Phagocytic Activity Research Articles

The Immunomodulatory Effect of Intraperitoneal Injection of Licorice Extract on Giant Salamander

The effect of licorice extract on non-specific immune function of giant salamanders was studied by intraperitoneal injection. The results showed that the lysozyme activity in the drug group increased, and the lysozyme activity increased with the increase of dose. From the 7th day onwards, each sampling of the drug group showed significant differences compared to the control group (P<0.05). The phagocytic activity of macrophages in the drug group showed some fluctuations, but there was no significant difference compared to the control group. The white blood cell volume value of the high-dose group gradually increased in the first three samplings, and showed a significant difference compared to the control group at 14 days (P<0.05). It decreased slightly in the last two samplings, but was still higher than the control group at the same time. The spleen organ coefficient of the low-dose group was (0.7 ± 0.01) % at 28 days, higher than that of the control group (0.4 ± 0.05) %, and the high-dose group was (0.7 ± 0.07) % at 14 days, higher than that of the control group (0.4 ± 0.02) %. Both differences were significant (P<0.05). After the last sampling (28 days), artificial infection with <i>Aeromonas hydrophila</i> bacteria resulted in a mortality rate of 80% in the control group, 50% in the low-dose group, and 30% in the high-dose group, all lower than the control group. The drug group also had higher immune protection rates than the control group. The results indicate that intraperitoneal injection of licorice extract can improve the immune function and disease resistance of giant salamanders to a certain extent.

IMMU-38. LEVERAGING INNATE IMMUNE SENSORS TO GENERATE DURABLE ANTI-GLIOMA ADAPTIVE IMMUNE RESPONSES

Abstract Glioblastoma harbors a tumor microenvironment (TME) that is mostly devoid of T cell effector infiltration but is dominated by immune suppressive microglia, macrophages, and myeloid-derived suppresser cells. The innate immune cells in the TME have impaired phagocytosis and antigen presentation and are inadequate for triggering the subsequent immune activating signals needed for anti-tumor effector responses. Phagocytic activity can also be blocked by the upregulation of CD47 on the tumor cells. In the immune checkpoint resistant Qki-/- Pten-/- P53-/- (QPP) glioma model, intratumoral treatment with the synthetic cyclic di-nucleotide STING agonist IACS-8803 (8803) induces global immunological reprogramming and long-term survival resistant to tumor rechallenge. More specifically, in the infiltrating myeloid stroma 8803 increases costimulatory protein CD86 while reducing the expression of suppressive markers CD163 and CD206. Conventional type 1 dendritic cells are increased in both the TME (p<0.05) and the draining cervical lymph node (p<0.01). Intratumoral infiltration of both CD8 T (p<0.05) and NK (p<0.001) cells increases. However, only T cells, but not NK cells, are necessary for 8803 therapeutic activity when assessed with in vivo NK cell depletion studies and treatment in the RAG-/- background. STING activation increases microglial phagocytic capacity, but also induces a compensatory increase in SIRPα expression (p<0.001) – the ligand for CD47. 8803 synergizes with anti-CD47 blockade to enhance microglia phagocytosis of QPP glioblastoma cells in vitro. Finally, the combination of 8803 and anti-CD47 synergizes to extend both median survival and the fraction of long-term survivors in orthotopic QPP-bearing mice (QPP8v; vehicle control MS=39d, anti-CD47 MS=40d, 8803 MS=67d 20% long-term survival, 8803 + anti-CD47 MS=107.5d 40% long-term survival). These data demonstrate that 8803 induces proinflammatory conversion of the glioblastoma TME, generates T cell-dependent survival benefits in QPP-bearing mice, and synergizes with anti-CD47 blockade to enhance microglia phagocytosis of glioblastoma cells and extend survival in QPP-bearing mice.

