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#28166930   2017/02/07 Save this To Up

Polymeric antigen BLSOmp31 in aluminium hydroxide induces serum bactericidal and opsonic antibodies against Brucella canis in dogs.

Polymeric antigen BLSOmp31 is an immunogenic vaccine candidate that confers protection against Brucella canis in mice. In this preliminary study, the immunogenicity and safety of BLSOmp31 adsorbed to aluminum hydroxide gel (BLSOmp31-AH) were evaluated in Beagle dogs. In addition, the potential to elicit serum antibodies with complement-dependent bactericidal activity and/or to enhance phagocytosis by neutrophils were analyzed. Dogs were immunized three times with BLSOmp31-AH by subcutaneous route, followed by an annual booster. The vaccine elicited specific antibodies 3 weeks after the first immunization. Annual booster induced comparable antibody response as the primary series. Humoral immune response stimulated by BLSOmp31-AH did not interfere with routine agglutination test for canine brucellosis. Antibodies demonstrated a high complement-dependent bactericidal activity against B. canis. Moreover, opsonization by immune serum not only stimulated binding and uptake of the bacteria by neutrophils but effectively enhanced the destruction of B. canis. Specific IgG was detected in 3/4 immunized dogs in preputial secretions. The antibody profile corresponded to a marked Th2 response, since IgG1 prevailed over IgG2 and cellular immune response was not detected in vitro or in vivo. These results require further evaluation in larger field studies to establish the full prophylactic activity of BLSOmp31 against canine brucellosis.

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#24901209   2016/02/12 Save this To Up

Immunosafety and chronic toxicity evaluation of monomethoxypoly(ethylene glycol)-b-poly(lactic acid) polymer micelles for paclitaxel delivery.

To investigate the physicochemical properties, immunosafety and chronic toxicity of monomethoxypoly(ethylene glycol)-b-poly(lactic acid) (mPEG-PLA), a copolymer used as a carrier for paclitaxel (PTX) delivery. The H-Nuclear Magnetic Resonance (H-NMR), dynamic light scattering and fluorescence probe technique were conducted to determine the physicochemical properties of mPEG-PLA copolymer. PTX-loaded polymeric micelles were characterized regarding their particle size, entrapment efficiency (EE), drug loading (DL), in vitro drug release and hemolysis rate. The complement activation in human serum and mast cells degranulation were performed by ELISA and RBL-2H3 cell line in vitro, respectively. The chronic toxicity study was carried out on beagle dogs. The optimized PTX-loaded mPEG-PLA (40/60) micelles showed a particle size of 37 nm and EE of 98.0% with a DL of 17.0% w/w. Transmission electron microscopy (TEM) analyses showed that mPEG-PLA (40/60) micelles have spherical shape with dense core. In vitro release study showed a sustained release for 24 h, and the hemolysis study revealed that mPEG-PLA (40/60) was a safe nanocarrier for intravenous administration. mPEG-PLA (40/60) showed a lower complement activation ability compared to mPEG-PLA (50/50) and Cremophor® EL (Cr EL). Furthermore, the chronic toxicity of PTX-loaded mPEG-PLA (40/60) micelles was significantly lower than those of mPEG-PLA (50/50) and Cr EL.

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#23497041   2013/03/20 Save this To Up

Specific humoral and cellular immunity induced by Trypanosoma cruzi DNA immunization in a canine model.

