Differential White Blood Cell Counts Research Articles

Korean Red Ginseng affects ovalbumin-induced asthma by modulating IL-12, IL-4, and IL-6 levels and the NF-κB/COX-2 and PGE2 pathways

BackgroundAsthma is an incurable hyper-responsive disease of the pulmonary system that is caused by various allergens, including indoor and outdoor stimulators. According to the Global Asthma Network, 339 million people suffered from asthma in 2018, with particularly severe forms in children. Numerous treatments for asthma are available; however, they are frequently associated with adverse effects such as growth retardation, neurological disorders (e.g., catatonia, poor concentration, and insomnia), and physiological disorders (e.g., immunosuppression, hypertension, hyperglycemia, and osteoporosis). MethodsKorean Red Ginseng has long been used to treat numerous diseases in many countries, and we investigated the anti-asthmatic effects and mechanisms of action of Korean Red Ginseng. Eighty-four BALB/c mice were assigned to 6 treatment groups: control, ovalbumin-induced asthma group, dexamethasone treatment group, and 3 groups treated with Korean Red Ginseng water extract (KRGWE) at 5, 25, or 50 mg/kg/day for 5 days. Anti-asthmatic effects of KRGWE were assessed based on biological changes, such as white blood cell counts and differential counts in the bronchoalveolar lavage fluid, serum IgE levels, and histopathological changes in the lungs, and by examining anti-asthmatic mechanisms, such as the cytokines associated with Th1, Th2, and Treg cells and inflammation pathways. ResultsKRGWE affected ovalbumin-induced changes, such as increased white blood cell counts, increased IgE levels, and morphological changes (mucous hypersecretion, epithelial cell hyperplasia, inflammatory cell infiltration) by downregulating cytokines such as IL-12, IL-4, and IL-6 via GATA-3 inactivation and suppression of inflammation via NF-κB/COX-2 and PGE2 pathways. ConclusionKRGWE is a promising drug for asthma treatment.

Piperine ameliorates insulin resistance via inhibiting metabolic inflammation in monosodium glutamate-treated obese mice

BackgroundMetabolic inflammation is an essential event in obesity-induced diabetes and insulin resistance. In obesity, an increasing number of macrophages recruited into visceral adipose tissues undergo significant M1-like polarization, secreting variable amounts of pro-inflammatory cytokines and causing insulin resistance. Piperine has excellent anti-inflammatory activities and may be used in the treatment of a variety of inflammatory diseases. In this study, we investigated the effect of piperine on adipose tissue inflammation and insulin resistance in obese mice.MethodsNewborn mice were subcutaneously (s.c.) injected with monosodium glutamate (MSG) to establish a diabetes model. After 24 weeks, the MSG obese mice were divided into three groups and treated with piperine (40 mg/kg/day), metformin (150 mg/kg/day) and vehicle for 10 successive weeks, respectively.ResultsThe obesity model was successfully established, as the body weight, insulin resistance, fasting blood glucose (FBG) and dyslipidemia were significantly increased. The 10-week administration of piperine to the obese mice not only significantly decreased the elevated FBG (Model: 6.45 ± 0.41 mM; Piperine: 4.72 ± 0.44 mM, p < 0.01), serum TC (Model: 5.66 ± 0.66 mM; Piperine: 3.55 ± 0.30 mM, p < 0.01) and TG (Model: 1.41 ± 0.08 mM; Piperine: 0.94 ± 0.05 mM, p < 0.001), but also enhanced the glucose infusion rate in the hyperglycemic clamp experiment. Meanwhile, piperine improved glucose intolerance and insulin resistance in MSG obese mice. Piperine markedly decreased the total and differential white blood cell (WBC) count, the serum levels of lipopolysaccharide (LPS) and pro-inflammatory cytokines such as galectin-3 (Gal-3) and interleukin-1β (IL-1β). Furthermore, piperine clearly down-regulated the mRNA levels of pro-inflammatory cytokines and the protein levels of M1-like polarization marker CD11c and Gal-3 in adipose tissues. The in vitro study showed that piperine inhibited LPS-stimulated polarization of RAW 264.7 cells toward the M1 phenotype.ConclusionsPiperine served as an immunomodulator for the treatment of obesity-related diabetes through its anti-inflammatory effects, which might be achieved by inhibiting macrophages M1 polarization in adipose tissues.

