Immature Neutrophils Research Articles

Microbe capture by splenic macrophages triggers sepsis via T cell-death-dependent neutrophil lifespan shortening

The mechanisms linking systemic infection to hyperinflammation and immune dysfunction in sepsis are poorly understood. Extracellular histones promote sepsis pathology, but their source and mechanism of action remain unclear. Here, we show that by controlling fungi and bacteria captured by splenic macrophages, neutrophil-derived myeloperoxidase attenuates sepsis by suppressing histone release. In systemic candidiasis, microbial capture via the phagocytic receptor SIGNR1 neutralizes myeloperoxidase by facilitating marginal zone infiltration and T cell death-dependent histone release. Histones and hyphae induce cytokines in adjacent CD169 macrophages including G-CSF that selectively depletes mature Ly6Ghigh neutrophils by shortening their lifespan in favour of immature Ly6Glow neutrophils with a defective oxidative burst. In sepsis patient plasma, these mediators shorten mature neutrophil lifespan and correlate with neutrophil mortality markers. Consequently, high G-CSF levels and neutrophil lifespan shortening activity are associated with sepsis patient mortality. Hence, by exploiting phagocytic receptors, pathogens degrade innate and adaptive immunity through the detrimental impact of downstream effectors on neutrophil lifespan.

Hematological, biochemical and histopathological changes in cattle submitted to emergency slaughter

This study aimed to evaluate the effects of stress in bovines submitted to emergency slaughter caused by lengthy transportation to slaughterhouses in the state of Tocantins, Brazil. Blood collections were performed individually during exsanguinations. Tissue samples were obtained after evisceration on the slaughter line. The animals were divided into the Experimental Group (EG), with 19 bovines destined for immediate emergency slaughter, and the Control Group (CG), with 24 bovines slaughtered normally at the abattoir flow. The EG showed lower values of erythrocytes, hemoglobin, hematocrit, platelets, neutrophilia with band neutrophils, and lymphopenia. Hyperalbuminemia, high levels of total proteins, glucose, and creatinine, and lower urea levels were observed in the EG. The globulins were equal between the groups. Histopathology revealed hepatocytes with vacuolated cytoplasm (87.5% of the cases) and liver congestion (83.33%). Interstitial emphysema (95.65%), alveolar emphysema (65.21%), and congestion (52.17%) were observed in the lungs. The kidneys showed congestion (80.95%) and hyaline casts (100%). The spleens showed rarefaction in the white (70%) and red pulp (65%), in addition to hemosiderosis (10%). This suggests that the animals transported over long distances are submitted to severe stress, with water and food deprivation, resulting in hematological and biochemical changes, with histological lesions and serious bruises affecting the musculoskeletal system. There is loss of homeostasis, severe debilitation, with destination for emergency slaughter with the conditional use of meat for consumption.

Preliminary investigation of age analysis in crustacean species from the Indian coast, using growth bands

Abstract The present study represents the first information on direct age determination based on band counts in the mesocardiac ossicle (gastric mill) and eyestalk, in Aristeus alcocki, Penaeus monodon, Penaeus indicus, Panulirus homarus, and Portunus sanguinolentus from India. The determination of growth bands is important in understanding the accurate age of a specimen for the effective management strategies for stock sustainability of the natural resource. Longitudinal sections of the eyestalk and meso-cardiac ossicle of the gastric mill were observed for growth band counts in all species except for P. homarus, where the growth bands were recorded only in the ossicle. Growth bands count five in females and four in males of A. alcocki, respectively. In the case of inshore prawns, two growth bands were observed in P. monodon and one growth band in P. indicus, five growth bands in P. homarus, and three in P. sanguinolentus.

