Neutrophils Research Articles

Is there a faster and easier method for assessing the severity of acute mesenteric ischemia?

Acute mesenteric ischemia (AMI) is a potentially fatal abdominal emergency. The estimation of the severity of AMI is of great importance since changes in disease severity may have different impacts on the treatment options. This study aims to define laboratory and radiological parameters that can successfully predict the severity of AMI. Data from 100 patients who were treated conservatively and underwent an operation with a diagnosis of AMI between the years 2010 and 2019 were reviewed. The patients were divided into 3 groups as those treated with a conservative approach (group 1), those with partial intestinal ischemia (group 2), and those with complete intestinal ischemia (group 3) according to the pathology results. Laboratory findings of the patients were recorded and matched with radiological findings. The white blood cell (WBC) count, neutrophil (NEUT) count, neutrophil/lymphocyte ratio, and C-reactive protein/albumin ratios were the considered distinctive parameters for distinguishing the third group from the first group. However, the same result cannot be applied to the first and the second groups since only the WBC and NEUT counts showed distinctive performance. The measurement of neutrophil/lymphocyte ratio, WBC, NEUT, and albumin levels can be used to predict the severity of AMI. We believe that evaluating these laboratory parameters will greatly prevent possible morbidity and mortality in the patient. Also, we were able to observe that the parameters used in predicting AMI severity can be verified with rapid and low-cost radiological imaging techniques.

Empyema associated with peritoneal dialysis peritonitis.

A 65-year-old woman on peritoneal dialysis (PD) was admitted due to abdominal pain with cloudy PD effluent. The white blood cell count in PD effluent was 5860/µL with 85% polymorphonuclear neutrophils. Therefore, she was clinically diagnosed with peritonitis. The cultures of PD effluent were negative. Initial abdominal computed tomography did not find suggest any intraabdominal pathology. The patient was treated with empirical intraperitoneal antibiotics. Because abdominal pain with cloudy PD effluent persisted, the PD catheter was removed eventually. The culture of the removed PD catheter grew Klebsiella pneumoniae. However, intermittent fever was noted over the following days and empyema developed approximately 2 weeks after PD catheter removal. The culture of pleural fluid also grew K. pneumoniae. Another computed tomography revealed multiple intraabdominal abscesses that was assumed to come from a complication of PD-associated peritonitis. We postulate that the empyema might be caused by transdiaphragmatic extension of the intraabdominal abscesses into the pleural space.

The predictive value of LGR for distant metastasis-free survival in locally advanced rectal cancer patients

Studies show that inflammation induced by cancer is a key factor in carcinogenesis. Here, we sought to assess the relationship between patients with locally advanced rectal cancer (LARC) and the lymphocyte to neutrophil granulocyte ratio (LGR) prior to neoadjuvant chemoradiotherapy (nCRT) and distant metastasis-free survival (DMFS). Using a receiver operating characteristic (ROC) analysis of 326 LARC patients who underwent total mesorectal excision (TME) surgery and neoadjuvant chemoradiotherapy, we were able to determine the ideal LGR cutoff value. We used the Kaplan-Meier method and univariate and multivariate Cox regression to study the clinical characteristics of LARC patients in comparison between the low LGR group and the high LGR group. DMFS analysis was one of the primary clinical variables examined. We discovered that the low LGR group of LARC patients had a longer DMFS than the high LGR group. The median duration of follow-up for LARC patients was 89.4 months, with a significantly lower DMFS observed in the high LGR group compared to the low LGR group. Multivariate Cox regression analysis revealed that LARC patients with low LGR levels, early ypTNM stages, and BRAF wild had longer DMFS. LGR prior to nCRT was a critical prognostic indicator that contributed extra predictive value beyond conventional clinicopathological characteristics to predict the outcome of LARC patients receiving neoadjuvant chemoradiotherapy followed by TME surgery.