CD47 in Osteosarcoma: Correlation with Metastasis and Macrophage-Mediated Phagocytosis

CD47 is expressed on cell surfaces and acts as a “don’t eat me” signal by interacting with signal-regulatory protein-α on the macrophage surface. Some cancer cells express CD47 protein and can evade macrophage phagocytosis. Herein, we evaluated the feasibility of targeting CD47 for osteosarcoma by analyzing its expression patterns, clinicopathological correlations, and immunotherapeutic potential. We performed a retrospective analysis on 24 biopsy samples from patients with osteosarcoma to investigate correlations between CD47 protein positivity and clinicopathological characteristics. CD47 protein expression was detected in 20.8% of the biopsy samples. CD47 positivity correlated with metastasis at diagnosis. Patients with CD47-positive tumors were older than those with CD47-negative tumors. However, CD47 protein expression was not associated with sex, tumor size, or histologic response to preoperative chemotherapy. In vitro, CD47 antibody (B6H12) did not affect osteosarcoma cell viability or apoptosis. In a wound-healing assay, CD47 inhibited the migration of osteosarcoma cells. Differentiated macrophages exhibited higher phagocytic activity against osteosarcoma cells when pretreated with B6H12 compared with the isotype control. Our preliminary data suggest a possible interaction between CD47 protein and macrophage phagocytosis in osteosarcoma metastasis. A better understanding of the role of CD47 is necessary to develop an innovative immunotherapeutic approach against osteosarcoma.

Functional characterization of macrophages and change of Th1/Th2 balance in patients with pythiosis after Pythium insidiosum antigen immunotherapy

There has been limited research into the role of the Pythium insidiosum antigen (PIA) in modulating immune response in patients with pythiosis. This study investigated the balance of T helper type 2 (Th2) and T helper type 1 (Th1) responses after receiving PIA immunotherapy in patients with pythiosis. Next, the phagocytic activity and phagocytic index of IFN-γ primed PIA-treated macrophages were examined. Furthermore, the phagocytosis of infective P. insidiosum zoospores by macrophages was investigated. This work showed that the PIA vaccine induced Th1 response and M1 macrophages in patients with vascular pythiosis who survived and those with localized pythiosis. Phagocytic activity and phagocytic index were increased considerably in localized pythiosis patients compared to vascular pythiosis patients with hematological diseases. IFN-γ priming of PIA-treated macrophages against P. insidiosum zoospores enhanced the phagocytic activity and phagocytic index in vascular and localized pythiosis patients. Macrophages engulfed P. insidiosum zoospores, but the zoospores continued germination, resulting in macrophage death. Overall, our results suggest that PIA can modulate the immune responses, contributing to higher levels of Th1-type cytokine and potentially improving the survival of patients with vascular pythiosis. This study is the first to uncover that P. insidiosum zoospores can survive within macrophages.

Diverse Epithelial Lymphocytes in Zebrafish Revealed Using a Novel Scale Biopsy Method.

Zebrafish (Danio rerio) are a compelling model for studying lymphocytes because zebrafish and humans have similar adaptive immune systems, including their lymphocytes. Antibodies that recognize zebrafish proteins are sparse, so many investigators use transgenic, lymphocyte-specific fluorophore-labeled lines. Human and zebrafish lymphocyte types are conserved, but many aspects of zebrafish lymphocyte biology remain uninvestigated, including lymphocytes in peripheral tissues, like epidermis. This study is, to our knowledge, the first study to focus on zebrafish epidermal lymphocytes, using scales. Obtaining zebrafish blood via nonlethal methods is difficult; scales represent a source to longitudinally sample live fish. We developed a novel biopsy technique, collecting scales to analyze epithelial lymphocytes from several transgenic lines. We imaged scales via confocal microscopy and demonstrated multiple lymphocyte types in scales/epidermis, quantifying them flow cytometrically. We profiled gene expression of scale, thymic, and kidney-marrow (analogous to mammalian bone marrow) lymphocytes from the same animals, revealing B- and T-lineage signatures. Single-cell quantitative real-time PCR and RNA sequencing show not only canonical B and T cells but also novel lymphocyte populations not described previously. To validate longitudinal scale biopsies, we serially sampled scales from fish treated with dexamethasone, demonstrating epidermal lymphocyte responses. To analyze cells functionally, we employed a bead-ingestion assay, showing that thymic, marrow, and epidermal lymphocytes have phagocytic activity. In summary, we establish a novel, nonlethal technique to obtain zebrafish lymphocytes, providing the first quantification, expression profiling, and functional data from zebrafish epidermal lymphocytes.

Alveolar epithelial cells shape lipopolysaccharide-induced inflammatory responses and reprogramming of alveolar macrophages.