T helper cell subsets and related target cells in acute COVID-19

Current review presents a brief overview of the immune system dysregulation during acute COVID-19 and illustrates the main alterations in peripheral blood CD4+ T-cell (Th) subsets as well as related target cells. Effects of dendritic cell dysfunction induced by SARS-CoV-2 exhibited decreased expression of cell-surface HLA-DR, CCR7 as well as co-stimulatory molecules CD80 and CD86, suggesting reduced antigen presentation, migratory and activation capacities of peripheral blood dendritic cells. SARS-CoV-2-specific Th cells could be detected as early as days 24 post-symptom onset, whereas the prolonged lack of SARS-CoV-2-specific Th cells was associated with severe and/or poor COVID-19 outcome. Firstly, in acute COVID-19 the frequency of Th1 cell was comparable with control levels, but several studies have reported about upregulated inhibitory immune checkpoint receptors and exhaustion-associated molecules (TIM3, PD-1, BTLA, TIGIT etc.) on circulating CD8+ T-cells and NK-cells, whereas the macrophage count was increased in bronchoalveolar lavage (BAL) samples. Next, type 2 immune responses are mediated mainly by Th2 cells, and several studies have revealed a skewing towards dominance of Th2 cell subset in peripheral blood samples from patients with acute COVID-19. Furthermore, the decrease of circulating main Th2 target cells basophiles and eosinophils were associated with severe COVID-19, whereas the lung tissue was enriched with mast cells and relevant mediators released during degranulation. Moreover, the frequency of peripheral blood Th17 cells was closely linked to COVID-19 severity, so that low level of Th17 cells was observed in patients with severe COVID-19, but in BAL the relative number of Th17 cells as well as the concentrations of relevant effector cytokines were dramatically increased. It was shown that severe COVID-19 patients vs. healthy control had higher relative numbers of neutrophils if compared, and the majority of patients with COVID-19 had increased frequency and absolute number of immature neutrophils with altered ROS production. Finally, the frequency of Tfh cells was decreased during acute COVID-19 infection. Elevated count of activated Tfh were found as well as the alterations in Tfh cell subsets characterized by decreased regulatory Tfh1 cell and increased pro-inflammatory Tfh2 as well as Tfh17 cell subsets were revealed. Descriptions of peripheral blood B cells during an acute SARS-CoV-2 infection werev reported as relative B cell lymphopenia with decreased frequency of nave and memory B cell subsets, as well as increased level of CD27hiCD38hiCD24 plasma cell precursors and atypical CD21low B cells. Thus, the emerging evidence suggests that functional alterations occur in all Th cell subsets being linked with loss-of-functions of main Th cell subsets target cells. Furthermore, recovered individuals could suffer from long-term immune dysregulation and other persistent symptoms lasting for many months even after SARS-CoV-2 elimination, a condition referred to as post-acute COVID-19 syndrome.

Genetic architecture of band neutrophil fraction in Iceland

The characteristic lobulated nuclear morphology of granulocytes is partially determined by composition of nuclear envelope proteins. Abnormal nuclear morphology is primarily observed as an increased number of hypolobulated immature neutrophils, called band cells, during infection or in rare envelopathies like Pelger-Huët anomaly. To search for sequence variants affecting nuclear morphology of granulocytes, we performed a genome-wide association study using band neutrophil fraction from 88,101 Icelanders. We describe 13 sequence variants affecting band neutrophil fraction at nine loci. Five of the variants are at the Lamin B receptor (LBR) locus, encoding an inner nuclear membrane protein. Mutations in LBR are linked to Pelger-Huët anomaly. In addition, we identify cosegregation of a rare stop-gain sequence variant in LBR and Pelger Huët anomaly in an Icelandic eight generation pedigree, initially reported in 1963. Two of the other loci include genes which, like LBR, play a role in the nuclear membrane function and integrity. These GWAS results highlight the role proteins of the inner nuclear membrane have as important for neutrophil nuclear morphology.

A CLINICAL STUDY OF NEONATAL SEPTICEMIA CONDUCTED IN KURNOOL MEDICAL COLLEGE/GOVT.GENERAL HOSPITAL,KURNOOL

Neonatal septicemia is a major cause of morbidity and mortality in new born infants. To study theOBJECTIVES: incidence,predisposing factors,clinical prole,outcome. The study was conductedMATERIALS AND METHODS: over a period of November-2017-february 2019 ,50 neonates who are showing well documented signs are included in this study,blood investigations like hb,tc,dc with band neutrophil ratio,peripheral smear,micro esr,crp,blood culture,csf,urine,pus,rectal swab will be subjected to c&s and radiological investigations when indicated. culture was bacteriologically positive in 34% cases. ciprooxacin had maximumRESULTS: sensitivity of 88.2%.Leucopenia< 5000/cmm had sensitivity of 47%,specicity of 66.67% and PPA of 42.11%,toxic granulation had 70.56 sensitivity,63.65%specicity and 50%PPA.B/N>0.2 had 88.2%sensitivity 62.4%PPA. m-ESR had sensitivity of 70.56%,specicity of 84.84%,PPA of 70.5%, C-reactive protein had 88.2 sensitivity and 87.8% specicity and 78.95%PPA ,case fatality rate was 28% C O N C L U S I O N : C l i n i c a l f e a t u r e s o f n e o n a t a l s e p t i c a e m i a a r e n o n s p e c i  c a n d v a g u e . s e p s i s s c r e e n h a s g o o d sensitivity,specicity,PPA.combination of tests increase the specicity and PPA. As an individual test C-reactive protein has highest sensitivity.

