Phagocytosis Index Research Articles

THE CHARACTERISTICS OF IMMUNE REGULATIONS IN INDIVIDUALS WORKING IN CONDITIONS OF COMBINED IMPACT OF HARMFUL PHYSICAL AND CHEMICAL FACTORS

The article presents results of the examination of workers of phthalic anhydride manufacture with work experience more than 10 years underwent impact of complex of harmful factors of production including chemical substances, noise, difficulty and intensity of working process. The group of comparison consisted of employees of administration department. The study established exceeding of both maximum permissible concentration (MPC) of phthalic anhydride (2.2-6.3 mg/m3 at MPC 1.0 mg/m3) and level of total and local vibration and thermal load at work places that corresponds to class 3.2-3.3 of work conditions on indicator of harmfulness and danger of industrial environment. The equivalent levels of noise with maximal values at work places of machinery operators of oxidation (84.6 dBA) are identified. The study established decreasing of phagocytic activity (relative phagocytosis, phagocytic number and phagocytic index in 1.13, 1.39 and 1.26 times correspondingly) relative to group of comparison and also decreasing of level of serum immunoglobulin G in relation of standard. On the basis of flow cytometry, the reliable decreasing of expression of CD95 T-cell receptors is established. The results of allergen-sorbent testing demonstrated increasing of the level of specific sensitization to phthalic anhydride by its metabolite of phthalic acid in 79.2% of workers. The indices of immune regulation CD95, phagocytic activity, specific to anhydride IgE, reflecting characteristics of immune regulation in workers of phthalic anhydride manufacture are recommended to be applied as markers of effect during assessment of health risk in people working in conditions of combined impact of physical and chemical factors.

Immunomodulating effect of magnetoinfrared laser therapy in children with chronic pyelonephritis

Aim. To determine the influence of magnetoinfrared laser therapy on the status of immunological reactivity in children with chronic pyelonephritis (CP).
 Materials and methods. One hundred thirty children aged 8 to12 years, suffering from relapsing course of chronic pyelonephritis, were observed. Group 1 (43 patients) received complex conventional therapy, group 2 (27 patients) – complex treatment, associated with magnetoinfrared laser therapy. The blood lymphocyte population and subpopulation, blood serum immunoglobulin and circulating immune complexes content, phagocytosis indices, blood serum cytokine content was studied in children with chronic pyelonephritis at the active stage of disease, at the partial and full clinicolaboratory remission stages.
 Results. The CP patients of group 1, who received complex conventional therapy, demonstrated a full, but not long clinicolaboratpory remission with preservation of marked changes in immunological reactivity parameters. In CP patients of group 2, who experienced complex treatment associated with magnetoinfrared laser therapy courses, there occurred full clinicolaboratory remission and normalization of immunological reactivity indices.
 Conclusions. The results of clinical observations and special studies demonstrate high clinical, immunomodulating and antirelapsing effects of complex treatment combined with magnetoinfrared laser therapy in children with chronic pyelonephritis.

Effect of Antibiotics Growth Promoters Alternatives on Growth Performance, Hematobiochemical and Immunological Profile of Healthy Broiler Chickens

A study was designed to explore the effect of Sanguinarine phytobiotic and Sodium butyrate on growth performance, the hematobiochemical and immunological profile of healthy Broiler Chickens. Ninety (90), one-day old healthy broiler chicks were used in this study and were divided into three equal groups (30 each). Group 1 healthy chicks was kept as control group, group 2 broilers received 2 ml/liter Sodium butyrate and group 3 broilers received 1 gm/10 liter phytobiotic (sangrovite). Administration of drugs in drinking water was continued for 5 successive days, from 19th to 24th day of age. On the first day post administration, two blood samples were collected for hemato-biochemical and immunological analysis. The obtained results revealed that, Broiler chicks received Sodium butyrate and Sanguinarine phytobiotic showed a significant increase in body weight (BW), weight gain (WG), feed consumption (FC) and improvement in feed conversion rate (FCR), total erythrogram and leukogram, phagocytosis and phagocytic index, nitric oxide, HI titre against ND virus, total protein, albumin with non-significant increase in total globulin, a, b, g globulin, albumin/globulin ratio, liver enzymes; aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase (AST ALT and ALP), uric acid and creatinine levels. From the obtained results it could be concluded that, the use of Sodium butyrate and Sanguinarine phytobiotic induced improvement in growth performance, hematobiochemical and immunological profile in healthy broiler chickens and they could be safely used as alternatives to antibiotics growth promoters (AGPs) in the commercial broiler diet to lower the risk of antibiotics residues in the meat and to avoid the antibiotic resistance to the consumers.

