Polymorphonuclear Neutrophil Count Research Articles

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.

Use of Bacterial DNA Concentration in Ascites as a Marker for Spontaneous Bacterial Peritonitis

Background & AimsSpontaneous bacterial peritonitis (SBP) is a common and serious complication in patients with decompensated cirrhosis. Precise quantification of bacterial DNA (bactDNA) and the related inflammatory response might add further information on the course of disease. The aim of the study was to evaluate the association between bactDNA, cytokine levels and clinical outcome. MethodsAscites and serum samples of 98 patients with decompensated liver cirrhosis (42 with SBP and 56 without SBP) as well as serum samples of 21 healthy controls were collected. BactDNA in ascites and serum was detected and quantified by 16S rRNA PCR. Concentrations of IL-1β, TNF-α, IL-6, IL-8 and IL-10 were measured by a LEGENDplexTM multi-analyte flow assay. Clinical data were collected and analyzed retrospectively. ResultsBactDNA was detected more frequently in ascites of patients with SBP (n=24/42; 57.1%) than in ascites of patients without SBP (n=5/56; 8.9%; p < 0.001). Additionally, IL-6 levels in both ascites and serum were significantly higher in patients with SBP (ascites p < 0.001, serum p = 0.036). The quantity of bactDNA in ascites was strongly correlated with polymorphonuclear neutrophil count in ascites (r = 0.755; p < 0.001) as well as ascites IL-6 levels (r = 0.399; p < 0.001). Receiver operating characteristic (ROC) curve analysis to diagnose SBP provided an AUC of 0.764 (95% CI: 0.661 – 0.867) for serum IL-6 levels, an AUC of 0.810 (95% CI: 0.714 - 0.905) for ascites IL-6 levels, and an AUC of 0.755 (95% CI: 0.651 – 0.858) for bactDNA levels in ascites. ConclusionsThe correlation between the amount of bactDNA and IL-6 confirms the pathophysiological relevance of bactDNA and IL-6 as potential biomarkers for the diagnosis of SBP.

Single Cell Analysis and Functional Features of Circulating CD34 + Cells from Patients with Sickle Cell Anaemia Unveil a Defect in the Myeloid Lineage

Polymorphonuclear neutrophils (PMN) have long been suspected to be involved in the pathophysiology of Sickle cell anemia (SCA) since early studies have reported a higher neutrophil count in patients at steady-state and its positive correlation with SCA severity. This high leukocyte count has long be considered as a peripheral phenomenon, driven by RBC hemolysis causing systemic inflammation and neutrophils recruitment. Nevertheless, several lines of evidence suggest that hematopoietic stem/progenitor cell (HSPC) function might be perturbed in SCA. Given that recent report showed an increased apoptosis during terminal erythroid maturation of SCA cells and that erythroid and myeloid differentiation derive from the same common myeloid progenitor (CMP), we investigated a possible enhanced myelopoiesis in SCA and its causes. First, to better understand when and how hyperleukocytosis and chronic inflammation begin in these patients, we analyzed longitudinal biological data from 1250 infants with SCA aged 1-24 months. We observed that the increase in neutrophils count began as early as the first months of life, and was statistically correlated with the drop in HbF rate and with the increase in reticulocytes count, much more than with the increase of the LDH level. These results seem to contradict the hypothesis of chronic inflammation driven by intravascular hemolysis. To assess whether the abnormal hematopoiesis is associated with a defect on lineage commitment of Hematopoietic Stem/Progenitor Cells (HSPCs), we evaluated the phenotype of myeloid and erythroid progenitors isolated from peripheral blood (PB) of untreated patients with SCA. While the portion of the common myeloid progenitor (CMP) was similar among both groups, we observed a significant increase of granulocyte-monocyte progenitors (GMP) in SCA patients (n=6) compared to health donors (HDs) (n=9), as well as a significant decrease of erythroid progenitors (MEP). Interestingly, HSPCs from SCA PB produced more myeloid colonies in vitro than those in HD HSPCs. In addition, colony-forming assay of sorted SCA CMP displayed an increase of granulo-monocytes colonies CFU-GM and decrease of erythroid colonies BFU-E compared to HDs, indicating an increased myeloid commitment of SCA HSPCs. To comprehensively characterize HSPC composition, we performed 10´ scRNA-seq on CD34+ HSPCs isolated from HDs (n=3) and patients with SCA (n=4). We confirmed that SCA HSPCs have a high proportion of neutrophil progenitors. More interestingly, MPP/HSC populations from SCA patients highly express several genes involved in myeloid differentiation like SELL (L-Selectin) and CSF3R (G-CSF receptor) genes, indicating an enhanced myeloid commitment of early progenitor in SCA. To better characterize the myeloid features of these progenitors, we investigated ex- vivo granulopoiesis of PB HSPCs from SCA patients (n=10) and HDs (n=9) in presence of G-CSF as CSF3R is highly expressed in early SCA progenitors. While cell proliferation was similar among both groups, unexpectedly SCA myeloid cells presented a significant delayed expression of CD11b (p&amp;lt;0.01). Interestingly, we also observed an overexpression of monocytic markers HLA-DR and CD36 in a subset of differentiated cells, suggesting a potential commitment of part of the myeloid progenitors to a monocyte lineage , confirmed by MGG staining. To further understand this defect, we performed RNAseq analysis of the GMP isolated from PB of SCA patients (n=4) and HD (n=4). We identified a total of 420 differentially expressed genes (DEGs). Among these DEGs, significantly downregulated genes in SCA included granulocyte differentiation program such as GFI1, C/EBPa, and JAK3. In return, significantly upregulated genes included KLF4, supporting the monocytic bias observed in the early myeloid progenitors. In conclusion, these data reveal the myeloid-primed signature in PB HSPCs of patients with SCA, likely responsible for the elevated PMN and monocyte count in these patients. This initial breakthrough in the understanding of the development of hyperleukocytosis and chronic inflammation in these patients will open up new early therapeutic options. Moreover, since CD34+ cells of the peripheral blood are the main source of HSPCs for the development of gene therapy in SCA, deciphering their functional feature and myelo-monocytic skewing could be of interest to guide the gene editing strategy.