Short-term biological variation of differential count in healthy subjects in a South Asian population

Estimates of Within-Subject and between subject biological variation for the white blood cell differential count (DC) have not been reported in South Asia. Therefore, we attempted to measure the short-term biological variation estimates for DC. The study was conducted on 28 healthy volunteers (15 males and 13 females). Blood from the volunteers was collected in the morning in K3-EDTA vials and analyzed in triplicate on the Sysmex XN-1000 analyzer, for six consecutive days. The Within subject, between subject and analytical coefficient of variation of the DC was calculated from the results by nested repeated measures ANOVA after outlier exclusion. The Reference change values (RCV) were also calculated. The within-subject variation for eosinophil Count and between subject variation for basophils in our study from South Asia was greater than the published European and American studies. Males and females showed similar biological variation for DC. The within-subject variation of other parameters (Neutrophils, Lymphocytes, Monocytes and Basophils) were similar or showed only mild differences to the published studies. The markedly different within-subject variation for Eosinophil counts suggest that the RCV for DC in South Asians need to be different from the published data in order to have clinical relevance. The Within-subject variation values of the other parameters seem transportable from the published European and American studies, but the small differences found mean that further regional estimates need to be reported for robust evidence of the same.

Treadmill exercise ameliorates memory deficits and hippocampal inflammation in ovalbumin-sensitized juvenile rats

The behavioral changes, including spatial learning and memory impairment as well as depressive- and anxiety-like behaviors in an animal model of asthma were demonstrated previously. On the other hand, there is increasing evidence that the anti-inflammatory actions of exercise are related to their neuroprotective properties against different insults in the brain. This study was aimed to explore the effects of moderate treadmill exercise on cognitive deficits and possible anti-inflammatory mechanisms in ovalbumin (OVA)-sensitized rats. The exercise groups were trained to run on the treadmill 30 min/day with an intensity of 12 m/min, 5 days/week for 4 weeks. Animals in the OVA groups were sensitized by two intraperitoneal (i.p.) injections of OVA (10 μg/injection) and challenged with OVA by inhalation during the treadmill running exercise period. Passive avoidance (PA) memory, levels of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in the hippocampus, total and differential white blood cell (WBC) count in the blood as well as pathological changes of the lung were then evaluated. OVA-sensitization was resulted in cognitive deficits in the PA task, along with increased total and differential WBC in blood and TNF-α in the hippocampus. However, exercise ameliorated these changes and increased the IL-10 level in the hippocampus, suggesting that moderate treadmill exercise can improve memory impairment in OVA-sensitized rats due to its anti-inflammatory properties.

PREVALENCE OF HAEMOPARASITES OF POLO HORSES AND THEIR EFFECT ON SOME HAEMATOLOGICAL PARAMETERS IN ZARIA NIGERIA

Haemoparasites are either intracellular or extracellular organisms that inhabit the blood stream of vertebrate hosts. This study was conducted in Zaria Nigeria inorder to investigate the prevalence of haemoparasites in polo horses and their effects on haematological parameters. Blood samples were collected from a total of 100 randomly selected horses from polo stables across Zaria. The blood samples were screened and examined using thin blood smear, buffy coat technique, and differential white blood cell count. From the results thirteen horses 100(13%) were found infected with haemoparasites. The prevalence of Theileria equi in local and exotic breeds was 5(9.4%) and4 (8.5%) while for microfilaria it was 3(5.3%) and1 (2.1%). There were no Babesia or Trypanosomes found in the samples 0%. Among sexes the prevalence of Theileria equi and microfilaria in males was 1(7.7%) while in females it was (9.3%) and 3(3.4%).A high prevalence of Theileria equi and microfilaria was seen among old horses 6(9.3%) and4 (3.4%) compared to young 3(7.7%) and1 (7.7%).There was no significant association between the presence of the infection and its effect on haematological values (p&gt;0.05). In conclusion some haemoparasites were found in polo horses in Zaria. It is hereby recommended that routine vector control and prophylactic treatment should be carried out in stables

Immuno-enhancement effects of Korean Red Ginseng in healthy adults: a randomized, double-blind, placebo-controlled trial