NaRNA-LL37 composite DAMPs define sterile NETs as self-propagating drivers of inflammation

Neutrophil extracellular traps (NETs) are a key antimicrobial feature of cellular innate immunity mediated by polymorphonuclear neutrophils (PMNs). NETs counteract microbes but are also linked to inflammation in atherosclerosis, arthritis, or psoriasis by unknown mechanisms. Here, we report that NET-associated RNA (naRNA) stimulates further NET formation in naive PMNs via a unique TLR8-NLRP3 inflammasome-dependent pathway. Keratinocytes respond to naRNA with expression of psoriasis-related genes (e.g., IL17, IL36) via atypical NOD2-RIPK signaling. In vivo, naRNA drives temporary skin inflammation, which is drastically ameliorated by genetic ablation of RNA sensing. Unexpectedly, the naRNA-LL37 ‘composite damage-associated molecular pattern (DAMP)’ is pre-stored in resting neutrophil granules, defining sterile NETs as inflammatory webs that amplify neutrophil activation. However, the activity of the naRNA-LL37 DAMP is transient and hence supposedly self-limiting under physiological conditions. Collectively, upon dysregulated NET release like in psoriasis, naRNA sensing may represent both a potential cause of disease and a new intervention target.

Application of metagenomic next-generation sequencing in optimizing the diagnosis of ascitic infection in patients with liver cirrhosis

BackgroundMetagenomic next-generation sequencing (mNGS) is an emerging technique for the clinical diagnosis of infectious disease that has rarely been used for the diagnosis of ascites infection in patients with cirrhosis. This study compared mNGS detection with conventional culture methods for the on etiological diagnosis of cirrhotic ascites and evaluated the clinical effect of mNGS.MethodsA total of 109 patients with ascites due to cirrhosis were included in the study. We compared mNGS with conventional culture detection by analyzing the diagnostic results, pathogen species and clinical effects. The influence of mNGS on the diagnosis and management of ascites infection in patients with cirrhosis was also evaluated.ResultsAscites cases were classified into three types: spontaneous bacterial peritonitis (SBP) (16/109, 14.7%), bacterascites (21/109, 19.3%) and sterile ascites (72/109, 66.1%). In addition, 109 patients were assigned to the ascites mNGS-positive group (80/109, 73.4%) or ascites mNGS-negative group (29/109, 26.6%). The percentage of positive mNGS results was significantly greater than that of traditional methods (73.4% vs. 28.4%, P < 0.001). mNGS detected 43 strains of bacteria, 9 strains of fungi and 8 strains of viruses. Fourteen bacterial strains and 3 fungal strains were detected via culture methods. Mycobacteria, viruses, and pneumocystis were detected only by the mNGS method. The mNGS assay produced a greater polymicrobial infection rate than the culture method (55% vs. 16%). Considering the polymorphonuclear neutrophil (PMN) counts, the overall percentage of pathogens detected by the two methods was comparable, with 87.5% (14/16) in the PMN ≥ 250/mm3 group and 72.0% (67/93) in the PMN < 250/mm3 group (P > 0.05). Based on the ascites PMN counts combined with the mNGS assay, 72 patients (66.1%) were diagnosed with ascitic fluid infection (AFI) (including SBP and bacterascites), whereas based on the ascites PMN counts combined with the culture assay, 37 patients (33.9%) were diagnosed with AFI (P < 0.05). In 60 (55.0%) patients, the mNGS assay produced positive clinical effects; 40 (85.7%) patients had their treatment regimen adjusted, and 48 patients were improved. The coincidence rate of the mNGS results and clinical findings was 75.0% (60/80).ConclusionsCompared with conventional culture methods, mNGS can improve the detection rate of ascites pathogens, including bacteria, viruses, and fungi, and has significant advantages in the diagnosis of rare pathogens and pathogens that are difficult to culture; moreover, mNGS may be an effective method for improving the diagnosis of ascites infection in patients with cirrhosis, guiding early antibiotic therapy, and for reducing complications related to abdominal infection. In addition, explaining mNGS results will be challenging, especially for guiding the treatment of infectious diseases.

Does COVID-19 infection alter serum biochemical and hematological biomarkers in deceased dementia patients?