Alveolar macrophages (AMs) are sentinels in the airways, where they sense and respond to invading microbes and other stimuli. Unlike macrophages in other locations, AMs can remain responsive to Gram-negative lipopolysaccharides (LPS) after they have responded to LPS in vivo (they do not develop "endotoxin tolerance"), suggesting that the alveolar microenvironment may influence their responses. Although alveolar epithelial cells (AECs) normally limit AMs' innate responses, preventing inflammation induced by harmless antigens in the lung, how AECs influence the innate responses of AMs to infectious agents has been uncertain. Here we report that (1) after exposure to aspirated (intranasal instillation) LPS, AMs increase their responses to TLR agonists and elevate their phagocytic and bactericidal activities in mice; (2) Aspirated LPS pre-exposure increases host resistance to pulmonary infection caused by Gram-negative bacteria and the protection effect lasts for at least 35 days; (3) LPS stimulation of AECs both increases AMs' innate immune responses and prevents AMs from developing tolerance in vitro; (4) Upon LPS stimulation, AMs secreted TNF-α induces AECs to release GM-CSF, which potentiates AMs' response. These experiments have revealed a previously unappreciated role that AECs may play in boosting the innate responses of AMs and promoting resistance to pulmonary infections.

CDC42 enhances phagocytic activity and exhibits antimicrobial functions through the recruitment of IQGAP1 in Apostichopus japonicus

Cell Division Cycle 42 protein (CDC42) is a pivotal member of the Rho family of small GTPases, integral to cell signaling and the regulation of cell polarity. CDC42 is extensively documented to participate in host immunity, particularly in processes such as phagocytosis and cell migration. Nevertheless, the precise mechanisms by which CDC42 affects host-bacteria interactions remain unclear in Echinoderms. In this study, we have isolated and characterized the CDC42 gene from Apostichopus japonicus, referred to as AjCDC42. The complete cDNA sequence of AjCDC42 extends 1282 base pairs, including a typical classical RHO domain, spanning amino acids from 6 to 179. Spatial expression analysis demonstrated that AjCDC42 is ubiquitously expressed across all examined tissues. Following infection with Vibrio splendidus and stimulation with LPS, the protein and mRNA levels of AjCDC42 were significantly upregulated in coelomocytes. Moreover, the knockdown of AjCDC42 significantly decreased V. splendidus-induced phagocytic activity in coelomocytes, leading to a reduced intracellular load of V. splendidus, as evidenced by flow cytometry and bacterial plate counting assays. Mechanistically, AjCDC42 was identified as an interacting partner of IQ Motif Containing GTPase Activating Protein 1 (AjIQGAP1) through the use of co-immunoprecipitation, immunofluorescence, and GST pull-down assays combined with mass spectrometry techniques. Additional functional analyses revealed that the knockdown of AjIQGAP1 similarly decreased the phagocytic activity and the intracellular load of V. splendidus in coelomocytes. Furthermore, we detected that decreased levels of AjCDC42 and AjIQGAP1 impaired cytoskeletal rearrangement and delayed lysosomal clearance of V. splendidus. In summary, our findings underscore the pivotal role of the AjCDC42-AjIQGAP1 axis in regulating coelomocytes phagocytosis in A. japonicus during V. splendidus infection.

Rab5 regulates the antimicrobial phagocytosis of hemocytes through class B scavenger receptor in Procambarus clarkii

The crucial involvement of Rab5 in regulating phagocytosis through mediating receptors, such as the Class B scavenger receptor (SRB), has been documented in higher animal species. However, little is known about the molecular mechanisms of Rab5-mediated phagocytosis in invertebrates. The present study aims to investigate whether the phagocytic function of hemocytes in Procambarus clarkii can be regulated by Rab5 through SRB. We first cloned the open reading frame of SRB from P. clarkii (defined as PcSRB), encoding a putative protein containing a typical CD36 domain. PcSRB was widely expressed in all tested tissues of P. clarkii and induced by viral and bacterial stimulation. The extracellular segment of PcSRB (PcSRB-EX) can agglutinate as well as bind with both Gram-positive (G+) and Gram-negative (G−) bacteria. Knockdown of PcSRB resulted in down-regulation of the phagocytic capacity and bactericidal activity against Aeromonas hydrophila, along with impaired expression of several phagocytosis-related genes and antimicrobial peptide (AMP) genes. PcSRB interference also affected the mRNA expression level of Tolls, indicating that PcSRB might function as a recognition receptor or co-receptor in the Toll signaling pathway. Furthermore, Rab5 interference significantly impaired the expression of PcSRB at both gene and protein levels, which confirmed its significant regulatory effect on PcSRB. Subsequently, we observed a significant decrease in the phagocytic activity and bacteria clearance of hemocytes when Rab5 and PcSRB were simultaneously knocked down. Additionally, there was a notable down-regulation in the expression levels of genes related to phagocytosis and AMPs. Overall, these findings provide preliminary evidence that Rab5 mediates PcSRB to influence the phagocytosis of P. clarkii hemocytes against A. hydrophila.