The association between oral inflammatory load and semen and sperm functional analysis: A cross-sectional study.

Studies have suggested a correlation between periodontitis and reduced male fertility. Inflammation has been described as the link between these ailments. Oral inflammatory load (OIL) can be measured through oral polymorphonuclear neutrophil (oPMN) count, which is associated with periodontal diseases. This cross-sectional study assessed the possible correlation between OIL and the functional parameters of sperm cells. In 229 volunteers, oral rinse and semen samples were assessed for oPMN, semen polymorphonuclears (sPMNs), sperm concentration, total sperm count, motility, morphology, and sperm DNA fragmentation index (SDFi). A multiple linear regression model was conducted to evaluate the relationships between oPMN and semen parameters. The effect of elevated oPMN counts on total motility rate, progressive rate, and percentage of sperm cells with normal morphology was significant (P<0.001), with an inverse relationship, i.e., with every unit increase in oPMN count, the mentioned parameters would decline by 0.573, 0.367, and 0.407 units, respectively. oPMN counts also correlated positively with sPMN counts and SDFi (P<0.001), i.e., with every unit increase in the oPMN measures, sPMN counts would increase by 0.126 million/mL, with an 0.733% increase in SDFi. However, there was no significant association between oPMN counts and sperm concentration. OIL, as represented by oPMN counts, might affect male fertility as there is a positive correlation between the levels of these inflammatory cells and decreased sperm motility, abnormal morphological changes, increased sPMN counts, and increased SDFi.

Effects of a stabilized stannous fluoride dentifrice on clinical, immunomodulatory, and microbial outcomes in a human experimental gingivitis model.

Stannous fluoride dentifrice is well established for its beneficial clinical effects. In this study, we evaluated the effects of stannous fluoride on inflammation and oral microbiome. In this randomized, parallel-arm, double-blind, controlled clinical trial, we compared clinical resolution of experimental gingivitis by evaluating bleeding on probing, gingival index, and plaque index between stannous fluoride stabilized with zinc phosphate (test) and sodium fluoride (control) dentifrices. Further, these groups were compared for oral neutrophil counts, systemic priming of neutrophils, gingival crevicular fluid (GCF) expression of inflammatory markers, and the oral microbiome. We found significant reduction in bleeding on probing in the test group compared to the control group in experimental gingivitis when participants used the test dentifrice prior to induction of experimental gingivitis. The test group also showed significant reductions in GCF levels of inflammatory markers (matrix metalloproteinase 8 [MMP8], receptor activator of nuclear factor kappa-Β ligand [RANKL]), oral polymorphonuclear neutrophil (PMN) counts, and systemic neutrophil priming (CD11b expression) during experimental gingivitis. Further, significant reductions in the gram-negative genera Porphyromonas, Tannerella, and Treponema were noted in the test group. The stannous fluoride stabilized with zinc phosphate dentifrice formulation demonstrated clinical reduction in gingival inflammation and a beneficial effect on microbiome and immune markers. This intervention should be explored as a preventive aid in the progression of plaque-induced gingivitis to periodontitis.

