVEL (conditionally called BI-V) as a nonspecific immunoprevention of tuberculosis (TB) among contacted children from the focies of chemodrug-resistant tuberculosis infection on the basis of clinical and immunological researches. Research metods. The object of study was 120 contacted from multidrug-resistant tuberculosis infection (FsMDR-TBI) including 75 children and 45 adolescents. The first contact group included 95 children/adolescents who did not receive BI-V and the second one ‒ 25 infected children/adolescents who received BI-V. Immunological studies determining the state of the phagocytic reactivity of immunity (phagocytic number and index, testes redox activity of neutrophils and non-enzymatic cationic lysosomal proteins of granulocytic leukocytes; cellular (CD3+, CD56+, CD3+HLA-DR+, CD4+, CD8+, CD4+/CD8+, CD4+45RA, CD16/56+) and humoral (CD19+, IgA, IgM, IgG, CIC) immunity; interleukins (TNF-α, IL-6, IL-10) and specific immunity (tuberculin diagnostics, Quantiferon test). Statistical analysis of the results obtained based on the software package Excel Results and discussion. On the background of non-specific preventive with the BI-V immunomodulator among contact infected children, normalization of blood phagocytic activity indicators was noted in the form of an increase in phagocytic number, phagocytic index, non-enzymatic cationic lysosomal proteins of granulocytic leukocytes and a decrease in HST test compared to children who had not yet received preventive BI-V. In infected children/adolescents from FsMDR-TBI, while maintaining normal values of CD3+, CD3+CD4+, CD3+CD8+, insignificant functional disorders of the cell response were revealed at the level of immunoregulatory processes due to the dominance of suppressor-cytotoxic reactions were revealed (decrease by 1.3 times IRI CD3+CD4+/CD3+CD8+, p<0.05, relative to donors), balance shift in the regulatory system of pro- and anti-inflammatory cytokines towards pro-inflammatory cytokines (increase by 2.0 times TNF-α/IL-10, p<0.01, relative to normal). The existing deviations in the regulatory system and the cellular response system disappeared after the completion of the autumn-spring BI-V course. Among the children from FsMDR-TBI who took a BI-V preventive course 8% people fell ill with various forms of primary pulmonary TB, among the children who BI-V was not prescribed ‒ 22.1%, p<0.05 during the 4-year observation. The maximum TB level occurred in the first two years of observation in both groups. Conclusions. The results of the appointment of a non-specific immunomodulator BI-V confirmed the feasibility of using this immunomodulator for contacted children and adolescents from FsMDR-TBI.

Correlation between elevated HCLS1 levels and heart failure: A diagnostic biomarker.

The correlation between hematopoietic cell-specific lyn substrate 1 (HCLS1) expression levels and heart failure (HF) remains unclear. HF datasets GSE192886 and GSE196656 profiles were generated from GPL24676 and GPL20301 platforms in gene expression omnibus (GEO) database and differentially expressed genes (DEGs) were obtained, which was followed by weighted gene co-expression network analysis, protein-protein interaction (PPI) networks, functional enrichment analysis and comparative toxicogenomics database (CTD) analysis. Heatmaps of gene expression levels were plotted. TargetScan was used to screen miRNAs regulating central DEGs. A total of 500 DEGs were found and mainly concentrated in leukocyte activation, protein phosphorylation, and protein complexes involved in cell adhesion, PI3K Akt signaling pathway, Notch signaling pathway, and right ventricular cardiomyopathy. PPI network identified 15 core genes (HCLS1, FERMT3, CD53, CD34, ITGAL, EP300, LYN, VAV1, ITGAX, LEP, ITGB1, IGF1, MMP9, SMAD2, RAC2). Heatmap shows that 4 genes (EP300, CD53, HCLS1, LYN) are highly expressed in HF tissue samples. We found that 4 genes (EP300, CD53, HCLS1, LYN) were associated with heart diseases, cardiovascular diseases, edema, rheumatoid arthritis, necrosis, and inflammation. HCLS1 is highly expressed in HF and maybe its target.