Abstract Objectives The elderly population is categorized as a risk group for COVID-19 infection, and dementia is the primary cause of disability in elderly individuals and affects 70 % of the elderly population. In this study, we evaluated the blood and serum biomarkers of deceased dementia patients infected by COVID-19 compared to the survived dementia and non-dementia patients. Methods Laboratory biomarkers of 11 dementia patients infected by COVID-19 have been used for this study. The five patients’ serum biochemistry and blood data were compared with the six patients who died because of COVID-19. Additionally, data from nine patients aged 85–96 infected with COVID-19 without dementia have been used to compare the difference between dementia and non-dementia individuals. Results D-dimer, C-reactive protein (CRP), glucose, blood urea nitrogen (BUN), alanine transaminase (ALT), aspartate aminotransferase (AST), troponin, procalcitonin, red cell distribution width (RDW), white blood cell (WBC), neutrophil (NEU) and %NEU levels significantly increased in the deceased dementia patients compared to the survived and non-dementia individuals. Calcium (Ca), hematocrit (HCT), red blood cells (RBC), lymphocyte (%LYM), monocyte %MONO, and basophil (%BASO) levels significantly decreased in the deceased dementia patients compared to the survived and non-dementia individuals infected by COVID-19. Conclusions Serum biochemistry and hematological biomarkers, including D-dimer, CRP, glucose, ALT, AST, BUN, troponin, procalcitonin, RDW, RBC, WBC, NEU, %NEU, Ca, HCT, %LYM, %MONO, and %BASO were significantly altered in deceased dementia patients infected by COVID-19 compared to the survived individuals.

Analysis of Risk Factors for Recurrent Pregnancy Loss in Patients Undergoing in vitro Fertilization-Embryo Transfer

Recurrent pregnancy loss (RPL) presents a formidable challenge for individuals undergoing in vitro fertilization-embryo transfer (IVF-ET), forming both a clinical dilemma and a focal point for scientific inquiry. This study endeavors to investigate the intricate interplay between clinical features, such as age, body mass index (BMI), and waist-to-hip ratio (WHR), and routine laboratory parameters, including sex hormones, blood composition, liver and thyroid functions, thyroid antibodies, and coagulation indicators, in RPL patients undergoing IVF-ET. By meticulously analyzing these variables, we aim to uncover the latent risk factors predisposing individuals to RPL. Identifying potential factors such as advanced maternal age, obesity, and insulin resistance will provide clinicians with vital insights and empirical evidence to strengthen preventive strategies aimed at reducing miscarriage recurrence. This retrospective case-controlled study included RPL patients who underwent IVF-ET treatment at Sun Yat-sen Memorial Hospital, Sun Yat-sen University, between January 2012 and March 2021 as the case cohort, compared with women receiving assisted reproductive treatment due to male infertility as the control cohort. The fasting peripheral blood was collected 5 days before the first menstrual cycle at least 12 weeks after the last abortion. The clinical characteristics and relevant laboratory indexes of the two groups were compared. Employing both univariate and multivariate logistic regression analyses, we sought to unearth potential high-risk factors underlying RPL. Additionally, a linear trend analysis was conducted to assess the linear relationship between total testosterone (TT) levels and the number of miscarriages. In contrast to the control cohort, the RPL cohort exhibited significant increases in age, BMI, and WHR (P<0.05). Notably, TT levels were markedly lower in the RPL cohort (P=0.022), while no significant differences were observed between the two groups concerning basal follicle-stimulating hormone, luteinizing hormone, estradiol, progesterone, prolactin levels, and anti-Müllerian hormone levels (P>0.05). Moreover, fasting insulin (FINS) levels and HOMA-IR index were notably elevated in the RPL cohort relative to the control cohort (P<0.001), although no significant differences were observed in fasting blood glucose levels (P>0.05). Furthermore, the neutrophil (NEU) count and NEU-to-lymphocyte ratio were notably higher in the RPL cohort (P<0.01). Univariate logistic regression analysis identified several factors, including age≥35 years old, BMI≥25 kg/m2, WHR>0.8, FINS>10 mU/L, HOMA-IR>2.14, NEU count>6.3×109 L－1, and an elevated NEU/lymphocyte ratio (NLR), as significantly increasing the risk of RPL (P<0.05). Although TT levels were within the normal range for both cohorts, higher TT levels were associated with a diminished RPL risk (odds ratio [OR]=0.67, 95% confidence interval [CI]: 0.510-0.890, P=0.005). After adjustments for confounding factors, age≥35 years old (OR=1.91, 95% CI: 1.06-3.43), WHR>0.8 (OR=2.30, 95% CI: 1.26-4.19), and FINS>10 mU/L (OR=4.50, 95% CI: 1.30-15.56) emerged as potent risk factors for RPL (P<0.05). Conversely, higher TT levels were associated with a reduced RPL risk (OR=0.59, 95% CI: 0.38-0.93, P=0.023). Furthermore, the linear trend analysis unveiled a discernible linear association between TT levels and the number of miscarriages (P trend=0.003), indicating a declining trend in TT levels with escalating miscarriage occurrences. In patients undergoing IVF-ET, advanced maternal age, lower TT levels, increased WHR, and elevated FINS levels emerged as potent risk factors for RPL. These findings provide clinicians with valuable insights and facilitate the identification of patients who are at high risks and the formulation of preventive strategies to reduce the recurrence of miscarriages.