КЛИНИЧЕСКОЕ ЗНАЧЕНИЕ ПОЛИМОРФНОЯДЕРНЫХ НЕЙТРОФИЛОВ В СТРАТИФИКАЦИИ ДЕТЕЙ С ОСТРОЙ БОЛЬЮ В ЖИВОТЕ

Background. Acute abdominal pain in children is defined as acute pain of non-traumatic genesis lasting up to 5-7 days, and presents a serious diagnostic and tactical problem. Many aspects of diagnosis and treatment remain controversial. Aim. To determine the clinical significance of polymorphonuclear neutrophil count in the stratification of children with acute abdominal pain. Material and Methods. A prospective randomized blind clinical trial was performed. The results of examination and treatment of 633 children admitted to Grodno Regional Children's Clinical Hospital with acute abdominal pain were analyzed. The number of polymorphonuclear neutrophils was analyzed in relative (PNRV) and absolute (PNAV) values. Statistics included evidence-based criteria, ROC and precision-recall curve analysis, Kullback's measure of informativeness (Jxi), Bayes' theorem. Results. Separation points were determined for the PNRV and PNAV tests. It was 65% for the PNRV test and 8.952×109/L for the PNAV test. For PNRV values less than 65%, the probability of acute appendicitis (AA) as a cause of acute abdominal pain was 1.8% (CI 0.2 to 3.4); the overall informative value was 2.94. With PNAV test values less than 6.5×109/L, the probability of AA was 1.2% (CI 0.03 to 2.4%); the overall informative value was 5.24. It was demonstrated that an increase in test values above the indicated values did not significantly increase the probability of AA as a cause of acute abdominal pain in children. Conclusion. The tests studied are highly informative and can be used to stratify patients with acute abdominal pain. It is necessary to validate the results obtained taking into account the specific capabilities of a given medical institution and the type of equipment used.

Injection of intravenous immunoglobulin (IVIg) for the treatment of Coronavirus induced cytokine storm

Background. COVID-19 disease is the cause of daily morbidity and mortality worldwide due to its high transmissibility and pathogenicity. To date, intravenous immunoglobulin (IVIg) injection has been used as one of the various treatments for this disease. This study aimed to determine the effect of IVIg injection on the treatment of patients with Coronavirus-induced cytokine storm. Methods. A total of 174 patients with COVID-19 were included in this study based on their clinical characteristics and laboratory findings and were divided into two groups of IVIg recipients and non-recipients according to the treatment they received. Statistical analysis was performed using SPSS version 22.0. A p-value less than 0.05 was considered statistically significant. Results. IVIg was used to treat merely 20 patients and Kaletra and hemoperfusion drugs were used more among IVIg recipient patients (P=0.003 and P=0.001, respectively). COVID-19-positive PCR tests were significantly more frequent among IVIg recipients (P=0.026). The polymorphonuclear neutrophil (PMN) count (P=0.007) and sodium level (P=0.007) were significantly higher in the IVIg recipient group on the first admission day. Moreover, INR levels in the IVIg recipient group were significantly lower on the seventh admission day (P=0.020). The median of total intensive care unit (ICU) duration of hospitalization among IVIg recipients was significantly higher (P=0.001). Conclusion. It seems that the use of IVIg in COVID-19 patients should be further investigated. Practical Implications. IVIg injection could decrease mortality and slightly increase the survival rate among COVID-19 patients.

A Primed Neutrophil Subset Predicts the Risk of Bloodstream Infections in Allogenic Bone Marrow Transplant Patients: A Prospective Study

Background: Blood stream infections (BSIs) are the most common infectious complications in patients receiving allogenic hematopoietic stem cell transplants (allo-HSCT). Timing of polymorphonuclear neutrophil (PMN) recovery following allo-HSCT varies widely among patients. PMN counts are monitored to assess susceptibility of patients to BSIs. Even though allo-HSCT patients undergo a phase of severe neutropenia, some of these patients develop BSIs while others don't. Recent studies have emerged describing the heterogeneity of PMNs and the different functional phenotypes. This raises questions on whether susceptibility to BSIs is related to levels of specific PMN phenotypes rather than total PMN counts. Our previous work has identified a primed PMNs (pPMN) phenotype as a steady-state subset representing (~10%) of PMNs in the circulation. These pPMNs display enhanced transmigration and activation, making them the first cells to respond to bacterial insults. In this study, we explore the potential use of the levels of pPMNs as a biomarker for assessing BSI risk in allo-HSCT patients. Methods: This study was a prospective longitudinal assessment of PMNs in consecutive blood and oral samples collected from 76 patients who underwent an allo-HSCT at Princess Margaret Cancer Hospital between August 2020 and August 2021. Patients received the transplant on day 0 and blood and oral samples were collected from the patients on days (-5, +5, +7, +9, +11, +13, +15, +17, +19 and +21). Blood and oral samples were immediately processed, PMNs were counted, phenotyped with a seven CD marker panel of antibodies to identify pPMNs. Flow cytometry was performed, and data was analyzed by FlowJo software. Results: While all allo-HSCT patients displayed a decline in PMN counts during the early post-transplant phase, two patient subsets were identified by their opposing trends in pPMN frequency. Patients were divided into a high pPMN group (n=36) and a low pPMN group (n=40) based on having above or below the 10% threshold of average blood pPMN percentage on day +5 post-transplant. Patients in the low group had increased susceptibly to BSIs (Hazards ratio= 4.814, 95% CI= 2.086-11.11, P= 0.0013). This was not affected after adjusting for sex, age at transplant, diagnosis, conditioning regimen, mucositis, and the stem cell source and counts and remained statistically significant (P<0.01). We also show that patients in the low pPMN group had delayed oral repopulation of PMNs (19.81 days post-transplant) compared to patients in the high group (15.95 days post-transplant). Oral PMN repopulation helps in maintaining equilibrium between the host immune response in the oral cavity and the colonizing microorganisms which is crucial to preventing infections. Therefore, delays in oral repopulation might be one of the means by which deficiencies in pPMNs contribute to BSIs. Conclusion: In patients receiving an allo-HSCT, having less than 10% pPMNs early in the post-transplant phase can be used as an independent early predictor of BSI in allo-HSCT patients