BackgroundMost clinical studies of immune responses activated by Korean Red Ginseng (KRG) have been conducted exclusively in patients. However, there is still a lack of clinical research on immune-boosting benefits of KRG for healthy persons. This study aims to confirm how KRG boosts the immune system of healthy subjects. MethodsA total of 100 healthy adult subjects were randomly divided into two groups that took either a 2 g KRG tablet or a placebo per day for 8 weeks. The primary efficacy evaluation variables included changes in T cells, B cells, and white blood cells (WBCs) before and after eight weeks of KRG ingestion. Cytokines (TNF-α, INF-γ, IL-2 and IL-4), WBC differential count, and incidence of colds were measured in the secondary efficacy evaluation variables. Safety evaluation variables were used to identify changes in laboratory test results that incorporated adverse reactions, vital signs, hematological tests, blood chemistry tests, and urinalysis. ResultsCompared to the placebo group, the KRG intake group showed a significant increase in the number of T cells (CD3) and its subtypes (CD4 and CD8), B cells, and the WBC count before and after eight weeks of the intake. There were no clinically significant adverse reactions or other notable results in the safety evaluation factors observed. ConclusionThis study has proven through its eight-week intake test and subsequent analysis that KRG boosts the immune system through an increase in T cells, B cells, and WBCs, and that it is safe according to the study's safety evaluation.

Effect of hydro-methanol stem bark extract of Burkea africana on erythrocyte osmotic fragility and haematological parameters in acetaminophen-poisoned rats

BackgroundBurkea africana is a widely used medicinal plant in folkloric medicine in many developing countries of the world. It is useful in the treatment of various ailments including hepatitis, jaundice, diarrhea, stomach aches, abscesses, oedema, epilepsy, bloody diarrhea, gonorrhea, syphilis, toothaches and poisoning. Nevertheless, there are little or no scientific evidence to substantiate this medicinal claim by traditional healers. Burkea africana stem bark was therefore, investigated for its protective or stabilizing effect on erythrocyte membrane in acetaminophen-treated rats. B. africana stem bark was extracted using 80% methanol. Erythrocyte stabilizing effect was studied using erythrocyte osmotic fragility (EOF) test. Thirty (30) male rats were randomly assigned into five (5) groups of six (6) rats each. Groups 1 and 2 served as normal control and negative control (acetaminophen-treated group) respectively. Groups 3, 4 and 5 were pretreated with methanol stem bark extract of Burkea africana (MSBEBA) at doses of 200, 400 and 600 mg/kg body weight respectively once daily for seven (7) days. Blood samples were collected from the animals in all the groups on the 8 day for evaluation of packed cell volume, haemoglobin, red blood cell, white blood cell counts, and differential white blood cell count as well as erythrocyte osmotic fragility.ResultsThe erythrocyte osmotic fragility test showed that there was a significantly (p < 0.05) low percentage hemolysis in the groups pre-treated with the extract when compared with the negative control. The percentage hemolysis was least at 600 mg/kg body weight of the extract. There was also a significant (p < 0.05) increase in the packed cell volume, haemoglobin, red blood cell count at all the doses of the extract used. Neutrophils were significantly (p < 0.05) decreased while lymphocytes were significantly increased in the groups administered MSBEBA 400 and 600 mg/kg body weight.ConclusionMethanol stem bark extract of Burkea africana had protective effect on the red blood cells and also improved haematological parameters. This indicates that Burkea africana may be useful in the treatment of disease conditions that results in hemolytic anemia by stabilizing red erythrocyte membranes and enhancing erythropoiesis.

Haematological and Histopathological Effects of Artemisinin-Based Combination Therapy in Healthy Mice

Artemisinin-based combination therapy had been recommended for the treatment of uncomplicated malaria in Africa. &#x0D; Aim: This study investigated the effects of artesunate/amodiaquine (A/A) and dihydroartemisinin/ piperaquine (D/P) on some blood parameters and histopathology of the liver and kidney of mice.&#x0D; Materials and Methods: A Complete randomized design was applied. Fifty mice were randomly assigned to five treatment groups of ten animals each. Therapeutic doses of the drugs were orally administered to the animals. Group A received normal saline (control), group B received 10/14mg/kg body weight of artesunate/amodiaquine for 3 days, group C received 10/18mg/kg of dihydroartemisinin/piperaquine for 3 days, group D received similar treatment with group B, but were followed up to 28 days, group E received similar treatment with group C but were also followed up to 28 days. After experimental period, blood samples were collected for determination of white blood cell count, white blood cell differential count, red blood cell count, packed cell volume and haemoglobin concentration. Liver and kidney were also harvested for histopathological analysis.&#x0D; Results: Result showed that therapeutic doses of artesunate/amodiaquine and dihydroaretmisinin/ piperaquine had no significant (P&gt;0.05) effects on heamatological parameters accessed. Mild inflammation and degeneration of hepatocyte were observed in the liver of the group treated with D/P while fatty change was found in the group treated with A/A. Venous congestion was observed in the kidney of the group treated with D/P. After 28 days, degeneration of hepatocyte and inflammatory cells were observed in the liver. Shrunken glomerulus was found in the kidney of the group treated with D/P.&#x0D; Conclusion: These drugs are detrimental to the liver and kidney even at therapeutic dose therefore, they should be used with caution.

