Cell Phagocytic Activity Research Articles

A Novel Multifunctional 5,6-Dimethoxy-Indanone-Chalcone-Carbamate Hybrids Alleviates Cognitive Decline in Alzheimer's Disease by Dual Inhibition of Acetylcholinesterase and Inflammation.

BackgroundsAlzheimer’s disease (AD) is a multifactorial neurodegenerative disease. The treatment of AD through multiple pathological targets may generate therapeutic efficacy better. The multifunctional molecules that simultaneously hit several pathological targets have been of great interest in the intervention of AD.MethodsHere, we combined the chalcone scaffold with carbamate moiety and 5,6-dimethoxy-indanone moiety to generate a novel multi-target-directed ligand (MTDL) molecule (E)-3-((5,6-dimethoxy-1-oxo-1,3-dihydro-2H-inden-2-ylidene)-methyl)phenylethyl(methyl) carbamate (named AP5). In silico approaches were used to virtually predict the binding interaction of AP5 with AChE, the drug-likeness, and BBB penetrance, and later validated by evaluation of pharmacokinetics (PK) in vivo by LC-MS/MS. Moreover, studies were conducted to examine the potential of AP5 for inhibiting AChE and AChE-induced amyloid-β (Aβ) aggregation, attenuating neuroinflammation, and providing neuroprotection in the APP/PS1 model of AD.ResultsWe found that AP5 can simultaneously bind to the peripheral and catalytic sites of AChE by molecular docking. AP5 exhibited desirable pharmacokinetic (PK) characteristics including oral bioavailability (67.2%), >10% brain penetrance, and favorable drug-likeness. AP5 inhibited AChE activity and AChE-induced Aβ aggregation in vivo and in vitro. Further, AP5 lowered Aβ plaque deposition and insoluble Aβ levels in APP/PS1 mice. Moreover, AP5 exerted anti-inflammatory responses by switching microglia to a disease-associated microglia (DAM) phenotype and preventing A1 astrocytes formation. The phagocytic activity of microglial cells to Aβ was recovered upon AP5 treatment. Importantly, chronic AP5 treatment significantly prevented neuronal and synaptic damage and memory deficits in AD mice.ConclusionTogether, our work demonstrated that AP5 inhibited the AChE activity, decreased Aβ plaque deposition by interfering Aβ aggregation and promoting microglial Aβ phagocytosis, and suppressed inflammation, thereby rescuing neuronal and synaptic damage and relieving cognitive decline. Thus, AP5 can be a new promising candidate for the treatment of AD.

Adipose-derived stem cells decolonize skin Staphylococcus aureus by enhancing phagocytic activity of peripheral blood mononuclear cells in the atopic rats.

Staphylococcus aureus (S. aureus) is known to induce apoptosis of host immune cells and impair phagocytic clearance, thereby being pivotal in the pathogenesis of atopic dermatitis (AD). Adipose-derived stem cells (ASCs) exert therapeutic effects against inflammatory and immune diseases. In the present study, we investigated whether systemic administration of ASCs restores the phagocytic activity of peripheral blood mononuclear cells (PBMCs) and decolonizes cutaneous S. aureus under AD conditions. AD was induced by injecting capsaicin into neonatal rat pups. ASCs were extracted from the subcutaneous adipose tissues of naïve rats and administered to AD rats once a week for a month. Systemic administration of ASCs ameliorated AD-like symptoms, such as dermatitis scores, serum IgE, IFN-γ+/IL-4+ cell ratio, and skin colonization by S. aureus in AD rats. Increased FasL mRNA and annexin V+/7-AAD+ cells in the PBMCs obtained from AD rats were drastically reversed when co-cultured with ASCs. In contrast, both PBMCs and CD163+ cells bearing fluorescent zymosan particles significantly increased in AD rats treated with ASCs. Additionally, the administration of ASCs led to an increase in the mRNA levels of antimicrobial peptides, such as cathelicidin and β-defensin, in the skin of AD rats. Our results demonstrate that systemic administration of ASCs led to decolonization of S. aureus by attenuating apoptosis of immune cells in addition to restoring phagocytic activity. This contributes to the improvement of skin conditions in AD rats. Therefore, administration of ASCs may be helpful in the treatment of patients with intractable AD.

Bioelectronic Sensor with Magnetic Modulation to Quantify Phagocytic Activity of Blood Cells Employing Machine Learning.