Hluboké krční záněty u dětí – přístup k diagnostice a léčbě

Summary Introduction: In the treatment of deep throat infections, it is necessary to make a correct decision about surgical treatment. The aim was to observe trends and characteristics of patients requiring surgical treatment and to identify factors that predict the type of treatment. Materials and methods: This is a retrospective study including pediatric patients hospitalized in the Children‘s Hospital of The University Hospital Brno with deep throat infections between 2010–2019. Studied parameters: age, sex, duration of problems before hospitalization, presence of dysphagia, odynophagia, impaired neck mobility, bulging of the pharyngeal wall, ankylostomia, respiratory problems, dysphonia, drooling, cervical lymphadenitis, sore throat, length of hospitalization, conservative or surgical approach, and well-being day. Laboratory investigations included CRP, neutrophil granulocyte count, and microbiological findings. Imaging examinations monitored the presence of phlegm or abscess, side, position relative to the great vessels and oppression of the great vessels, and size of abscess. Results: This study presents the clinical characteristics of 78 deep throat infections. Observed trends: patients with abscesses were younger, had higher CRP, and were hospitalized longer. Larger abscess statistically resulted in surgical treatment. Patients with phlegm had a shorter time to achieve comprehensive improvement. The incidence of an abscess is not related to gender. Boys had statistically significantly higher CRP and neutrophil granulocyte counts. The most common microbiological finding was Streptococcus pyogenes. Conclusion: Patients with abscesses were younger, had longer hospital stays, and had higher CRP. Key words children – deep neck infection – abscess

INFECTION CONTROL IN COVID-19 PATIENTS BASED ON POLYMORPHISMS OF TMPRSS2 (rs12329760), FGB (rs1800790), AND NOS3 (rs2070744) GENES

Objective of this study is to investigate the potential of anti-infective protection in patients with COVID-19 by analyzing the absolute and relative number of the main populations of immunocompetent peripheral blood cells depending on the polymorphism of the genes of transmembrane serine protease 2 (TMPRSS2, rs12329760), fibrinogen beta (FGB, rs1800790) and endothelial nitric oxide synthase (NOS3, T-786C, rs2070744). Materials and methods. A total of 204 patients with mild, moderate, and severe COVID-19-associated pneumonia were included in the single-center study. Among the patients were 51.97% (106) women and 48.03% (98) men. Among the patients, there were 51.97% (106) women and 48.03% (98) men, with an average age of 55.93±8.75 years. Anti-infective protection was assessed based on an extended complete blood count (CBC) with the calculation of the main populations of immunocompetent cells. The polymorphism of the TMPRSS2 (rs12329760), FGB (rs1800790) and NOS3 (rs2070744) genes was investigated by real-time polymerase chain reaction (Real Time PCR). Results. There were no differences in the absolute and relative number of most populations of immunocompetent peripheral blood cells between the genotypes of the TMPRSS2 (rs12329760) gene. The absolute and relative number of immunocompetent peripheral blood cell populations between the genotypes of the FGB (rs1800790) gene also did not differ significantly. However, in AA genotype carriers, there was a tendency to decrease the number of neutrophilic granulocytes due to mature segmented nucleated forms against the background of an increase in eosinophilic granulocytes by 27.27% (p=0, 038) and 55.55% (p=0.007) and agranulocytes due to lymphocyte sprouting and monocytes by 8.87-20.09% that implies a more severe course of the disease and a stronger stress of non-specific immunity than in G-allele holders. Conclusions. Inflammatory changes in the complete blood count of the main populations of immunocompetent cells in patients with COVID-19 do not show a consistent dependence on the genotypes of the TMPRSS2 (rs12329760) and NOS3 (rs2070744) genes. However, the presence of the AA genotype of the FGB gene (rs1800790) in patients with COVID-19 is associated with a more severe course of the disease and increased stress on the monocyte-macrophage system.