Glutathione S-transferase P1-1 induction influences the cellular proteostasis and apoptotic response to uremic proteins in peripheral blood leukocytes.

Glutathione S-transferase P1-1 induction influences the cellular proteostasis and apoptotic response to uremic proteins in peripheral blood leukocytes.

Effects of Euphorbia characias subsp. characias flower extracts on nociceptive pain and acute inflammatory models in mice

Pain and inflammation are major health issues worldwide, leading to negative consequences. Despite several drugs being available to manage these conditions, their effectiveness can be limited by cost, adverse reactions, and potential tolerance and dependence with long-term use. Euphorbia characias traditionally used in folk medicine for its diverse biological activities - including antiproliferative, antimicrobial, and anti-inflammatory effects - has not been extensively studied in vivo for its analgesic and anti-inflammatory properties. In this study, the antinociceptive and anti-inflammatory properties of the water and ethanolic extracts of E. characias flowers (ECAEFl and ECEEFl) were evaluated using various models. Both extracts significantly reduced paw licking time in a formalin-induced paw licking model, with ECAEFl specifically targeting and ECEEFl affecting both the neurogenic and inflammatory phases. Additionally, in the carrageenan-induced cell migration model, both extracts showed a significant decrease in leukocyte migration, protein extravasation and nitric oxide levels, further demostrating their anti-inflammatory activity. High-Resolution HPLC-ESI-QTOF-MS-MS and HPLC-PDA analysis characterized the chemical composition of the extracts, identifying a significant presence of phenolic compounds, particularly quercetin and its derivatives, which likely contribute to the observed biological activities. These findings highlight the potential of E. characias extracts as natural sources of compounds with antinociceptive and anti-inflammatory properties. Further investigations are needed to elucidate the underlying mechanisms and explore their therapeutic potential in pain and inflammation-related disorders.

Non-Classical Swine Leukocyte Antigens SLA-6, -7, and -8, Are Xenoantigens for Some Waitlisted Patients.

Attack of donor tissues by pre-formed anti-pig antibodies is well known to cause graft failure in xenotransplantation. Genetic engineering of porcine donors to eliminate targets of these pre-formed antibodies coupled with advances in immunosuppressive medicines have now made it possible to achieve extended survival in the pre-clinical pig-to-non-human primate model. Despite these improvements, antibodies remain a risk over the lifetime of the transplant, and many patients continue to have pre-formed donor-specific antibodies even to highly engineered pigs. While therapeutics exist that can help mitigate the detrimental effects of antibodies, they act broadly potentially dampening beneficial immunity. Identifying additional xenoantigens may enable more targeted approaches, such as gene editing, to overcome these challenges by further eliminating antibody targets on donor tissue. Because we have found that classical class I swine leukocyte antigens are targets of human antibodies, we now examine whether related pig proteins may also be targeted by human antibodies. We show here that non-classical class I swine leukocyte proteins (SLA-6, -7, -8) can be expressed at the surface of mammalian cells and act as antibody targets.

Influence of lymphopenia on long-term mortality in septic shock, a retrospective observational study

Background and objectiveThe diagnosis of infection, to diagnose septic shock, has been qualified by leukocyte counts and protein biomarkers. Septic shock mortality is persistently high (20%–50%), and rising in the long term. The definition of sepsis does not include leukocyte count, and lymphopenia has been associated with its mortality in the short term. Immunosuppression and increased mortality in the long term due to sepsis have not been demonstrated. The aim is to relate the occurrence of lymphopenia and its lack of recovery during septic shock with mortality at 2 years. Patients and methodsCohort of 332 elderly patients diagnosed with septic shock. Mortality at 28 days and 2 years was analysed according to leukocyte, neutrophil, and lymphocyte counts, and the ability to recover from lymphopenia (LRec). ResultsA total of 74.1% of patients showed lymphopenia, and 73.5% did not improve during ICU stay. Mortality was 31.0% and 50.3% at 28 days and 2 years, respectively. Lymphopenia was a predictor of early mortality (OR 2.96) and LRec of late mortality (OR 3.98). Long-term mortality was associated with LRec (HR 1.69). ConclusionsIn elderly patients with septic shock, 28-day mortality is associated with lymphopenia and neutrophilia, and LRec with 2-year mortality; this may represent 2 distinct phenotypes of behaviour after septic shock.