Microbial infections result in activating an immune response in the human body, which triggers inflammatory pathways resulting in recognition and subsequent killing of the pathogens. Quantifying the blood cells' natural ability to kill pathogens, i.e., phagocytosis, is critical to demonstrating the effectiveness of an individual's response in combating pathogens. Current laboratory processes and equipment that can be used to monitor phagocytic activity are costly and time-consuming and require significant technical expertise to run such assays. Here, we design and develop a novel biosensing platform capable of quantifying the phagocytic ability of blood cells. The sensor design is composed of electronic sensing and magnetic modulation sub-systems that work in conjunction to monitor phagocytic activity in microfluidic channels. The phagocytes internalize the IgG-coated magnetic beads, and when infused into the sensor, their speed will be modulated using the quadrupole magnetic field configuration as they pass through microfluidic channels where microfabricated electrodes are placed. The electronic sensor will generate the voltage pulse for each passage of the phagocyte, whose distinct features are correlative to the phagocytic activity. We experimentally tested this device using 17 blood samples collected from patients at Robert Wood Johnson Medical Hospital. Further, we developed artificial neural networks (ANN) to improve the accuracy of the phagocytic activity detection. ANN model detected the phagocytic activity with 88.2% accuracy. This novel sensing platform can potentially be used to triage high risk patients and develop personalized theranostics for the septic patients in the future.

Phytochemical Analysis and Immunomodulatory Potential on Diabetic-Infected Tuberculosis by Fruit Etlingera rubroloba A.D. Poulsen.

&lt;b&gt;Background and Objective:&lt;/b&gt; &lt;i&gt; Etlingera rubroloba&lt;/i&gt; (&lt;i&gt;E. rubroloba&lt;/i&gt;) A.D. Poulsen is an endemic plant in South-East Sulawesi and is a newly discovered species. This plant is expected to have the potential as an immunomodulator in patients with diabetes mellitus (DM), which can prevent tuberculosis infection by increasing the phagocytic function of macrophage cells and interleukin-12 (IL-12) levels. &lt;b&gt;Materials and Methods:&lt;/b&gt; Phytochemical analysis of the ethanolic extract of the fruit of &lt;i&gt;E. rubroloba&lt;/i&gt; A.D. Poulsen using Liquid Chromatography-Mass Spectrometry (LC-MS/MS) was carried out. The immunomodulatory potential &lt;i&gt;in vivo&lt;/i&gt; on BALB/c mice model DM was carried out by oral induction of TB antigen with extract dose, control positive, negative and normal groups. Furthermore, the phagocytic activity of macrophage cells can be seen with a microscope and the levels of IL-12 with the Elisa kit. &lt;b&gt;Results:&lt;/b&gt; The results showed the ethanol extract of the fruit of &lt;i&gt;E. rubroloba&lt;/i&gt; contained eight chemical compounds and had potential as immunomodulators in BALB/c DM mice induced by TB antigen by increasing the phagocytic activity of macrophage cells and levels of IL-12, which were significantly different from the negative control (p<0.05). &lt;b&gt;Conclusion:&lt;/b&gt; The chemical composition of the ethanol extract of the fruit of &lt;i&gt;E. rubroloba&lt;/i&gt; has the potential as an immunomodulator in TB antigen-induced DM &lt;i&gt;in vivo&lt;/i&gt;.

Abstract 246: Application of the 3d fresh patient tumoroid platform 3d-explore to assess the immunomodulatory and phagocytic activity of the receptor tyrosine kinase inhibitor, Sunitinib

Abstract Background: The receptor tyrosine kinase inhibitor (TKI) Sunitinib has proven useful in the treatment of a variety of tumors. Sunitinib has shown therapeutic activity in patients with metastatic renal cell carcinoma and currently represents a frontline therapy for this disease. The immune modulatory effect of Sunitinib on immune cells within the tumor microenvironment has been well documented. Here, we developed a novel 3D ex-vivo platform (3D-EXplore) using fresh patient tumor samples with intact stromal components and tumor immune microenvironment to assess the therapeutic efficacy of Sunitinib. Methods: All tumor samples were obtained with patient consent and relevant IRB approval. Unpropagated 3D tumoroids with intact TME measuring 150 µm in size were prepared from fresh tumor samples of renal cell carcinoma using a proprietary technology developed at Nilogen Oncosystems. Tumoroids prepared from each patient’s tumor sample were pooled to represent the tumor heterogeneity and treated ex vivo with Sunitinib for 48h to detect treatment-mediated changes in tumor immune cell composition including CD4 and CD8 T-cells, NK cells, and macrophages. Additionally, we analyzed treatment-mediated changes in T-cell activation and phagocytic activity of myeloid cells. Results: Multiparameter flow analysis revealed that Sunitinib treatment led to an increase in CD8+CD25+ central memory-like phenotype and increased granzyme B expression in CD4 and CD8+ T cells indicating T cell activation. Furthermore, multiparameter flow cytometry analysis showed increased EdU uptake by CD8 and NK cells with sunitinib treatment, while cytokine release assays demonstrated increased IFN-γ and IL-2 production accompanied by downregulation of IL-10 and IL-6 cytokines. We observed Sunitinib mediated phagocytotic activity by tumor resident myeloid macrophages, monocytes, and dendritic cells using a quantitative phagocytosis assay. Conclusions: These results demonstrate that 3D-EXplore using fresh tumor samples provides a clinically relevant platform to assess drug response and serves as a critical tool for reflecting the true intact tumor microenvironment for novel immune-oncology drug development. Citation Format: Brittany Bunch, Krithika N. Kodumudi, Jared Ehrhart, Matt Weitzman, Olivia MacIntosh, Kelly Sussman, Soner Altiok. Application of the 3d fresh patient tumoroid platform 3d-explore to assess the immunomodulatory and phagocytic activity of the receptor tyrosine kinase inhibitor, Sunitinib [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 246.

