The potential benefits of orange peels derived pectin on serum and skin mucus immune parameters, antioxidant defence and growth performance in common carp (Cyprinus carpio).

This study was performed to determine the effects of pectin derived from orange peel (PDOP) on growth performance, antioxidant enzyme activity and serum and skin mucus immune response of common carp (Cyprinus carpio). Common Carp (16.94 ± 0.03 g) were distributed into 12 tanks representing four treatments repeated in triplicates. Four diets were prepared to contain four levels of PDOP as follows: 0 (control), 0.5, 1, and 2% PDOP. Growth and immunological parameters as skin mucus lysozyme activity (SMLA) and total immunoglobulin (SMTIg), serum total immunoglobulin (STIg), serum peroxidase activities (SPA), Catalyse activity (CAT), DPPH radical scavenging activity, specific growth rate (SGR), weight gain (WG), final weight (FW), and feed conversion ratio (FCR) were assessed. Fish fed diets supplemented with PDOP showed an improvement of SGR, WG, FW, and FCR (P < 0.05). In terms of skin mucus immunological parameters, dietary inclusion of pectin significantly (P < 0.05) increased SMTIg. Likewise, carps fed either 1 or 2% PDOP showed notable enhancement of SMLA. In the case of serum immune parameters and antioxidant defence, carps in 1% PDOP treatment showed significantly (P < 0.05) higher SPA and CAT compared to fish fed either control diet or 0.5% OPDP. Additionally, no significant change (P > 0.05) was found in SPA and CAT of fish fed either 1% PDOP or 2% PDOP. Also, no significant (P > 0.05) difference was noticed between treated groups and control in the case of STIg. Furthermore, no significant differences were observed in DPPH radical activity among treatments (P > 0.05). Overall, these results suggested that inclusion of PDOP in common carp diet can beneficially affect growth and immune response.

Temporal changes in postprandial blood transcriptomes reveal subject-specific pattern of expression of innate immunity genes after a high-fat meal.

White blood cells are among the first responders to dietary components and their metabolites absorbed from the gut. The objective of this study was to determine the whole blood transcriptome response to high-fat challenge meals. A total of 45 fasting and postprandial (3-h and 6-h) whole blood transcriptomes from 5 subjects in a crossover intervention trial of a high-fat meal supplemented with placebo, blueberry powder or docosahexaenoic acid (DHA) were analyzed using RNA sequencing. Select target genes were validated by quantitative reverse-transcription polymerase chain reaction in 180 samples from 20 subjects. The largest contributor to variance was the subject (13,856 genes differentially expressed), followed by the subject on a specific day (2276 genes), followed by the subject's postprandial response (651 genes). After determining the nonsignificance of individual dietary treatments (blueberry, DHA, placebo), treatments were used as replicates to examine postprandial responses to a high-fat meal. The universal postprandial response (95 genes) was associated with lipid utilization, fatty acid beta-oxidation and circadian rhythms. Subject-specific postprandial responses were enriched for genes involved in the innate immune response, particularly those of pattern recognition receptors and their downstream signaling components. Genes involved in innate immune responses are differentially expressed in a subject-specific and time-dependent manner in response to the high-fat meals. These genes can serve as biomarkers to assess individual responsiveness to a high-fat diet in inducing postprandial inflammation. Furthermore, the dynamic temporal change in gene expression in postprandial blood suggests that monitoring these genes at multiple time points is necessary to reveal responders to dietary intervention.

Enrichment of rainbow trout (Oncorhynchus mykiss) fingerlings diet with microbial lysozyme: Effects on growth performance, serum and skin mucus immune parameters.

A two-month study was conducted to determine the influence of different levels of microbial lysozyme (LZ) contents (0, 0.5, 1.0, and 1.5 g kg-1 of diet) on growth performance, serum and skin mucus immune parameters as well as intestinal immune-related genes expression in rainbow trout Oncorhynchus mykiss fingerlings (5.5 ±â€¯0.1 g). Growth performance and feed utilization were not affected significantly by dietary LZ. Fish fed LZ-supplemented diets had higher serum total immunoglobulins concentration than the control group. In addition, fish fed 1.5 g LZ kg-1 diet had the highest skin mucosal total protein and immunoglobulin contents compared to other experimental groups. Furthermore, skin mucosal lysozyme and alkaline phosphatase activities as well as intestinal tumor necrosis factor-α and interlukine-1ß relative genes expression were higher in fish fed 1.0 and 1.5 g LZ kg-1 diets than the other groups. Overall, the present results clearly showed that LZ powder can be considered as a potential immunostimulant in O. mykiss fingerlings, but in the long term period it may result in negative effects on intestinal health as a consequence of inducing pro-inflammatory cytokines gene expression in the intestine.