Immune regulation features in workers engaged into underground mining of chromite ores

The study of immune regulation features in workers engaged into underground mining of chromite ores demonstrated that high serum chromium levels were associated with increased phagocytic activity, if compared to physiologic normal values, in 25.7% of the workers. Findings also are phagocytic activity reliably changed by phagocytic index criterion, and significantly (1.2–1.33 times) increased absolute and relative phagocytosis and phagocytic index vs. the reference group values. IgM levels appeared 1.2 times lower vs. the reference group values, in association with IgG deficiency and increased IgA production vs. the reference values. The authors also revealed changes in cellular subpopulations balance and T-cell receptors transformation, with depressed regulatory marker CD95 in 90.0% in the main group workers, and higher number of CD4+CD25+CD127 lymphocytes, increased CD3+ and CD4+ cells 1.5- and 1.2-fold respectively vs. the reference group values. Other findings are disordered launching and regulation of apoptosis via lower Fas-receptor expression and changed balance of regulatory proteins Bcl–2 and Bax vs. the normal value in 50.0–62.5% of the workers, with reliable 2.6-fold increase of Bax level vs. the reference group.

Phagocytosis of monosodium urate crystals by human synoviocytes induces inflammation.

Gout is distinguished by an inflammatory process that is mediated by phagocytosis of monosodium urate (MSU) crystals in synoviocytes by regulation of unknown mechanisms. Here we suggest that the synovial cells play a crucial role in gouty arthritis by activating inflammation by MSU uptake and increasing the secretion of pro-inflammatory cytokines IL-1β, IL-6, IL-8, TNF-α, MCP-1, and the growth factors NGF and HGF. We discuss some co-existing features in synoviocytes, including anomalous morphologies of the cells, and microvesicle formation, dysregulation in VEGF gene expression. We provide evidence that phagocytosis of MSU crystals triggers an inflammatory cellular state in synoviocytes in the pathogenesis of crystal-induced arthritis.

Oral immunization with recombinant Lactobacillus casei expressing flaB confers protection against Aeromonas veronii challenge in common carp, Cyprinus carpio

Aeromonas veronii is an important type of gram-negative pathogen of human-livestock-aquatic animal and causes great economic losses in the aquaculture industry. Vaccination is an effective method of defence against A. veronii. There are many factors that restrict the use of vaccination, and the development of new oral vaccines is urgently needed. The selection of suitable antigens is of great significance for the development of aquaculture vaccines. Bacterial flagellin can specifically bind to TLR5 and induce the release of cytokines from the organism, which could be used in the development of vaccines. In this study, we constructed two recombinant Lactobacillus casei (L. casei) (surface-displayed or secretory) expressing the flaB of A. veronii and evaluated the effect of immune responses in common carp. The flaB gene (900 bp) of A. veronii was subcloned into the L. casei expression plasmids pPG-1 (surface-displayed) and pPG-2 (secretory). Western blot and immunofluorescence assays confirmed the expression of the recombinant flaB protein. Common carp immunized with Lc-pPG-1-flaB and Lc-pPG-2-flaB via oral administration route exhibited induction of antibody expression and innate immune responses. The results indicated that Lc-pPG-1-flaB and Lc-pPG-2-flaB can induce high levels of IgM, ACP, AKP, LZM and SOD activity in organisms, and Lc-pPG-1-flaB can induce even higher levels. The recombinant L. casei may effectively induce humoral immunity and increase the serum immunological index. Furthermore, leukocytes phagocytosis percentage and index of the recombinant L. casei were enhanced. The results of qRT-PCR showed that recombinant L. casei can significantly increase the expression of IL-10, IL-β, IFN-γ and TNF-α in the tissues of immunized common carp, compared with control groups. Viable recombinant L. casei strains, which were delivered directly survived throughout the intestinal tract. Common carp that received Lc-pPG-1-flaB (66.7%) and Lc-pPG-2-flaB (53.3%) exhibited higher survival rates than the controls after challenge with the pathogen A. veronii. Our work indicated that Lc-pPG-1-flaB and Lc-pPG-2-flaB had beneficial effects on immune response and enhanced the disease resistance of common carp against A. veronii infection. The combination of flaB delivery and the Lactic acid bacteria (LAB) approach may be a promising method for the development of oral vaccines for treating A. veronii. In future research, we will focus on the colonization ability of LAB in the intestines and on the impact of these bacteria on intestinal flora.