Role for Granulocyte Colony-Stimulating Factor in Neutrophilic Extramedullary Myelopoiesis in a Murine Model of Systemic Juvenile Idiopathic Arthritis.

Systemic juvenile idiopathic arthritis (JIA) is a systemic inflammatory disease with childhood onset. Systemic JIA is associated with neutrophilia, including immature granulocytes, potentially driven by the growth factor granulocyte-colony stimulating factor (G-CSF). This study was undertaken to investigate the role of G-CSF in the pathology of systemic JIA. Injection of Freund's complete adjuvant (CFA) in BALB/c mice induces mild inflammation and neutrophilia in wild-type (WT) mice and a more pronounced disease, reminiscent to that of JIA patients, in interferon-γ-knockout (IFNγ-KO) mice. Extramedullary myelopoiesis was studied in CFA-immunized mice by single-cell RNA sequencing, and the effect of G-CSF receptor (G-CSFR) blockage on neutrophil development and systemic JIA pathology was evaluated. Additionally, plasma G-CSF levels were measured in patients. Both in systemic JIA patients and in a corresponding mouse model, plasma G-CSF levels were increased. In the mouse model, we demonstrated that G-CSF is responsible for the observed neutrophilia and extramedullary myelopoiesis and the induction of immature neutrophils and myeloid-derived suppressor-like cells. Administration of a G-CSFR antagonizing antibody blocked the maturation and differentiation of neutrophils in CFA-immunized mice. In IFNγ-KO mice, treatment was associated with almost complete inhibition of arthritis due to reduced neutrophilia and osteoclast formation. Disease symptoms were ameliorated, but slight increases in interleukin-6 (IL-6), tumor necrosis factor, and IL-17 were detected upon G-CSFR inhibition in the IFNγ-KO mice, and were associated with mild increases in weight loss, tail damage, and immature red blood cells. We describe the role of G-CSF in a mouse model of systemic JIA and suggest an important role for G-CSF-induced myelopoiesis and neutrophilia in regulating the development of arthritis.

Characterization of Zymosan-Modulated Neutrophils With Neuroregenerative Properties.

Recent studies using advanced techniques such as single cell RNA sequencing (scRNAseq), high parameter flow cytometry, and proteomics reveal that neutrophils are more heterogeneous than previously appreciated. Unique subsets have been identified in the context of bacterial and parasitic infections, cancer, and tissue injury and repair. The characteristics of infiltrating neutrophils differ depending on the nature of the inflammation-inciting stimulus, the stage of the inflammatory response, as well as the tissue microenvironment in which they accumulate. We previously described a new subpopulation of immature Ly6Glow neutrophils that accumulate in the peritoneal cavity 3 days following intraperitoneal (i.p.) administration of the fungal cell wall extract, zymosan. These neutrophils express markers of alternative activation and possess neuroprotective/regenerative properties. In addition to inducing neurite outgrowth of explanted neurons, they enhance neuronal survival and axon regeneration in vivo following traumatic injury to the optic nerve or spinal cord. In contrast, the majority of neutrophils that accumulate in the peritoneal fluid 4 hours following i.p. zymosan injection (4h NΦ) have features of conventional, mature Ly6Ghi neutrophils and lack neuroprotective or neuroregenerative properties. In the current study, we expand upon on our previously published observations by performing a granular, in-depth analysis of these i.p. zymosan-modulated neutrophil populations using scRNAseq and high parameter flow cytometry. We also analyze cell lysates of each neutrophil population by liquid chromatography/mass spectrometry. Circulating blood neutrophils, harvested from naive mice, are analyzed in parallel as a control. When samples were pooled from all three groups, scRNAseq revealed 11 distinct neutrophil clusters. Pathway analyses demonstrated that 3d NΦ upregulate genes involved in tissue development and wound healing, while 4h NΦ upregulate genes involved in cytokine production and perpetuation of the immune response. Proteomics analysis revealed that 3d NΦ and 4h NΦ also express distinct protein signatures. Adding to our earlier findings, 3d NΦ expressed a number of neuroprotective/neuroregenerative candidate proteins that may contribute to their biological functions. Collectively, the data generated by the current study add to the growing literature on neutrophil heterogeneity and functional sub-specialization and might provide new insights in elucidating the mechanisms of action of pro-regenerative, neuroprotective neutrophil subsets.