Chagas disease has a high incidence in Mexico and other Latin American countries. Because one of the most important known methods of prevention is vector control, which has been effective only in certain areas of South America, the development of a vaccine to protect people at risk has been proposed. In this study, we assessed the cellular and humoral immune response generated following immunization with pBCSP and pBCSSP4 plasmids containing the genes encoding a trans-sialidase protein (present in all three forms of T. cruzi) and an amastigote specific glycoprotein, respectively, in a canine model. Thirty-five beagle dogs were divided randomly into 5 groups (n=7) and were immunized twice intramuscularly with 500 μg of pBCSSP4, pBCSP, pBk-CMV (empty plasmid) or saline solution. Fifteen days after the last immunization the 4 groups were infected intraperitoneally with 500,000 metacyclic trypomastigotes. The fifth group was unimmunized/infected. The parasitaemia in the immunized/infected dogs was for a shorter period (14 vs. 29 days) and the parasite load was lower. The concentration of IgG1 (0.612±0.019 O.D.) and IgG2 (1.167±0.097 O.D.) subclasses was measured (absorbance) 15 days after the last immunization with both recombinant plasmids, the majority of which were IgG2. The treatment of parasites using the serum from dogs immunized with pBCSP and pBCSSP4 plasmids produced 54% (±11.8) and 68% (±21.4) complement-mediated lysis, respectively. At 12 h post immunization, an increase in cytokines was not observed; however, vaccination with pBCSSP4 significantly increased the levels of IFN-γ and IL-10 at 9 months post-infection. The recombinant plasmid immunization stimulated the spleen cell proliferation showing a positive stimulatory index above 2.0. In conclusion, immunization using both genes effectively induces a humoral and cellular immune response.

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#23159336   2013/01/16 Save this To Up

Complement activation associated with polysorbate 80 in beagle dogs.

Polysorbate 80 (Tween® 80) is the most extensively used surfactant in parenteral drug formulation. Its application as an adjunct for intravenous drug administration is approved by the Food and Drug Administration. However, severe hypersensitive reactions, which are typical non-immune anaphylactic reactions (pseudoallergy) characterized by the release of histamine and unvaried IgE antibodies, have been associated with Tween® 80. In order to explore the non-immune anaphylactic mechanisms of Tween® 80, we performed in vivo experiments to assess the changes in physiological and hematologic indicators after intravenous injection of Tween® 80 into dogs. Tween® 80 induced the release of histamine, and a 2-fold increase in SC5b-9, 2.5-fold increase in C4d, 1.3-fold increase in Bb, while IgE remained unchanged. It also produced changes in pulmonary pressure, systemic pressure and ECG. In in vitro experiments, Tween® 80 was incubated with dog serum in the presence of an inhibitor of complement activation (EGTA/Mg(2+)). Under these conditions, Tween® 80 increased the contents of C4d and Bb. The results of this study reveal that Tween® 80 can cause cardiopulmonary distress in dogs and activate the complement system through classical and alternative pathways as indicated in both in vivo and in vitro preparations. Moreover, they demonstrate the utility of the beagle dog as an animal model for the study of complement activation-related pseudoallergy. These findings raise concerns with regard to the indiscriminate use of Tween® 80 in clinical applications.

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#20815217   2010/09/06 Save this To Up

[Material and mechanisms induced pseudo allergic reactions of Yuxingcao injection].

To investigate the characteristics, sensitizin and the mechanism of pseudo allergic reaction induced by Yuxingcao injection.

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#21351726   2011/02/28 Save this To Up

Pharmacokinetics and plasma protein binding of rutin deca (H-) sulfate sodium.

Rutin deca (H-) sulfate sodium (RDS) possesses very good activity as an inhibitor of the complement system of warm-blooded animals and HIV. An ion-pair coupled with solid-phase extraction technique (IP-SPE) was developed to extract RDS from rat plasma, urine, bile and protein solution samples. The assay was applied to pharmacokinetics of RDS, including plasma pharmacokinetics, excretion and protein binding studies. After i.v. 5, 20 and 100 mg x kg(-1) RDS via tail vein in rats, the plasma concentration-time profiles were fitted using 3P97 software. The average terminal half-life (t(1/2)) was 3.432 +/- 0.185 2 h. The relationship of dose and AUC of RDS was linear within the dosage range. This suggested that the disposition of RDS in rats belong to linear kinetics and the pharmacokinetic parameters of RDS were dose independent. After iv RDS 20 mg x kg(-1) in rats, the biliary excretion amount of parent drug amount was only 0.3181% +/- 0.2087% of given dosage, and the urinary excretion was 86.0% +/- 6.1% in 36 h. Ultrafiltration techniques were applied to determine the protein binding of RDS in plasma (from SD rat, Beagle dog and human), human serum albumin (HSA) and human alpha1-acid glycoprotein (AGP). The mean protein binding rate in plasma of SD rat, Beagle dog and human plasma of RDS were 80%-90%, in which the range of concentration of RDS was 5 to 100 microg x mL(-1). The protein binding to HSA was 85.7% +/- 1.3% and 14.0% +/- 3.2% to AGP.