CHITOSAN NANOPARTICLE AS A DELIVERY SYSTEM FOR POLYPHENOLS FROM MENIRAN EXTRACT (PHYLLANTHUS NIRURI L.): FORMULATION, OPTIMIZATION, AND IMMUNOMODULATORY ACTIVITY

Objective: This study aims to formulate meniran extract into polymeric nanoparticles. Better stability of active substances in formulas compared to unformulated extracts is expected to increase immunomodulatory activity.
 Methods: Nanoparticles were formulated using ionic gelation method with chitosan and tripolyphosphate polymers. Optimize the mixture of nanoparticles using simplex lattice design (SLD) with the help of Design-Expert (DX) software. Evaluation of particle size and potential zeta using dynamic light scattering (DLS). Interactions between components were analyzed using Fourier transform infrared spectrophotometry-attenuated total reflectance (FTIR-ATR) and morphology of the lyophilization results observed using scanning electron microscopy (SEM). Immunomodulatory tests using the latex assay method. The parameters tested included phagocytosis index, phagocytic activity, and nitric oxide secretion.
 Results: The optimum mixture of the formulation process was obtained in the composition of chitosan 0.270 %, extract 0.626 %, and tripolyphosphate 0.074 % with desirability value of 0.841. Optimal response with particle size 434.7±3.90 d. nm, polydispersity index 0.285±0.03 and entrapment efficiency 62.98±0.65 %. The zeta potential value in the optimum formula is 11.9±0.1 mV with a positive charge. Phagocytosis index and phagocytic activity of nanoparticles differed significantly (p<0.05) compared with unformulated extracts.
 Conclusion: Meniran extract was successfully formulated into polymeric nanoparticles using chitosan-tripolyphosphate polymer. The developed nanoparticles have the immunomodulatory activity that is better than unformulated extract.

Influence of vitamins E and C on the indices of pseudoeozinofiles fagocytosis in chickens broilers blood of cross ROSS-308

The purpose of the research was to clarify the effect of chicken broilers feeding in the composition of mixed fodder with Vitamins E and C on the cellular link of non-specific resistance of the organism and, in particular, the parameters of pseudoiesinophils phagocytosis in blood. The research was carried out on chickens broiler – 308 grown in the farm “Fedyuk M” in the Zolochiv district of Lviv region. The keeping of the chickens was cellular with free access to feed and water. Experiments were conducted in 4 groups of broiler chickens of 100 chicks in each according to the scheme: the control group was fed with the standard feed (SC) according to the existing norms recommended for the cross ROSS-308; the first experimental group in addition to the SC received vitamin Е 1g/10 kg of mixed fodder; the second experimental group received vitamin C 2.5 g/10 kg of mixed fodder; the third experimental groups in addition to the ration received Vitamin C 2.5 g/10 kg and Vitamin E 1 g/10 of mixed fodder. For carrying out of immunological researches in chickens blood was taken in different age periods: 27-, 34- and 41-day-old age. In the whole blood, the phagocytic activity of pseudoiesinophils (FA), phagocytic index (FI) and phagocytic number (FF) were determined. For research, the daily culture of Escherichia coli (strain VKM-125) was used. The stimulatory effect of vitamins E and C in the composition of mixed fodder for chicken broiler broilers on the indices of pseudoiesinophils phagocytosis of blood was determined. As evidenced by a significantly higher phagocytic activity, phagocytic index and phagocytic count of pseudoiselinophils in blood of chickens in the experimental groups by comparison with the control group. At the same time there was a direct correlation between phagocytic activity and phagocytic index and index in blood of chickens broiler in experimental groups. At the same time, the ability of blood pseudoiesinophils to phagocytosis of microbial cells was higher in chicken broilers, which were fed with vitamin E and C.