Statistical methodology for the evaluation of leukocyte data in wild reptile populations: A case study with the common wall lizard (Podarcis muralis).

The leukocyte profile has the potential to be a reliable method to measure health conditions and stress in wild animals, but limitations occur because current knowledge on reference intervals is largely incomplete, especially because data come from studies on captive animals involving few individuals from single populations. Here we propose a general framework for achieving reliable leukocyte reference intervals, encompassing a set of internal and external factors, potentially affecting the leukogram. To do so, we present a systematic survey of the hematology of the common wall lizard, Podarcis muralis, involving 794 lizards from 54 populations over the whole geographic range of the species in Italy. Reference intervals for white blood cell (WBC) and leukocyte differential count were obtained by using linear mixed models in a Bayesian framework. The application of the procedure clearly showed that both internal (sex and size) and external (latitude and season) factors are a source of variation of leukocyte profile. Furthermore, the leukogram of common wall lizard has a strong variability among populations, which accounts for more than 50% of the whole variation. Consequently, some common assumptions used in studies on captive individuals are no longer supported in wild populations, namely, i) any group of individuals is a representative sample, ii) any population is representative of all others, iii) geographic clines do not occur over the species range, and iv) seasonal variation has limited effects. We encourage researchers aimed at the definition of leukocyte reference intervals for wild populations of reptiles to involve a large number of populations over a wide geographic range in ad hoc statistical models to disentangle local and geographic effects on leukocyte profile variation.

Haematological parameters of apparently healthy neonates at the Federal Medical Centre Bida, North Central Nigeria

Background: Haematological parameters are used as screening tools for patient’s care and treatment. Objective: To determine the haematological parameters of apparently healthy term Nigerian babies and generate normative data. Methods: This was a descriptive, longitudinal study of consecutively recruited babies. The haematological parameters were determined using venous blood samples drawn at delivery and on the 14th day of life using a haematology auto-analyser. Results: Of the 309 babies recruited on the first day of life, 160 (51.8%) were males while 149 (48.2%) were females. Two hundred and seventy-nine (90.3%) were delivered vaginally. Most (75%) of the neonates were of Nupe origin. The mean haematocrit, haemoglobin concentration, Mean Corpuscular Volume (MCV), Mean Corpuscular Haemoglobin (MCH), Mean Corpuscular Haemoglobin Concentration (MCHC), Red Blood Cell and reticulocyte count on Day 1 were 48.8±8.0%, 15.0±2.6g/dl, 104.1±8.8fl, 31.9±3.3pg, 30.7±1.7g/dl, 4.7±0.9 (×109/l) and 2.0±1.4% respectively. Corresponding Day 14 values were 41.2±5.2%, 12.8±1.9g/dl, 97.2±9.8fl, 30.3±3.8pg, 31.1±1.7g/dl, 4.5±0.6 (×109/l) and 1.0±0.7% respectively. The mean White Blood Cell (WBC) and platelet counts were 13.3±4.8 (×109/l) and 228.3±81.2 (×109/l) respectively, on the 1st day of life and 10.1±2.4 (×109/l) and 370.1±120.7 (×109/l) respectively on the 14th day. The mean differential WBC counts were 49.4±12.0% for neutrophils and 37.4±11.3% for lymphocytes on the 1st day and 32.4±10.0% and 52.8±9.3% respectively on the 14th day. Conclusion: Neonates from Bida have haematological parameters comparable with previously reported values from other parts of Nigeria.

Age-dependent changes of total and differential white blood cell counts in children.