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#11061633   2001/02/27 Save this To Up

Establishment of a canine monocyte cell line.

A canine monocyte cell line was established from the peripheral blood sample collected from a healthy young male Beagle dog. The cloned cells grew easily and were serially passaged in vitro in the medium, a slight modification further made on Dulbecco's modified Eagle medium, supplemented with 10% fetal bovine serum. The morphology of single cell was shown in triangular or round form, however, it became epithelioid in a densely grown monolayer. Non-specific esterase was detected in all cells by a cytochemical examination. The cells reacted rapidly to the addition of a small amount of LPS and differentiated to the cells of morphologically typical macrophages. Both complement receptor (CD35) and Fc gamma receptor (CD64) were demonstrated on the cell membrane.

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#8850363   1997/02/04 Save this To Up

Biochemical characterization of x-ray contrast media.

Eleven ionic and nonionic contrast media were compared in parallel regarding their effects on various biochemical parameters in vitro. Partition coefficient, protein binding, release of histamine, hemolysis inhibition and complement activation were determined as well as inhibition of various enzymes. Additionally, incompatibilities between contrast media and intravascular drugs that often are coadministered were determined.

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#7650902   1995/09/27 Save this To Up

Analysis of a polyacrylamide gel electrophoretogram of beagle serum protein by laser densitometer.

Beagle serum proteins were separated by polyacrylamide gel electrophoresis (PAGE) and the electrophoretograms were examined by one- and two-dimensional analyses with a laser densitometer. In order from the anodic side of the PAGE pattern, pre-albumin, hexokinase, tyrosinase, alkaline phosphatase, urease, and aldehyde dehydrogenase were assumed to be present based on Rf and Mw. Serum albumin, lactate dehydrogenase, and catalase appeared to be present based on a comparison of their electrophoretic mobility with that of protein standards of known Mw. Verification of beagle serum protein fractions by immunofixation electrophoresis and western blotting electrophoresis, with rabbit anti-human serum, indicated alpha 1-antitrypsin, albumin, haptoglobin, ceruloplasmin, C3c complement, IgG, and IgA. Serum protein fraction values (%) obtained by one- and two-dimensional analyses were similar.

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#3876298   1985/11/04 Save this To Up

Glucocorticosteroid effects on dog and rat serum complement.

Inhibiting properties of glucocorticosteroids in vivo on the complement system are not fully recognized. Dexamethasone and prednisolone (10(-7)-10(-3)mol/l.) have no direct effects on serum complement of the dog, rat, human and guinea pig. Yet after subcutaneous application of dexamethasone (0.4 mg/kg) on two consecutive days to healthy beagle dogs the total functional complement is reduced to about 60% of the normal level within the next three days. This effect could be shown to be dose-dependent in male beagle dogs. Partial recovery of the depressed complement level (CH50 U/ml) occurs after the 6th day without treatment. In parallel a decrease in serum C3 is observed. The qualitative serum protein pattern documented by cellulose acetate film electrophoresis (CAF) reflects the decrease of C3 by a reduced beta 1-beta 2 level with a concomitant increase of the alpha 2 serum proteins. In contrast, serum complement, C3 and the serum CAF pattern in rats are not much altered by prednisolone (2 X 15 mg/kg s.c.) or dexamethasone (2 X 2 mg/kg s.c.). Thus the rat appears relatively insensitive to humoral changes induced by steroid alone. Altered liver protein synthesis of complement and other acute phase proteins appear responsible for the profound steroid effects observed in the dog.

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