Combination of Methotrexate and Resveratrol Reduces Pro-Inflammatory Chemokines in Human THP-1 Cells.

Methotrexate (MTX) is a widely used anti-metabolite drug in cancer therapy, but its efficacy is often hindered by reactive oxygen species (ROS)-induced cellular toxicity. Resveratrol, a natural polyphenol, possesses antioxidant and anticancer properties. Therefore, this in vitro study aimed to investigate the synergistic anti-proliferative and anti-inflammatory effects of MTX and resveratrol in human THP-1 cells. THP-1 cells were differentiated into macrophage-like cells using phorbol 12-myristate 13-acetate. In vitro experiments assessed the impact of various concentrations of MTX and resveratrol on cell viability and proliferation using the MTT assay. Concentration-effect curves were generated to elucidate their relationship. Gene expression was analyzed by RT-qPCR, while chemokine expression was measured via ELISA. Phagocytic and migratory activities were also evaluated. Differentiated THP-1 cells were treated with MTX and resveratrol and stimulated with dimethyl sulfoxide (DMSO) and lipopolysaccharide (LPS) as inflammatory stimuli. The combination of MTX and resveratrol exhibited an anti-proliferative effect in THP-1 cells (p = 0.03). The concentration-effect curve revealed IC50 values of 49.15 µg at 24hours (R = 0.8236) and 2.029e-005 µg at 48hours (R = 0.97) for MTX, and 20.17 µg at 48hours (R = 1.000) and 55.38 µg at 96hours (R = 0.9666) for resveratrol. Co-treatment with MTX and resveratrol significantly reduced mRNA and chemokine expression of CCL2, CCL3, CCL4, CCL5, and CXCL10 (p < 0.05). Moreover, decreased phagocytic and migratory activities were confirmed by phagocytosis and migration assays (p < 0.05). The combination of MTX and resveratrol effectively attenuated pro-inflammatory activity in THP-1 cells, as evidenced by the downregulation of mRNA and chemokine expression. These findings suggest that the synergistic effects of MTX and resveratrol hold promise for enhancing cancer therapeutics.

Cancer-associated fibroblasts shape early myeloid cell response to chemotherapy-induced immunogenic signals in next generation tumor organoid cultures

BackgroundPatient-derived colorectal cancer (CRC) organoids (PDOs) solely consisting of malignant cells led to major advances in the understanding of cancer treatments. Yet, a major limitation is the absence of cells...

Possibility of short synthetic peptides with activities of suppressing amyloid β aggregation and resolving its aggregated form as therapeutic drugs for Alzheimer's disease

Lecanemab is a new anti-amyloid antibody being developed as a treatment for Alzheimer's disease. It is expected to delay the progression of the disease by reducing the accumulation of amyloid beta (Aβ) in the brain. However, no drug has been developed that can completely eliminate Aβ and improve symptoms. A representative Catalytide, JAL-TA9 (YKGSGFRMI), cleaves Aβ42 and improves symptoms in an Alzheimer's disease mouse model, suggesting that JAL-TA9 is a promising candidate for treating Alzheimer's disease by effectively eliminating Aβ. The catalytic center of JAL-TA9 is GSGFR. To identify better Catalytides for Alzheimer's treatment, we analyzed the structure-activity relationship of 21 point-mutated GSGFR derivatives. In this process, we discovered two peptides, GSGFK and GSGNR, that not only inhibit Aβ25-35 aggregation but also dissolve aggregated Aβ25-35. Intracerebroventricular administration of GSGFK protected mice against Aβ25-35-induced short-term memory deficits and promoted microglial phagocytic activity. Like Lecanemab, GSGFK targets Aβ, but it has advantages such as safety, administration method, and cost. In this talk, we will discuss the potential of GSGFK as a therapeutic candidate for Alzheimer's disease.