BackgroundTotal and differential white blood cell counts are important for the diagnostic evaluation of suspected diseases. To facilitate the interpretation of total and differential white blood cell counts in pediatric patients, the present study investigated age-dependent changes in total and differential white blood cell counts in healthy reference children.MethodsData were obtained from the Pediatric Reference Intervals in China study (PRINCE), which aims to establish and verify pediatric reference intervals for Chinese children based on a nationwide multicenter cross-sectional study from January 2017 to December 2018. Quantile curves were calculated using the generalized additive models for location, shape, and scale method. The 2.5th, 50th, and 97.5th quantile curves were calculated for both total and differential white blood counts. Percents of stacked area charts were used to demonstrate the proportions of differential white blood cells. All statistical analyses were performed using R software.ResultsBoth 50th and 97.5th quantiles of total white blood cell count and monocyte count were highest at birth, then rapidly decreased in the first 6 months of life; relatively slow reduction continued until 2 years of age. The lymphocyte count was low during infancy and increased to its highest level at 6 months of age; it then exhibited moderate and continuous reduction until approximately 9 years of age. The pattern of neutrophil count changed with age in a manner opposite to that of lymphocyte count. Besides, there were two inter-sections of lymphocyte count and neutrophil count during infancy and at approximately 5 years of age, based on locally weighted regression (LOESS) analysis. There were no apparent age-related changes in eosinophil or basophil counts.ConclusionThese data regarding age-related changes in total and differential white blood cell counts can be used to assess the health of pediatric patients and guide clinical decisions.

Abstract 6164: Small-molecule targeting of the NPM1/Rad51 repair pathway for the radiosensitization of cancer

Abstract Nucleophosmin1 (NPM1) is rapidly recruited to sites of DNA double-strand breaks and is required for radiation-induced Rad51 foci formation. Inhibition of NPM1 or Rad51 is a radiosensitizing event. YTR107 is a small molecule, identified in a forward chemical genetics functional structure/activity screening, that binds to the amino terminus of NPM1 and prevents NPM1 recruitment to damaged chromatin. In silico docking studies of YTR107 and the crystal structure of the NPM1 N-terminal oligomerization domain revealed that YTR107 docked to monomeric NPM1 in a pocket crucial for forming an interface between neighboring subunits of the NPM1 pentamer. YTR107 inhibited radiation-induced Rad51 foci formation and repair of DNA DSBs, as measured by a neutral comet assay. Colony formation assays revealed that YTR107 radiosensitized 7 NSCLC lines, HT29 colorectal cancer cells, D54 glioblastoma cells, PANC1 pancreatic cancer cells, and 2 breast cancer cell lines (dose modifying factors of &amp;gt; 1.5; cells irradiated with 300 kVp/10mA X-rays @ 2 Gy/min). Radiosensitization was found to be NPM1-dependent as YTR107 did not radiosensitize an NPM1 null mouse embryo fibroblast cell line. YTR107 produced a 4.7 fold increase in HT29 xenograft growth delay (time required for the tumor to increase 4-fold in volume (p = 0.001) following 10 mg/kg YTR107 administered 30 min prior to administering 3 Gy to the tumor for 7 consecutive days). YTR107 increased the survival of A549 tumor-bearing mice determined 70 days after the last treatment. Survival of tumor-bearing mice was 60% for mice treated with YTR107 and 3 Gy vs 20% for mice treated with 3 Gy alone, p = 0.0012 (0 or 20 mg/kg YTR107 administered 1 hr prior to 0 or 2.2 Gy being administered to the tumor for 7 consecutive days). Female C57BL/6J mice were administered 0 or 10 mg/kg i.p. YTR107 for 5 consecutive days. Thirty five days after injection mice were euthanized and subjected to necropsy by a veterinarian trained in veterinary pathology. Gross and histological examination of liver, lung, thymus, heart, spleen, cerebellum, pancreas, small intestine, kidney, and adrenal gland did not reveal evidence of toxicity. Both a complete blood count and a white blood cell differential count were performed. No significant differences were noted. Mice were subjected to five daily injections of 30 mg/kg YTR107. This dose level was also tolerated without overt toxicity after 35 days. Mice were injected with 0 or 30 mg/kg YTR107 (i.p.) and 30 min later administered 0 or 16-Gy to the thorax. Pulmonary fibrotic lesions were quantified using respiratory-gated micro CT 136 days later. YTR107 did not increase the severity of radiation-induced lung fibrosis and did not affect body weights or life span of unirradiated or irradiated mice (p &amp;gt; 0.05) over the 136-day study. Thus, this study provides support for development of YTR107 as a therapeutic radiation sensitizer and was supported in part by R44CA228756-02 Citation Format: Konjeti R. Sekhar, Geri Traver, Peter A. Crooks, Michael L. Freeman. Small-molecule targeting of the NPM1/Rad51 repair pathway for the radiosensitization of cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6164.