Quantitative Analysis of the Transcriptome of Two Commonly Used Human Monocytic Cell Lines—THP-1 and Mono Mac 6—Reveals Their Arrest during Early Monocyte/Neutrophil Differentiation

Cell lines of monocyte/macrophage origin are often used as model systems to study monocyte/macrophage biology. A relevant question is how similar these cell lines are to their in vivo counterparts? To address this issue, we performed a detailed analysis of the transcriptome of two commonly used human monocyte/macrophage cell lines, Mono Mac 6 and THP-1. Both of these cell lines originate from leukemic cells with myelo-monocytic characteristics. We found that both Mono Mac 6 and THP-1 represent cells of very immature origin. Their transcriptomes show more similarities to immature neutrophils than cells of the monocyte/macrophage lineage. They express significant levels of N-elastase, proteinase 3, cathepsin G, and azurocidin but very low levels of CD14, ficolin, and complement factor P. All major MHC class II genes are also expressed at low levels. They show high levels of lysozyme and low levels of one of the immunoglobulin Fc receptors, FCGRIIA, which is characteristic of both neutrophils and monocytes. THP-1, but not Mono Mac 6, also expresses the high-affinity receptor for IgG, FCGRIA. Both cell lines lack the expression of the connective tissue components fibronectin, proteoglycan 4, and syndecan 3, which are characteristics of tissue macrophages but are absent in blood monocytes, indicating that they originate from bone marrow precursors and not yolk sac-derived hematopoietic cells. Both of these cell lines seem, therefore, to represent cells arrested during early myelo-monocytic development, at a branch point between neutrophil and monocyte differentiation. Their very immature phenotype indicates that great care should be taken when using these cell lines as models for normal monocyte/macrophage biology.

Alveolar rhabdomyosarcoma with massive bone marrow infiltration and disseminated intravascular coagulation mimicking acute leukemia.

A 27-year-old man was admitted with a 2-month history of general fatigue, appetite loss, and back pain, leading to a worsening of his general condition. On admission, laboratory tests showed an increased white blood cell count (15.9 × 109/L), with leucoerythroblastosis (myeloblasts, 1.0%; promyelocytes, 0%; myelocytes, 2.5%; metamyelocytes, 1.5%; band neutrophils, 3.5%; segmented neutrophils, 67.0%; lymphocytes, 18%; erythroblasts, 48%), severe anemia (hemoglobin, 45.0 g/L), and thrombocytopenia (platelet count, 21.0 × 109/L). Biochemical tests revealed elevated lactate dehydrogenase (2582 U/L) level, and coagulation tests showed a prolonged prothrombin time (20.3 s; reference interval: 10.0–13.0), normal fibrinogen (2.24 g/L), elevated fibrinogen-fibrin degradation product (331.9 mg/L), plasmin inhibitor-plasmin complex (9.05 mg/L), thrombin-antithrombin complex (41.2 μg/L), and decreased antithrombin (75.3%) levels. A computed tomography scan revealed systemic lymphadenopathy, splenomegaly, bilateral pleural effusions, and massive ascites. A bone marrow aspirate revealed massive infiltration by large abnormal cells with a basophilic cytoplasm and cytoplasmic vacuoles (Figure 1A). Flow cytometry analysis showed that the abnormal cells expressed only CD56, but no myeloid or lymphoid cell markers. Because he suffered severe disseminated intravascular coagulation (DIC) and worsening hypoxemia due to increased pleural effusions, initial treatment for acute leukemia comprised prednisolone and cytarabine. On Day 2 postadmission, biopsy of the right axillary lymph node was performed. Immunohistological findings from a bone marrow clot and biopsy tissue from the right axillary lymph node revealed diffuse proliferation of abnormal round cells, which expressed desmin (Figure 1B), myogenin, vimentin, and CD56 (Figure 1C), but not cytokeratin and leukocyte antigen. Therefore, a diagnosis of alveolar rhabdomyosarcoma was made. Subsequently, treatment with vincristine, doxorubicin, and cyclophosphamide was initiated, which ameliorated the lymphadenopathy, ascites, and DIC. Although a leukemic phase or bone marrow infiltration is an uncommon manifestation of alveolar rhabdomyosarcoma, flow cytometry and immunohistological analyses of bone marrow specimens can facilitate correct diagnosis. The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported. The authors received no specific funding for this work. AS, HK, HU, and SK contributed to patient's care. SK performed pathological diagnosis. All authors wrote and reviewed the manuscript. Written informed consent for publication was obtained from the patient. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Written informed consent for publication was obtained from the patient. All data can be assessed by contacting the corresponding author.