The Spectrum of Still's Disease: A Comparative Analysis of Phenotypic Forms in a Cohort of 238 Patients.

Still's disease (SD) is a heterogeneous autoinflammatory disorder for which several phenotypes have been described. We conducted a retrospective study to re-evaluate the dichotomous view of the disease, to compare the juvenile and adult forms, and to look for prognostic factors. We collected data from ten French centers, seeking patients with a diagnosis of adult-onset SD (AOSD) or systemic juvenile idiopathic arthritis (sJIA). We identified 238 patients, 152 (64%) of whom had AOSD while 86 (36%) had sJIA. The median age at SD onset was 26.6 years. In patients with identifiable patterns, the course of SD was systemic in 159 patients (74%), chronic in 55 (26%). Sore throat and myalgia were more frequent in patients with AOSD. Abnormal liver tests, serum ferritin and C-reactive protein levels were higher in AOSD group. Fever and skin rash were predictive of complete remission or recovery and high lactate dehydrogenase level was a poor prognosis factor. Symptoms such as splenomegaly, skin rash, high polymorphonuclear neutrophils count and macrophage activation syndrome were predictive of a systemic phenotype. Overall, there were no major differences between sJIA and AOSD. Our results are consistent with the "biphasic" model of an autoinflammatory disease that can progress to chronic arthritis if not treated early.

Oxidative Stress Induces Bovine Endometrial Epithelial Cell Damage through Mitochondria-Dependent Pathways

Simple SummaryPolymorphonuclear neutrophil (PMN) count is the main diagnostic method of bovine endometritis. High neutrophil PMN counts in the endometrium of cows affected by endometritis suggest the involvement of oxidative stress among the causes of impaired fertility. The damage mechanism of oxidative stress on bovine endometrial epithelial cells (BEECs) is still unelucidated. The objective of this experiment was to investigate the relationship between oxidative stress and graded endometritis in dairy uteri and the molecular mechanism of oxidative stress injury to BEECs. Our research showed that there was an imbalance of antioxidant stress in dairy cow uterine with endometritis, oxidative stress damaged dairy cow endometrial epithelial cells through mitochondria-dependent pathways. These findings may provide new insight into the therapeutic target of bovine endometrial cell injury.Bovine endometritis is a mucosal inflammation that is characterized by sustained polymorphonuclear neutrophil (PMN) infiltration. Elevated PMN counts in the uterine discharge of dairy cows affected by endometritis suggest that oxidative stress may be among the causes of impaired fertility due to the condition. Nevertheless, the effects of oxidative stress-mediated endometritis in dairy cows largely remain uninvestigated. Therefore, fresh uterine tissue and uterine discharge samples were collected to diagnose the severity of endometritis according to the numbers of inflammatory cells in the samples. Twenty-six fresh uteri were classified into healthy, mild, moderate, and severe endometritis groups based on hematoxylin and eosin stain characteristics and the percentage of PMNs in discharge. BEECs were treated with graded concentrations of H2O2 from 50 μM to 200 μM in vitro as a model to explore the mechanism of oxidative stress during bovine graded endometritis. The expressions of antioxidant stress kinases were detected by quantitative fluorescence PCR to verify the oxidative stress level in uteri with endometritis. Reactive oxygen species were detected by fluorescence microscope, and inflammation-related mRNA expression increased significantly after H2O2 stimulation. Moreover, mRNA expression levels of antioxidant oxidative stress-related enzymes (glutathione peroxidase, superoxide dismutase, and catalase) and mitochondrial membrane potential both decreased. Further investigation revealed that expression of the apoptosis regulator Bcl-2/Bax decreased, whereas expression of the mitochondrial apoptosis-related proteins cytochrome c and caspase-3 increased in response to oxidative stress. Our results indicate that an imbalance exists between oxidation and antioxidation during bovine endometritis. Moreover, apoptosis induced in vitro by oxidative stress was characterized by mitochondrial damage in BEECs.