COLD INHIBITION OF BIOACTIVITY OF PROBIOTIC MYCOBACTERIA AND AEROCOCCI IN THE MICROBIOME

The organism of farm animals is constantly under the influence of various factors of the external environment, and one of the negative factors of violation of the technology of keeping and creation of unsatisfactory conditions of existence with deterioration of well-being is cold injury. Mostly it manifests itself as a permanent effect of positive temperatures well below the physiological norm or periodic fluctuations of an uncomfortable temperature. Such a stress factor of low intensity, but permanent over a long period of time, causes negative pathophysiological changes in the macroorganism with the development of immune-depression, metabolic syndrome, perversions of the qualitative composition of the microbiome and, as a result, disruption of native physiological processes of stimulation of cellular mechanisms of nonspecific innate immunity, induced by immune-biological mechanisms of the functioning of neutrophil granulocytes. The negative effect of low-intensity permanent cold stress was studied on a model object - murchakаs, which were kept for two weeks at a temperature of 4-6 °C under conditions of light and movement restriction. Before the start of the experiment, initial microbiological studies of the microbiome of the large intestine were conducted using generally accepted methods. Resident indigenous probiotic prokaryotes - M. vaccae &amp; A. viridans, which possessed typical species characteristics, were isolated. Pathogenic variants of enterobacteria were not isolated, isolated cultures of E. coli were apathogenic and belonged to the usual normal flora. A two-week cold injury resulted in the disappearance of the indigenous probiotic microbiota M. vaccae &amp; A. viridans, which are indicator prokaryotes of the physiological well-being of the macroorganism, from the microbiome. At the same time, the number of potentially pathogenic enteral microorganisms, coccal and rod-shaped, increased. This was correlated with a decrease in the phagocytic activity of native neutrophil granulocytes and their preformed filamentous spatial nucleoprotective mesh structures, which inhibit the de-fragmentary function in relation to genetically non-syngenic objects.

INFLUENCE OF FEED ADDITIVE "ENZAKTIVE MIX" ON THE HEMATOLOGICAL PROFILE OF THE BLOOD OF SOWS AND THEIR PIGLETS

The dynamics of the haematological profile changes and the productivity of sows and piglets obtained from them when feeding the feed additive "EnzActive Mix" were described in the article. The feed additive contains yeast of the genus Saccharomyces cerevisiae with an activity of at least 1.5 x 10^10 CFU/g and a six enzymes complex with protease, cellulase, xylanase, ɑ-amylase, β-gluconase, phytase, which are located under the layer of yeast cells. The results of the study showed that the use of "EnzActive Mix" in sows and their piglets’ diet has a stimulating effect on the oxygen transport function of blood. This is evidenced by a higher number of erythrocytes and platelets, a higher concentration of haemoglobin and haematocrit in the blood of sows of the experimental group compared to the control group on 21st day of lactation (p&lt;0.05). Addition of the feed additive "EnzActive Mix" to the diet caused changes in certain forms of leukocytes ratio in the sows’ blood. Thus, during this period, a higher (p&lt;0.05) relative lymphocytes number and a tendency to decrease of segmented neutrophils number in the blood of experimental group sows compared to the control group were found. Similar changes were noted in the haematological parameters study of piglets obtained from these sows. In particular, the leukocytes number in the blood of the experimental group piglets at 5-, 14- and 28-day age was higher than in their peers of the control group. At the same time, on the 5th day of life, the differences relatively to the control group animals were significant (p&lt;0.05). Feeding of the feed additive led to a decrease in the segmented neutrophils relative number in the blood of the experimental group piglets on the 14th and 28th day of life (p&lt;0.05) and an increase in the lymphocytes number (p&lt;0.05) during the indicated periods of the study. The positive effect of the feed additive "EnzActive Mix" on the productivity parameters of the studied animals was established. In particular, the body weight of experimental group piglets at 5-, 14- and 28-day age was 7.8%, 13.7% (p&lt;0.05) and 8.4% (p&lt;0.01) higher, respectively, than that of the control group.

PROSTHETIC GRAFT INFECTION: WHEN ECHOCARDIOGRAPHY IS NOT ENOUGH!