Alteration of CD47 Expression in Human and Murine Tumor Models Changes M1/M2 Phagocytosis Effect in Vitro and In Vivo

IntroductionCD47 expression in human tumor cell represents a resistance mechanism escaping the macrophage surveillance system. To reinstate macrophage function in recognizing and engulfing tumor cells, unblocking CD47-SIRPalpha interaction and thereby re-activating macrophage activity has been an important strategy in immune-oncology therapy. To validate the strategy, we profiled a large panel of tumor models including tumor cell lines, PDX models in Crown tissue bank and clinical samples for CD47 expression. Despite of its expression in the majority of tumor cell models, the expression level of CD47 vary in flow cytometry examination. Interestingly a portion of tumor cells do not express CD47 at all.To facilitate in vitro screening for effective macrophage therapy, we established an in vitro assay platform using CD14+ monocytes as starting material. We also engineered human and murine cell models to overexpress human CD47 at one end and delete endogenous CD47 at the other end, and observed robust phagocytosis effect in these models. Finally, the findings in the in vitro assays were validated in MC38-hCD47 homograft in vivo.Material and MethodsTumor cell lines were CFSE-labeled and incubated with human monocyte-derived M1 or M2 macrophages (T:M 1:1) in the presence of 20μg/ml IgG1 isotype control, or anti-CD47 blocking antibodies B6H12 and BRIC126 and non-blocking antibody 2D3, for 2hrs. Cells were then harvested and stained with a macrophage marker (CD14+). Phagocytosed tumor cells were identified by flow cytometry as CD14+CFSE+. The phagocytosis index was calculated by dividing the number of macrophages engulfing tumor cells by the total number of macrophages. CD47 overexpression and deletion clones (KARPAS299, SK-OV3, and MC38 background) were developed in house. The CD14+ cell isolation kit was from Miltenyi. M1/M2 differentiation reagents were purchased from R&D and Stem Cell. Anti-CD47 antibodies B6H12 and 2D3 and mouse IgG1 were purchased from Thermo Fisher, and BRIC126 from GeneTex. B6H12 for animal study was sourced from Bioxcell.ResultsTo facilitate in vitro screening for effective macrophage therapy, we sought to establish a robust yet reproducible assay platform using CD14+ monocytes as starting material. Through macrophage differentiation we were able to achieve > 80% of M1 and M2 populations, respectively, though variation exists. Differentiated M1/M2 were then co-cultured with CFSE-labeled target tumor cells for subsequent flow cytometry analysis. Strong phagocytosis effect was observed with CD47 expression tumor cells when anti-CD47 antibody BRICK126, which blocks the CD47-SIRPalpha interaction, was introduced. In contrast, aCD274 (PD-L1) antibody didn't yield more than background phagocytosis effect in the same assay suggesting the specific activation effect of BRICK126 triggering M1/M2 functionality.To prove the CD47-specific effect, we engineered CARPAS299 and SK-OV3 tumor cell line models and MC38 murine cell model to overexpress human CD47 at one end and delete endogenous CD47 at the other end. The recombinant and isogenic models were then subject to the CD47 biology examination and more profound changes in phagocytosis responses were observed.To validate our findings in vivo, we introduced engineered MC38-hCD47 clone in to C57/BL6 mice to measure homograft tumor growth kinetics and pharmacological responses to B6H12, another CD47 blocking agent. Details of results will be presented.ConclusionDisrupting the CD47-SIRPa anti-phagocytic axis is a novel immunotherapy approach against cancer. This study reveals that CD47 expression level varies in different tumor cell models. Our in vitro screen demonstrated that M1 and M2 macrophages resulted in comparable anti-CD47 phagocytic activities, and suggests that Karpas299 with higher CD47 expression level may be a better model than SK-OV3 in cell engulfing study. We also developed Karpas299-hCD47, SK-OV-3-hCD47 and MC38-hCD47 stable clones to facilitate in vitro and in vivo screening towards macrophage therapy, and discovered strong positive correlation between CD47 expression level and phagocytic activity in in vitro and in vivo systems. DisclosuresNo relevant conflicts of interest to declare.

β-Glucan successfully stimulated the immune system in different jawed vertebrate species.

Several reports have shown the positive effects of β-glucans on the immune. Howeverthese studies have a broad experimental design including β-glucans compounds. Consequently, a study using the same β-glucan molecule, administration route and experimental design is needed to compare the effects of β-glucan across vertebrate species. For this end, during 28 days we fed four different vertebrate species: mice, dogs, piglets and chicks, with two β-glucan molecules (BG01 and BG02). We measured the serum interleukin 2 as an indicator of innate immune response, the neutrophils and monocytes phagocytosis index as a cellular response and antibody formation as an adaptive response. The results clearly showed that the different β-glucan molecules exhibited biologically differently behaviors, but both molecules stimulate the immune system in a similar pattern in these four species. This finding suggests that vertebrates shared similar mechanisms/patterns in recognizing the β-glucans and confirms the benefits of β-glucans across different vertebrate species.