Usefulness of Haematological Scoring System in Early Diagnosis of Neonatal Sepsis

Background: Neonatal sepsis is one of the major causes of morbidity and mortality of the new born, especially in developing countries. The major concerns of the clinicians are its non-specific presentation and unavailability of tests with a high predictive value. Therefore, the need is for an infallible test that can be easily performed, quick, simple and cost-effective. Objectives: To assess the reliability of Haematological scoring system in diagnosis of suspected neonatal sepsis at early stage. Materials and Methods: A total of 84 suspected cases of neonatal sepsis were included in this prospective cross-sectional study carried out in Khulna City Medical College Hospital during the period from July 2021 to June 2022. The peripheral blood smears from all 84 patients were analysed using HSS of Rowell et al by pathologists. HSS assigns a score of 1 for each of seven findings significantly associated with sepsis such as total leukocyte count, total polymorphonuclear neutrophil count, elevated immature PMN count, elevated immature: total PMN ratio, Immature: mature PMN ratio &gt;0.3, platelet count &lt;150000/cmm, and pronounced degenerative or toxic changes in PMNs. The HSS score of &gt;5 was interpreted as sepsis, blood culture was done as gold standard for diagnosis of sepsis. Each haematological parameter was assessed for its individual performance and sensitivity, specificity, positive and negative predictive values were calculated. Results: Among the total 84 suspected cases, 20 cases were diagnosed neonatal sepsis, (23.8%), 31 cases had probable infection (36.9%), and 33 cases were normal (39.3%). Elevated immature: total PMN count ratio (&gt;0.2) and Immature: Mature PMN ratio (&gt;0.3) were the most sensitive and specific parameter for sepsis. It was also seen that with increasing score, the likelihood of sepsis also increases. Conclusion: The sensitivities and specificities of the various parameters of HSS were found to be satisfactorily significant in identifying ........

Are Leukocyte Esterase (LE) strip and Alpha-Defensin kit reliable enough to diagnose peri-prosthetic joint infection, intra-operatively?

Background: Despite improvement in sterilization techniques, peri-prosthetic joint infection (PJI) is the most fearsome complication after hip and knee arthroplasties. Various strategies have been devised from time to time to diagnose and treat PJI. Recently, Leukocyte esterase and alpha-defensin are identified as markers of active infection in synovial fluid. Therefore, kits are designed to detect these two markers during surgery. If found reliable, these tests will increase the confidence of surgeons in situations, where the diagnosis of PJI is not established. Material and methods: This study was conducted on 132 patients in Lahore General Hospital; from August 2020 to December 2021. Leukocyte Esterase strip and Alpha-Defensin kit were used to detect infection in synovial samples taken just before performing arthrotomy, intra-operatively. Patients were divided into 2 groups, Group-A had 31 patients with peri-prosthetic joint infection (PJI) diagnosed as per Musculoskeletal Infection Society (MSIS) criteria, whereas in Group B 101 patients were present in whom PJI was not present. Synovial tissue and fluid samples were also sent to a laboratory for culture and histopathology; so that sensitivity and specificity of LE strips and Alpha-Defensin Kits could find out. Results: The mean age of patients in our study was 59.6 SD 11.90 years with a male to female ratio of 85:47. 31 patients were diagnosed as PJI after primary hip or knee arthroplasty based on serological investigations and culture of joint aspiration. The sensitivity and specificity of LE strips were 90.32% and 95.04%, whereas that of Alpha defensin was 93.54% and 100% respectively. The correlation coefficient between the LE strip test and synovial fluid polymorphonuclear neutrophils (PMN) counts was 0.811 and it was even higher when the Alpha-Defensin kit was used (0.845). Conclusion: Both LE and Alpha-Defensin kits are highly specific and sensitive in diagnosing PJI. Though Alpha-defensin is more accurate the cost-effectiveness of the LE strip makes it a more feasible option in diagnosing PJI, intra-operatively.

Myeloperoxydase and CD15 With Glycophorin C Double Staining in the Evaluation of Skin Wound Vitality in Forensic Practice.

