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Objective: Recent advances in hematology analyzers have generated cell population data (CPD), which quantify features of cells. The characteristics of CPD in pediatric systemic inflammatory response syndrome (SIRS) and sepsis were evaluated with 255 patients., Methods: The ADVIA 2120i hematology analyzer was used for measurement of the delta neutrophil index (DN) including DNI and DNII. The XN-2000 was used for measurement of immature granulocytes (IG), neutrophil reactivity intensity (NEUT-RI), neutrophil granularity intensity (NEUT-GI), reactive lymphocytes (RE-LYMP), antibody synthesizing lymphocytes (AS-LYMP), RBC hemoglobin equivalent (RBC-He), and difference between RBC and reticulocyte hemoglobin equivalent (Delta-He). Measurement of high-sensitivity C-reactive protein (hsCRP) was performed using the Architect ci16200., Results: The area under the receiver operating characteristic curve (AUC) values with confidence interval (CI) of IG (0.65, CI 0.58-0.72), DNI (0.70, CI 0.63-0.77), DNII (0.69, CI 0.62-0.76), and AS-LYMP (0.58, CI 0.51-0.65) were significant for diagnosis of sepsis. The levels of IG, NEUT-RI, DNI, DNII, RE-LYMP, and hsCRP exhibited gradual increasing trends from control to sepsis. In Cox regression analysis, the highest hazard ratio was observed for NEUT-RI (39.57, CI 4.87-321.75), higher than those for hsCRP (12.33, CI 2.49-61.12) and DNII (16.13, CI 1.98-131.08). IG (10.34, CI 2.47-43.26), DNI (11.60, CI 2.34-57.49), and RE-LYMP (8.20, CI 1.96-34.33) also showed high hazard ratios., Conclusion: NEUT-RI along with DNI and DNII can provide additional information regarding the diagnosis of sepsis and prediction of mortality in the pediatric ward., (© 2023 by the Association of Clinical Scientists, Inc.)

Although portal vein thrombosis (PVT) commonly occurs in patients with hepatocellular carcinoma (HCC), the hypercoagulability mechanism in patients with HCC is not entirely clear. Recently, tumor-induced formation of neutrophil extracellular traps (NET) has been shown to trigger contact system activation, and contact system activation has been shown to be a new mechanism of thrombosis. Therefore, we investigated whether contact system activation and NET formation occurred in relation to PVT in HCC patients. The circulating levels of NET formation markers (DNA-histone complex, double-stranded DNA, neutrophil elastase) and contact system activation markers (factor XIIa and high-molecular-weight kininogen) were measured in 177 patients who had been diagnosed with HCC and 48 healthy controls. Presence of PVT was confirmed in 77 HCC patients. The levels of NET formation and contact system activation markers were significantly higher in patients than in healthy controls and they increased significantly with the increase in the model for end-stage liver disease (MELD) scores. Of note, these markers were significantly higher in HCC patients with PVT than in those without PVT. These NET formation markers and the contact system activation markers were significant thrombotic risk factors in HCC patients. The well-known liver injury markers (alanine transaminase, prothrombin time) significantly contributed to factor XIIa level. Contact system activation and NET formation are well correlated with liver disease severity and the markers of these can be used as thrombotic risk factors in HCC patients. In addition, therapeutics inhibiting the contact system can be potentially used to manage PVT in HCC patients.

This study aims to evaluate the diagnostic and prognostic significance of white blood cell (WBC) and platelet parameters measured by automated hematology analyzers in pediatric sepsis. The study included 232 pediatric patients whose C-reactive protein test had been requested for evaluation of infection or inflammation at a pediatric tertiary hospital. The patients were allocated to control, systemic inflammatory response syndrome (SIRS) without infection, and sepsis groups. An ADVIA 2120i hematology system was used to measure the delta neutrophil index (DN) and platelet parameters including platelet count, mean platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT), and immature platelet fraction (IPF). A Coulter LH750 hematology analyzer was used to measure the neutrophil volume distribution width (NDW) and monocyte volume distribution width (MDW). The levels of DN, NDW, MDW, MPV, and IPF each gradually increased from the control to SIRS without infection to sepsis. For sepsis, the highest area under the curve (AUC) value was observed for DN. In addition, NDW, MDW, platelet count, PCT, and IPF showed higher AUC values than C-reactive protein. WBC, DN, NDW, MDW, MPV, PDW, PMDW, and IPF were valuable markers for predicting mortality. Of note, the combination of DN and MPV showed a significantly higher prognostic value when compared with individual DN or MPV. Therefore, considering that DN and MPV are each included in a routine complete blood cell count, their combination can be an alternative parameter to assist in achieving a fast and accurate diagnosis and predict the prognosis of sepsis in pediatric patients., (© 2018 by the Association of Clinical Scientists, Inc.)