Effects of Three Types of Inactivation Agents on the Antibody Response and Immune Protection of Inactivated IHNV Vaccine in Rainbow Trout.

Infectious hematopoietic necrosis virus (IHNV) infects salmonid fish, resulting in high mortality and serious economic losses to salmonid aquaculture. Therefore, an effective IHNV vaccine is urgently needed. To select an inactivation agent for the preparation of an effective IHNV vaccine, rainbow trout were immunized with mineral oil emulsions of IHNV vaccines inactivated by formaldehyde, binary ethylenimine (BEI), or ß-propiolactone (BPL). The fish were challenged 8 weeks after vaccination, and their IgM antibody response and relative percent survival (RPS) were evaluated. The results show that formaldehyde, BEI, and BPL abolished IHNV HLJ-09 infectivity within 24, 48, and 24 h at final concentrations of 0.2%, 0.02%, and 0.01%, respectively. The mean levels of specific IgM, both in serum and mucus (collected from the skin surface and gills), for the three immunized groups (from high to low) ranked as follows: the BPL group, BEI group, and formaldehyde group. From weeks 5 to 9, the mean log2 serum titers of IgM in the BPL group were significantly higher compared with those of the other groups (p < 0.05) during the 9 weeks of observation after vaccination (immunized at weeks 0 and6). Mucus OD490 values of the BPL group were significantly higher compared with those of the other groups (p < 0.05) when reaching their peak at weeks 5 and 8, but the difference between the formaldehyde and BEI groups was not significant (p > 0.05). The BPL-inactivated whole-virus vaccine had the greatest protective effect on the rainbow trout after challenge by an intraperitoneal injection of live IHNV, with an RPS rate of 91.67%, which was significantly higher compared with the BEI (83.33%) and formaldehyde (79.17%) groups. These results indicate that the BPL-inactivated IHNV oil-adjuvant vaccine was more effective than the formaldehyde- or BEI-inactivated vaccines. The results of this study provide an important foundation for further studies on inactivated IHNV vaccines.

Synthesis of Leucas Aspera Extract Loaded Gold-PLA-PEG-PLA Amphiphilic Copolymer Nanoconjugates: In Vitro Cytotoxicity and Anti-Inflammatory Activity Studies.

Gold nanoparticles (GNPs) are synthesized using the medicinal plant Leucas Aspera extract (LAE) and poly lactic acid-co-poly ethylene glycol-co-poly lactic acid (PLA-PEG-PLA) copolymer by water-in-oil (W/O) emulsion method. The proposed method of W/O emulsion technique involves synthesis of GNPs and loading of Leucas Aspera extract on to the PLA-PEG-PLA copolymer matrix simultaneously. The synthesized GNPs are characterized by Fourier transform infra-red (FTIR) spectroscopy, dynamic light scattering (DLS), X-ray diffractometry (XRD) and transmission electron microscopy (TEM). The GNPs-LAE loaded polymer NPs are examined for the in vitro cytotoxicity on South African green monkey's kidney cells. The GNPs-LAE loaded polymer nanoconjugates exhibit maximum up to 95% of cell viability with 100 µg concentration of GNPs in the sample. The GNPs-LAE loaded polymer NPs exhibit better anti-inflammatory activity when compared to the pure LAE.

Fermented Yupingfeng polysaccharides enhance immunity by improving the foregut microflora and intestinal barrier in weaning rex rabbits.