BackgroundThe determination of skin wound vitality based on tissue sections is a challenge for the forensic pathologist. Histology is still the gold standard, despite its low sensitivity. Immunohistochemistry could allow to obtain a higher sensitivity. Upon the candidate markers, CD15 and myeloperoxidase (MPO) may allow to early detect polymorphonuclear neutrophils (PMN). The aim of this study was to evaluate the sensitivity and the specificity of CD15 and MPO, with glycophorin C co-staining, compared to standard histology, in a series of medicolegal autopsies, and in a human model of recent wounds.MethodsTwenty-four deceased individuals with at least one recent open skin wound were included. For each corpse, a post-mortem wound was performed in an uninjured skin area. At autopsy, a skin sample from the margins of each wound and skin controls were collected (n = 72). Additionally, the cutaneous surgical margins of abdominoplasty specimens were sampled as a model of early intravital stab wound injury (scalpel blade), associated with post-devascularization wounds (n = 39). MPO/glycophorin C and CD15/glycophorin C immunohistochemical double staining was performed. The number of MPO and CD15 positive cells per 10 high power fields (HPF) was evaluated, excluding glycophorin C—positive areas.ResultsWith a threshold of at least 4 PMN/10 high power fields, the sensitivity and specificity of the PMN count for the diagnostic of vitality were 16 and 100%, respectively. With MPO/glycophorin C as well as CD15/glycophorin C IHC, the number of positive cells was significantly higher in vital than in non-vital wounds (p < 0.001). With a threshold of at least 4 positive cells/10 HPF, the sensitivity and specificity of CD15 immunohistochemistry were 53 and 100%, respectively; with the same threshold, MPO sensitivity and specificity were 28 and 95%.ConclusionWe showed that combined MPO or CD15/glycophorin C double staining is an interesting and original method to detect early vital reaction. CD15 allowed to obtain a higher, albeit still limited, sensitivity, with a high specificity. Confirmation studies in independent and larger cohorts are still needed to confirm its accuracy in forensic pathology.

Serum 25-hydroxyvitamin D and C-reactive protein and plasma von Willebrand concentrations in 23 dogs with chronic hepatopathies.

BackgroundSerum concentrations of 25‐hydroxyvitamin D (25(OH)VD) and C‐reactive protein (CRP) and von Willebrand's factor (vWF) concentration correlate with histopathologic disease grade and stage in chronic inflammatory and fibrotic hepatopathies (CH) in humans.ObjectivesTo evaluate serum 25(OH)VD and serum CRP concentrations and plasma vWF concentration and determine if they correlate with histopathologic and biochemical variables in dog with CH.AnimalsTwenty‐three client‐owned dogs with a histopathologic diagnosis of CH were prospectively enrolled.MethodsBlood samples were collected before liver biopsy. Correlations between biomarkers and clinical pathological and histopathologic variables were evaluated using Pearson's or Spearman's test.ResultsSerum 25(OH)VD concentration (median, 213 nmol/L; range, 42‐527 nmol/L) was negatively correlated with serum aspartate aminotransferase activity (AST; rho = −0.59, P < .01), polymorphonuclear neutrophil count (PMN; r = −0.46, P < .05), and positively correlated with serum albumin concentration (r = 0.69, P < .001). Serum CRP concentration (median, 7.4 μg/L; range, 1‐44.9 μg/L) was positively correlated with overall histopathologic necroinflammatory activity (r = 0.78, P < .001) and fibrosis score (rho = 0.49, P < .05). Plasma vWF concentration (median, 73.3%; range, 15‐141%) was positively correlated with fibrosis score (r = 0.53, P < .05) and prothrombin time (rho = 0.67, P < .01), and negatively correlated with serum albumin concentration (r = −0.73, P < .001).Conclusion and Clinical ImportanceIn dogs with CH, serum 25(OH)VD concentration was negatively correlated with disease activity, whereas serum CRP concentration and plasma vWF concentration were positively correlated with histopathologic grade and stage. Our results provide preliminary evidence that these biomarkers may be useful to assess grade and stage of CH in dogs in the absence of liver biopsy.

Sevoflurane Dampens Acute Pulmonary Inflammation via the Adenosine Receptor A2B and Heme Oxygenase-1.