Gastric Ulcers in Alpacas- Clinical, Laboratory, and Pathological Findings.

Gastric ulcers are a common finding in post-mortem examinations of South American camelids (SAC), but diagnosis in living animals is often difficult. The aim of this study was to provide an overview of the incidence of gastric ulcers in alpacas, common concomitant diseases, and clinical as well as laboratory findings to facilitate diagnosis for veterinarians. For this purpose, a total of 187 necropsy reports of alpacas were evaluated, including clinical and laboratory findings on the living animal. A total of 23.5% of the animals (n = 44) were found to have gastric ulcers, nine were perforated. Compartment 3 was most frequently affected by gastric ulcers. No sex predilection could be detected, but animals 1 year of age and older were more frequently affected by gastric ulcers than animals under 1 year of age. Alpacas with gastric ulcers were presented to the clinic due to different non-specific symptoms. In alpacas with gastric ulcers, significantly more organs or organ systems besides the stomach revealed clinical findings than in animals without gastric ulcers. Of the 44 animals with gastric ulcers, a total of 21 alpacas (47.7%) had a poor nutritional status, but cachexia was not significantly more frequent in animals with gastric ulcers than in other dissected animals without ulcers. Hematologic investigations revealed a significantly lower white blood count and significantly lower segmented neutrophils than in deceased animals without ulcers. Compared to animals discharged after treatment, alpacas that died with gastric ulcers had significantly higher levels of band neutrophils and fewer eosinophils and basophils. Occult blood in feces was found in three of 12 animals with gastric ulcers examined for occult blood. In summary, gastric ulcers are a common problem in SAC, which is difficult to diagnose clinically or by laboratory investigations. As these are often chronic processes involving other organ systems, regular monitoring of the animals' nutritional status and early detection of disease symptoms may help to prevent gastric ulcers.

First Estimates of Age and Growth of the Lusitanian Cownose Ray (Rhinoptera marginata) from the Mediterranean Sea

Within the eastern Mediterranean, increased fishing pressure coupled with a lack of basic biological information is threatening the sustainability of the rare Lusitanian cownose ray (Rhinoptera marginata). To provide baseline life-history data for this species, age estimates were obtained from vertebral band counts of 224 rays (size range: 210–998 mm disc width and 140 to 13,600 g weight) collected from Iskenderun Bay. Females ranged in size from 210 to 998 mm disc width and 238 to 13,600 g weight, while males ranged from 130 to 866 mm disc width and 140 to 8250 g weight. The index of average percent error (2.8%) and age-bias plot suggest that the aging method used represents a precise and non-biased approach. Marginal increment analysis indicated that a single opaque band is deposited annually between August and September. The oldest ages obtained for R. marginata were 9 years for males and 19 years for females, which corresponded to total lengths of 866 and 998 mm disc width, respectively. For males, limited samples prevented the accurate calculation of growth rates; however, for combined sexes, observed and disc width-at-age data resulted in the following von Bertalanffy growth parameters: DW∞ = 1102.16 mm, k = 0.148 and t0 = −0.2167. Although additional samples are necessary to determine growth rates in males, the results of the present study indicate that R. marginata females exhibit life history characteristics similar to other Rhinoptera species.

Utility and prognostic significance of leukocyte ratios in dogs withPrimary Immune-Mediated Hemolytic Anemia.