Monocyte and neutrophil activation occur during microvascular disturbance of disseminated intravascular coagulation (DIC). This study investigated the diagnostic and prognostic value of circulating neutrophil elastase (NE) and neutrophil volume distribution width (NDW) as neutrophil activation markers and circulating soluble CD163 (sCD163) and monocyte volume distribution width (MDW) as monocyte activation markers in 168 patients suspected of having DIC. The sCD163 provided significant diagnostic value. The prognostic value of sCD163 was comparable to that of D-dimer, but was dependent on other coagulation markers. In vitro, thrombin significantly induced sCD163 from monocytes upregulated with IL-10 or dexamethasone. NDW was an independent and powerful prognostic marker. MDW and NE did not provide diagnostic and prognostic power. Excessive thrombin during ongoing DIC induces florid secretion of CD163; sCD163 might therefore be a potential diagnostic and prognostic marker for DIC. NDW, a convenient parameter measured by an automated hematology analyzer, may be an independent prognostic parameter for DIC., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Introduction: Although neutrophil can be activated by microcirculatory disturbances in DIC, its relation with the severity and prognosis of DIC has not been elucidated. We investigated the relationship between neutrophil activation and DIC severity and determined the prognostic value of neutrophil activation markers in patients with DIC., Materials and Methods: We measured two neutrophil activation markers, neutrophil CD64 expression and plasma neutrophil elastase, in 94 patients with suspected DIC and 55 healthy controls. Neutrophil CD64 expression and plasma neutrophil elastase were measured using CD64 quantitation kit, and PMN Elastase enzyme immunoassay kit., Results: Neutrophil CD64 expression and plasma neutrophil elastase level were significantly increased in patients with overt and non-overt DIC compared to normal controls (P<0.001, each). Neutrophil CD64 expression showed a significant linear trend of increase with increasing DIC score (P<0.001). In DIC patients, significant differences were observed between those with and without infection both in overt (P=0.003) and non-overt DIC (P=0.004). Neutrophil CD64 expression was significantly increased in 28-day non-survival group compared to survival group in both overt (P<0.001) and non-overt DIC (P=0.032). The 28-day survival rate showed a stronger association with the neutrophil CD64 expression (P<0.001) than with DIC score (P=0.028) or plasma neutrophil elastase level (P=0.246)., Conclusions: Neutrophil CD64 expression is significantly correlated with DIC severity and has a good predictive value for the 28-day mortality in patients suspected of having DIC. Neutrophil CD64 expression might be useful in monitoring the disease course and in predicting mortality in DIC patients.

The chemokine receptor CXCR4 and its unique ligand, stromal-derived factor 1 (SDF-1), play critical roles in the retention of hematopoietic cells within bone marrow and in their mobilization into the circulation. Surface CXCR4 down-regulation in hematopoietic cells is associated with a loss of retention of the cells in bone marrow. Lipopolysaccharide (LPS), commonly referred to as endotoxin, induces neutrophilia in vivo, but the mechanism of mobilization related to neutrophilia has not been fully clarified. We show that LPS reduces CXCR4 surface expression in a dose- and time-dependent manner in neutrophils and monocytes, but not in lymphocytes. Polymyxin B neutralization of LPS in culture supernatants still induced this down-modulation, and LPS-stimulated neutrophils released interferon gamma and interleukin 1beta. These results provide evidence that CXCR4 down-regulation can be attributed to soluble factors released by neutrophils upon LPS treatment. Moreover, LPS treatment increased CXCR4 messenger RNA in neutrophils, suggesting that the down-regulation of surface CXCR4 is caused by a posttranslational mechanism, and the chemotactic migration of neutrophils in response to SDF-1 was reduced by LPS pretreatment. Thus, the present study has shown that by down-regulating neutrophil CXCR4 expression and attenuating neutrophil responsiveness to SDF-1, LPS can mobilize neutrophils from bone marrow to the peripheral blood through reducing neutrophil retention in bone marrow.

Introduction: We sought to determine whether the delta neutrophil index (DNI), a marker that is reported to be used to predict the diagnosis, prognosis, and disease severity of bacteremia and sepsis, is useful in differentiating bacterial infection without bacteremia (BIWB) from viral infections (VI) in pediatric febrile patients in the emergency department (ED). Method: We conducted a retrospective analysis of febrile patients' medical records from the pediatric ED of the teaching hospital. The patients with BIWB and those with VI were identified with a review of medical records. The primary outcome was the diagnostic performance of DNI in differentiating BIWB from VI. The secondary outcome was a comparison of the diagnostic performances of DNI, CRP, WBC, and neutrophil count between the two groups. Results: A total of 151 (26.3%) patients were in the BIWB group, and 423 (73.7%) were in the VI group. There was no significant difference in DNI between the two groups (3.51 ± 6.90 vs. 3.07 ± 5.82, mean ± SD, BIWB vs. VI). However, CRP levels were significantly higher in the BIWB group than in the VI group (4.56 ± 5.45 vs. 1.39 ± 2.12, mean ± SD, BIWB vs. VI, p < 0.05). The AUROCs of DNI, WBC count, neutrophil levels, RDW, and CRP levels were 0.5016, 0.5531, 0.5631, 0.5131, and 0.7389, respectively, and only CRP levels were helpful in differentiating BIWB from VI. Conclusion: In the absence of bacteremia, DNI would not be helpful in differentiating BIWB from VI in pediatric febrile patients. [ABSTRACT FROM AUTHOR]