Yupingfeng (YPF) is a kind of Astragali radix-based ancient Chinese herbal supplemented with Atractylodis Macrocephalae Rhizoma and Radix Saposhnikoviae. Increasing evidence has proven the beneficial immunomodulating activity of YPF. However, the action mechanism(s) of it is not known. Here, we explored the immunomodulatory activity of unfermented Yupingfeng polysaccharides (UYP) and fermented Yupingfeng polysaccharides (FYP) obtained using Rhizopus oligosporus SH in weaning Rex rabbits. The results showed that both UYP and FYP exhibited notable growth-promoting and immune-enhancing activities, improvement of the intestinal flora homeostasis, and maintenance of intestinal barrier integrity and functionality. Notably, compared with UYP, FYP effectively enhanced average daily gain, organ indices, interleukin-2 (IL-2), IL-4, IL-10, tumor necrosis factor-alpha (TNF-α), TLR2, and TLR4 mRNA levels in spleen, IL-1, IL-2, IL-4, IL-6, IL-10, IL-12, TNF-α, and IFN-γ protein concentrations in serum, and TLR2 and TLR4 mRNA expressions in the gastrointestinal tract (GIT). Moreover, FYP exhibited greater beneficial effects in improving the intestinal flora, including augment flora diversity and the abundance of cellulolytic bacteria, reduction the abundance of Streptococcus spp. and Enterococcus spp. in the GIT, particularly the foregut and maintaining the intestinal barrier integrity and functionality by upregulating zonula occludens 1, claudin, polymeric immunoglobulin receptor, trefoil factor, and epidermal growth factor mRNA levels in the jejunum and ileum. Our results indicated the immunoenhancement effect of FYP is superior over that of UYP, which is probably related with the amelioration of the intestinal microflora and intestinal barrier in the foregut.

Effects of Fuzheng Paidu tablet on peripheral blood T lymphocytes, intestinal mucosa T lymphocytes, and immune organs in cyclophosphamide-induced immunosuppressed mice.

OBJECTIVE: The aim of this study is to investigate the effects of a Fuzheng Paidu tablet on peripheral blood T lymphocytes, intestinal mucosa T lymphocytes, and immune organs in cyclophosphamide (CY)-induced immunosuppressed mice. METHODS: The experimental mice (but not the control mice) were intraperitoneally injected with 80 mg/kg of CY solution every day for 3 consecutive days. Meanwhile, each mouse was administered with corresponding drugs for 7 continuous days. Then, 1 h after the last administration, each index was detected. RESULTS: The Fuzheng Paidu tablet significantly increases the CD4+/CD8+ ratio (P < 0.01) and the number of CD3+ and CD4+ cells in immunosuppressed mice (P < 0.01). In addition, the tablet apparently enhances the CD3+, CD4+, and CD8+ levels in the intestinal mucosal immune system (P < 0.01) as well as reverses the reduction of spleen lymphoid nodules and lymphocytes (P < 0.01). It also significantly improves intestinal inflammation, thymic atrophy, and sparse thymocytes in immunosuppressed mice (P < 0.01). DISCUSSION: The Fuzheng Paidu tablet greatly increases the levels of peripheral blood T lymphocytes and intestinal mucosal T lymphocytes as well as improves atrophied thymuses and spleens in CY-induced immunosuppressed mice.

Use of black vulture (Coragyps atratus) in complementary and alternative therapies for cancer in Colombia: a qualitative study.

BACKGROUND: Although Coragyps atratus has been used as a traditional therapy for patients with cancer, the scientific literature does not contain enough information on how this therapy is used or the mechanisms that explain this therapeutic practice. OBJECTIVES: To understand the methods of use and the reasons given by patients and caregivers for the use of Coragyps atratus in cancer treatment. METHODS: This study used a qualitative design based on twenty in-depth interviews of patients with cancer or caregivers of patients with the disease. The analysis of the text was based on an inductive thematic approach. RESULTS: Resistance to disease and immune enhancement are properties attributed to Coragyps atratus when used for cancer treatment. The most recommended method of use is fresh blood ingestion, and the associated mechanism of action is transfer of immune factors to the individual who consumes it. CONCLUSIONS: Use of Coragyps atratus as a treatment for cancer is a popular alternative therapy in Colombia. More studies are needed to understand the clinical effects of this intervention in cancer patients.

Specific inhibition of spleen tyrosine kinase suppresses leukocyte immune function and inflammation in animal models of rheumatoid arthritis.