Abstract A 68–year–old Caucasian male, post–Bentall procedure with a bioprosthetic valve following extensive aortic dissection involving the ascending aorta and aortic arch, presented to the Emergency Department with persistent fever for over a month. During his hospitalization, he experienced ventricular fibrillation in the setting of hypokalemia, successfully managed with Advanced Cardiovascular Life Support (ACLS) and DC shock, leading to the restoration of sinus rhythm. Subsequently, he was admitted to the Cardiovascular Intensive Care Unite. Blood tests revealed neutrophilic leukocytosis with elevated inflammatory markers, anemia, and a troponin curve consistent with myocardial injury in the context of sepsis and anemia. Echocardiography demonstrated a left ventricle of normal dimensions with globally depressed function (EF 45%), attributed to apical akinesia. The prosthetic valve was functioning normally, and there were no signs of endocarditic vegetations or pericardial involvement. A transesophageal echocardiogram confirmed the absence of vegetations. Given echocardiographic signs of myocardial ischemia, coronary angiography was performed, revealing a coronary circulation free from significant stenosis. Cultural exams showed positivity for Enterococcus faecalis. A PET–CT revealed the presence of radioisotope uptake around the vascular prosthesis situated in the aortic arch. An Infectious Disease specialist recommended antibiotic therapy with Piperacillin/Tazobactam and Teicoplanin. At the one–month follow–up, the patient remained asymptomatic, free from new febrile episodes. The follow–up echocardiogram indicated improved global systolic function, and inflammatory markers remained consistently negative. Repeat blood cultures were negative. This case illustrates a suspected endocarditis on a prosthetic valve where both transthoraci and transesophageal echocardiography yielded negative results. The use of PET–CT, on the other hand, revealed the presence of vascular graft infection, proving to be a valuable method in this clinical setting.

A rare cause of acute abdomen in a young immunocompetent woman

Streptococcus pyogenes is ahuman pathogenic, gram positive bacterium that primarily leads to pharyngitis or soft tissue infections. Primary peritonitis caused by S.pyogenes infection is rare and there are only afew published cases worldwide. Primary peritonitis due to other pathogens occurs in immunosuppressed conditions such as HIV or other chronic diseases. However, younger, healthy women are more likely to be affected by S.pyogenes peritonitis. At present, the underlying molecular mechanisms can only be speculated on. One possibility is that, similar to the clinical picture of streptococcal toxic shock syndrome (STSS), aspecific serotype of the Mprotein in combination with inhibition of the cell response of neutrophil granulocytes could play arole. In addition to peritonitis, the clinical picture may include other organ manifestations such as acute kidney damage or circulatory dysregulation. In terms of treatment, rapid pathogen-directed empirical antibiotic therapy is the treatment of choice. If there is no indication of secondary peritonitis, diagnostic laparoscopy can be dispensed with in the further diagnostic work-up.

Regulation of vascular remodeling by immune microenvironment after the establishment of autologous arteriovenous fistula in ESRD patients.

Autogenous arteriovenous fistula (AVF) is the preferred dialysis access for receiving hemodialysis treatment in end-stage renal disease patients. After AVF is established, vascular remodeling occurs in order to adapt to hemodynamic changes. Uremia toxins, surgical injury, blood flow changes and other factors can induce inflammatory response, immune microenvironment changes, and play an important role in the maintenance of AVF vascular remodeling. This process involves the infiltration of pro-inflammatory and anti-inflammatory immune cells and the secretion of cytokines. Pro-inflammatory and anti-inflammatory immune cells include neutrophil (NEUT), dendritic cell (DC), T lymphocyte, macrophage (Mφ), etc. This article reviews the latest research progress and focuses on the role of immune microenvironment changes in vascular remodeling of AVF, in order to provide a new theoretical basis for the prevention and treatment of AVF failure.

Idiopathic Immune Mediated Haemolytic Anaemia (IMHA) in a 2.5-Month-old Puppy: A Case Report

Background: A 2.5-month-old puppy was presented to the Veterinary Clinical Complex of the College of Veterinary Sciences and AH, Jalukie, Nagaland with a complaint of emesis for one week, inappetence, icterus of the lower abdominal area and other visible mucous membrane. Detailed clinical history was collected and clinical examination was conducted. Methods: 4 ml blood peripheral blood was collected from the cephalic vein in the Lev-Lock blood clot activator vacutainer to harvest serum for biochemical analysis. Another 2 ml of whole blood was also collected in Lev-Lock K3 EDTA vacutainer for complete blood count, blood smear examination for blood parasites, morphological changes in the RBC and IDEXX SNAP 4Dx Plus test kit was also used for diagnosis of the blood parasites test. Result: Peripheral blood smear examination and IDEXX SNAP 4Dx Plus test were found to be negative for blood parasites, but spherocytosis could be observed in the blood smears. A complete blood count (CBC) examination revealed severe anaemia of macrocytic normochromic type, neutrophilic leucocytosis and thrombocytopenia. Serum biochemistry revealed a decrease in blood urea nitrogen (BUN) and creatinine with normal alanine transaminase (ALT). Treatment with whole blood transfusion, corticosteroid and other supportive therapy was successful in this case. Based on the clinical presentation, laboratory findings and its response to the treatment with prednisolone, it was diagnosed as an idiopathic immune-mediated haemolytic anaemia (IMHA) with an extravascular mechanism of haemolysis. A follow-up review after 1 year and 4 months post-treatment reported that the dog was still alive and healthy.