Abstract 2115: Glutamine blockade via prodrug JHU083 reprograms immunosuppressive tumor associated macrophages (TAMs) and drives tumor immunity in urologic cancers

Abstract Introduction: Metastatic prostate cancer is currently incurable and kills more than 30,000 men in the United States each year, making it the second leading cancer related cause of death and an area of unmet need. Previously, we reported increased infiltration of immunosuppressive CD206-positive tumor associated macrophages (TAMs) with disease progression in prostate adenocarcinoma.[1] Glutamine metabolism has been implicated in immunosuppressive TAMs as well as metastatic castration resistant prostate cancer (mCRPC)[2] and radioresistant urothelial carcinoma.[3] As such, we hypothesize that rational metabolic interventions could be a promising strategy to simultaneously target both TAMs and cancer cells, resulting in anti-tumor immunity. Methods: We utilized a novel pro-drug (JHU083) derived from 6-Diazo-5-oxo-L-norleucine (DON) which has significantly lowered toxicity to treat two urological syngeneic immunogenic mouse tumor models in vivo; B6CaP (prostate cancer) and MB49 (bladder cancer). We studied the direct effect on tumor cells, as well as the required immune compartment for drug efficacy in vivo using antibody targeted depletions or adoptively transferred TAMs. Moreover, we used RNA-sequencing and multi-parameter flow cytometry to characterize the effect of JHU083 on TAMs transcriptional programming and proteomic expression profiles in vivo. Lastly, we assessed the phagocytotic capacity of macrophages after treatment with JHU083 with flow cytometry and immunofluorescence (IF) microscopy. Results: JHU083 treatment showed significant tumor regression in both urologic cancer models. Using in vivo depletion of CD4 or CD8 T cells, or adoptively transferring previously in-vivo JHU083 treated TAMs we established a direct anti-tumor role of TAMs. These TAMs were more inflammatory as shown by the increased TNF-positivity. Strikingly, in the TME we also observed an increase of Glut1 and Hexokinase II indicating metabolic reprogramming of these TAMs towards glycolytic phenotype. Importantly, JHU083-treated TAMs showed significantly increased phagocytic activity of cancer cells, providing direct evidence of functional reprogramming. Conclusion: We found that JHU083 has two distinct functions in vivo; first, it directly impairs cancer cells which are glutamine dependent and second, it reprograms TAMs from an immunosuppressive to an inflammatory state via antagonism of glutamine metabolism in vivo. These macrophages convert to a highly glycolytic state, have increased TNF production, and have improved phagocytic activity against tumor cells. As urologic cancers are heavily infiltrated with immunosuppressive TAMs, JHU083 is an excellent preclinical candidate for these diseases. It remains to be seen if JHU083 treatment can be effectively combined with checkpoint therapy to elicit durable anti-tumor immunity. Citation Format: Monali Praharaj, Fan Shen, Liang Zhao, Thomas R. Nirschl, Laura Sena, Alok K. Singh, Debebe Theodros, Xiaoxu Wang, Raekwon A. Williams, Elizabeth Thompson, Ada Tam, Srinivasan Yegnasubramanian, Robert D. Leone, Jesse Al, Rana Rais, Barbara S. Slusher, Drew M. Pardoll, Jonathan D. Powell, Jelani C. Zarif. Glutamine blockade via prodrug JHU083 reprograms immunosuppressive tumor associated macrophages (TAMs) and drives tumor immunity in urologic cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2115.