Antiinflammatory and antioxidant evaluation of Maytenus senegalensis hydroalcoholic roots extract fractions in allergic asthma

The present work aims to evaluate the antiinflammatory and antioxidant activities of the Maytenus senegalensis (Lam.) roots hydroethanolic extract and to determine its active fraction. The total extract is fractioned in cold alcohol 75%. Phytochemical studies and in vitro antioxidant test are performed on the extract and obtained fractions. During 28 days, mice were sensitized and challenged with OVA to induce allergic airway inflammation. the anti-inflammatory effect of the plant extract and its fractions is assessed by differential white blood cell counts and histopathological examinations. Them antioxidant effect is evaluated in vivo by MDA and glutathione dosage. Qualitative and quantitative phytochemical tests showed that the extract and its fractions contain phenolics compounds. In vitro, both extract and its fractions expressed antioxidant activities. Treatment with M. senegalensis extract and its fractions significantly decreased inflammatory cells number in BALF, reduced MDA concentration and enhanced glutathione level (p&lt;0.05; p&lt;0.01; p&lt;0.001) in lung homogenate. Histological observations confirmed the anti-inflammatory effect of the extract and its fractions. Comparatively, the supernatant fraction showed the best effect. This study may land up in establishing the scientific evidences for the traditional claims anti-asthmatic and anti-inflammatory activity of Maytenus senegalensis (Lam.). It also suggests that the phytochemical groups involved in this activity would be more concentrated in the supernatant fraction.

C-Kit+ progenitors restore rat asthmatic lung function by modulation of T-bet and GATA-3 expression.

What is the central question of this study? The aim of the experiment was to highlight the regenerative capacity of bone marrow Kit+ cells in the restoration of asthmatic pulmonary function in the rat model. What is the main finding and its importance? Data showed that these cells were recruited successfully to the asthmatic niche after intratracheal administration and accelerated the regeneration of asthmatic lungs by the modulation of inflammation via the control of Gata3 and Tbx21 expression, leading to decreased tracheal responsiveness to methacholine and reduction of pathological remodelling. Allergic asthma is a Thelper (Th)2 immunological disorder with consequential uncontrolled inflammatory responses. There is an increasing demand to use new methods for the treatment of asthma based on modulation of the Th2-to-Th1 ratio in favour of the Th1 population. Accordingly, we decided to evaluate the effects of intratracheal administration of Kit+ bone marrow cells on tracheal responsiveness and the expression of Gata3 and Tbx21 genes. Forty male Wistar rats were allocated randomly into four experimental groups: healthy rats (control group), sensitized rats (OVA group), sensitized rats receiving Kit- cells (OVA+Kit- group) and sensitized rats receiving Kit+ cells (OVA+Kit+ group). Total and differential white blood cell counts, tracheal responsiveness to cumulative methacholine concentrations and histopathological analysis were evaluated. The results showed a statistically significant increase in total white blood cell, eosinophil and neutrophil counts, tracheal contractility, Gata3 expression and prototypical histopathology of asthma. Along with these conditions, we found that the number of lymphocytes was decreased and expression of Tbx21 diminished in sensitized rats compared with control animals. Monitoring of labelled tagged cells confirmed successful engraftment of transplanted cells in pulmonary tissue. Juxtaposition of Kit+ cells changed the blood leucogram closer to the control values. Kit+ cells increased the expression of Tbx21 and suppressed Gata3 (P<0.05). In the OVA+Kit+ group, tracheal responsiveness was improved coincident with increased pulmonary regeneration. In conclusion, this study showed that intratrachealadministration of bone marrow-derived Kit+ cells, but not Kit- cells, could be effective in the alleviation of asthma, presumably by the modulation of Gata3 and Tbx21.