Agranulocytosis: from diagnosis to selecting treatment tactics

Neutropenia is a condition accompanied by a decrease in an absolute neutrophil count (of band and segmented neutrophils) in the peripheral blood to less than 1.0 × 109/l in children under 1 year of age and less than 1.5 × 109/l in children over one year of age and adults. Among neutropenia types, congenital (genetically determined), acquired, and chronic benign neutropenia of childhood are distinguished. Agranulocytosis is a decrease in the level of neutrophils in peripheral blood to less than 0.5 × 109/l.

IRG1/ACOD1 Promotes Neutrophil Reverse Migration and Alleviates Local Inflammation.

Polymorphonuclear neutrophil (PMN) infiltration at inflammatory site plays a critical role in inflammation. PMN reverse migration (rM) describes the phenomenon that PMNs migrate away from inflammatory site back into the vasculature, and its role within inflammatory scenarios remains to be fully determined. This study aimed to investigate the mechanism underlying PMN rM and its role in inflammation. First, we demonstrated PMN rM in a mouse model of LPS-induced acute lung inflammation. By single-cell RNA sequencing (scRNA-seq), we demonstrated that reverse migrated (rM-ed) PMNs in blood expressed high level of immuneresponsive gene 1 (Irg1), the encoding gene of cis-aconitate decarboxylase (ACOD1). Using a mouse air pouch model, which enables us to directly track rM-ed PMNs in vivo, we detected higher expression of ACOD1 in the rM-ed PMNs in circulation. Furthermore, mice with Irg1 knockout exhibited decreased PMN rM and higher levels of inflammatory cytokine in inflammatory site. Mechanistically, we found that itaconate, the product of ACOD1 catalyzation, decreased PMN ICAM-1 expression at the inflammation site. Furthermore, inflammatory site showed a high level of shed CD11a, the ligand of ICAM-1. Neutralization of either ICAM-1 or CD11a leading to increased PMN rM. These findings suggest that the binding of ICAM-1 and shed CD11a serves as a retaining force to hold PMNs in the site of inflammation, and ACOD1-decreased PMN surface expression of ICAM-1 weakens the retaining force, so promoting PMNs to leave the inflammatory site. These results indicate a regulatory role of IRG1 in PMN rM and subsequent contributions to inflammation resolution.

Protein Signature Differentiating Neutrophils and Myeloid-Derived Suppressor Cells Determined Using a Human Isogenic Cell Line Model and Protein Profiling.

Myeloid-derived suppressor cells (MDSCs) play an essential role in suppressing the antitumor activity of T lymphocytes in solid tumors, thus representing an attractive therapeutic target to enhance the efficacy of immunotherapy. However, the differences in protein expression between MDSCs and their physiological counterparts, particularly polymorphonuclear neutrophils (PMNs), remain inadequately characterized, making the specific identification and targeting of MDSCs difficult. PMNs and PMN-MDSCs share markers such as CD11b+CD14-CD15+/CD66b+, and some MDSC-enriched markers are emerging, such as LOX-1 and CD84. More proteomics studies are needed to identify the signature and markers for MDSCs. Recently, we reported the induced differentiation of isogenic PMNs or MDSCs (referred to as iPMNs and iMDSCs, respectively) from the human promyelocytic cell line HL60. Here, we profiled the global proteomics and membrane proteomics of these cells with quantitative mass spectrometry, which identified a 41-protein signature ("cluster 6") that was upregulated in iMDSCs compared with HL60 and iPMN. We further integrated our cell line-based proteomics data with a published proteomics dataset of normal human primary monocytes and monocyte-derived MDSCs induced by cancer-associated fibroblasts. The analysis identified a 38-protein signature that exhibits an upregulated expression pattern in MDSCs compared with normal monocytes or PMNs. These signatures may provide a hypothesis-generating platform to identify protein biomarkers that phenotypically distinguish MDSCs from their healthy counterparts, as well as potential therapeutic targets that impair MDSCs without harming normal myeloid cells.

