Number Of Blood Monocytes Research Articles

Protocolo diagnóstico de las leucocitosis afebriles

Leukocytosis, defined as an increase in the white blood cell count, can occur in a multitude of clinical situations. This protocol will focus on afebrile leukocytosis. To assess leukocytosis, the type of cell with increased values, the degree of leukocytosis, and the patient's symptoms must be evaluated and the morphology of cells observed under the microscope must be taken into account.Neutrophilia refers to an increase of at least two standard deviations above the mean in the number of neutrophils in peripheral blood. The most frequent causes of neutrophilia are reactive forms (secondary to another condition) and the other group are those of primary origin when there is autonomous medullary hyperproduction.Lymphocytosis is defined as an absolute lymphocyte count greater than 4000/μl, although up to 8000/μl may be normal in newborns and children. Reactive causes are more common in children or young adults, notable among which are mononucleosis syndromes. Other important causes are those secondary to clonal disorders; therefore, a blood smear study for cell morphology and flow cytometry study are required.Monocytosis is defined as an increase in the number of monocytes in peripheral blood above 1000/μl. The most frequent causes are infections. Other important causes to consider, especially if monocytosis is sustained and secondary causes have been ruled out, are primary hematologic malignancies, especially in older adults or in patients with unexplained cytopenias.

Diethylhexyl phthalate induces immune dysregulation and is an environmental immune disruptor

Diethylhexyl phthalate (DEHP) is the most abundant phthalate compound in the environment, and has been linked with multiple human diseases. The immune system is closely associated with the occurrence and progression of various diseases. However, minimal research has addressed the impact of DEHP on the immune system. In this study, single-cell RNA sequencing was performed using spleen tissue of mice to comprehensively determine alterations of the immune system in response to DEHP. The results showed that DEHP exposure reduced the absolute number of peripheral white blood cells (WBCs), including lymphocytes, monocytes, eosinophils, basophils, and neutrophils in mice. In addition, scRNA-seq analyses showed that inflammatory signaling and the expression of heat shock proteins (HSPs) were reduced in all peripheral immune cell populations. Furthermore, we established a mice cecal ligation and puncture (CLP) model, and showed that DEHP exacerbated sepsis-induced immunosuppression and organ damage. These results suggest that DEHP is an environmental immune disruptor that undermines the immune system, exacerbating acute infections and organ damage. Our findings offer a novel perspective on the hazards of DEHP to human health.

Glucose metabolism controls monocyte homeostasis and migration but has no impact on atherosclerosis development in mice

Monocytes directly contribute to atherosclerosis development by their recruitment to plaques in which they differentiate into macrophages. In the present study, we ask how modulating monocyte glucose metabolism could affect their homeostasis and their impact on atherosclerosis. Here we investigate how circulating metabolites control monocyte behavior in blood, bone marrow and peripheral tissues of mice. We find that serum glucose concentrations correlate with monocyte numbers. In diet-restricted mice, monocytes fail to metabolically reprogram from glycolysis to fatty acid oxidation, leading to reduced monocyte numbers in the blood. Mechanistically, Glut1-dependent glucose metabolism helps maintain CD115 membrane expression on monocytes and their progenitors, and regulates monocyte migratory capacity by modulating CCR2 expression. Results from genetic models and pharmacological inhibitors further depict the relative contribution of different metabolic pathways to the regulation of CD115 and CCR2 expression. Meanwhile, Glut1 inhibition does not impact atherosclerotic plaque development in mouse models despite dramatically reducing blood monocyte numbers, potentially due to the remaining monocytes having increased migratory capacity. Together, these data emphasize the role of glucose uptake and intracellular glucose metabolism in controlling monocyte homeostasis and functions.