Leukocytes differential under effect of ciprofloxacin in experimental colibacillosis in chickens

This study was aimed at evaluation of white blood cells of domestic birds over time using a broad-spectrum chemotherapeutic drug Ciprofloxacin. Ciprofloxacin more actively inhibits gram-negative bacteria. In order to evaluate the dynamics, three groups of chickens were formed (1st group – control; 2nd and 3rd groups – experimental ones), which consisted of Hisex Brown cross-bred cockerels. Within the framework of this experiment chickens in the 2nd and 3rd groups were contaminated with Escherichia coli. A day before and for four days after the contamination chickens in the 3rd group received 200 mg/L of ciprofloxacin with water. Blood samples were drawn from all experimental chickens at Day 1, 5, 10, 15 and 20 after the contamination. White blood cell differential count was performed. Analysis of the data obtained showed significant changes in some blood cell parameters in the 2nd and 3rd groups in comparison to the control group. Inflammatory response was detected, which was characterized by significant increase in absolute count and percentage of pseudoeosinophils, monocytes and basophils in the 2nd and 3rd group in comparison to the control group. Nevertheless, white blood cells differential count in the 3rd group was relatively closer to analogous count in the control group.

Phagocytosis and inflammation in crystal-induced arthritis: a synovial fluid and in vitro study

The aim of this study was to investigate the relationship between the degree of crystal phagocytosis and the magnitude of the local inflammatory process using fresh synovial fluid (SF) collected from patients with crystal-induced arthritis. In parallel, an in vitro model of crystal-induced inflammation was used to assess the effect of cell priming on crystal phagocytosis and IL-1ß production. SF was collected from 20 patients with gout and 20 with pyrophosphate crystal-induced arthritis and examined under ordinary and polarised light microscopy for total and differential white blood cell (WBC) count and crystal search. The total phagocytosis index was determined in SF along with IL-1β, IL-8, IL-10, and TGFβ levels. The in vitro studies were performed using primed or unprimed THP-1 cells stimulated with calcium pyrophosphate (CPP) crystals, monosodium urate (MSU) crystals and/or cytochalasin D. We demonstrated that the phagocytosis index calculated on the total number of cells was independent from the inflammatory local indices such as WBC and the percentage of polymorphonuclear cells but showed a positive correlation with pro-inflammatory cytokines. By contrast, the local inflammatory indices (WBC and IL-1ß) showed a strong positive correlation with the percentage of polymorphonuclear cells with crystals internalised and a negative correlation with the percentage of mononuclear cells with crystals internalised. The in vitro study showed that phagocytosis represents a fundamental step in the induction of the inflammatory response to MSU and CPP crystals, but it also occurs in absence of cell priming. The results of this study indicate a possible role of non-inflammatory phagocytosis in limiting the acute attack of crystal-induced arthritis.

Performance of the Sysmex White Precursor Channel to discover circulating leukemic blast cells.

Circulating immature precursor cells indicate malignant diseases like acute myeloid or lymphoid leukemia, and blast cells are key finds for disease management. Automatized cell counters are an essential contemporary appliance for blast detection, but false-positive samples remain challenging in terms of time and resources. To reduce this issue, the White Precursor Channel (WPC) was introduced to Sysmex XN series; however, sensitivity may reduce when accommodating low specificity. Therefore, our aim was to evaluate WPC blast alarm flag performance with regard to detecting blast cells. At two major Danish hospitals, random blood samples were collected from the routine setting in a four-week period and analyzed on WPC XN20 (Sysmex, Japan). Results were compared with manual differential white blood cell count (Manual WBCC) assisted by CellaVisionDM96. In 117 samples, we found 0.2 to 34.4% blasts, WPC blast flag specificity=82% and a low sensitivity=40%. However, other XN alarm flags forwarded samples to Manual WBCC, so blast cells were detected despite missing a specific blast flag: With all alarm flags, combined sensitivity increased to 88%. Overall, the WPC application stopped 18% of the 117 samples going to Manual WBCC (three false negatives). Q values are arbitrary probability measurements for the blast flag, and in five samples (0.5 to 47.3% blasts) imprecision ranged from 5.3 to 122 CV%. WPC blast alarm flags are imprecise and inaccurate, especially when blast counts are low. However, the XN20 will alarm samples with other flags so that most samples containing blast cells will be manually reviewed after all. Hence, the presented flag types should not bias the decisions of manual reviewers.

Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics.