Effect of Sanguinarine phytobiotic, Sodium butyrate compared to ampicillin on controlling necrotic enteritis in broiler chickens.

This study aimed to investigate the efficacy of Sanguinarine phytobiotic, Sodium butyrate compared to ampicillin in treatment of Clostridium perfringens infection in broiler chickens with a special reference to their effects on growth performance, hematobiochemical and immunological profile. A total of 150 one-day old Cobb broiler chicks were used in this study. On day 14 th of age all chicks were divided into 5 equal groups (30 each). Group (G1) non-infected, non-treated (control), G2 infected with C. perfringens , non treated, G3 infected, treated with Sodium butyrate, G4 infected and treated with Sanguinarine phytobiotic and G5 infected and treated with ampicillin. Administration of drugs in drinking water was continued for 5 days from 18-23 day of age. The results revealed that the infected broiler chickens with C. perfringens and non treated(G2) showed clinical signs of necrotic enteritis represented by loss of appetite, diarrhea, dehydration, anorexia in addition to a recorded mortality rate 30% and lesion score 80%. Moreover, significant reduction (P<0.05) in body weight, weight gain, erythrogram, phagocytosis and phagocytic index, Nitric oxide, HI titers, total protein, albumin and total globulin beside significant increase (P<0.05) in feed conversion rate (FCR), leukogram, liver enzymes, aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase (AST-ALT and ALP), uric acid and creatinine associated with non-significant increase (P<0.05) in globulin fractions were observed. Treatment of the infected chickens with these drugs led to improvement in clinical signs, mortality rate, lesion score, growth performance, hematobiochemical and immunological profile compared with infected non treated birds (G2), but ampicillin had superior effect against Costridium perfringes infection. It could be concluded that C. perfringens infection in broiler chickens induced adverse effects on growth performance, hemato-biochemical and immumological profile of birds which could be reversed or ameliorated by using Sanguinarine phytobiotic, Sodium butyrate or ampicillin . So, the study recommended the use of such treatments hand in hand with antibiotics for controlling necrotic enteritis in broilers. Key words: necrotic enteritis; Sodium butyrate; phytobiotic; ampicillin

Biological Actions, Electrical Conductance and Silicon-Containing Microparticles of Arsenicum Album Prepared in Plastic and Glass Vials.

According to the "silica hypothesis" formulated to explain homeopathy, the information of starting materials would be transferred to cells by silica nanoparticles detached from the glassware walls by serial dilution and agitation through epitaxy. We compared the biological activity, electrical current and silicon microparticle content (by means of scanning electron microscopy/energy-dispersive X-ray spectroscopy) of high dilutions (HDs) of arsenic prepared in plastic and glass vials to investigate the role of silica in their biological effects in vitro. Co-cultures of macrophages and yeast (Saccharomyces cerevisiae) were treated with different HDs of arsenic prepared in plastic and glass vials. Macrophage morphology, phagocytosis index, nitric oxide (NO), and cytokine production were evaluated. Measurable amounts of silicon microparticles were detected only in the HDs prepared in glass vials, but ultra-centrifugation eliminated them. Specific and non-specific results were observed. Non-specific pro-inflammatory effects were seen in all dilutions prepared in plastic vials, including elevation of pro-inflammatory cytokines, NO and macrophage phagocytic index. Only the 200th centesimal dilution of arsenic produced specific decrease in interleukin-6 production in macrophages, and it was independent of the vial type or the presence of microparticles of silica in the medicine samples. The nature of the vials had an impact on the electric flow in the respective fluids. The non-specific, pro-inflammatory effects might be attributed to organic residuals detached from the vials' plastic walls during manipulation. Instead, specific silica-independent effects of the homeopathic medicine can be attributed to the decrease of interleukin-6 after treatment with the 200th centesimal dilution of arsenic.