Acute respiratory distress syndrome is a life-threatening disease associated with high mortality. The adenosine receptor A2B (Adora2b) provides anti-inflammatory effects, which are also associated with the intracellular enzyme heme oxygenase-1 (HO-1). Our study determined the mechanism of sevoflurane’s protective properties and investigated the link between sevoflurane and the impact of a functional Adora2b via HO-1 modulation during lipopolysaccharide (LPS)-induced lung injury. We examined the LPS-induced infiltration of polymorphonuclear neutrophils (PMNs) into the lung tissue and protein extravasation in wild-type and Adora2b−/− animals. We generated chimeric animals, to identify the impact of sevoflurane on Adora2b of hematopoietic and non-hematopoietic cells. Sevoflurane decreased the LPS-induced PMN-infiltration and diminished the edema formation in wild-type mice. Reduced PMN counts after sevoflurane treatment were detected only in chimeric mice, which expressed Adora2b exclusively on leukocytes. The Adora2b on hematopoietic and non-hematopoietic cells was required to improve the permeability after sevoflurane inhalation. Further, sevoflurane increased the protective effects of HO-1 modulation on PMN migration and microvascular permeability. These protective effects were abrogated by specific HO-1 inhibition. In conclusion, our data revealed new insights into the protective mechanisms of sevoflurane application during acute pulmonary inflammation and the link between sevoflurane and Adora2b, and HO-1 signaling, respectively.

Increasing Age Affected Polymorphonuclear Neutrophils in Prognosis of Mycoplasma pneumoniae Pneumonia.

PurposeIt is well known that age is related to the incidence of Mycoplasma pneumoniae pneumonia (MPP), and how age and other factors contribute to MPP remains unclear. In this study, we investigate how age affects the prognosis of MPP.Patients and MethodsA total number of 1875 hospitalized children with pneumonia were enrolled in this study, including 52 children with refractory M. pneumoniae pneumonia (RMPP) and 298 children with non-RMPP. We used multiple logistic regression analysis to further identify the risk factors of RMPP, and found that age and polymorphonuclear neutrophils (PMNs) count were the key independent risk factors for the occurrence of RMPP. In order to improve specificity, 4.5 years old was taken as the cut-off value. Then, according to the cut-off value of age, 76 participants were recruited and divided into four groups: <4.5y MPP group, ≥4.5y MPP group, <4.5y health control (<4.5yHC) and ≥4.5y HC group. We explored the diverse functions of primary PMNs from children of different ages with MPP at cellular level. Besides, we studied the relationship between lung injury and PMNs in mice model with MPP of different ages.ResultsWe found that the age and PMNs count of RMPP group were significantly higher than those of the non-RMPP group. Importantly, there is a linear correlation between the age of patients with RMPP and the percentage of PMNs. Further analysis showed that elderly patients infected with M. pneumoniae had more active PMNs function. Meanwhile, proteomics showed that children with M. pneumoniae infection in different age groups have differences in PMNs apoptosis, nicotinamide adenine dinucleotide phosphate, mitochondrial function and oxidative stress. Finally, we found that age is also involved in the pathogenesis of mouse model with MPP.ConclusionWe speculate that age may contribute to the development of RMPP.

Variants of Ascitic Fluid Bacterial Infection in Children with Chronic Liver Disease in a tertiary care hospital in Bangladesh

Introduction: Ascitic fluid bacterial infection is an infection of preexisting ascites in the absence of abdominal source. Ascitic fluid bacterial infection may presents in both symptomatic and asymptomatic children with chronic liver disease (CLD). Culture negative neutrocytic ascites(CNNA) is the commonest type of infection in children with CLD. Objective: To observe the variants of ascitic fluid bacterial infection in children with CLD. Methods: Thirty five consecutive children with clinical features suggestive of CLD, aged between 2-14 years of both sexes with clinically detected ascites and admitted at the Department of Paediatric Gastroenterology and Nutrition, Bangabandhu Sheikh Mujib Medical University (BSMMU) from January 2013 through June 2013 were enrolled in this cross sectional study. Children who had features of ascitic fluid infection like l fever, abdominal pain with tenderness were categorized as symptomatic and children who had no features of ascitic fluid infection were categorized as asymptomatic children. Demographic, clinical, haematological, biochemical and ascitic fluid study were done for both symptomatic and asymptomatic children. Variants of ascitic fluid bacterial infection was identified by ascitic fluid polymorphonuclear leucocyte count and culture report. Results: Among 35 children, majority 17 (48.6%) were between 6-10 years of age group with male predominance. About 12 (34.3%) were symptomatic and among the symptomatic children 7 (58.4%) were infected and 5 (41.6%) were non-infected, 16 (45.7%) had culture negative neutrocytic ascites (CNNA) variant of ascitic fluid bacterial infection as evident by ascitic fluid polymorphonuclear leucocyte count of  250/mm3 and negative culture report. Mean ascitic fluid polymorphonuclear neutrophil count was 515177.82/HPF among infected children and 85.4770.60/HPF among non-infected children, which is statistically significant (p-0.001). Wilson’s disease was the commonest 11(30) cause of the ...

Inflammation-Related Markers and Thyroid Function Measures in Pediatric Patients: Is the Grade of Obesity Relevant?