Evaluation of plant commercial feed additives for equine cyathostomin control

The increasing emergence of anthelmintic-resistant parasitic isolates prompts us to reassess the management of intestinal strongylosis in horses. Additionally, societal demand is shifting toward reducing the use of chemical treatments, aligning with environmentally-friendly practices and the exploration of natural alternatives. In this context, we provide an initial view of the antiparasitic activity and the effect on immune circulating blood cells of three commercialized plant-based feed additives in ponies. Three treatments, based either on mugwort (Artemisia vulgaris), echinacea (Echinacea purpurea) or curcumin (Curcuma longa) were administrated to 18 (six per treatment) Welsh female ponies naturally infected with cyathostomins to mimic their practical use in farming conditions. Another group of six untreated ponies was used as a control. Fecal egg count (FEC), the larval development percentage and the number of red blood cells, lymphocytes, monocytes, neutrophils, eosinophils and basophils were measured the first and the last day of each treatment, and compared with those characterizing the control group. None of the three treatments showed a significant effect on the studied parameters. Moreover, the efficacy of treatments, measured from the FEC reduction compared to the control group, was weak (≤ 38.6 %). Therefore, these results do not support the practical use of these additives in equine farming, even if the determination of Cohen's d values associated with the three treatments revealed some incidences on FEC and blood immune cell counts, as well as on larval development for mugwort.

Prognostic Value of LMR and CD163+TAM for Patients with Diffuse Large B Cell Lymphoma

To investigate the prognostic value of lymphocyte-to-monocyte ratio (LMR) and CD163+tumor-associated macrophages (TAM) in patients with diffuse large B cell lymphoma (DLBCL). Peripheral blood and lymph node tissues were collected from 63 newly diagnosed DLBCL patients. LMR was calculated by the number of lymphocytes and monocytes in peripheral blood from the result of blood routine examination. The level of CD163+TAM in lymph nodes was detected by immunohistochemistry. The cut-off values of LMR and CD163+TAM were determined by ROC curves, and the prognostic value of LMR and CD163+TAM in DLBCL patients was analyzed. The LMR level of 63 newly diagnosed DLBCL patients was 3.69±1.71, and the median value of CD163+TAM was 26/HPF. The number of CD163+TAM was negatively correlated with LMR (r =-0.58) and positively correlated with monocyte count (r =0.46). The cut-off values of LMR and CD163+TAM determined by ROC curve were 2.95 and 29/HPF, respectively, and based on this, the patients were divided into low LMR group and high LMR group, as well as low CD163+TAM group and high CD163+TAM group. The proportion of patients with clinical stage III-IV, IPI score 3-5 and bone marrow infiltration in the low LMR group were higher than those in the high LMR group (P < 0.05). The proportion of patients with clinical stage III-IV, IPI score 3-5, elevated LDH level and bone marrow infiltration in the high CD163+TAM group were higher than those in the low CD163+TAM group (P < 0.05). There was a positive correlation between LMR and OS (r =0.43) and a negative correlation between CD163+TAM and OS (r =-0.65). DLBCL patients with low LMR and high CD163+TAM had shorter OS (P < 0.05). Low LMR and high CD163+TAM can be used as biological markers for poor prognosis of DLBCL patients.

Functional analysis of airway remodeling is related with fibrotic mediators in asthmatic children

Background Asthmatic children present variable degrees of airway inflammation, remodeling, and resistance, which correlate with disease control and severity. The chronic inflammatory process of the airway triggers airway remodeling, which reflects the degree of airway resistance. Pro-inflammatory and pro-fibrotic mediators are centrally involved in this process. Objective To investigate whether the levels of pulmonary and systemic pro-inflammatory and pro-fibrotic mediators present a correlation with the resistance of the respiratory system and of the proximal and distal airways. Methods 39 Asthmatic children (persistent mild and moderate) and 39 non-asthmatic children (both between 6 and 13 years old) were evaluated for anthropometric characteristics, lung function and mechanics, and pulmonary and systemic immune responses. Results Asthmatic children showed an increased number of blood eosinophils (p < 0.04), basophils (p < 0.04), monocytes (p < 0.002) and lymphocytes (p < 0.03). In addition, asthmatic children showed impaired lung function, as demonstrated by FEV1 (p < 0.0005) and FEV1/FVC (p < 0.004), decreased total resistance of the respiratory system (R5Hz; p < 0.009), increased resistance of the proximal airways (R20Hz; p < 0.02), increased elastance (Z5Hz; p < 0.02) and increased reactance (X5Hz; p < 0.002) compared to non-asthmatic children. Moreover, the following inflammatory factors were significantly higher in asthmatic than non-asthmatic children: GM-CSF in the breath condensate (BC) (p < 0.0001) and in the serum (p < 0.0001); TGF-beta in the BC (p < 0.0001) and in the serum (p < 0.004); IL-5 in the BC (p < 0.02) and in the serum (p < 0.01); IL-4 in the serum (p < 0.0002). Conclusions Impulse oscillometry is a sensitive method to detect airway resistance in persistent mild and moderate asthmatic children, an event followed by increased levels of pro-inflammatory and pro-fibrotic mediators.