A multiplexed droplet PCR (mdPCR) workflow and detailed protocol for determining epigenetic-based white blood cell (WBC) differential count is described, along with a thermoplastic elastomer (TPE) microfluidic droplet generation device. Epigenetic markers are used for WBCsubtyping which is of important prognostic value in different diseases. This is achieved through the quantification of DNA methylation patterns of specific CG-rich regions in the genome (CpG loci). In this paper, bisulfite-treated DNA from peripheral blood mononuclear cells (PBMCs) is encapsulated in droplets with mdPCR reagents including primers and hydrolysis fluorescent probes specific for CpG loci that correlate with WBC sub-populations. The multiplex approach allows for the interrogation of many CpG loci without the need for separate mdPCR reactions, enabling more accurate parametric determination of WBC sub-populations using epigenetic analysis of methylation sites. This precise quantification can be extended to different applications and highlights the benefits for clinical diagnosis and subsequent prognosis.

Correlating immune toxicity, blood cell counts, and overall survival in cancer patients receiving immune therapy.

3043 Background: Baseline circulating white blood cell differential counts have been proposed as possible markers of response to Immune therapy (ICI) and immune toxicity (irAE), but have not been validated for clinical practice. Methods: 214 patients with various cancers receiving ICI had clinical lab results and overall/organ specific irAE analyzed. Absolute lymphocyte (ALC), eosinophil (AEC) and neutrophil (ANC) counts &lt;7 days of starting and 3-12 weeks after ICI were correlated with survival and development of irAE. The association between overall irAE and cell count changes was evaluated using multivariate logistic regression, adjusting for patients’ age, gender, race, and including all 3 types of cell count changes in one model. The association between overall survival and cell counts changes was tested using multivariate Cox proportional hazard model, adjusting for age, gender, race, and overall irAE (yes/no). Results: The combination of MEDIAN rise or fall in ALC, ANC and AEC after ICI combined with development of irAE was associated with altered overall survival (OS). Higher statistically significant (p=&lt;0.05) OS was seen in patients with AEC↑/irAE+(n=94; median OS 14.9 months), ANC↓/irAE+(60; 17.9 mo), ALC↑/irAE+(65; 11.5 mo), ALC↓/irAE+(92; 12.2 mo). Lower OS was seen with ANC↑/irAE-(21; 5.4 mo), ALC↑/irAE-(9; 5.8 mo), ALC↓/irAE-(24; 6.1 mo). All these relationships were unchanged if patients were grouped into ANY rise during the 3-12 weeks rather than median. After multivariable adjustment for age/sex/race there was statistically significant association between higher baseline ALC and development of any irAE (p=0.05) and grade ≥2 irAE (p=0.02). No association was seen between overall toxicities and ANC or AEC. Organ-specific irAE’s with statistically significant associations included pneumonitis:↓AEC (p=0.05), hepatitis:↓ANC (p=0.05), hypothyroidism:↓ANC (p=0.02), colitis:↑ALC (p=0.02). Skin rash, hypophysitis and adrenalitis were not associated with any cell count changes. Conclusions: In patients with cancer receiving immune therapy, the predictive value of serial monitoring in ALC, AEC and ANC is greatly enhanced by the addition of irAE detection. irAEs appear to be a greater predictor of survival than changes in blood counts. In contrast to prior reports we found ANC/ALC/AEC in isolation were less reliable in predicting improved survival. Lack of standardized methods of assessing irAE’s confound identifying strong associations with clinical lab-derived cell counts.

Cardiogenic shock elicits acute inflammation, delayed eosinophilia, and depletion of immune cells in most severe cases

Patients with cardiogenic shock (CS) display systemic inflammation and a high rate of infections, suggesting important immune disturbances. To explore the immune response to CS, we prospectively measured, in 24 consecutive CS patients, differential white blood cell (WBC) counts and the cytokines IL-1β, IL-5, IL-6, IL-10, TNFα, IFNγ, MCP-1 and eotaxin (CCL11), at Day 1 (T1), day 3 (T2) and day 6-8 (T3). Secondary infections and their influence on cytokines and WBCs were determined. CS induced early (T1) neutrophilia and elevated levels of IL-6, IL-10 and MCP-1, correlating with shock severity. The eosinophil chemoattractant eotaxin was elevated at T1 and decreased thereafter, and a progressive rise of blood eosinophils was noted over time. Patients with the most severe shock had reduced lymphocytes and monocytes at T2 and T3. Sixty-two percent of patients developed an infection, which did not alter the profile of immune response, except from higher IL-6 levels at T2. Therefore, CS elicits an acute pro-inflammatory response, followed by a delayed increase in blood eosinophils, consistent with the development of a tissue repair response, as well as depletion of immune cells in the most severely affected patients, which might predispose to secondary infections.