SYNLAC prime probiotics enhances growth performance, and resistance of white shrimp, Penaeus vannamei to Enterocytozoon hepatopenaei and Vibrio alginollyticus: insights into immune and metabolic pathway modulations

This study explores the impact of SYNLAC Prime probiotics on the growth performance, health status, and metabolic profile of white shrimp, Penaeus vannamei. Shrimp fed with the experimental diets, including the control diet without probiotic supplementation, and the diets supplemented with SYNLAC Prime probiotics at concentrations of 105 CFU (g diet)-1 (P5) and 106 CFU (g diet)-1 (P6) for 56 days. Results indicated a significant enhancement in growth performance in probiotic-treated shrimp relative to the control group, attributed to structural improvements in the digestive tract, particularly the increased abundances of B cells in the hepatopancreas. The administration of dietary probiotics markedly reduced the severity of Enterocytozoon hepatopenaei (EHP) infection and decreased cumulative mortalities following Vibrio alginolyticus challenge. Shrimp in the P6 group exhibited significant elevations in phenoloxidase activity, respiratory burst, lysozyme activity and phagocytic activity compared to control group. Furthermore, there was an upregulation of several immune-related genes in hepatopancreas, including serine protease (SP), prophenoloxidase (proPO) I, proPO II, and penaeidin 3a. Additionally, the expression of β-1, 3-glucan binding protein and SP mRNA was significantly increased in hemocytes. Untargeted metabolomics analysis using LC-MS/MS revealed significant changes in the hepatopancreas metabolic profile, highlighting alterations in energy metabolisms pathways, such as citrate cycle and nicotinate and nicotinamide metabolism, as well as amino acid metabolisms pathways including arginine and proline metabolism, taurine and hypotaurine metabolism, and histidine metabolism. These findings underscore the potential of SYNLAC Prime probiotics in enhancing shrimp growth, immune function, and metabolic pathways, offering valuable insights for advancing health management strategies in shrimp aquaculture.

Phospholipase D Family Member 4 Regulates Microglial Phagocytosis and Remyelination via the AKT Pathway in a Cuprizone-Induced Multiple Sclerosis Mouse Model.

Remyelination is an endogenous repair process that is often deficient in multiple sclerosis (MS). Stimulation of remyelination is thought to help limit the progression of MS. This study aimed to investigate the expression pattern and function of a microglial phagocytosis-related gene, phospholipase D family member 4 (PLD4), in a cuprizone (CPZ)-induced MS mouse model. The extent of remyelination was assessed using LFB staining. Myelin phagocytosis assay was used to investigate the effect of Pld4 on microglial phagocytic activity. Pld4 was upregulated in the corpus callosum during demyelination and remyelination. AAV9-mediated Pld4 deficiency impaired remyelination and reduced the number of Olig2-positive cells. In the corpus callosum of Pld4-deficient mice, the microglial phagocytosis marker MAC2 was reduced, accompanied by inhibition of TrkA/AKT signaling. Similarly, the phagocytosis assay showed that Pld4 knockdown significantly inhibited myelin debris phagocytosis by BV2 cells. The AKT activator SC79 reversed the Pld4 deficiency-induced inhibition of microglial phagocytic activity and rescued the impaired remyelination in Pld4-deficient mice. PLD4 is upregulated in CPZ-induced MS and modulates microglial phagocytosis and remyelination via the AKT pathway. Our findings provide experimental evidence for a better understanding of the molecular mechanism of MS.

Serotonin and adrenaline as inhibitors of neutrophil extracellular traps formation (experimental study)