Uncommon bilateral orbital lesion in leukemic patient: is it cutis leukemia, celullitis or both?

Introduction : Acute Myeloblastic Leukemia can infiltrate any parts of the body including the orbit and periocular area. This article was aimed to report uncommon sign of cutis leukemia representing as orbital cellulitis.
 Case Illustration : Male, 35 yo with chief complained progressive lesion in his bilateral upper and lower eyelid since 4 days before admission. He felt pain and stingy sensation around lesion. There was history of toothache. He was diagnosed with acute myeloblastic leukemia since 2 month before. Ophthalmology examination showed his visual acuity was 6/6 in both eyes with good ocular motility.His bilateral upper and lower eyelid were edematous and spasm with multiple vesicles, papule and pustule. Conjunctival injection was found in both eyes. The reminder of ophthalmology examination was unremarkable. There were other hyperemic nodules in left mandibulae and right mastoid bone. Laboratory examination showed increased leukocyte, procalcitonin and C-Reactive Protein level. Orbital CT scan revealed edema in cutis and subcutis area in both superior and inferior bilateral palpebra. Board spectrum and anaerobe antibiotic along with steroid were administered intravenously. Biopsy was performed. After two weeks medication, lesion around both palpebra was decreased. Unfortunately, because of his poor systemic condition, he was readministered to our hospital and hospitalized in ICU. He was passed away after 8 days of hospitalization.
 Discussion : Male patient is more likely to develop orbital infiltration of leukemia, and it necessitates significant morbidity and mortality condition.
 Conclusion : Leukemia with orbital invasion should bring more awareness from both ophthalmologist and hematooncologist as it indicates more mortality and morbidity.

DYSREGULATION OF AUTOPHAGY IN PERIPHERAL BLOOD LEUCOCYTES IS A FACTOR IN THE DEVELOPMENT OF INFLAMMAGING IN IMMUNOCOMPROMISED PERSONS ON THE EXAMPLE OF THE SERVICEMEN OF THE DEFENSE FORCES OF UKRAINE AND CLEAN-UP WORKERS OF THE CHORNOBYL ACCIDENT.

To assess the functional state and age-related characteristics of autophagy in peripheral blood leukocytes as a risk factor for the development of inflammaging using the example of the servicemen of the DefenseForces of Ukraine and clean-up workers of the Chornobyl accident. A total of 103 male patients aged 28-77 (56,48 ∓ 9,05) years were examined. They included: the main group - 23 servicemen of the Defense Forces of Ukraine aged 44-59 (50,21 ∓ 5,13) years; the comparison group - 57 clean-up workers of the Chornobyl accident aged 56-63 (60,31 ∓ 1,78) years; and the control group -23 civilians aged 28-77 (53,26 ∓ 15,98) years. The individuals in the main and control groups were divided according to age into subgroups under 50 years and over 50 years. Clean-up workers were divided into 3 subgroups depending on the radiation dose: І - D < 100 mSv, ІІ - 100 < D < 500 mSv and ІІІ - D > 500 mSv. Analysis of autophagyparameters in peripheral blood leukocytes (PB) was performed using flow cytometry and polymerase chain reaction. In patients of the main group, the autophagy activity factor (AAF) of granulocytes and the expression of theSQSTM1 gene in PB leukocytes decreased. A decrease in chloroquine-induced accumulation of LC3B protein in leukocytes, AAF in PB monocytes and the expression of the MTOR, RB1CC1 and MAP1LC3B genes was revealed in servicemenof the Defense Forces of Ukraine under 50 years of age. The spontaneous levels of LC3B protein and AAF in monocytesand the expression level of PIK3C3, ULK1 and MAP1LC3B genes in PB leukocytes were increased in servicemen of theDefense Forces of Ukraine over 50 years of age. The clean-up workers of different dose groups showed a decrease inthe AAF in lymphocytes and granulocytes, the LC3B level in monocytes after incubation with chloroquine, the expression of the MTOR, RB1CC1, SQSTM1, ULK1, MAP1LC3B, BECN1 and PIK3C3 genes in PB leukocytes, and the AAF of monocytes was higher. Similar changes were revealed in the indices of chloroquine-induced LC3B accumulation in lymphocytes and monocytes of the clean-up workers and servicemen of both age groups, as well as the spontaneous LC3B protein level in PB monocytes of the clean-up workers irradiated at doses above 100 mSv and civilians over 50 years old. Unidirectional dysregulation of autophagy was established in the servicemen of the Defense Forces ofUkraine and the clean-up workers of the Chornobyl accident. The existing changes in autophagy parameters can leadto disruption of the functioning of the autophagic apparatus of leukocytes at the level of mRNA and protein, as wellas signaling pathways, and be associated with age-related changes at both the cellular and organismal levels. Theemergence of new and persistent earlier stress factors as a result of the war creates an additional load on the mechanisms of maintaining homeostasis, which is observed in individuals exposed to ionizing radiation more than 30years later. The found intergroup differences and similarities can activate the same or similar mechanisms of pathological processes, which will ultimately increase the risks of developing age-associated chronic somatic pathologyin younger age groups.