Ecotoxicological Assessment of Ponds Contaminated by Wastewater of Livestock Enterprises by Fish Biomarkers

The morphometric characteristics of internal organs, the morphological composition of blood, and indicators of metabolic processes in the blood plasma and hepatopancreas of the two-year-old carp (&lt;i&gt;Cyprinus carpio&lt;/i&gt; L.) from the conditionally clean pond and contaminated with sewage from a livestock enterprise pond were examined. It was established that the considered contamination caused increase of the spleen mass and its index respectively 1.63 and 1.65 times, but did not affect live weight, body length and height, body structure index, hepatopancreas mass and its index. The hemoglobin concentration, number of erythrocytes, thrombocytes and monocytes in blood of carps from the contaminated pond was lower than in control, whereas number of the segmented neutrophils and ESR were higher, and the number of leukocytes, eosinophils, lymphocytes and rod-nucleated neutrophils did not differ. Wastewater from the livestock enterprise affected metabolic processes in carp, particularly caused increase of the glucose and total protein concentration, the lactate dehydrogenase, aspartate aminotransferase and creatine kinase activity in blood plasma and hepatopancreas. At the same time, there were no differences between specimens from the conditionally clean and contaminated pond in terms of content of triglycerols, cholesterol, urea, creatinine, calcium, inorganic phosphorus, AlAT and AsAt activity in blood plasma and hepatopancreas and total protein content and LHD activity in hepatopancreas, content of total protein, glucose, creatinine, urea, the glucose content in hepatopancreas. The established changes in internal organs, morphological composition of blood and metabolic parameters, particularly concentration of hemoglobin and the erythrocytes number in blood, glucose and total protein content, and activity of lactate dehydrogenase and creatine kinase in carp enabled to propose to use these parameters as supplementary over assessment of the ecotoxicological state of ponds contaminated by wastewater from livestock enterprises.

Eosinophils at diagnosis are elevated in amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a rare, devastating neurodegenerative disease that affects upper and lower motor neurons. To date, no effective treatment or reliable biomarker for ALS has been developed. In recent years, many factors have been proposed as possible biomarkers of ALS; however, no consensus has been reached. Therefore, a reliable biomarker is urgently needed. Eosinophils may play a crucial role in healthy humans and diseases, and serve as a biomarker for many chronic diseases. Routine blood test results were collected from 66 healthy controls and 59 patients with ALS. The percentages and total numbers of each cell population were analyzed, and the correlation between these indicators and patient ALS functional rating scale-revised (ALSFRS-R) score or disease progression rate (ΔFS score) was analyzed. Compared to healthy controls, the number of blood leukocytes, neutrophils, monocytes, and basophils was significantly decreased in patients with ALS (p = 0.002, p = 0.001, p = 0.049, and p < 0.0001, respectively). There was an increase in the number of eosinophils (p < 0.0001), but no difference in the number of lymphocytes between patients with ALS and healthy controls was found (p = 0.563). Compared to healthy controls, the percentage of neutrophils was decreased and the percentage of lymphocytes and eosinophils was increased in patients with ALS (p = 0.01, p = 0.012, and p = 0.001, respectively). There was no difference between patients with ALS and healthy controls in the percentage of monocytes and basophils (p = 0.622 and p = 0.09, respectively). However, only the percentage and number of eosinophils had a correlation with the ΔFS score. Further multivariate analysis revealed a significant correlation between the disease duration, eosinophil count and percentage, and the disease progression rate (p < 0.0001, p = 0.048, and p = 0.023, respectively). The neutrophil-to-eosinophil ratio (NER), lymphocyte-to-eosinophil ratio (LER), and monocyte-to-eosinophil ratio (MER) were significantly lower in patients with ALS than in healthy controls. However, only the LER was significantly correlated with the ΔFS score. These observations implicate neutrophils, lymphocytes, and eosinophils as important factors, and increasing eosinophil counts were negatively correlated with the ΔFS score in patients with ALS.

