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#27941713   2016/12/12 Save this To Up

Pathogenic Effects of Biofilm on Pseudomonas Aeruginosa Pulmonary Infection and Its Relationship to Cytokines.

BACKGROUND An animal (Sprague-Dawley rat) model of Pseudomonas aeruginosa biofilm associated with chronic pulmonary infection in vivo was established and the effects of the biofilm on P. aeruginosa and its relationship to cytokines were investigated. MATERIAL AND METHODS Biofilm of P. aeruginosa in alginate beads and planktonic PA0725 were purified by anion-exchange chromatograph. Sprague-Dawley (SD) rats were immunized with the biofilm and then inhaled the same strain of P. aeruginosa. Anti-biofilm antibody titer was detected using the enzyme linked immunosorbent assay (ELISA) method. The cell count and differential count in the bronchoalveolar lavage fluid (BALF) were measured. The levels of cytokines (IL-17, IL-1β, MIP-2, and G-CSF) and tumor necrosis factor (TNF)-α in sera were also measured using an ELISA kit. RESULTS The sera anti-biofilm IgG antibody titer of immunized SD rats was increased significantly on the 5th and 8th days after inhalation. The IL-17 concentration was significantly higher on the 8th day after inhalation. The results indicated that when biofilm-pre-immunized rats were challenged with inhalation of PA0725 of P. aeruginosa, the biofilm acted as an antigen substance and mediated the antibody reaction of the antigen, which might cause serious airway inflammatory response and lung tissue injury. This effect may be related to IL-17. CONCLUSIONS P. aeruginosa biofilm protected the bacterium from antibiotics and might induce host immune damage in lung tissue and facilitate bacterium evading the host barrier.

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#27363620   2016/08/16 Save this To Up

N-Acetyl-l-cysteine exacerbates generation of IL-10 in cells stimulated with endotoxin in vitro and produces antipyresis via IL-10 dependent pathway in vivo.

N-Acetyl-l-cysteine (NAC) is a well-known medication, primarily used as a mucolytic agent in pulmonary disease. Recently, we have found that NAC possesses antipyretic properties. The aim of the present study was to investigate the mechanism by which NAC attenuates fever. The concentration of interleukin (IL)-10 and prostaglandin (PG) E2 were measured using ELISA kit in the supernatants aspirated after stimulation of peripheral blood mononuclear cells (PBMCs) with lipopolysaccharide (LPS, 1μg/mL) and NAC (10mM). The body temperature of the Wistar rats was measured using biotelemetry system. To inhibit endotoxic fever, NAC (200mg/kg; i.p.) was injected into the rats one hour prior to the LPS administration (50μg/kg; i.p.). The pre-treatment of LPS-stimulated PBMCs with NAC resulted in a significant decrease in PGE2 concentration in comparison to the cells treated with LPS alone (PGE2 level was 386.1±61.9pg/mL vs. 2078.9±157.9pg/mL, respectively, p<0.001). Furthermore, in these cells we observed a significant increase in IL-10 level (142.1±2.62pg/mL in NAC+LPS stimulated cells vs. 54.4±0.6pg/mL in LPS stimulated cells, p<0.001). The injection of anti-IL-10 antibody into the rats abolished antipyretic properties of NAC. Body temperature in animals treated with anti-IL-10+NAC/LPS was 38.28±0.12°C vs. 37.73±0.06°C in IgG+NAC/LPS rats (p<0.001) and 38.31±0.20°C in NaCl/LPS-treated animals (n.s.). Based on these data, we conclude that NAC acts as an antipyretic via IL-10 stimulation. This finding provides a new insight into the immunopharmacology of NAC, and we believe that in a future it will contribute to the new and/or more accurate application of NAC in medicine.

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#27072237   2016/04/13 Save this To Up

Dendritic cell vaccination enhances antiangiogenesis induced by endostatin in rat glioma.

It has been verified that dendritic cell (DC) vaccination can improve the prognosis of malignant glioma. However, recent evidence suggests the problems with DC vaccines lies, at least in part, with the cancers ability to induce an immunosupressive response that suppresses any vaccine-mediated active immunity. Our previous studies indicate that subcutaneous vaccine can restrain the cancer cells implanted in the brain, but the effect is limited on vascularized tumor in the brain. Furthermore, vascular endothelial growth factor (VEGF) and vascular cell adhesion molecule (VCAM) play an important role in immunoevasion.

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Rat Anti-Mouse Dendritic Jurkat Cell Extract (Indu Jurkat Cell Extract (Indu Jurkat Cell Extract (Indu Jurkat Cell Extract (Indu GLP 1 ELISA Kit, Rat Gluc GLP 2 ELISA Kit, Rat Prog Glucagon ELISA KIT, Rat G Leptin ELISA Kit, Rat Lep Rat anti mouse 33D1 (a de Rat anti mouse CD205 (a d OxiSelect™ Cellular UV-

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#24238666   2013/11/28 Save this To Up

Age-related and regional differences in the prevalence of hepatitis E virus-specific antibodies in pigs in Germany.