Regulation of the NETosis can provoke cancer, inflammatory and autoimmune diseases and become a basis for effective targeted therapy. The aim: to study the modifying effects of serotonin and adrenaline on NETosis during phagocytosis in rats.Materials and methods.Wistar rats were divided into control (solvent, 0.9% NaCl solution) and experimental (serotonin and adrenaline 1.43 and 0.143 mg/kg body weight, respectively) groups. Intraperitoneal injection of test substances was carried out 90 minutes before blood sampling, in which phagocytic activity (PhA), phagocytic index (PhI) and degree of suicidal neutrophil NETosis were determined: with total and partial chromatin decondensation (TCD, PCD, accordingly). A paired Student’s t test was used to compare groups with numerical values and McNemar’s test (for dichotomous variables). Pearson’s linear correlation coefficient (rxy) was assessed to measure a linear relationship between time points.Results.The administration of adrenaline contributed to NETosis inhibition by 41.5% compared to control group (p 0.001), serotonin — by 27.6% (p 0.001). The ratio of neutrophils with total and partial chromatin decondensation for control group was 1.0:0.9; for serotonin and adrenaline it changed to 1.0:1.7 and 1.0:1.4, respectively. Relative to control group, phagocytic index after serotonin administration increased by 4.9%, in case of adrenaline, on the contrary, it decreased by 12.4% (p 0.01). A high positive correlation was revealed between NETosis and phagocytic index for control group and experimental group with adrenaline: rxy= 0.823–0.997. For phagocytic index and NETosis in these groups there was a high negative (rxy= –0.714–(–0.871)) and a noticeable positive correlation for serotonin (rxy= 0.638).Conclusion.Serotonin and adrenaline in therapeutic dosages have a pronounced inhibitory effect on NETosis, which may pave the avenue for increasing the effectiveness of therapy for immunoinflammatory and oncological diseases. The use of serotonin is preferable because it has additional immunomodulatory effects and cause no direct stimulation ofα- andβ-adrenergic receptors.

The microglial innate immune receptor TREM2 participates in fear memory formation through excessive prelimbic cortical synaptic pruning.

Fear memory formation has been implicated in fear- and stress-related psychiatric disorders, including post-traumatic stress disorder (PTSD) and phobias. Synapse deficiency and microglial activation are common among patients with PTSD, and induced in animal models of fear conditioning. Increasing studies now focus on explaining the specific mechanisms between microglia and synapse deficiency. Though newly-identified microglia regulator triggering receptor expressed on myeloid cells 2 (TREM2) plays a role in microglial phagocytic activity, its role in fear-formation remains unknown. We successfully constructed a fear- formation model by foot-shock. Four days after foot-shock, microglial capacity of synaptic pruning was investigated via western blotting, immunofluorescence and Golgi-Cox staining. Prelimbic chemical deletion or microglia inhibition was performed to detect the role of microglia in synaptic loss and neuron activity. Finally, Trem2 knockout mice or wild-type mice with Trem2 siRNA injection were exposed to foot-shock to identify the involvement of TREM2 in fear memory formation. The results herein indicate that the foot-shock protocol in male mice resulted in a fear formation model. Mechanistically, fear conditioning enhanced the microglial capacity for engulfing synapse materials, and led to glutamatergic neuron activation in the prelimbic cortex. Prelimbic chemical deletion or microglia inhibition improved fear memory formation. Further investigation demonstrated that TREM2 regulates microglial phagocytosis, enhancing synaptic pruning. Trem2 knockout mice showed remarkable reductions in prelimbic synaptic pruning and reduced neuron activation, with decreased fear memory formation. Our cumulative results suggest that prelimbic TREM2-mediated excessive microglial synaptic pruning is involved in the fear memory formation process, leading to development of abnormal stress-related behavior.

Effect of a pre-calving injectable trace mineral supplement on white blood cell function in seasonally calving pastoral dairy cows