We aimed to investigate the effect of weight status on inflammation-related markers and thyroid function tests in overweight and obese pediatric patients. Children and adolescents diagnosed between January 2017 and January 2019 with overweight or obesity were included in the study. Neutrophil-to-lymphocyte ratio (NLR), platelet-to lymphocyte ratio (PLR) and systemic immune-inflammation index (SII) were calculated for the groups defined according to Body Mass Index (BMI)-for-age z-score: overweight (≥1 BMI-for-age z-score), obese (≥2 BMI-for-age z-score) and severely obese (≥3 BMI-for-age z-score). Severely obese patients had significantly higher value of white blood cells (WBC) counts (median = 7.92) compared with overweight patients (7.37, p = 0.014). Absolute lymphocyte count was significantly associated with obesity degree in children (Spearman’s Rho coefficient ρ = 0.228. p = 0.035), whereas absolute polymorphonuclear neutrophils (PMNCs) count was significantly higher in severely obese adolescents than overweight adolescents (overweight: 4.04 vs. severely obese: 5.3 (p = 0.029)). In 8.19% of patients an elevated thyroid-stimulating hormone (TSH) level was found, and 3.36% of patients had a low level of free thyroxine with an elevated level of TSH. Total absolute WBC count may be a reliable inflammation-related marker in obese pediatric patients without metabolic syndrome, but needs to be validated in the context of all possible covariates. Subclinical and overt hypothyroidism may develop from an early age in overweight or obese patients.

Ribosomal Protein SA-Positive Neutrophil Elicits Stronger Phagocytosis and Neutrophil Extracellular Trap Formation and Subdues Pro-Inflammatory Cytokine Secretion Against Streptococcus suis Serotype 2 Infection.

Streptococcus suis serotype 2 (SS2), an important zoonotic pathogen that causes septicemia, arthritis, and irreversible meningitis in pigs and humans, can be transmitted to humans from pigs. S. suis causes huge economic losses to the swine industry and poses a serious threat to public health. Previously, we found that the brain tissues of mice with SS2-induced meningitis showed disrupted structural integrity and significantly enhanced polymorphonuclear neutrophil (PMN) infiltration. We showed that the brain tissues of SS2-infected mice had increased ribosomal protein SA (RPSA)-positive PMN counts. However, the inflammatory responses of RPSA+ PMNs to SS2 and their effects on the blood-brain barrier (BBB) remain unclear. Therefore, in studying the pathogenesis of SS2-induced meningitis, it is essential that we explore the functions of RPSA+ PMNs and their effects on the BBB. Herein, using flow cytometry and immunofluorescence microscopy analyses, we found that RPSA expression enhances PMN-induced phagocytosis and PMN-induced formation of neutrophil extracellular traps (NETs), which facilitate further elimination of bacteria. PMN surface expression of RPSA also alleviates local inflammation and tissue injuries by inhibiting secretion of the pro-inflammatory cytokines, TNF-α and IL-6. Moreover, the single-cell BBB model showed that RPSA disrupts BBB integrity by downregulating expression of tight junction-associated membrane proteins on PMNs. Taken together, our data suggest that PMN-surface expression of RPSA is a double-edged sword. RPSA+ PMN owns a stronger ability of bacterial cleaning and weakens inflammatory cytokines release which are useful to anti-infection, but does hurt BBB. Partly, RPSA+ PMN may be extremely useful to control the infection as a therapeutic cellular population, following novel insights into the special PMN population.

Determining the Risk of Spontaneous Bacterial Peritonitis due to increase use of Proton Pump Inhibitors among cirrhotic patients with ascites.

Objectives:The current study aimed to determine the Spontaneous Bacterial Peritonitis (SBP) risk due to increased use of Proton Pump Inhibitors (PPIs) among cirrhotic patients with ascites.Methods:This retrospective case-control study was conducted at Chandka Medical College & Hospital, Larkana from March 2013 to February 2014, involving 215 cirrhotic patients with ascites. Paracentesis was performed to distinguish cirrhotic patients with SBP and Polymorphonuclear Neutrophil (PMN) count ≥ 250 neutrophils/mm3 (cases) and non-SBP with PMN count < 250 neutrophils/mm3 (controls). The demographic details, history of PPIs use before admission and duration of Chronic Liver Disease (CLD) were inquired and statistical analysis was carried through SPSS Version 23.0.Results:Increased pre-hospital PPI intake was observed among cirrhotic patients with SBP (69.8%) as compared to those without SBP (48.8%; p = 0.014). The mean duration of PPI use was 19.16 ± 4.772 days, and it was more significant among older cirrhotic patients (p < 0.05). Increased duration of CLD was observed among PPI users, i.e. 20.47 ± 6.305 months vs. 18.95 ± 5.527 months among non-PPI users (p < 0.05).Conclusions:Our results show that cirrhotic patients with ascites consuming PPIs are more likely to develop SBP as compared to non-PPI users.