An increasing number of acute autochthonous human hepatitis E virus (HEV)-infections was noticed in Germany and other developed countries, most likely the result of a zoonotic virus transmission from pig, wild boar and deer. Currently there is still a lack of profound data concerning the actual prevalence of HEV-specific antibodies in domestic pig herds in Germany, in particular for regions with high pig density, and its age-dependency. 2273 domestic pig sera were collected in 2011 mainly from Bavaria, North Rhine-Westphalia and Lower Saxony from areas having a high pig density. Initially, 420 randomly selected pig sera were tested in three commercially available and in two in-house HEV-antibody ELISAs. 43.6% (183/420) to 65.5% (275/420) of the sera were demonstrated to be reactive against human pathogenic HEV genotypes 1 and/or 3. The majority of sera reacted only weakly or not at all with the rat HEV antigen with very few sera showing a stronger reactivity to this antigen compared to the genotype 3 antigen. The results of all three HEV-IgG tests, i.e. the PrioCHECK(®) HEV Ab porcine ELISA kit, the ID Screen(®) Hepatitis E Indirect Multi-species ELISA kit and the genotype 3 in-house ELISA were in good accordance. Therefore, the remaining sera were tested using the PrioCHECK(®) HEV Ab porcine ELISA kit. Samples with a borderline result were finally determined by application of the conjugate-modified recomLine HEV IgG assay. A total of 1065 of the 2273 sera (46.9%) were found to be anti-HEV IgG-positive. While 38.4% (306/796) of fatteners (age between 3 and 9 months) exhibited HEV-specific antibodies, 51.4% (759/1477) of sows (age older than 9 months) exhibited anti-HEV antibodies (P<0.001). Fatteners kept in Southern Germany had a significantly higher HEV IgG prevalence compared to fatteners kept in the high pig density federal states North Rhine-Westphalia and Lower Saxony but also in German federal states with a low pig density. In conclusion, the present study clearly demonstrates that a high percentage of domestic pigs in Germany have had contact with HEV. Seroprevalence depends on the pig's age and herd origin with the most significant regional variations for fatteners. The presence of anti-HEV-free herds may indicate that it is feasible to establish and sustain HEV-free pig herds. HEV seroprevalence still depends on the assay used for testing. This demonstrates an urgent need for test validation.

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#21341161   2011/02/22 Save this To Up

[Tularemia seroprevalence in the risky population living in both rural and urban areas of Erzurum].

Tularemia which is a zoonotic infection, caused by Francisella tularensis, has become a re-emerging disease in Turkey. Infection is often transmitted to human by handling animal tissues and products, but it is also possible to acquire the disease from contaminated water or food. Recently several cases and epidemics of tularemia have been reported in the northwest areas of Turkey, particularly in Marmara and West Black Sea regions. Erzurum is a city in Eastern Anatolia Region, Turkey and animal husbandry is the main agricultural activity in that area. However, neither tularemia cases were reported from this province nor seroprevalence studies were performed. In this study we aimed to determine F.tularensis antibody seropositivity in the risky population living at both rural and urban area of Erzurum. Blood samples from 240 volunteer subjects (134 male with mean age: 36.2, age range: 17-75 years and 106 female with mean age: 39.1, age range: 16-77 years) whose occupations were farming and animal husbandry, were included in the study. Serum samples were screened for the presence of F.tularensis antibodies by slide agglutination method (BD, USA) and Serazym ELISA kit (anti-F.tularensis IgG/IgA/IgM, Seramun, Germany). The positive samples with those tests were also retested by microagglutination test (MAT) in National Tularemia Reference Laboratory of Refik Saydam Hygiene Center, using antigen prepared in the same laboratory from the local strain. The serum samples were also searched for the presence of Brucella and Salmonella antibodies in terms of cross-reactivity. Seropositivity was detected in 71 (29.6%) out of 240 subjects by slide agglutination test (SAT), whereas only 5 (2.1%) gave positive result for total antibody by ELISA. Twenty-five of the 71 SAT positive samples yielded F.tularensis antibodies by MAT, of which 21 were between 1/20-1/40 and four were between 1/80-1/160 titers. However, all of the MAT positive samples (n= 25) were found reactive in Brucella and/or Salmonella antibody tests. One of the four MAT positive samples with 1/40 titer and all of the four MAT positive samples with ≥ 1/80 titer yielded positive results in ELISA. Since MAT gave very high cross reactive results, the five subjects (2.1%) found positive with ELISA were evaluated as seropositive for tularemia. Of those subjects (four were female, one was male; age range: 27-38 years), four were the inhabitants of the same village, and one from another neighboring village. All of the seropositive subjects were dealing with raising livestock and two were also farming. No history of contact with rat and wild animals or tick bite were detected, however it was noted that non-chlorinated fountain water has been used in both of these villages. In conclusion, our data emphasized that, populations inhabiting especially in rural area and dealing with farming and stock raising in our region are at risk for tularemia.