Depletion of muscularis macrophages ameliorates inflammation-driven dysmotility in murine colitis model

Previously, the presence of a blood-myenteric plexus barrier and its disruption was reported in experimentally induced colitis via a macrophage-dependent process. The aim of this study is to reveal how myenteric barrier disruption and subsequent neuronal injury affects gut motility in vivo in a murine colitis model. We induced colitis with dextran sulfate sodium (DSS), with the co-administration of liposome-encapsulated clodronate (l-clodronate) to simultaneously deplete blood monocytes contributing to macrophage infiltration in the inflamed muscularis of experimental mice. DSS-treated animals receiving concurrent l-clodronate injection showed significantly decreased blood monocyte numbers and colon muscularis macrophage (MM) density compared to DSS-treated control (DSS-vehicle). DSS-clodronate-treated mice exhibited significantly slower whole gut transit time than DSS-vehicle-treated animals and comparable to that of controls. Experiments with oral gavage-fed Evans-blue dye showed similar whole gut transit times in DSS-clodronate-treated mice as in control animals. Furthermore, qPCR-analysis and immunofluorescence on colon muscularis samples revealed that factors associated with neuroinflammation and neurodegeneration, including Bax1, Hdac4, IL-18, Casp8 and Hif1a are overexpressed after DSS-treatment, but not in the case of concurrent l-clodronate administration. Our findings highlight that MM-infiltration in the muscularis layer is responsible for colitis-associated dysmotility and enteric neuronal dysfunction along with the release of mediators associated with neurodegeneration in a murine experimental model.

Correlation between Blood Monocytes and CSF Tau in Alzheimer's Disease: The Effect of Gender and Cognitive Decline.

Neuroinflammation is one of the main mechanisms contributing to the pathogenesis of Alzheimer's disease (AD), although its key role and the immune cells involved have not yet been identified. Blood monocytes appear to play a role in the clearance of AD-related amyloid-β (Aβ) and tau protein. This retrospective study evaluated a possible correlation between blood monocytes; the concentrations of Aβ, total tau (t-Tau), and phosphorylated tau (p-Tau) in the cerebrospinal fluid (CSF); and cognitive decline assessed according to the Montreal Cognitive Assessment (MoCA). We collected data from 33 patients with AD or mild cognitive impairment (MCI) due to AD (15 men and 18 women) and found, along with a significant reduction in the concentration of blood monocytes in women (p-value = 0.083),significant correlations between the number of blood monocytes and the concentration of t-Tau in CSF (p-value = 0.045) and between blood monocytes and MoCA score (p-value = 0.037). These results confirm the role of blood monocytes in the pathogenesis of AD, provide further evidence of a gender difference in the neuroinflammatory process underlying AD, and show that blood monocyte count may reflect the cognitive impairment of AD patients.

Hematological and biochemical indicators of lambs’ blood after the use of chemotherapeutic agents for spontaneous eimeriosis

Eimeria infection is one of the protozoal diseases of animals caused by various species of Eimeria and causes reduced productivity and raised mortality in ruminants, especially in young ones. To study the effect of anti-emeriosis drugs on the morphological and biochemical parameters of lambs’ blood infected with Eimeria, 3 groups of Romanov breed animals, two months old, affected by the Eimeria pathogen were formed. The first group of animals was treated with a drug containing the active ingredient decoquinate; the second group of animals with a drug containing the active ingredient toltrazuril; the third group of lambs were given a combined drug containing 80 mg of norsulfazole, 70 mg of sulgin, 30 mg of trimethoprim, 45 mg of oxytetracycline hydrochloride and 25 mg of tylosin tartrate per 1 g); the fourth group of lambs was the control group, the animals were free from the Eimeria pathogen and were kept separately. The blood of the animals was analysed on days 7 and 14 of treatment. In the blood of the experimental lambs, an increase in the number of red blood cells, neutrophils, lymphocytes, and monocytes was noted. There was also a decrease in the total number of leukocytes, eosinophils, young and segmented neutrophils. According to the results of the treatment, there was an increase in the content of total, albumin, hemoglobin and glucose, as well as the activity of gamma-glutamate transferase. At the same time, there was a decrease in the content of globulins and the activity of aspartate aminotransferase and alanine aminotransferase compared to the biochemical parameters of healthy lamb blood serum of the control group. According to treatment results, the restoration of sick lambs’ blood parameters to those of healthy animals was noted; however, the most stable effect was using the combined drug.