ECRG4 mediates host response to cutaneous infection by regulating neutrophil recruitment and adhesion receptor expression.

Rapid neutrophil recruitment is critical for controlling infection, with dysfunctional neutrophil responses in diseases like diabetes associated with greater morbidity and mortality. We have shown that the leukocyte protein ECRG4 enhances early neutrophil recruitment to cutaneous wounds and hypothesized that ECRG4 regulates the early host response to infection. Using a cutaneous infection model, we found that ECRG4 KO mice had decreased early neutrophil recruitment with persistent larger lesions, increased bacterial proliferation and systemic dissemination. Although previous work identified ECRG4 as a negative regulator of CD44 on neutrophils, the mechanism regulating neutrophil recruitment remained unknown. We demonstrated that pro-inflammatory responses were intact in ECRG4 KO mice, but found decreased neutrophil mobilization from bone marrow and decreased migration to chemokines. ECRG4 KO mouse neutrophils demonstrated an increase in adhesion molecules that regulate recruitment, including enhanced induction of integrin CD11b and increased L-selectin and CD44 on bone marrow neutrophils. Analysis of gene expression in leukocytes from diabetic patients found decreased ECRG4 expression with similar increased L-selectin and CD44. We propose a previously unrecognized mechanism governing neutrophil recruitment, whereby ECRG4 mediates neutrophil surface adhesion molecules that determine both recruitment and outside-in signaling that modulates neutrophil response to pro-inflammatory stimuli.

BACTERIAL INFECTIONS-INDUCED ORAL ERYTHEMA MULTIFORME IN A SEPTIC CHILD: A RARE CASE REPORT

Background: Sepsis is a medical emergency condition that triggers an immune response due to an infectious process. Causative organisms in sepsis including viruses, bacteria, fungi, and parasites. Microorganism infection is one of the etiologies of erythema multiforme. Objective: This case report was aimed to describe the bacterial infection as a trigger for oral erythema multiforme in a septic child. Case: A 12-years-old boy was referred to the Oral Medicine Department. Extraoral showed a symmetric face and he was using a nasogastric tube. There were erosive lesions and tend to bleed, hemorrhagic crusts on the upper and lower lips. Laboratory test results revealed increasing in leukocyte (15,880/µL), procalcitonin (24.58 ng/mL), and C-reactive protein (3.67 mg/L). The identified microorganisms in pus specimens including gram-positive coccus and gram-negative rods as well as the isolated bacterial colonies were Enterococcus faecalis, Klebsiella pneumoniae, and Citrobacter koseri. The diagnosis was oral erythema multiforme induced by bacterial infections. Case management: The medications given by the Pediatric Department were cefotaxime vial 1 gram, metronidazole 500 mg/100 ml, and paracetamol 1 gr/100 ml. The Oral Medicine Department gave the instructions for compressing the lips with gauze soaked in 0.9% Sodium Chloride solution four times a day to remove crusts and accelerate wound healing. Oral lesions showed significant improvement after 7 days of therapy. Conclusion: Oral erythema multiforme in a pediatric patient could be induced by sepsis of bacterial infection. The microbial infection causes the release of endotoxins that trigger erythema multiforme. Keywords: Bacterial infection, Oral erythema multiforme, Sepsis.