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#20133791   2010/02/10 Save this To Up

Evidence for a functional role of IgE anticitrullinated protein antibodies in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a systemic autoimmune disease involving inflammation of the joints. Among the autoantibodies described in RA, anticitrullinated protein antibodies (ACPAs) are highly specific and predictive for RA. In addition, ACPAs have been implicated in the pathogenesis of RA. However, a direct functional response of immune cells from ACPA(+) RA patients toward citrullinated proteins has not been demonstrated. In this study, we show that exposure to citrullinated antigens leads to activation of basophils from ACPA(+) RA patients within 20 minutes. This was not observed after exposure of basophils to noncitrullinated control antigens or after stimulation of basophils from ACPA(-) RA patients and healthy controls. Basophil activation was correlated with the binding of citrullinated proteins to basophils. Furthermore, serum from ACPA(+) RA patients in contrast to that from ACPA(-) RA patients could specifically sensitize human FcepsilonRI expressing rat basophil cells (RBL), enabling activation by citrullinated proteins. Mast cell degranulation products such as histamine levels were enhanced in synovial fluid of ACPA(+) RA patients as compared with ACPA(-) RA and osteoarthritis patients. In addition, histamine levels in synovial fluid from ACPA(+) RA patients correlated with IgE levels, suggesting degranulation of mast cells by cross-linking IgE. Immunohistochemistry on synovial biopsies demonstrated an increased number of degranulated CD117(+) mast cells in ACPA(+) RA patients; IgE and FcepsilonRI expression in synovial mast cells from ACPA(+) RA patients was increased. In conclusion, our results show an immunological response of immune cells from ACPA(+) RA patients in a citrulline-specific manner. Moreover, these data indicate a role for IgE-ACPAs and FcepsilonRI-positive cells in the pathogenesis of RA.

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#19890359   2009/11/05 Save this To Up

Good Manufacturing Practices production and analysis of a DNA vaccine against dental caries.

To prepare a clinical-grade anti-caries DNA vaccine pGJA-P/VAX and explore its immune effect and protective efficacy against a cariogenic bacterial challenge.

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#16563283   2006/03/27 Save this To Up

[Distribution and property of anti-beta3-adrenoceptor autoantibody in patients with heart failure].

To investigate the biological effects of anti-beta(3) adrenoceptor (beta(3)-AR) autoantibody in the serum of patients with heart failure, which may contribute to a new therapeutic clue for heart failure.

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#16149151   2005/09/08 Save this To Up

Studies on BN rats model to determine the potential allergenicity of proteins from genetically modified foods.

To develop a Brown Norway (BN) rat model to determine the potential allergenicity of novel proteins in genetically modified food.

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#14604537   2003/11/07 Save this To Up

Immunogenicity of venoms from four common snakes in the South of Vietnam and development of ELISA kit for venom detection.

The antigenicity and antigenic relationship between venoms of four common snakes in the South of Vietnam-Trimeresurus popeorum, Calloselasma rhodostoma, Naja naja and Ophiophagus hannah-were studied. Most of venom components expressed antigenicity and produced high titre antivenoms. The venoms share common components and antivenoms cross-reacted along them. Furthermore, cross-reactions were observed among non-common antigens, indicating that they share common epitopes. Hence, using single component as immunogen for species diagnosis of snakebites can reduce cross-reaction, perhaps may not be totally specific. A three-step affinity purification protocol was set up for preparation of species-specific antivenom antibodies. The steps involved affinity chromatography of IgG from hyper-immunized rabbit sera with protein A columns, immuno-affinity chromatography of monovalent antivenom antibodies with respective homologous venom columns, and immuno-absorption of cross-species reacting antibody molecules with heterologous venom columns. The antibodies were then used for construction of an enzyme-linked immunosorbent assay (ELISA) test kit. The kit can differentiate among the four common snake venoms in various types of samples with the detection limit of 0.2-1.6 ng/ml, depending on the type of samples and species of the snake. The efficacy of this kit for snake venom detection was successfully demonstrated in experimental envenomation in rats. Preliminary evaluation with 140 samples taken from 88 human snakebite victims in Vietnam showed that the kit could detect venom in human samples and would be a very useful tool for fast identification of snakebites in clinics.

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