DIVERSITY OF BACTERIA AND FUNGI AND PHAGOCYTIC PROPERTIES OF NEUTROPHILS IN SOME RESPIRATORY DISEASES

Respiratory diseases are considered serious diseases that affect all age groups and can lead, in most cases, to death. This study aimed to investigate the bacterial and fungal causes of some respiratory infections and the relationship between the phagocytic activity of polymorphonuclear cells and respiratory tract infection.146 clinical samples of sputum were collected from 50 patients with chronic tuberculosis, and pharyngeal swabs from 96 patients with respiratory infections other than tuberculosis, including chronic bronchitis (22 patients), allergic (66 patients), and asthma (5patients) pneumonia (3 patients). These samples were collected from patients referred to the Specialized National Centre for Chest and Respiratory Diseases in Baghdad for the period between 2018and 2019. It has been found that out of the total 146 patients, the number of infected males is 76 (52.1 %) and 70 females (47.9%). 174 bacterial isolates were isolated and identified, as well as it was noted that the percentage of respiratory infections caused by gram-positive bacteria was higher (66.1%) compared to the respiratory infections caused by gram-negative bacteria (33.9%), As for fungal infections, from 140 isolates showed that the percentage of respiratory infections caused by fungi was the highest (70.9%) compared to respiratory infections caused by yeasts (29%). The current study indicated that respiratory tract infections stimulated the phagocytic activity of polymorphonuclear cells for all age groups. Keywords:

The pharmaceutical applications of total flavonoids extract from Isatis tinctoria L. leaves

Isatis tinctoria L. leaves (Folium isatidis), as dried leave of the isatis indigotica in the cruciferous family, has a variety of pharmacological effects, including regulation of immune function, anti-inflammatory, bacteriostasis, antipyretic effect. In this study, total flavonoids extract from Folium isatidis was systematically studied about extractive process, main components, antibacterial, antioxidant and anti-inflammatory activities. The total flavonoids extract yield was (7.65±0.07)% under the optimal extraction parameters of ethanol concentration 54.37%, ratio of solvent volume to material weight 40:1 ml/g, and ultrasonic extraction time 20 min. Nineteen chemical components were inferred and identified by UPLC-QTOF-MS method. Although the extract showed mild antibacterial and antioxidant effects with MIC values 31.25, 62.5, 125 and 125 mg/ml against Streptococcus, Salmonella, Staphylococcus aureus and Escherichia coli, and IC50 DPPH 12.64 µg/ml, IC50 ABTS 16.72 µg/ml and FRAP value at 0.11 mmol Fe2+/100 µg/ml compared with the positive control butylated hydroxytoluene with IC50 DPPH 8.13 µg/ml, IC50 ABTS 2.22 µg/ml and FRAP value at 0.16 mmol Fe2+/100 µg/ml. It exhibited better anti-inflammatory effects in LPS induced RAW 264.7 cell (50, 100 and 200 µg/ml of TFE) and ALI mice (100, 250, or 500 mg/kg•b•w of TFE), which was demonstrated by reducing cell phagocytic activity, decreasing the content of pro-inflammatory cytokine and inflammatory mediator, and improving lung tissue injury. Furthermore, the mechanism of anti-inflammatory action was further revealed to be related to the regulation of TLR4-TRIF-NF-κB signaling pathway. These studies revealed that total flavonoids extract could be used as a natural and green candidate for anti-inflammatory drugs and enrich the variety of clinical anti-inflammatory drugs.

Lifelong Exposure to a Low-Dose of the Glyphosate-Based Herbicide RoundUp® Causes Intestinal Damage, Gut Dysbiosis, and Behavioral Changes in Mice.

RoundUp® (RUp) is a comercial formulation containing glyphosate (N-(phosphono-methyl) glycine), and is the world’s leading wide-spectrum herbicide used in agriculture. Supporters of the broad use of glyphosate-based herbicides (GBH) claim they are innocuous to humans, since the active compound acts on the inhibition of enzymes which are absent in human cells. However, the neurotoxic effects of GBH have already been shown in many animal models. Further, these formulations were shown to disrupt the microbiome of different species. Here, we investigated the effects of a lifelong exposure to low doses of the GBH-RUp on the gut environment, including morphological and microbiome changes. We also aimed to determine whether exposure to GBH-RUp could harm the developing brain and lead to behavioral changes in adult mice. To this end, animals were exposed to GBH-RUp in drinking water from pregnancy to adulthood. GBH-RUp-exposed mice had no changes in cognitive function, but developed impaired social behavior and increased repetitive behavior. GBH-Rup-exposed mice also showed an activation of phagocytic cells (Iba-1–positive) in the cortical brain tissue. GBH-RUp exposure caused increased mucus production and the infiltration of plama cells (CD138-positive), with a reduction in phagocytic cells. Long-term exposure to GBH-RUp also induced changes in intestinal integrity, as demonstrated by the altered expression of tight junction effector proteins (ZO-1 and ZO-2) and a change in the distribution of syndecan-1 proteoglycan. The herbicide also led to changes in the gut microbiome composition, which is also crucial for the establishment of the intestinal barrier. Altogether, our findings suggest that long-term GBH-RUp exposure leads to morphological and functional changes in the gut, which correlate with behavioral changes that are similar to those observed in patients with neurodevelopmental disorders.