Canine immune-mediated hemolytic anemia (IMHA) is a life-threatening condition that is commonly associated with neutrophilia and monocytosis. Leukocyte ratios have been found to have prognostic value in humans and animals affected by a range of inflammatory, infectious, and neoplastic disorders. We hypothesized that in primary IMHA, neutrophil to lymphocyte (NLR), neutrophil to monocyte (NMR), band neutrophil to segmented neutrophil (BNR) and monocyte to lymphocyte (MLR) ratios would be higher in dogs that did not survive to discharge. Medical records of dogs diagnosed with IMHA at two veterinary teaching hospitals were retrospectively reviewed. Twenty-three of the 72 included dogs did not survive to discharge. NLR, NMR, BNR and MLR ratios were compared between dogs that survived to discharge and dogs that died or were euthanized. None of the ratios were significantly different between survivors and non-survivors (P = 0.14-0.99). Area under the Receiver Operating Characteristic (ROC) curve for prediction of non-survival ranged from 0.5 (95% confidence interval 0.38-0.62) for MLR to 0.61 (0.49-0.72) for NMR and was not significantly different from 0.5 for any ratio (P = 0.29-0.99). After exclusion of 31 dogs that received one or both immunosuppressive medications and blood transfusion before presentation, the area under the ROC curve for prediction of survival was significantly different from 0.5 for MLR (0.78, P = 0.01) and NMR (0.78, P = 0.0002). This study suggests that lower MLR and higher NMR may predict poorer prognosis in untreated dogs with IMHA.

Androgen exposure alters the neutrophil response to pyelonephritis

Abstract Urinary tract infections (UTIs) in men are uncommon but carry increased risk for severe upper-tract UTI (pyelonephritis) and complications including renal scarring. In preclinical models of Escherichia coli UTI, male C3H/HeN mice uniformly develop high-titer pyelonephritis (most with renal abscesses), while the majority of female C3H/HeN mice resolve renal infection within 7–14 days. We previously showed that this difference in outcomes is testosterone dependent. Here, we find that androgen exposure in female C3H/HeN mice alters the neutrophil response to UTI, increasing the propensity for severe renal inflammation and abscess formation as opposed to bacterial clearance and resolution of infection. Concordant with our prior published results, androgen exposure in C3H female mice given Escherichia coli UTI led to 3-log higher bacterial loads in the kidneys by 14 days post infection. We found that the kidneys of androgen-exposed mice harbored more neutrophils than standard females at baseline and throughout infection, correlated with higher levels of the neutrophil-recruiting chemokine CXCL1 (murine KC) in kidney tissue. In addition, the kidneys of androgen-exposed females were enriched for aged, immature neutrophils (Ly6G+, CD49d+, CD101−). Compared to their mature (CD101+) counterparts, these cells exhibited markers of increased degranulation, altered phagocytic activity, and diminished capacity for efflux from kidney tissue. These data support a model in which androgen-exposed mice, despite an apparently more robust neutrophil presence, fail to control renal bacterial infection due to altered neutrophil functions. Supported by grants from NIH (R01 DK111541, R01 DK126697, R01 AI158418)

A Distinct Immature Low-Density Neutrophil Population Characterizes Acute Generalized Pustular Psoriasis

Generalized pustular psoriasis (GPP) is a severe disease featured by neutrophilic pustules and enhanced IL-36 inflammatory pathway in the skin (Hoegler et al., 2018). PBMCs reflecting the systemic inflammatory activation can provide pathogenetically relevant information (Swindell et al., 2016). Recently, a transcriptomic analysis of PBMCs and neutrophils in patients with GPP has been performed in a stable disease state (Haskamp et al., 2021). However, transcriptomic profiling of PBMCs during an acute flare of GPP is unclear.

Disseminated Talaromyces marneffei in newly diagnosed HIV patient.