Background Neutrophils play a role in xenograft rejection. When neutrophils are stimulated, they eject the DNA-histone complex into the extracellular space, called neutrophil extracellular traps (NET). We investigated whether NET formation actively occurs in the xenograft and contributes to coagulation and endothelial activation. Methods Human whole blood was incubated with porcine aortic endothelial cells (pEC) from wild-type or α1,3-galactosyltransferase gene-knockout (GTKO) pigs. In the supernatant plasma from human blood, the level of the DNA-histone complex was measured by ELISA, and thrombin generation was measured using a calibrated automated thrombogram. Histone-induced tissue factor and adhesion molecule expression were measured by flow cytometry. Results pEC from both wild-type and GTKO pigs significantly induced DNA-histone complex formation in human whole blood. The DNA-histone complex produced shortened the thrombin generation time and clotting time. Histone alone dose-dependently induced tissue factor and adhesion molecule expression in pEC. Aurintricarboxylic acid pretreatment partially inhibited pEC-induced DNA-histone complex formation. Conclusions DNA-histone complex actively generated upon xenotransplantation is a novel target to inhibit coagulation and endothelial activation. To prevent tissue factor and adhesion molecule expression, a strategy to block soluble histone may be required in xenotransplantation. [ABSTRACT FROM AUTHOR]

Introduction: When leukocytes are stimulated by reactive oxygen species (ROS), they release nuclear contents into the extracellular milieu, called by extracellular traps (ET). The nuclear contents are mainly composed of the histone–DNA complex and neutrophil elastase. This study investigated whether leukemic cells could release ET and the released histone could induce endothelial activation, eventually resulting in leukemic progression. Methods: The circulating ET were measured in 80 patients with hematologic diseases and 40 healthy controls. ET formation and ROS levels were investigated during leukemic cell proliferation in vitro. Histone-induced endothelial adhesion molecules expression and cell survival were measured by flow cytometry. Results: Acute leukemia patients had high levels of ET, which correlated with peripheral blast count. Leukemic cells produced high ROS levels and released extracellular histone, which was significantly blocked by antioxidants. Histone significantly induced 3 endothelial adhesion molecules expression, and promoted leukemic cell adhesion to endothelial cells, which was inhibited by histone inhibitors (heparin, polysialic acid, and activated protein C), neutralizing antibodies against these adhesion molecules, and a Toll like receptor(TLR)9 antagonist. When leukemic cells were co-cultured with endothelial cells, adherent leukemic cells showed better survival than the non-adherent ones, demonstrating that histone-treated endothelial cells protected leukemic cells from both spontaneous and chemotherapy-induced death. Conclusion: Our data demonstrate for the first time that extracellular histone can be released from leukemic cells through a ROS-dependent mechanism. The released histone promotes leukemic cell adhesion by inducting the surface expression of endothelial adhesion molecules and eventually protects leukemic cells from cell death. [ABSTRACT FROM AUTHOR]

Abstract: The CXC chemokine receptor CXCR4 and its unique ligand SDF-1 (stromal-derived factor-1) play critical roles for the retention of hematopoietic cells within the bone marrow (BM) and for their mobilization into the circulation. Lithium often produces neutrophilia in psychiatric patients, but the mechanism of mobilization related to neutrophilia has not been fully clarified. We showed here that lithium dose-dependently reduces the levels of surface CXCR4 protein and mRNA in neutrophils, but not in lymphocytes. The chemotactic migration of neutrophils in response to SDF-1 was reduced after a pre-incubation with lithium. We provide evidence that lithium down-regulates the CXCR4 expression of neutrophils and it attenuates their responsiveness to SDF-1. Our studies support the concept that down-regulation of CXCR4 is one of the mechanisms by which causes neutrophilia. [Copyright &y& Elsevier]

Abstract: Background: Neutrophil gelatinase-associated lipocalin (NGAL) exists as monomers, homodimers, and NGAL/matrix metalloproteinase-9 (MMP-9) complexes. Circulating neutrophils are activated in ongoing disseminated intravascular coagulation (DIC); therefore, plasma NGAL levels are likely to be increased in DIC. We investigated the diagnostic performance of the plasma NGAL level in predicting acute kidney injury (AKI) in patients with DIC and determined the prognostic value of NGAL. Methods: A total of 126 patients with suspected DIC were enrolled. The plasma free NGAL was analyzed with a point-of-care immunoassay. Plasma total NGAL and NGAL/MMP-9 complex levels were measured using commercial kits. Results: Median free and total NGAL levels were markedly higher in patients with AKI than those without AKI. However, no significant difference in the NGAL/MMP-9 complex level was found between the groups. In patients without AKI, the plasma free and total NGAL levels were significantly higher in those with overt-DIC than in those without overt-DIC. Of note, the free NGAL level showed a significant prognostic value in DIC. Conclusion: Plasma free and total NGAL proved to be powerful markers for AKI in DIC patients and plasma free NGAL also has a prognostic relevance in DIC patients without AKI. [Copyright &y& Elsevier]