Based on genetic studies that establish the role of spleen tyrosine kinase (Syk) in immune function, inhibitors of this kinase are being investigated as therapeutic agents for inflammatory diseases. Because genetic studies eliminate both adapter functions and kinase activity of Syk, it is difficult to delineate the effect of kinase inhibition alone as would be the goal with small-molecule kinase inhibitors. We tested the hypothesis that specific pharmacological inhibition of Syk activity retains the immunomodulatory potential of Syk genetic deficiency. We report here on the discovery of (4-(3-(2H-1,2,3-triazol-2-yl)phenylamino)-2-((1R,2S)-2-aminocyclohexylamino) pyrimidine-5-carboxamide acetate (P505-15), a highly specific and potent inhibitor of purified Syk (IC50 1-2 nM). In human whole blood, P505-15 potently inhibited B cell antigen receptor-mediated B cell signaling and activation (IC50 0.27 and 0.28 µM, respectively) and Fcε receptor 1-mediated basophil degranulation (IC50 0.15 µM). Similar levels of ex vivo inhibition were measured after dosing in mice (Syk signaling IC50 0.32 µM). Syk-independent signaling and activation were unaffected at much higher concentrations, demonstrating the specificity of kinase inhibition in cellular systems. Oral administration of P505-15 produced dose-dependent anti-inflammatory activity in two rodent models of rheumatoid arthritis. Statistically significant efficacy was observed at concentrations that specifically suppressed Syk activity by ∼67%. Thus specific Syk inhibition can mimic Syk genetic deficiency to modulate immune function, providing a therapeutic strategy in P505-15 for the treatment of human diseases.

[Observation on therapeutic effect of acupoint injection desensitization with autoblood on chronic urticaria].

OBJECTIVE: To compare the effect of acupoint injection desensitization with autoblood and routine combined therapy for treatment of chronic urticaria. METHODS: Two hundred patients with chronic urticaria were randomly divided into an acupoint injection with autoblood (AJA) group and a medicine group, 100 cases in each group. The AJA group was treated by acupoint injection desensitization with autoblood and Dazhui (GV 14), Fengfu (GV 16), Feishu (BL 13), Neiguan (PC 6) and etc. were selected, 3-5 acupoints each time, once every three days, 30 days for a course. The therapeutic effect was assessed after one course. The medicine group was treated with external application of Dexamethasone Acetate cream, twice a day, and oral administration of Setastine Hydrochloride, twice a day, 1 mg each time and the treatment duration was the same as that in the AJA group. RESULTS: The clinical cured rate was 66.0% (66/100) in the AJA group, which was superior to that of 0 (0/100) in the medicine group (P < 0.05). CONCLUSION: The acupoint injection desensitization with autoblood has obvious therapeutic effect on chronic urticaria with no apparent dependence and rebound problem.

Clinical and immunobiochemical characterization of airborne Peltophorum pterocarpum (yellow gulmohar tree) pollen: a dominant avenue tree of India.

BACKGROUND: Peltophorum pterocarpum (yellow gulmohar, PP) pollen is an important aeroallergen for type I hypersensitivity in the tropics. OBJECTIVE: To isolate and characterize the IgE-binding proteins of PP pollen for the first time. METHODS: Pollen extract was fractionated by a combination of Sephacryl S-200 column and diethylaminoethyl-Sephadex column. Allergen characterization was done by sodium dodecyl sulphate polyacrylamide gel electrophoresis, periodic acid-Schiff staining, enzyme-linked immunosorbent assay, and western blotting. Allergenic activities were determined by in vivo (skin prick test) and in vitro (enzyme-linked immunosorbent assay and histamine release) analyses. To determine whether the carbohydrate chains are involved in immunoreactivity, deglycosylation of PP pollen proteins was performed. RESULTS: SPT results on the respiratory allergic patients of Calcutta showed that 32.77% showed positivity with PP pollen. Eight IgE-reactive protein components were found in crude extract. Optimum IgE-reactive fraction 1 was resolved into five subfractions. The subfraction 1a showed maximum IgE reactivity containing the 28 kDa IgE-reactive component. Periodate oxidation showed that protein component was involved in its IgE binding. Twenty-eight kilodalton IgE reactive protein component was recognized by 75% of PP-sensitive patients in Western blotting. It also induced significant histamine release in sensitive patient sera. CONCLUSIONS: The purified 28 kDa protein is a clinically relevant allergen with a potential for diagnosis and therapy of patients susceptible to PP pollen.