ABSTRACT Aims To investigate the effect of injection of trace mineral supplement (TMS) 14–28 days before calving on white blood cell count (WBCC) and function, serum antioxidant capacity (SAC) and reactive oxygen species (ROS) in pasture-fed cattle after calving. Methods On each of two South Island, seasonally calving, pastoral dairy farms,1 month before dry-off, a random sample of 150 multiparous cows predicted to calve within 7 days of the herd’s planned start of calving (PSC) were stratified on individual somatic cell count, age, breed and expected calving date. On each farm, 14–24 days before PSC, 60 selected cows were randomly assigned for TMS (Zn, Mn, Se, Cu) injection, and 60 were controls. All 240 cows were contemporaneously injected with hydroxocobalamin, and controls with Se. Blood samples were collected pre-injection and 3, 12 and 40 days after calving. Phagocytic activity, count and proportion of neutrophils, lymphocytes and monocytes, WBCC, ROS, SAC were measured. Plasma concentrations of Se, Cu and glutathione peroxidase (GPx) were monitored from a random subset of animals. Differences attributable to TMS were estimated using mixed-multivariable Bayesian analysis, expressed as mean and highest density interval (HDI). Results Three and 40 days after calving, TMS-treated cows had 0.36 (90% HDI = 0.00–0.77) x 109 and 0.25 (90% HDI = 0.00–0.55) x 109 fewer neutrophils/L. Neutrophils comprised 6 (90% HDI = 0–11)% and 4 (90% HDI = 0–8)% less of the WBCC, and the neutrophil count was 14 (90% HDI = 0–27)% and 9 (90% HDI = 0–18)% less than controls. However, 3 days after calving, there were 7 (95% HDI = 2–12)% more cells phagocytosing and 2,900 (95% HDI = 2,600–3,200) more bacteria ingested/cell. Twelve and 40 days after calving, TMS-treated cows had 0.65 (95% HDI = 0.17–1.17) x 109 and 0.28 (95% HDI = 0.00–0.59) x 109 more lymphocytes/L. Lymphocytes comprised 10 (95% HDI = 3–18)% and 5 (95% HDI = 0–9)% more of the WBCC, and the lymphocyte count was 30 (95% HDI = 11–51)% and 9 (95% HDI = 0–9)% more than controls. There were no meaningful differences in ROS, SAC, ROS/SAC, other white blood cells, or WBCC. Plasma Cu, Se and GPx concentrations were above recommended thresholds. Conclusions Pre-calving TMS injection was associated with differences in white blood cell population and function that may reduce the risk of disease. Abbreviations BHOB: Beta-hydroxybutyrate; GPx: Glutathione peroxidase; HDI: Highest density interval; MESF: Molecules of equivalent soluble fluorophore; OSi: Oxidative stress index; PSC: Planned start of calving; ROPE: Region of probable equivalence; ROS: Reactive oxygen species; SAC: Serum antioxidant capacity; THI: Temperature humidity index; TMS: Trace mineral supplement; WAIC: Widely applicable information criterion; WBCC: White blood cell count.

Enriched Diet With Orange Essential Oil Citrus sinensis for Tambaqui Colossoma macropomum Promotes Growth Performance and Resistance Against Aeromonas hydrophila.

The objective of this study was to evaluate the effect of dietary supplementation with Citrus sinensis (L.) Osbeck essential oil on the growth, immune system, and resistance to Aeromonas hydrophila infection in Colossoma macropomum fingerlings. The experiment was conducted with five treatments (control diet, Tween80 diet, and diets supplemented with 200, 400, and 800 mg L-1 of C. sinensis essential oil) with four replicates. At the end of the experimental period, growth parameters were measured, and blood samples were collected for thrombogram, leukogram, and phagocytic activity analysis. A bacterial challenge with A. hydrophila was conducted for 96 h. C. macropomum fingerlings that were fed with 400 and 800 mg L-1 of C. sinensis essential oil had the highest growth parameters, with final weights of 533.18 ± 2.03 mg and 531.91 ± 2.67 mg, respectively, and an increase in the number of thrombocytes, lymphocytes, monocytes, and neutrophils, as well as higher phagocytosis rates compared to the control group. Regarding the challenge, fish in the 400 and 800 mg L-1 treatments also exhibited the lowest cumulative mortality rate (26.66% ± 3.33%). Therefore, supplementation with C. sinensis essential oil promotes growth, improves health, and enhances resistance to A. hydrophila in C. macropomum fingerlings.

Psilocin, the Psychoactive Metabolite of Psilocybin, Modulates Select Neuroimmune Functions of Microglial Cells in a 5-HT2 Receptor-Dependent Manner.

Neuroinflammation that is caused by microglia, the main immune cells of the brain, contributes to neurodegenerative diseases. Psychedelics, including psilocybin and lysergic acid diethylamide (LSD), possess certain anti-inflammatory properties and, therefore, should be considered as drug candidates for treating neuroinflammatory pathologies. When ingested, psilocybin is rapidly dephosphorylated to yield psilocin, which crosses the blood-brain barrier and exerts psychotropic activity by interacting with the 5-hydroxytryptamine 2A receptors (5-HT2ARs) on neurons. Since microglia express all three 5-HT2R isoforms, we hypothesized that, by interacting with these receptors, psilocin beneficially modulates select neuroimmune functions of microglia. We used microglia-like cell lines to demonstrate that psilocin, at non-toxic concentrations, did not affect the secretion of tumor necrosis factor (TNF) by immune-stimulated microglial cells, but significantly inhibited their phagocytic activity, the release of reactive oxygen species (ROS), and nitric oxide (NO) production. The inhibitory activity of psilocin on the latter two functions was similar to that of two selective 5-HT2R agonists, namely, 25I-NBOH and Ro60-0175. The role of this subfamily of receptors was further demonstrated by the application of 5-HT2R antagonists cyproheptadine and risperidone. Psilocin should be considered a novel drug candidate that might be effective in treating neuroimmune disorders, such as neurodegenerative diseases, where reactive microglia are significant contributors.