Essentials in saline pharmacology for nasal or respiratory hygiene in times of COVID-19

Purpose: Nasal irrigation or nebulizing aerosol of isotonic or hypertonic saline is a traditional method for respiratory or nasal care. A recent small study in outpatients with COVID-19 without acute respiratory distress syndrome suggests substantial symptom resolution. We therefore analyzed pharmacological/pharmacodynamic effects of isotonic or hypertonic saline, relevant to SARS- CoV-2 infection and respiratory care. Methods: Mixed search method. Results: Due to its wetting properties, saline achieves an improved spreading of alveolar lining fluid and has been shown to reduce bio-aerosols and viral load. Saline provides moisture to respiratory epithelia and gels mucus, promotes ciliary beating, and improves mucociliary clearance. Coronaviruses and SARS-CoV-2 damage ciliated epithelium in the nose and airways. Saline inhibits SARS- CoV-2 replication in Vero cells; possible interactions involve the viral ACE2-entry mechanism (chloride-dependent ACE2 configuration), furin and 3CLpro (inhibition by NaCl), and the sodium channel ENaC. Saline shifts myeloperoxidase activity in epithelial or phagocytic cells to produce hypochlorous acid. Clinically, nasal or respiratory airway care with saline reduces symptoms of seasonal coronaviruses and other common cold viruses. Its use as aerosol reduces hospitalization rates for bronchiolitis in children. Preliminary data suggest symptom reduction in symptomatic COVID-19 patients if saline is initiated within 48 h of symptom onset. Conclusions: Saline interacts at various levels relevant to nasal or respiratory hygiene (nasal irrigation, gargling or aerosol). If used from the onset of common cold symptoms, it may represent a useful add-on to first-line interventions for COVID-19. Formal evaluation in mild COVID-19 is desirable as to establish efficacy and optimal treatment regimens.

Effect of glyprolins on white blood cell parameters and phagocytic activity of neutrophils in conditions of experimental hyperthyroidism

This study investigated the effects of glyproline neuropeptide compounds (selank and Pro-Gly-Pro) on the white blood cell count and phagocytic activity of neutrophils in 40 nonlinear white male rats aged 68 months with experimental hyperthyroidism. Experimental hyperthyroidism was simulated by intragastric administration of L-thyroxine sodium salt pentahydrate at a dose of 150 g/kg/day for 21 days. All animals were equally divided into four groups: healthy rats (control), rats treated with L-thyroxine sodium pentahydrate (hyperthyroidism), rats treated with selank, and rats treated with Pro-Gly-Pro at doses of 200 g/kg/day intraperitoneally for 21 days, starting one day after the last administration of L-thyroxine sodium pentahydrate. After the animals were removed from the experiment, the white blood cell count and the percentage of lymphocytes, stick, and segmentonuclear neutrophils were calculated, and the phagocytic activity of neutrophils was evaluated. In the case of experimental hyperthyroidism, neutrophilic leukocytosis with a shift to the right, lymphopenia, and decreased phagocytic activity of neutrophils were observed. Glyproline neuropeptides contributed to the correction of observed changes in white blood cell indices and phagocytic processes, which indicates the immunocorrigating effect of the test compounds. Thus, glyproline neuropeptides demonstrated pronounced immunotropic activity, which manifested in the correction of changes arising from the leukocyte count and phagocytosis processes. However, further detailed study of the pharmacological effects of neuropeptide agents on experimental hyperthyroidism is necessary.

PSVII-2 Effects of Oxidized Beta-Carotene on Sow Reproduction and Piglet Growth Performance