Emodin reduces surgical wounding-accelerated tumor growth and metastasis via macrophage suppression in a murine triple-negative breast cancer model.

It has been suspected that tumor resection surgery itself may accelerate breast cancer (BC) lung metastasis in some patients. Emodin, a natural anthraquinone found in the roots and rhizomes of various plants, exhibits anticancer activity. We examined the perioperative use of emodin in our established surgery wounding murine BC model. Emodin reduced primary BC tumor growth and metastasis in the lungs in both sham and surgical wounded mice, consistent with a reduction in proliferation and enhanced apoptosis (primary tumor and lungs). Further, emodin reduced systemic inflammation, most notably the number of monocytes in the peripheral blood and reduced pro-tumoral M2 macrophages in the primary tumor and the lungs. Consistently, we show that emodin reduces gene expression of select macrophage markers and associated cytokines in the primary tumor and lungs of wounded mice. Overall, we demonstrate that emodin is beneficial in mitigating surgical wounding accelerated lung metastasis in a model of triple-negative BC, which appears to be mediated, at least in part, by its actions on macrophages. These data support the development of emodin as a safe, low-cost, and effective agent to be used perioperatively to alleviate the surgery triggered inflammatory response and consequential metastasis of BC to the lungs.

Demethyleneberberine alleviates Pseudomonas aeruginosa-induced acute pneumonia by inhibiting the AIM2 inflammasome and oxidative stress

BackgroundAcute pneumonia induced by Pseudomonas aeruginosa is characterized by massive infiltration of inflammatory cell and the production of reactive oxygen species (ROS), which lead to severe and transient pulmonary inflammation and acute lung injury. However, P.aeruginosa infection is resistant to multiple antibiotics and causes high mortality in clinic, the search for alternative prophylactic and therapeutic strategies is imperative. PurposeThis study was aimed to investigate the anti-inflammatory and antioxidant effects of DMB, a novel derivative of berberine, and explore the role of AIM2 inflammasome in P. aeruginosa-induced acute pneumonia. MethodsAcute pneumonia mice were established by tracheal injection of P. aeruginosa suspension. Pathological changes of lung tissue were observed by its appearance and H&E staining. The lung coefficient ratio was measured to evaluate pulmonary edema. Inflammatory factors were detected by qRT-PCR, western blotting and immunohistochemistry. ROS and other indicators of oxidative damage were analyzed by flow cytometry and specific kit. Proteins related to AIM2 inflammasome were detected by western blotting. ResultsCompared with the P. aeruginosa-induced group, DMB ameliorated pulmonary edema, hyperemia, and pathological damage based on its appearance and H&E staining in DMB groups. First, DMB attenuated the inflammatory response induced by P.aeruginosa. Compared with the P. aeruginosa-induced group, the lung coefficient ratio was decreased by 31.5%, the MPO activity of lung tissue was decreased by 44.0%, the mRNA expression levels of TNF-α, IL-1β and IL-6 were decreased by 64.8%, 51.2% and 64.0% respectively, and those protein expression levels were decreased by 40.1%, 42.8% and 47.8% respectively, and the number of white blood cells, neutrophils and monocytes were decreased by 53.5%, 29.4% and 13.7% in high dose (200 mg/kg) DMB group. Second, DMB alleviates oxidative stress in the lung tissue during P. aeruginosa-induced acute pneumonia. Compared with the P. aeruginosa-induced group, the level of GSH was increased by 42.5% and MDA was decreased by 49.5% in high dose DMB group. Moreover, the western blotting results showed that DMB markedly suppressed the expression of AIM2, ASC, Cleaved caspase1 and decreased the secretion of IL-1β. Additionally, these results were also confirmed by in vitro experiments using MH-S and BEAS-2B cell lines. ConclusionsTaken together, these results indicated that DMB ameliorates P. aeruginosa-induced acute pneumonia through anti-inflammatory, antioxidant effects, and inhibition of AIM2 inflammasome activation.