Structure and radio‐protective effects of sulfated Auricularia auricula polysaccharides

Radiation-protective effects of sulfated polysaccharides (SNAAP80) from Auricularia auricular were studied. Crude polysaccharides were extracted using ultrasound-assisted water extraction. Bioactive polysaccharide fragments were screened using ethanol precipitation with an in vitro antioxidant activity test. The chlorosulfonic acid-dimethylformamide method was used to obtain the sulfated polysaccharide. Monosaccharide composition and molecular weight were determined using GC-MS and HPLC-GPC, respectively. IR, UV, and SEM were used for structural characterization. The antiradiation effect of SNAAP80 was studied using a 60Co-γ radiation model in mice. The results showed that the SNAAP80, with a degree of substitution of 0.514, was composed of glucose and mannose. A bioactivity analysis showed that the SNAAP80 increased the organ indexes, phagocytosis index, stimulation index, and the superoxide dismutase, glutathione peroxidase, catalase, and lactate dehydrogenase activity. The SNAAP80 decreased TBARS, IL-2, IL-12, INF-γ, protein carbonyls, creatinine, and blood urea nitrogen. The results suggested that SNAAP80 had an effective radiation-protective effect. Practical applications Auricularia auricula is a commonly cultivated commercial mushroom in East Asia; it is also called “wood ear” in China and “black fungus” in Russia. Many published studies regarding polysaccharides from A. auricula have shown their beneficial health effects for both humans and animals. A. auricula is acknowledged for its pharmaceutical effects such as inhibiting lipid peroxidation, decreasing liver damage, hypolipidemic properties and so on. Functional activity of polysaccharides is related to their solubility. The activities of polysaccharide NAAP80 from A. auricular were restricted because of its poor solubility. Therefore, it was modified by chlorosulphonic acid-N,N dimethylformamide to obtain SNAAP80 which has a significantly higher solubility. Antiradiation effect of SNAAP80 has been studied in this paper, and its potential application in antiradiation may be relevant to space exploration, the military, daily health care, and therapeutic medications.

IMMUNOMODULATORY EFFECT TEST FROM MORINGA LEAF EXTRACT (MORINGA OLEIFERA L.) WITH CARBON CLEARANCE METHOD IN MALE WHITE MICE

Objective: Moringa oleifera leaf has chemical compounds that have been utilized by the community to cure health problems. One of its activities is an immunomodulator. The aim of this study is to determine the immunomodulatory effect from M. oleifera leaf using a carbon clearance method to measure the activity of phagocytic cells in exterminating pathogens that enter the body then followed by calculating the total leukocyte cells. The parameters of this test are phagocytosis index and total leukocyte cells.Methods: Twenty white male mice were divided into four groups. Group I (vehicle control) was treated with sodium-carboxymethyl cellulose (NaCMC) 0.5%, Group II-IV were treated by M. oleifera leaf extract given to the mice for six consecutive days orally in doses of 10, 30, 100 mg/kg. On the seventh day, white male mice were given with intravenous carbon suspension through their tails. The value of phagocytosis index (PI &gt; 1) indicated immunostimulant activity. The data were analyzed by using one-way analysis of variance and Duncan test.Results: The analysis of variance results showed that the groups treated with Moringa leaf extract are significantly different with the vehicle groups (NaCMC 0.5%) (p&lt;0.05). Increased doses of Moringa leaf extract are effective to improve the immunomodulator effect. It was included that Moringa leaf extract had the immunomodulatory capabilities as an immunostimulant.Conclusion: Immunomodulatory effect test of M. oleifera Lam. Based on the result of the research about immunomodulatory effect test from Moringa leaf extract (Moringa Oleifera L.) with carbon clearance method in male white mice, it can be concluded that Moringa leaf extract (Moringa Oleifera L.) has effect as an Immunomodulator.

The ethanol extract of honeysuckle stem modulates the innate immunity of Chinese mitten crab Eriocheir sinensis against Aeromonas hydrophila

Honeysuckle stem had been used as feed additives to modulate immunity in breeding industry, which was limited in the aquaculture field. In this study, the immunomodulation of honeysuckle stem ethanol extract (designed as HSE) on Chinese mitten crab Eriocheir sinensis was detected. The crabs fed with HSE diets for 30 days had higher level of the total haemocyte count (HTC), lysozyme activity and PO activity (P < 0.05), and had no obvious affect on the phagocytic activity, NO and TNF-α level. When challenged with Aeromonas hydrophila (1.0 × 107 colony-forming units), HSE exhibited weak antibacterial activity against A. hydrophila and increased survival rate of crabs. The decreasing of THC and the increasing of TNF-α concentration, EsCaspase and EsLITAF mRNA expression level were all inhibited significantly by HSE treatment (P < 0.05), when the crabs were challenged by A. hydrophila. Moreover, the following immune parameters of crabs were enhanced by HSE treatment after A. hydrophila infection, including the rising of phagocytosis index and phagocytic rate of haemocyte, the rising of lysozyme, PO, NOS activities and nitric oxide concentration (P < 0.05). Therefore, it was concluded that HSE had great potential to develop into feed additive of crabs, which could enhance the innate immunity of Chinese mitten crabs E. sinensis effectively after A. hydrophila infection.