Abstract As reproductive performance of sows in the commercial U.S. sow herd continues to improve, advancing the impact of gestation and lactation diet supplementation is critical. Beginning on d 60 of gestation through lactation, sows (blocked by parity) were assigned to a standard gestation diet (n = 88), or the standard diet supplemented with 80 ppm OxC-beta (n = 90) provided by Avivagen (Ottawa, ON, Canada) to determine if the supplementation of OxC-beta improves sow reproductive performance, litter growth performance, and alters the abundance of immune cells in sows and piglets. Blood samples were taken from a subset of sows (n = 54 per treatment) at the start of the study, at farrowing, and at weaning; and also, on a subset of piglets at birth and weaning. Also, at birth and weaning, 12 pigs (n = 6 per treatment) were euthanized, and livers were collected and subjected to flow cytometry to analyze phagocytic activity of the Kupfer cells. CD335, CD8, and CD4 populations within whole blood were assessed via flow cytometry. Colostrum at farrowing and milk at weaning were collected for analysis of IgG and IgA. Data were analyzed via SAS 9.4 using PROC GLIMMIX. No differences (P &amp;gt; 0.05) between dietary treatments were observed in sow reproductive performance, sow feed intake, wean to estrus interval, or piglet growth performance. No differences (P &amp;gt; 0.05) were observed for vitamin A in the plasma or liver. No immune differences (P &amp;gt; 0.05) were observed in the piglet’s liver and blood nor sow antibodies in colostrum and milk. Subsequent litter information was obtained from the sows, and no differences (P &amp;gt; 0.05) were observed in reproductive performance. Overall, there were no differences in sow reproductive performance, litter growth performance, and the immune system of sows and piglets of sows supplemented OxC-beta.

HCAR1-Mediated L-Lactate Signaling Suppresses Microglial Phagocytosis.

Microglia, the primary brain-resident immune cells, protect the brain from various harmful pathogens, insulting and maintaining its homeostasis by phagocytosing extracellular particles. How microglia are metabolically regulated by their microenvironment remains largely elusive. Here, we investigated how extracellular lactate, which is abundant in the brain and dynamically changes in pathological states, affects microglial phagocytotic ability. We show that L-lactate reduces microglia phagocytic capacity in a Hydroxycarboxylic Acid Receptor 1 but not Monocarboxylate transporter 1-dependent manner. Our findings point to a potential role for extracellular lactate in suppressing the phagocytic activity of microglial cells in homeostasis and inflammatory conditions.

Lyophilized Human Cord Blood Leukoconcentrate to Treat Brain Ischemia in Rats

The paper outlines the experimental data on the impact of lyophilization on preservation of structural and functional characteristics of cells from human cord blood leukoconcentrate (HCBL). Lyophilization of HCBL was shown to ensure the preservation of cells with immune modulating potential in a heterogeneous population of HCBL. Lyophilized HCBL (lHCBL), to the same extent as cryopreserved one (cHCBL), revealed an equal immune correcting effect during ischemic brain injury in vivo in the experimental model of ischemic stroke (IS). The inclusion of lHCBL in therapy of brain ischemia demonstrated the improved indices of IFN-ү+ and IL-10+ spleen cells, adhesive and phagocytic activity of peritoneal cavity cells in rats with IS. The efficiency of using both lHCBL and cHCBL during IS may be associated with the implementation of therapeutic effect by presented among them cells and mediators, including multi-vector regulatory systems that maintain homeostatic stability of the body (immune, endocrine, nervous ones).

PHAGOCYTIC ACTIVITY OF NEUTROPHILIC GRANULOCYTES OF ORAL FLUID IN CHILDREN WITH ACUTE FORMS OF LEUKEMIA

Relevance. In acute leukemia, the phagocytic activity of leukocytes changes. But the extend and details of these changes are still insufficiently studied.&#x0D; Objective: to assess the state of phagocytic activity of phagocytic cells (neutrophils and macrophages) in terms of phagocytic index, phagocytic number and oxygen-dependent phagocytosis in children with acute forms of leukemia.&#x0D; Materials and methods. We examined 35 children aged 6 to 16 years, who were at different stages of treatment of the underlying disease. The children were divided into two groups depending on the form of leukemia: Group I – nineteen children with acute lymphoblastic leukemia (ALL), nine of them at the stage of the first acute period, four at the stage of clinical remission, six at the stage of relapse; Group II - sixteen children with acute myeloid leukemia (AML), seven of them - at the stage of the first acute period, four -in the period of the clinical remission, five - at the stage of relapse. Control Group – fifteen practically healthy children. In oral fluid, the phagocytic activity of granulocytes was determined: phagocytic number (PN) – the percentage of phagocytic neutrophils; phagocytic index (PI) – the number (in conditional units) of phagocytosed microorganisms by one neutrophil; oxygen-dependent phagocytosis according to the NBT test (Nitroblue Tetrazolium) – the percentage of the maximum activity of neutrophils.&#x0D; Results. PN in the control group - 70.0±5.1%. In children with acute lymphoblastic leukemia at the stage of the first acute period of PN - 85.7±5.8%, at the stage of remission - 73.8 ±6.1%, during the relapse period - 88.3% ±4.8. In children with acute myeloblastic leukemia at the stage of treatment of the first acute period, PN was 87.2±6.0%, at the stage of remission - 72.6±5.9%, at the stage of relapse - 89.1±5.1%.&#x0D; PI in the control group – 3.9±0.6 In children with acute myeloid leukemia at the stage of treatment of the first acute period, PI was 21.1±0.7, at the stage of relapse – 25.3±0.4 In children with acute myeloblastic leukemia at the stage of treatment of the first acute period, PI was 16.1±0.5, during remission – 10.1±0.4, during the relapse period – 18.5±0.2.&#x0D; The indicator of oxygen-dependent cell activity, according to the NBT test, in practically healthy children was 17.6±1.23%. In children with acute myeloblastic leukemia at the stage of treatment of the first acute period, the percentage of cell activity was 40.4±1.86%, at the stage of relapse - 46.4±2.0%, during remission - 23.7±1.56%. In children with acute myeloblastic leukemia at the stage of treatment of the first acute period, the percentage of cell activity was 41.6±1.34%, during the period of relapse - 48.5±1.05%, at the stage of remission - 27.9±1.86%.&#x0D; Conclusion. The dependence of phagocytic activity of neutrophils in the oral fluid on the stage of treatment of acute leukemia in children was revealed. The phagocytic number, phagocytic index and oxygen-dependent phagocytosis are significantly increased at the stage of treatment of the first acute period and at the stage of relapse. At the stage of remission, the phagocytic number was within the normal range, but the phagocytic index and oxygen-dependent phagocytosis did not return to normal.&#x0D; No difference was found between the phagocytic activity of neutrophils in various forms of acute leukemia – acute lymphoblastic leukemia and acute myeloblastic leukemia.