Impact of Pretransplant C-Reactive Protein, Neutrophiles, Platelets, and Albumin Levels on Recipient Survival After Simultaneous Pancreas and Kidney Transplantation

BackgroundLong-lasting diabetes mellitus type 1 and end-stage renal disease induce severe metabolic and immunological deterioration. Pretransplant C-reactive protein (CRP) and albumin (ALB) levels have impact on kidney transplantation. We evaluated impact of preoperative CRP, ALB, neutrophiles (NEU) and platelets (PLT) count, on one- and five-year recipients survival after simultaneous pancreas and kidney transplantation (SPK). MethodsAmong 103 SPK recipients, parameters: CRP (mean: 4.5±4.97mg/L); NEU (mean: 5.12 ±2.13∗103/mm3); PLT (mean: 244±84∗103/mm3); ALB (mean 4.5±0.75 g/dL) were obtained before transplantation. Cox regression, uni-, multivariate analysis for one- and five-year survivals were performed with 95% Confidence Intervals (CI) and area under ROC curve (AUC) was assessed. ResultsIn Cox regression, ALB <3,65g/dL had significant impact for one-year and five-year survivors with hazard ratios: 8 (CI:1.5-38.28, p<0.05) and 3.13 (CI:1.45-6.73, p<0.05), respectively. In univariate analysis, we found significantly decreased one-year survival if: PLT<180∗103/mm3, ALB <3.65g/dL, NEU>5.8∗103/mm3 and CRP>2.25mg/L with odds ratios (OR): 6.75 (CI: 2.12-21.15); 4.05(CI:1.3-12.09); 2.97 (CI: 1.02-8.64) and 5.51 (CI:1.67-18.19), respectively. Independent factors for five-year survival were: CRP, ALB and PLT with OR: 4.72(CI:1.67–13.29); 3.31(CI:1.18–9.25) and 4.2(CI:1.39–12.68), respectively. In multivariate analysis, we built two models for one-year survival. Model 1 (ALB+PLT) with OR: 3.12 (CI:0.97-10.07) and 5.55 (CI:1.67-18.4); and model 2 (CRP+PLT) with OR: 5.51 (CI:1.5-17.3) and 4.3 (CI:1.2-15.06), respectively. AUC for models 1 and 2 were: 0.74 and 0.759, respectively. ConclusionsNEU, PLT, ALB and CRP levels assessed before transplantation are independent factors for one- and five-year SPK recipients survival.

Roles of mitofusin-2 in mouse neutrophil adhesion and development

Neutrophils play a critical role in inflammation and innate immunity. A crucial step in neutrophil recruitment is their adhesion to vascular endothelium. Previous research demonstrated the necessity of mitofusin-2 (MFN2) for integrin activation, adhesion, and the development of neutrophil-like HL60 cells. However, it remains unknown whether this holds true for primary neutrophils. Additionally, neutrophil adhesion is closely associated with their development, making it challenging to independently investigate the roles of MFN2 in regulating mouse neutrophil adhesion and development. In this study, we successfully addressed these challenges by utilizing two distinct conditional knockout mouse models. Our findings revealed that blood neutrophils from MFN2flox/flox LysM-cre mice exhibited approximately a 50% reduction in MFN2 expression. Remarkably, these MFN2 knockdown (KD) mice did not display any defects in neutrophil development. In contrast, blood neutrophils from MFN2flox/flox Vav-iCre mice showed a substantial (&gt;90%) reduction in MFN2 expression. These MFN2 knockout mice exhibited a notable defect in the maturation of band cells into polymorphonuclear neutrophils. We further investigated the neutrophil adhesion using MFN2 knockdown mice and identified the neutrophil adhesion defect using microfluidics, the integrin activation defect using soluble ligand binding, and the Ca2+ flux defect using time-lapse flow cytometry. These findings provide valuable insights into the role of MFN2 in leukocyte adhesion and development and offer potential new perspectives for understanding and treating MFN2-deficiency-related diseases. This research was supported by grants from the National Institutes of Health, National Heart, Lung, and Blood Institute, USA (R01HL145454), National Institute of General Medical Sciences (R35GM119787), National Institute on Aging (P30AG024832), a Career re-entry fellowship from the American Association of Immunologists (G602182), and a startup fund from UConn Health. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.