BIOLOGY OF THE ORAL CAVITY IN CHILDREN LIVING IN THE CITY OF CHIRCHIK

The study of microbiological and immunological parameters of the oral cavity among children living in the industrial city of Chirchik, the total number of anaerobes, aerobes, phagocytic activity of neutrophils, the level of lysozyme and the immunoglobulin A secretion fraction of the secretory fraction (sIgA) has been studied. To address this goal, we conducted microbiological and immunological studies in 57 children living in the town of Chirchik. All these children, depending on their age, were divided into 3 groups: Group 1 included 17 children aged 6-7 years. Group 2 consisted of 25 children aged 8-12 years. The third group consisted of 15 children aged 13-15. In children aged 6-7 years, it can be noted that the total number of the optional group of microbes is higher than in anaerobic. In this case, the quantitative indicators of the lactobacillus are especially affected among the anaerobes, so their number is Lg. 2.10 ± 0.1 cfu /ml. Alarming in this group of children is the sowing of pathogenic strains of staphylococcus. The remaining groups of microbes, although there are minor changes, but they are unreliable. At children in the age of 2 and 3 group the flora of the oral cavity in many respects stands close to normal indices. Although it should be noted a significant increase in the number of streptococci, especially strains of Str. mutans and mitis. In children in the age group 6-7 years, almost all indicators show immunodeficiency, so the lysozyme titer was 14 ± 0.41 mg /%, it was normally 19.180 ± 0.60 mg/%, the index of phagocytosis was 48.4 ± 1.45% significantly lower than the control indices, the level of secretory immunoglobulin of class A (sIgA) was 1.7 ± 0.1 g/l, which is much lower than normal values. It is interesting to note that in children older than 8-12 and 13-15 years, these indicators have improved significantly, although it should be noted that their immunodeficiency has worsened, only in terms of the fraction of secretory immunoglobulin A (sIgA). From the above, it follows: 1. According to the sanitary and hygienic and microbiological studies carried out, it is possible to state that the city of Chirchik is an ecologically unfavorable region. 2. The study of oral micro-ecology in children in Chirchik made it possible to establish that in children in the age group 6-7 years the most pronounced are dysbiotic conditions, the main feature of which is the decrease in the amount of lactobacillus, but the increase in the number of staphylococci and fungi of the genus Candida 3. When studying the local factors of oral protection in children living in Chirchik in the age aspect, it was found that immunodeficiency states are most pronounced in children of the I group. 4. The study of the state of colonization resistance of microbes in children living in the city of Chirchik established that the most pronounced violations in these issues were noted in the children of group I.

Preparation optimization of extract of Moringa oleifera leaves and its immune regulation effect on immunosuppressed mice

With total flavonoid content and dry extract yield as the observation indexes, the optimal extraction conditions of Moringa oleifera leaves were determined by using single factor test and orthogonal test, and cyclophosphamide modeling method was used to establish immunosuppressed mice models, so as to investigate the effects of M. oleifera leaves extract on immune regulation in mice. The results showed that the optimal preparation conditions were as follows: extraction with 70% ethanol, material-liquid ratio 1:15, extraction temperature 80 °C, three times, 1.5 hours for each time. Under these conditions, the content of total flavonoids from M. oleifera leaves was 15.64 mg·g⁻¹, which can significantly enhance macrophage phagocytosis and immune organ index, promote the synthesis of serum immunoglobulin IgG and hemolysin, and decrease AST activity, with regulation effect on immune dysfunction.

Effects of corncob derived xylooligosaccharide on innate immune response, disease resistance, and growth performance in Nile tilapia (Oreochromis niloticus) fingerlings