Focus on the anti-inflammatory effect of glucocorticosteroids in experimental acute lung injury

AIM: Assessment of the effect of various doses of dexamethasone as an inflammation modulator in experimental lipopolysaccharide-induced acute lung injury in rats.&#x0D; MATERIALS AND METHODS: Acute lung injury in rats was modeled by intratracheal administration of cell wall lipopolysaccharide from the Salmonella enterica. White male rats were divided into groups: a group of intact animals (n = 10); the control group (n = 40), in which the animals were simulated acute lung injury without further treatment and removed from the experiment on day 3; three experimental groups (n = 40), in which, 3 hours after modeling acute lung injury, and then daily once a day for 3 days, dexamethasone solution was administered intraperitoneally in the following doses: in group 1 0.52 mg/kg (equivalent to 6.0 mg/day for a person), in group 2 1.71 mg/kg (20.0 mg/day for a person), in group 3 8.0 mg/kg (94.0 mg/day, pulse therapy for humans). On the 3rd day, blood samples were taken from the caudal vena cava in surviving animals for clinical analysis and evaluation of the function of mitochondria of peripheral blood leukocytes. To determine the severity of local inflammatory reactions and pulmonary edema, bronchoalveolar lavage was performed with the study of an endopulmonary cytogram and an assessment of pathomorphological changes in the lung tissue.&#x0D; RESULTS: indicate that dexamethasone reduces the amount of lung tissue damage and animal mortality, dose-dependently reduces the functions of mitochondria and the number of lymphocytes and monocytes in peripheral blood, as well as neutrophils, lymphocytes and macrophages in bronchoalveolar lavage samples.&#x0D; CONCLUSION: The use of dexamethasone at a dose of 0.52 mg/kg (equivalent to 6.0 mg/day for humans) is accompanied by better survival, minimal effect on the viability and functional activity of inflammatory cells. Pulse therapy leads to a significant decrease in the number of immunocompetent cells in bronchoalveolar lavage, mitochondrial dysfunction in the form of a decrease in the ability of these cells to use the reserve power of mitochondrial respiration in response to the action of a stress factor. Excessive inhibition of immunocompetent cells can contribute to the activation of latent and opportunistic infections, which must be taken into account when choosing a dosing regimen for glucocorticosteroids.

The Concentration of PCSK9-Lp(a) Complexes and the Level of Blood Monocytes in Males with Coronary Atherosclerosis.

In this study we analyzed the concentration of lipoprotein(a) (Lp(a)), PCSK9-Lp(a) complexes and the circulating monocyte subsets in coronary atherosclerosis. For this study, 257 patients with coronary atherosclerosis and 68 patients without stenotic atherosclerosis in the coronary, carotid and lower extremity arteries (control group) were enrolled. The monocyte subpopulations (classical CD14++CD16-, intermediate CD14++CD16+ and non-classical CD14+CD16++) were analyzed by direct immunofluorescence and flow cytometry. The Lp(a) and PCSK9-Lp(a) complexes in the serum were detected by ELISA. The concentration of Lp(a) was higher in the coronary atherosclerosis group compared with the controls (23.0 (9.1; 73.3) mg/dL versus 10.7 (4.7; 25.0) mg/dL, p < 0.05). No correlations between the level of Lp(a) and the concentration of the PCSK9-Lp(a) complexes, nor between the level of Lp(a) or PCSK9 and the total number of monocytes, were observed in either group. A slight positive correlation between the concentration of PCSK9-Lp(a) complexes and the absolute level of monocytes was obtained (r = 0.20, p = 0.002) in the patients with atherosclerosis due to the intermediate monocyte subsets (r = 0.33, p = 0.04). According to regression analysis, both the PCSK9-Lp(a) complexes concentration and BMI were related to the absolute number of blood monocytes in patients with atherosclerosis. Further studies are required to determine the pathogenetic contribution of PCSK9-Lp(a) complexes to the development of atherosclerosis.

Clinical Significance of Nodal DCsign Expression in Non-small-cell Lung Cancer Patients.