Antibody responses in mice stimulated by various doses of the potato-derived major surface antigen of hepatitis B virus.

The ability of potato-derived major surface antigen of hepatitis B virus (P-HBsAg) to elicit antibody responses to different dosages of P-HBsAg ranging from 0.02 to 30 µg administered orally in mice was examined. All immunized groups produced specific serum IgG and fecal IgA antibodies against P-HBsAg, even at low levels (<5 µg), after administration of a 0.5-µg yeast-derived HBsAg (Y-HBsAg; LG Life Sciences, Republic of Korea) booster.

Immunogenicity of an autogenous Streptococcus suis bacterin in preparturient sows and their piglets in relation to protection after weaning.

Streptococcus suis is an important porcine pathogen causing meningitis and other invasive diseases in piglets of different ages. Application of S. suis serotype 2 bacterins to specific-pathogen-free (SPF) weaning piglets has been demonstrated to protect against the homologous serotype. However, autogenous S. suis bacterins are also applied to sows and suckling piglets in the field. Therefore, comparative evaluation of different bacterin immunization regimes, including sow vaccination, was performed in this study. The main objectives were to determine the immunogenicity of an S. suis bacterin in sows prepartum and its influence on active immunization of piglets. Experimental infection of 6- and 8-week-old weaning piglets was performed to elucidate protective efficacies. Humoral immune responses were investigated by an enzyme-linked immunosorbent assay (ELISA) measuring muramidase-released protein (MRP)-specific IgG titers and by opsonophagocytosis assays. Bacterin application elicited high MRP-specific IgG titers in the serum and colostrum of sows, as well as opsonizing antibodies. Piglets from vaccinated sows had significantly higher MRP-specific titers than respective piglets from nonvaccinated sows until 6 weeks postpartum. Vaccination of suckling piglets did not result in high MRP-specific titers nor in induction of opsonizing antibodies. Furthermore, neither vaccination of suckling nor of weaning piglets from immunized sows was associated with a prominent active immune response and protection at 8 weeks postpartum. However, protection was observed in respective 6-week-old weaning piglets, most likely because of protective maternal immunity. In conclusion, this study provides the first results suggesting protective passive maternal immunity for S. suis serotype 2 after bacterin vaccination of sows and a strong inhibitory effect on active immunization of suckling and weaning piglets, leading to highly susceptible growers.

Identifications of inhibitors of IgE production by human lymphocytes isolated from 'Cha Chuukanbohon Nou 6' tea leaves.

BACKGROUND: Tea (Camellia sinensis L.) is consumed all over the world and in especially large quantities in Japan and China, where it has been used not only as a daily beverage but also for medicinal purposes for thousands of years. Tea has been found to exhibit various bioregulatory activities, including antiallergic, anticarcinogenic, antimetastatic, antioxidative, antihypertensive, antihypercholesterolemic, anti-dental caries and antibacterial effects, and to influence intestinal flora. RESULTS: Cha Chuukanbohon Nou 6 is a tea cultivar improved by the National Institute of Vegetable and Tea Science (NIVTS) in Japan. On comparing chemical constituents of 11 varieties of tea leaves by high-performance liquid chromatography, we found two new major compounds in Cha Chuukanbohon Nou 6. Nuclear magnetic resonance spectroscopy revealed these compounds to be theogallin and 1,2-di-O-galloyl-4,6-O-(S)-hexahydroxydiphenoyl-beta-D-glucopyranose. The two were similar in chemical structure to strictinin, an inhibitor of immunoglobulin (Ig) production. Thus their effects on the production of Igs by peripheral blood lymphocytes were tested. Both compounds, like strictinin, inhibited IgE production. CONCLUSION: The results suggest Cha Chuukanbohon Nou 6 to be the basis of an antiallergic beverage.

Hemodynamic effects of cardiotomy suction blood.