Phagocytic activity and proinflammatory activation potential of smooth muscle cells of the Tunica intima of human aorta under experimental conditions

The aorta is the largest blood vessel in the body, which reaches a diameter of about 3 cm and is responsible for transporting oxygen-enriched arterial blood from the heart to tissues and organs. The aortic wall consists of three layers: the inner Tunica intima, the middle Tunica media and the outer Tunica adventitia. The layers of the aortic wall have a diverse cellular composition and include smooth muscle cells (SMCs), fibroblasts, endothelial cells, etc. Functional disorders in the cells of the intima of the human aorta can lead to various cardiovascular diseases (СVD), such as aneurysm and, as a result, dissection or rupture of the thoracic aorta. The cellular and molecular mechanisms of CVD development remain not fully understood, therefore, the study of the functional characteristics of various cell populations that make up the human aorta is an urgent task today. The aim is to evaluate the inflammatory response formed by cells that are part of the Tunica intima of the human aorta during phagocytosis of latex particles and internalization of low-density lipoproteins (LDL) to study their role in the development of aneurysms. The experiments were performed on smooth muscle cells (SMCs) isolated from the intima of the human aorta in patients with aneurysm. Phagocytic activity was studied by adding latex beads to the SMCs of Tunica intima, the ability to internalize LDL was evaluated using the BDP 630/650 dye and a biochemical method, the assessment of the ability to pro- and anti-inflammatory activation was studied using enzyme-linked immunosorbent assay (ELISA). Our results demonstrated that the absorption of latex beads and LDL stimulates the activation of interleukin secretion by smooth muscle cells that are part of the Tunica intima of the aorta, namely the proinflammatory cytokines IL-6 and IL-8. This fact may indicate that in the conditions of the body, the human aortic intima SMCs phenotype may switch from contractile to secretory or macrophage-like, which indicates the participation of this phenotypic cell transition in the process of aneurysm development.

Phagocytic activity of neutrophil blood granulocytes in children with chronic non-specific lung diseases with post-inflammatory pneumofibrosis

One of the reasons for the development of chronic forms of lung diseases and pneumofibrosis as a pathological outcome of this inflammation may be an insufficient immune response. The key position belongs to neutrophilic granulocytes. Given the importance and renewed interest in neutrophils as tools for regulating immunity, leading to the progression of the fibrous process in children with CKD is an urgent problem. The purpose of the study was to study the phagocytic activity of blood neutrophils as an important component of innate immunity in patients with CNLD. Seventy children with CNLD with focal or segmental post-inflammatory pulmonary fibrosis were examined. Of these, 27 were children with progression of PPF (main group) and 43 children with PPF without progression (comparison group). The controls were the indicators of 23 healthy children, comparable by gender and age. The studies were carried out during clinical remission. The phagocytic activity of neutrophils (PНAN), phagocytic number (PN), spontaneous and stimulated NBT test were determined. The stimulation index of the NBT test (NBTst-NBTsp) was calculated. The mitochondrial membrane potential was determined using a BD FACS Calibur cytometer (USA) (BD Pharmigen, USA). PN in children with chronic heart disease with progressive PF was significantly reduced in comparison with patients in the comparison group and with the control group. The PHAN in children with CKD in both groups was within the standard values, however, it was lower than in the control group. The activity of the spontaneous HBT test in children of the main group was 1.7 times higher than in children in the comparison group and 3 times higher than in children from the control group. However, the stimulation index in the main group was significantly lower than in the control group. An increase in cells with a reduced membrane potential of granulocyte mitochondria was revealed in children of the main group compared with the control group. Children with CKD with the progression of fibrosis processes develop changes in the functional and metabolic activity of peripheral blood neutrophils typical for an inflammatory reaction, characterized by an increase in oxygen-dependent and a decrease in phagocytic cell activity.