Dendritic cells (DCs) are difficult to evaluate in lung regional lymph nodes because of region-specific structures, such as abundant trabeculae connecting the medullary and subcapsular sinuses, the latter of which contains few anthracotic macrophages. Therefore, DC-specific intercellular adhesion molecule-3-grabbing non-integrin (DCsign)-positive DCs and CD68-positive macrophages are unlikely to show a typical distribution. The present study therefore explored quantitative factors connecting the nodal DC morphology to the patient outcome. Lymph nodes from 34 non-small-cell lung cancer patients who underwent complete resection were used for immunohistochemical assessments of DCsign and CD68 and terminal deoxynucleotidyl transferase dUTP nick-end labeling. Preoperative patient blood samples were used for the quantitative evaluation of monocytes. The nodal DCs showed a complementary distribution with macrophages, thus few DCs were seen in clusters of macrophages. DCs often presented as a mesh-like rosette that was solitary or connected to a DC cluster. DCs disappeared, and some macrophages were apoptotic when surrounded by cancer cells that have metastasized to lymph nodes. The proportional area of a DC cluster was significantly associated with the histological differentiation of cancer (p=0.013), with a higher ratio tending to lead to a better overall survival (p=0.059), and significantly so in adenocarcinoma (p=0.007). The rosette number was significantly correlated with the smoking index and blood monocyte number (p=0.013 and p=0.005, respectively). The nodal DC morphology appears useful as a prognostic factor and may lead to a new phase of clinicopathological studies of solid cancers.

Toll-like receptor 2 activation in monocytes contributes to systemic inflammation and alcohol-associated liver disease in humans.

In the context of gut leakiness and translocation of microbial products in alcohol-associated liver disease (ALD), it is possible that systemic and liver inflammation involve the activation of circulating monocyte through gut-derived factors. We explored the association between monocytes, microbial translocation, systemic inflammation, and ALD. Patients with alcohol use disorder following a rehabilitation program were compared with healthy controls. We determined the circulating number and proportion of monocyte subsets by FACS. The activation of signaling pathways by gut-derived microbes was analyzed by quantitative PCR in isolated monocytes. Cytokines secretion by monocytes and phagocytosis were assessed in vitro. Serum microbial translocation markers and cytokines were measured by ELISA and multiplex assay, respectively. ALD severity and liver inflammatory responses were analyzed in liver biopsies by various methods. In patients with alcohol use disorder, the number of blood monocytes increased compared with controls. Monocytes from patients with alcohol use disorder upregulated IL-1β and IL-8 together with toll-like receptor 2 and downstream AP-1, while fungal sensor CARD9 was downregulated. IL-1β and IL-8 were actively secreted upon stimulation in vitro with the toll-like receptor 2 ligand peptidoglycan. Exposure with Escherichia coli confirmed preserved bacterial phagocytic activity. In contrast, Candida albicans stimulation leads to downregulation of IL-1β and TNFα compared with controls. Systemic cytokines and monocyte changes correlated with microbial translocation. Hepatic IL-1β and IL-8 increased with ALD severity together with liver macrophage activation and upregulation of chemokines involved in monocyte attraction. Our results point to the contribution of activated monocytes to systemic inflammation and ALD. Monocytes likely infiltrate the liver, transform into monocyte-derived macrophages and release IL-1β and IL-8 in response to peptidoglycan and toll-like receptor 2 activation.

Effects of regorafenib on the mononuclear/phagocyte system and how these contribute to the inhibition of colorectal tumors in mice

BackgroundRegorafenib was previously shown to reduce tumor-associated macrophages and potently inhibit colony-stimulating factor 1 receptor (CSF1R), also known as CD115, in biochemical assays. The CSF1R signaling pathway is essential in the biology of the mononuclear/phagocyte system, which can promote the development of cancer.MethodsA deeper investigation of regorafenib’s effects on CSF1R signaling was performed using preclinical in vitro and in vivo studies with syngeneic CT26 and MC38 mouse models of colorectal cancer. Peripheral blood and tumor tissue were analyzed mechanistically by flow cytometry using antibodies against CD115/CSF1R and F4/80 and by ELISA for chemokine (C–C motif) ligand 2 (CCL2) levels. These read-outs were correlated with drug levels for the detection of pharmacokinetic/pharmacodynamic relationships.ResultsPotent inhibition of CSF1R by regorafenib and its metabolites M-2, M-4, and M-5 was confirmed in vitro in RAW264.7 macrophages. The dose-dependent growth inhibition of subcutaneous CT26 tumors by regorafenib was associated with a significant reduction in both the number of CD115hi monocytes in peripheral blood and the number of selective subpopulations of intratumoral F4/80hi tumor-associated macrophages. CCL2 levels were not affected by regorafenib in blood but increased in tumor tissue, which may contribute to drug resistance and prevent complete tumor remission. An inverse relationship between regorafenib concentration and the number of CD115hi monocytes and CCL2 levels was observed in peripheral blood, supporting the mechanistic involvement of regorafenib.ConclusionsThese findings may be clinically useful in optimizing drug dosing using blood-based pharmacodynamic markers and in identifying resistance mechanisms and ways to overcome them by appropriate drug combinations.