OBJECTIVE: Cardiac surgery induces a systemic inflammatory activation, which in severe cases is associated with peripheral vasodilation and hypotension. Cardiotomy suction blood contains high levels of inflammatory mediators, but the effect of cardiotomy suction blood on the vasculture is unknown. We investigated the effect of cardiotomy suction blood on systemic vascular resistance in vivo and whether cell-saver processing of suction blood affects the vascular response. METHODS: Twenty-five patients undergoing coronary surgery (mean age, 68 +/- 2 years; 80% men) were included in a prospective randomized study. The patients were randomized to retransfusion of cell-saver processed (n = 13) or cell-saver unprocessed (n = 12) suction blood during full cardiopulmonary bypass. Mean arterial blood pressure was continuously registered during retransfusion, and systemic vascular resistance was calculated. Plasma concentrations of tumor necrosis factor alpha, interleukin 6, and complement factor C3a were measured in suction blood. RESULTS: Retransfusion of cardiotomy suction blood induced a transient reduction in systemic vascular resistance in all patients. The peak reduction was significantly less pronounced in the group receiving cell-saver processed blood (-12% +/- 2% vs -28% +/- 3%, P = .001). There was a significant correlation between tumor necrosis factor alpha concentration in retransfused cardiotomy suction blood and peak reduction of systemic vascular resistance (r = 0.60, P = .002). CONCLUSIONS: The results suggest cardiotomy suction blood is vasoactive and might influence vascular resistance and blood pressure during cardiac surgery. The observed vasodilation is proportional to the inflammatory activation of suction blood and can be reduced by processing suction blood with a cell-saving device before retransfusion.

Ganoderma lucidum mycelia enhance innate immunity by activating NF-kappaB.

Ganoderma lucidum is a popular medicinal mushroom in China and Japan for its immunomodulatory and antitumor effects. The goal of this research is to investigate the effect of dried mycelia of Ganoderma lucidum produced by submerged cultivation on the enhancement of innate immune response. We found that Ganoderma lucidum mycelia (0.2-1.6 mg/ml) stimulated TNF-alpha and IL-6 production after 8h treatment in human whole blood. IFN-gamma release from human whole blood was also enhanced after 3 day-culture with Ganoderma lucidum mycelia (0.2-1.0mg/ml). However, Ganoderma lucidum mycelia did not potentiate nitric oxide production in RAW264.7 cells. To better understand the possible immuno-enhancement mechanisms involved, we focused on nuclear factor (NF)-kappaB activation. Electrophoretic mobility shift assay revealed that the Ganoderma lucidum mycelia (1.6 mg/ml) activated kappaB DNA binding activity in RAW264.7 cells. These results provide supporting evidences for the immunomodulatory effect of Ganoderma lucidum mycelia.

A reliable, rapid and inexpensive two-color fluorescence assay to monitor serum cytotoxicity in xenotransplantation.

Removal and/or neutralization of preformed anti-pig antibodies in non-human primate blood have been shown to prevent the hyperacute rejection of transplanted pig organs. The purpose of this study was to establish a suitable in vitro method that would allow for screening and comparison of various agents and methods potentially useful in the prevention of hyperacute rejection. The pig kidney cell line (PK15), pig aortic endothelial cell line (AG08472), and a primary culture of endothelial cells explanted from a pig aorta were incubated with either human or baboon sera. Complement-dependent cytotoxic activity of human and baboon sera was determined on all three types of pig cells using a two-color fluorescence assay and compared with the conventional 51Chromium (51Cr)-release assay. The assay was also performed on PK15 cells as a 2-chambered slide assay and compared with a microcytotoxicity assay performed in Terasaki trays. Using the microcytotoxicity assay, a 1-step assay utilizing endogenous complement was compared with a 2-step assay where rabbit complement was added. Of the three types of cells studied, PK15 cells were the most sensitive to cytotoxic injury, followed by AG cells and the primary endothelial culture. Good correlation between the 51Cr-release and the two-color fluorescence method was documented. There was good agreement between the results obtained using the 2-chambered slide method and the microcytotoxicity assay, as there was between the 1- and the 2-step assays. The 1- and 2-step assays provided information on the level and efficacy of endogenous complement. We conclude that the two-color fluorescence assay is suitable for the rapid and inexpensive screening of therapeutic interventions that might be useful in the prevention of hyperacute xenograft rejection, and that PK15 cells are suitable for use in this assay.