Effects of Social Hierarchy Establishment on Stress Response and Cell Phagocytosis in Gilt-Head Sea Bream (Sparus aurata)

Social stress can affect the ability of fish to respond to various stressors, such as pathogens or environmental variations. In this paper, the effects of social stress on gilt-head bream (Sparus aurata) were investigated. To study the effects of physiological stress, we evaluated biochemical and cellular parameters, such as cortisol, glucose, lactate, osmolarity, and phagocytosis, 24 h after the establishment of social hierarchy in a group of three fish. Social hierarchy was determined and characterized by behavioral observation (aggressive acts and feeding order) of the specimens (dominant: “α”; subordinate: “β” and “γ”). After the establishment of social hierarchy, we observed that, overall, levels of plasma cortisol and other biochemical plasmatic stress markers (glucose and lactate) were higher in subordinate individuals than in dominant individuals. In addition, the modulation of phagocytic activity of the peritoneal exudate cells (PECs) demonstrated that social stress appeared to affect immune response. Finally, principal component analysis clearly separated the subordinate fish groups from the dominant groups, based on stress markers and the phagocytic activity of peritoneal exudate cells. This study contributes to current knowledge on gilt-head sea bream, helping to understand the link between social stress, behavior, and physiology of this species, relevant in the aquaculture sector, where fish are subjected to several kinds of stress.

Extracellular Cardiolipin Modulates Select Immune Functions of Astrocytes in Toll-Like Receptor (TLR) 4-Dependent Manner.

Alzheimer's disease (AD) is characterized by chronic neuroinflammation, which is partially mediated by dysregulated functions of glial cells. Cardiolipin (CL) is a phospholipid normally confined to the inner mitochondrial membrane; however, it has been detected in human sera, indicating that it can exist in the extracellular space where it may interact with nearby cells. Although CL has been shown to modulate several functions of microglia in a toll-like receptor (TLR) 4-dependent manner, the effects of extracellular CL on astrocytes are unknown. In addition to their homeostatic functions, astrocytes participate in neuroimmune responses of the brain and express TLR 4. Therefore, we hypothesized that extracellular CL (1) modulates the secretion of cytokines and cytotoxins by astrocytes, as well as their phagocytic activity, and (2) acts by interacting with astrocyte TLR 4. We demonstrate that CL inhibits the lipopolysaccharide- (LPS-) induced secretion of cytotoxins and expression of glial fibrillary acidic protein (GFAP) by human U118 MG astrocytic cells. CL alone upregulates the phagocytic activity of human astrocytic cells and primary murine astrocytes. CL in combination with LPS upregulates secretion of interleukin (IL)-1β by astrocytic cells. Furthermore, CL alone increases the secretion of monocyte chemoattractant protein (MCP)-1 by astrocytic cells, which is blocked by the TLR 4-specific antagonist TAK-242. We demonstrate that CL upregulates MCP-1 secretion in the absence of its natural carrier protein, β2-glycoprotein 1, indicating that CL may be bioactive in the brain where this protein is not present. Lastly, we show that CL downregulates the expression of astrocytic TLR 4, implying that CL engages this receptor, as its activation has been shown to lead to its degradation. Overall, our study extends the list of cell type functions of which CL modulates and provides evidence that CL, or liposomes containing this phospholipid can be used to modulate specific neuroimmune functions of astrocytes.