Transplantation of fecal filtrate to neonatal pigs reduces post-weaning diarrhea: A pilot study.

Post-weaning diarrhea (PWD) remains a major source of mortality and morbidity in swine production. Transplantation of bacteria-free filtrate of feces (fecal filtrate transplant, FFT) has shown gut protective effects in neonatal pigs, and early postnatal establishment of the gut microbiome is suggested to determine later stability and robustness of the gut. We, therefore, hypothesized that early postnatal transplantation of bacteria-free feces would have a protective effect against PWD. Using fecal filtrates derived from healthy lactating sows, we compared oral administration of fecal filtrate transplantation (FFT, n = 20) and saline (CON, n = 18) in newborn piglets. We assessed growth, diarrhea prevalence, blood parameters, organ measurements, morphology, and gut brush border enzymes and analyzed luminal bacterial composition using 16S rRNA gene amplicon sequencing. The two groups showed similar average daily gain (ADG) during the suckling period, whereas in the post-weaning period, a negative ADG was observed in both groups. While diarrhea was largely absent in both groups before weaning, there was a lower diarrhea prevalence on days 27 (p = 2.07*10-9), 28 (p = 0.04), and 35 (p = 0.04) in the FFT group relative to CON. At weaning on day 27, the FFT group had higher numbers of red blood cells, monocytes, and lymphocytes, while on day 35, i.e., 1 week after weaning, the two groups were similar regarding hematology. The biochemical profile was largely similar between FFT and CON on days 27 and 35, except for a higher level of alanine aminotransferase and a lower level of Mg in the FFT group. Likewise, organ weights relative to body weight were largely similar on day 35, albeit with a lower stomach weight and more colon content in FFT relative to CON. Gut mucosal percentage and mucosal enzyme activity were similar between the two groups on days 27 and 35. Gut bacterial composition was slightly different on day 35 but not on day 27. In conclusion, early postnatal administration of FFT, showed positive clinical effects in post-weaning pigs, albeit with subtle effects on the gut mucosa and microbiome. Prophylactic treatment with FFT may offer a means to reduce morbidity, yet larger studies are required to document effect size.

Associations between Short-Term Air Pollution Exposure and the Peripheral Leukocyte Distribution in the Adult Male Population in Beijing, China.

The inflammatory effects of air pollution exposure may account for increased public health risk. However, evidence regarding the effects of air pollution on peripheral blood leukocytes in the population is inconsistent. We investigated the association between the short-term effects of ambient air pollution and the peripheral blood leukocyte distribution in adult men in Beijing, China. From January 2015 to December 2019, a total of 11,035 men aged 22-45 years in Beijing were included in the study. Their peripheral blood routine parameters were measured. The ambient pollution monitoring parameters (particulate matter ≤ 10 µm (PM10), PM2.5, nitrogen dioxide (NO2), sulfur dioxide (SO2), carbon monoxide (CO), and ozone (O3)) were collected daily. The potential association between ambient air pollution exposure and peripheral blood leukocyte count and classification was analyzed with generalized additive models (GAMs). After adjusting for confounding factors, PM2.5, PM10, SO2, NO2, O3, and CO were significantly correlated with changes to at least one peripheral leukocyte subtype. Short-term and cumulative air pollutant exposure dramatically increased the participants' peripheral blood neutrophil, lymphocyte, and monocyte numbers and decreased eosinophils and basophils. Our results demonstrated that air pollution induced inflammation in the participants. The peripheral leukocyte count and classification can be utilized to evaluate the inflammation induced by air pollution in the exposed male population.