A 31-year-old Chinese man referred to the haematology clinic, was presented with recurrent fever for 19 days and two enlarged lymph nodes on the left neck. Past medical history was unremarkable. Laboratory data revealed white blood cell (WBC) count 9.81 × 109/L (neutrophilic band granulocyte 67%, neutrophilic myelocyte 11%, neutrophilic metamyelocyte 7%, polymorphonuclear neutrophil 6%, lymphocyte 5%, eosinophil 1%, monocyte 2%, basophil 1% and nucleated red blood cell 18/100 WBC), haemoglobin 77 g/L and platelet count 41 × 109/L. The peripheral blood and bone marrow aspirate smear revealed small round-to-ovoid yeast cells located predominantly within or surrounding the neutrophil (top left, middle and right, 1000×, Wright-Giemsa staining). The blood culture showed the fungus grown in a mould form arranged in a mulberry-like pattern and produced a soluble red pigment that diffuses into the sabouraud medium (bottom left). Lactophenol cotton blue staining confirmed numerous yeast-like organisms by the microscopic morphology (bottom middle, 400×). Fungus staining also showed numerous blue yeast-like organisms by fluorescence microscope (bottom right, 400×). The patient was subsequently diagnosed with AIDS having HIV antibody, and the blood cultures showed the growth of Talaromyces marneffei, a dimorphic fungus that is an important opportunistic pathogen. After treatment with anti-fungus and anti-HIV drug, this patient had significantly improved and discharged. The authors would like to thank the encouragement and support of the members who take advanced study in the unit of cytology of the Department of Clinical Laboratory, Zhejiang Provincial People's Hospital. Yanxia Chen was supported by the Zhejiang Provincial Natural Science Fund (No. LY20H100008). Financing entity had no role in the study design, data collection or analysis, decision to publish or preparation of the manuscript. The authors declared that they have no conflict of interest. Zhejiang Provincial Natural Science Fund, Grant Number: LY20H100008. Yingcai Huang provided the haematology picture and wrote the manuscript. Haiyan Cui provided the microbiology picture. Yanxia Chen analysed the data. Jinlin Liu wrote the manuscript. All authors reviewed this manuscript and agreed to submit it. This study was approved by the ethical committee of Longgang District People's Hospital of Shenzhen & The Second Affiliated Hospital of the Chinese University of Hong Kong (No. 2022007). This study did not involve any personal information, but only reports the laboratory data. Therefore, the patient consent was waived, and this waive was approved by the ethical committee of Longgang District People's Hospital of Shenzhen & The Second Affiliated Hospital of the Chinese University of Hong Kong.

Hyperspectral-Based Classification of Managed Permanent Grassland with Multilayer Perceptrons: Influence of Spectral Band Count and Spectral Regions on Model Performance

Detailed knowledge of botanical composition is a key factor for the sustainable and site-specific management of permanent grassland and facilitates an improvement in the performance and efficiency of livestock feeding. Spectral-based data acquisition combined with machine learning has the potential to classify species groups and plant parts in permanent grassland with high accuracy. However, a disadvantage of this method is the fact that hyperspectral sensors with a wide spectral range and fine spectral and high spatial resolution are costly and create large amounts of data. Therefore, the question arises as to whether these parameters are necessary for accurate grassland classification. Thus, the use of sensors with lower spectral and spatial resolution and correspondingly lower data processing requirements could be a conceivable approach. Therefore, we investigated the classification performance with reduced predictor sets formed by different approaches in permanent grassland. For pixel-based classification, a cross-validated mean accuracy of 86.1% was reached using a multilayer perceptron (MLP) including all 191 available predictors, i.e., spectral bands. Using only 48 high-performing predictors, an accuracy of 80% could still be achieved. In particular, the spectral regions of 954 nm to 956 nm, 684 nm to 744 nm and 442 nm to 444 nm contributed most to the classification performance. These results provide a promising basis for future data acquisition and the analysis of grassland vegetation.

Data-Driven EEG Band Discovery with Decision Trees.

Electroencephalography (EEG) is a brain imaging technique in which electrodes are placed on the scalp. EEG signals are commonly decomposed into frequency bands called delta, theta, alpha, and beta. While these bands have been shown to be useful for characterizing various brain states, their utility as a one-size-fits-all analysis tool remains unclear. The goal of this work is to outline an objective strategy for discovering optimal EEG bands based on signal power spectra. A two-step data-driven methodology is presented for objectively determining the best EEG bands for a given dataset. First, a decision tree is used to estimate the optimal frequency band boundaries for reproducing the signal’s power spectrum for a predetermined number of bands. The optimal number of bands is then determined using an Akaike Information Criterion (AIC)-inspired quality score that balances goodness-of-fit with a small band count. This data-driven approach led to better characterization of the underlying power spectrum by identifying bands that outperformed the more commonly used band boundaries by a factor of two. Additionally, key spectral components were isolated in dedicated frequency bands. The proposed method provides a fully automated and flexible approach to capturing key signal components and possibly discovering new indices of brain activity.