Improving Rabbit Doe Metabolism and Whole Reproductive Cycle Outcomes via Fatty Acid-Rich Moringa oleifera Leaf Extract Supplementation in Free and Nano-Encapsulated Forms.

Simple SummaryUnder intensive rabbit production systems, due to the increased energy requirements of reproductive events, specifically pregnancy and lactation, rabbit does may confront several metabolic disorders as a result of energy imbalance. Moringa oleifera leaf ethanolic extract (ME) is one of the phytogenic extracts that has an impressive range of phytochemicals, specifically fatty acids (FAs). These phytochemicals may be biologically effective to support metabolism and reproductive functions of rabbit does during different reproductive cycle events. However, the high FAs content of ME makes them highly susceptible to lipid oxidation, diminishing their nutritional value and biological effects. In this study, we aimed to test the effects of FAs of ME either in a free-from or in a nano-encapsulated form on metabolism, immunity, milk production, milk composition, and reproductive performance of rabbit does during different physiological status (premating, mating, pregnancy, and lactation). The results showed that ME improved health, metabolism, immune functions, milk production and composition, and reproductive performance of rabbit does. These effects remained obvious even when a lower dose of ME was used in a nano-encapsulated form.The effects of free and nano-encapsulated ME supplementations on the metabolism, immunity, milk production and composition, and reproductive performance of rabbit does during premating, mating, pregnancy, and lactation were investigated. Multiparous rabbit does (n = 26 per group) received 50 mg of free ME (FME) daily, 25 mg of nano-encapsulated ME (HNME), or 10 mg of nano-encapsulated ME (LNME) per kilogram of body weight or were not supplemented (C) during a whole reproductive cycle. The ME contained 30 fatty acids with 54.27% total unsaturated fatty acids (USFAs). The fatty acid encapsulation efficiency of alginate nanoparticles was 70.46%. Compared with the C group, rabbits in all ME treatments had significantly increased body weight, feed intake, and glucose concentration and significantly decreased non-esterified free fatty acids and β-hydroxybutyrate concentrations. Rabbits supplemented with ME also had significantly increased white blood cell counts, phagocytic activity, lysozyme activity, and immunoglobulin G and decreased interleukin-1β concentrations. Moreover, ME supplementation significantly increased the concentrations of colostrum immunoglobulins, milk yield and energy content, and milk USFAs (omega-3 and 6). Rabbit does in the ME treatments had significantly higher conception and parturition rates and better litter characteristics than the C rabbit does. These results demonstrate the positive role of ME fatty acids on the health status and productive and reproductive performance of rabbit does at different physiological stages. Compared with the FME treatment, these parameters were further improved in rabbits that received nano-encapsulated ME at lower doses, illustrating how nano-encapsulation technology improves the bioavailability of ME.

Surface treatment of magnetron-sputtered tantalum pentoxide coatings and its effect on the phenotype and function of immune cells in vitro

The effect was analyzed of surface treatment by argon ions and electron beam on the properties of tantalum pentoxide coatings deposited by reactive magnetron sputtering and the treatment’s correlation with immune cells response. The characteristics of the coatings were determined by means of energy dispersive X-ray (EDX) scanning electron microscopy (SEM), X-ray photoemission spectroscopy (XPS), and atomic force microscopy (AFM). The results demonstrate that the electron-irradiation post-treatment process stimulates the adhesive potential and phagocytic activity of antigen-presenting cells.

POS-900 STUDY OF CLINICAL SPECTRUM AND OUTCOMES OF COVID RELATED MUCORMYCOSIS IN PATIENTS WITH RENAL INSUFFICIENCY

Coronavirus disease 2019(COVID-19) caused by Severe Acute Respiratory Syndrome Coronavirus 2 has been associated with a wide range of Opportunistic Bacterial and Fungal Infections. There are increased case reports of Rhino Orbital Mucormycosis in COVID -19 patients, especially from India. Hypoxia, High sugars (Diabetes, new-onset Hyperglycemia, Steroid-induced hyperglycemia),Acidic medium (metabolic acidosis, Diabetic Ketoacidosis [DKA]),High iron levels (increased ferritins) ,Decreased phagocytic activity of white blood cells (WBC) due to immunosuppression (SARS-CoV-2 mediated, steroid-mediated or background comorbidities) and Prolonged hospitalization with or without mechanical ventilators were associated with increased number of Mucormycosis.