An-eight week experiment was conducted to test efficacy of corncob derived xylooligosaccharides (CDXOS) on mucosal and serum immune, disease resistance, and growth performance of Nile tilapia (Oreochromis niloticus). Three hundred and twenty tilapia fingerlings (20.72 ± 0.02 g) were fed the following diets; 0 (Diet 1- control), added 5 – (Diet 2), 10 – (Diet 3), and 20 g kg−1 CDXOS (Diet 4). After 4 and 8 weeks feeding were mucosal, serum immune responses, and growth performance measured. A challenge test (15 days) against Streptococcus agalactiae was conducted after 8 weeks post-feeding. A significant (P < 0.05) stimulation of skin mucus lysozyme and peroxidase activities, as well as serum immune parameters (serum lysozyme, serum peroxidase, alternative complement activities, phagocytosis index, and respiratory burst activity) was noticed by feeding the fish CDXOS supplemented diets compared to the control group. Highest (P < 0.05) innate immune parameters were observed by feeding the fish 10 g kg−1 CDXOS vs. the other treatments. With regard to the challenge test, relative percent survival (RSP) of Nile tilapia fingerlings fed Diet 2, Diet 3, and Diet 4 was 34.78%, 60.87%, and 30.43%, respectively. Among the supplemented groups, dietary inclusion of 10 g kg−1 CDXOS revealed significant (P < 0.05) higher RPS and resistance towards S. agalactiae than the other groups. Regarding growth performance, final weight, weight gain, specific growth rate, and feed conversion ratio were remarkably (P < 0.05) improved in the CDXOS groups; highest improvement was observed in the 10 g kg−1 CDXOS treatment. In conclusion, inclusion of 10 g kg−1 CDXOS derived from corncob improved growth performance and health status of Nile tilapia fingerlings.

In Vitro study for hesperidin nanoparticles effect on phagocytic activity against Staphylococcus aureus

Hesperidin is one of the flavonoids from citrus peels and it recognized to possess various biological activities such as, anti-inflammatory, anti-carcinogenic, antioxidant and antimicrobial potentials. The present investigation studies the immunological adjuvant influence of hesperidin nanoparticles. Hesperidin nanoparticles were prepared by nano-precipitation technique by using Poly (D, L-lactic-co-glycolic acid) (PLGA) polymer and Poloxamer 407 was used as a stabilizer. This method was used because of their advantage of low setup cost and simplicity. Hesperidin nanoparticles were characterized by fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD) and particle size analysis (PSA) analytical methods. The effect of hesperidin nanoparticles was higher than the effect of pure hesperidin, and there was an obvious increase in phagocytosis index (PI 82%) of hesperidin nanoparticles when compared with pure hesperidin (PI 56%) and in comparison with the control samples (PI 22%). In conclusion we need further studies about if nano-hesperidin has therapeutic effects.

Correlation Between Total Flavonoid Contents and Macrophage Phagocytosis Activity of Fractions From Faloak (Sterculia quadrifida R.Br.) Barks Ethanolic Extract In Vitro

On Timor island, Nusa Tenggara Timur, faloak barks (Sterculia quadrifida R.Br.) has been used empirically to restore stamina. Faloak bark ethanolic extract proved to have immunomodulatory activity in vitro, which can increase macrophage phagocytosis activity. This research aimed: (i) to determine the immunomodulatory active fraction of faloak bark ethanolic extract, (ii) to determine the total flavonoid contents of faloak extract and fractions, and (iii) to evaluate the correlation of the total flavonoid contents of those extract and fractions with their macrophage phagocytosis activity. The simplisia powder is macerated with 96% ethanol. The extract was dissolved in methanol:water (9:1v/v) was then subsequently partitioned with n-hexane, ethyl acetate, and water to obtain n-hexane fraction, ethyl acetate fraction, water fraction, and insoluble fraction. Faloak extract and fractions at concentration 62,5; 125; 250; 500μg/mL were tested for their effect on the peritoneal macrophage phagocytosis of Balb/c mice in vitro by the latex beads method. Phagocytosis capacity and phagocytosis index were analyzed using one-way anova and post hoc Tukey HSD test with 95% confidence level. The results showed that ethyl acetate fraction had the highest macrophage phagocytosis capacity and the highest total flavonoid content compared to other fractions. The highest macrophage phagocytosis capacity of ethyl acetate fraction at concentration of 250 μg/mL was 51,94±4,67%, this value was significantly different from cell control (7,50±1,29%), negative controls of 0,0625% dimethylsulphoxide (6,25±0,36%), as well as positive control of 200 μg/mL echinaceae extract syrup® (9,97±0,33%). The total flavonoid content of ethyl acetate fraction determined by aluminum chloride method was 4,290±0.029 mg of quercetin equivalent/g fraction. There was a positive and strong correlation between the total flavonoid content of these extract and fractions with their macrophage phagocytosis capacity (Pearson correlation coefficient of 0,781) and showing linear relationship y=4,721x+19,663; R2=0,61.