Extracellular Traps Increase Burden of Bleeding by Damaging Endothelial Cell in Acute Promyelocytic Leukaemia.

The rate of complete remission of acute promyelocytic leukemia (APL) is currently over 90% because of the use of all-trans retinoic acid (ATRA) with arsenic trioxide (ATO). However, hemorrhagic mortality has emerged as the most significant barrier to APL-induced remission. Neutrophils extracellular traps (NETs/ETs) cause vascular leakage by damaging the integrity of endothelial cells. We have previously demonstrated that APL cells treated with ATRA/ATO undergo a cell death process, releasing extracellular chromatin, termed ETosis/NETosis. However, the mechanism underlying the involvement of ETs in endothelial injury in APL remain largely unknown. Here, we analysed the ability of mature and immature neutrophils to release ETs, and their interaction with platelets (PLTs) in APL. Importantly, the effect of ETs on vascular endothelium in APL was discussed. Our results showed that the ability of immature neutrophils to release ETs was impaired in APL, whereas mature neutrophils produced ETs, which were associated with activated PLTs. Moreover, ATRA+ATO induced immature neutrophil differentiation, as well as increased the release of ETs from mature neutrophils. The excessive ETs damaged endothelial cells, causing blood cell leakage. Removing ETs using DNase 1 alleviated endothelial damage and improved blood cells leakage. Our results indicate that vascular endothelial injury is at least partially associated with ETs in APL, and that targeting ETs production may be an effective approach for relieving vascular leakage and reducing the burden of bleeding in APL.

Diagnostic utility of combined immature and total neutrophil counts along with C-reactive protein in early detection of neonatal sepsis: A cross-sectional study

IntroductionA timely diagnosis is critical for management of Neonatal sepsis. Blood Culture is considered to be the “Gold Standard” for its diagnosis, but it has some limitations. In recent times, highly sensitive and specific inflammatory markers like interleukins, ELISA, counter immune-electrophoresis etc. have been in use for its diagnosis. But these are impractical for developing countries, due to their high cost and requirement of sophisticated equipments. A combination of haematological parameters like total leucocyte count (TLC), immature to total neutrophil ratio (I/T ratio), absolute neutrophil count (ANC), platelet count and C-reactive protein (CRP) estimation provide an early diagnosis of bacteremia. This study was undertaken to evaluate the usefulness of the above mentioned parameters as indicators for early diagnosis of neonatal sepsis. Material and methodsIn the present cross-sectional study, we intent to analyse various hematologic parameters in 160 neonates admitted in the neonatal care unit of a tertiary care hospital in Delhi. We obtained data from the records of blood culture and complete blood counts of neonates from pathology and microbiology departments of the hospital. Out of 160 admitted neonates, 80 were taken as cases and remaining 80 were taken as controls. Medical records were studied to identify infants born at ≥ 34 weeks gestation. CBCs was analysed, blood cultures and CRP were done in department of Microbiology. CBC, CRP and Blood culture was done as per standard protocols and clinical assesment by paediatrician. The statistical analyses were performed using SPSS version 22 for windows. ResultsAmong 80 neonates, who were in early neonatal sepsis, 44 cases (55%) were females, and 36 (45%) were males. The Microbiological profile of 80 septic neonates was analysed. The I/T value, ANC and CRP values were significantly higher in the neonates suffering from sepsis as compared to the control group. Among 80 septic neonates (cases), 30 (37.5%) were having normal ANC while 50 (62.5%) were having increased ANC and 34 (42.5%) were having normal I:T ratio while 46 (57.5%) were having increased I:T ratio. Out of 80 septic neonates (cases), 18 (22.5%) were having normal CRP while 62 (77.5%) were having increased CRP. ConclusionANC, I/T Ratio and CRP are quick, simple and cost-effective routine laboratory tests which help in early diagnosis of neonatal sepsis. Although there are many serological markers available, ANC and I/T Ratio serves as a reliable predictor of neonatal sepsis. With a good sensitivity, high specificity and a good negative predictive value these parameters can therefore help in timely and early identification of neonatal sepsis.