Relationship between catheter related cerebrospinal fluid infections and systemic immune-inflammation index.

This study investigated the relationship between the systemic immune inflammation index (SII) and catheter-related infections and their effects on prognosis in pediatric patients. A total of 56 pediatric patients diagnosed with ventriculoperitoneal (V-P) shunt infection between January 2017 and October 2019 were included. V-P shunt infection diagnosis was made based on clinical findings. All cerebrospinal fluid (CSF) samples were subjected to direct microscopic examination and culture. Protein, glucose, and sodium levels in CSF, CSF leukocytes, and hematological and biochemical parameters were measured. Fifty-six patients with growth in CSF culture were included in this study. 55.4% of the cases were female and 44.6% male. V-P shunt was detected in 82.1% of the cases and external ventricular drainage (EVD) catheter-related infection in 17.9%. The CSF/blood glucose ratio was significantly lower (p=0.046), and SII was significantly increased (p=0.002) in non-coagulase-negative staphylococci. Early and appropriate antibiotic therapy reduces morbidity and mortality in catheter-related infections. However, it is important to start empirical antibiotherapy until culture results are expected. Therefore, further research on the estimation of possible factors is needed.

Diagnostic Value of Ascitic Endocan Levels in the Diagnosis of the Spontaneous Bacterial Peritonitis

Abstract Background Spontaneous bacterial peritonitis (SBP) is a serious complication of liver cirrhosis and is defined as infected ascites in the absence of any recognizable secondary cause of infection. One of the possible mechanisms of developing Spontaneous bacterial peritonitis is bacterial translocation. Aim of the Work The aim of this study is to assess the role of ascitic endocan levels in the diagnosis of SBP in decompensated cirrhosis. Patients and Methods 90 cirrhotic patients with ascites were included in the current study and divided into 2 groups: SBP group with positive ascites culture or with an AF PMNL count ≥ 250/mm3 included 45 subjects. Non SBP group with negative ascites culture or with an AF PMNL count &amp;gt; 250/mm3 included 45 subjects. In the SBP group, the mean age was 58.77 years with male predominance (68.8%).As regard to Non SBP group, the mean age was 59.31 years with male predominance 57.8%. Results There was a statistical significant difference in between SBP group and Non-SBP group as regard to Platelets, ALT, ascetic Total leucocyte count, Protein and LDH. The most common presentation in SBP group was Upper GIT bleeding and Abdominal pain and the most common organism detected in the culture of ascitic fluid Escherichia coli (80%) followed by Streptococcus viridans (15.5%). Ascitic Endocan level was significantly higher in SBP group than Non-SBP group with P &amp;lt; 0.001. The sensitivity and specificity of Ascitic Endocan in detection of SBP were 84.85% and 82.69% respectively. There was a positive significant correlation in between Ascitic Endocan level and ALT, AST, bilirubin, CRP, Child class, MELD score, Blood Leukocytes and TLC (cell/mm3) in ascitic fluid in SBP group. The significant predictors of bad outcome in patients with SBP were higher ascitic fluid Endocan level, CRP and higher Glucose in ascitic fluid (mg/dl) levels Conclusion High Ascitic endocan level is common in cirrhotic patients with SBP than in Non-SBP and that means Ascitic endocan is related to the severity of the disease. Ascitic endocan level is an independent risk factor for SBP. High Ascitic endocan level is a significant predictor of bad outcome in patients with SBP. Therefore, measurement of Ascitic endocan level provide a new strategy to more aggressive treatments for high-risk groups. Ascitic endocan level could be used as an inflammatory biomarker for the prognosis of patients with SBP