Validation of eGFP fluorescence intensity for testing in vitro cytotoxicity according to ISO 10993-5.

ISO 10993-5 provides one of the accepted standards for testing the biotoxicity of new materials. All of the recommended test procedures rely upon the uptake or metabolism of dye by living cells. Results of direct contact tests can be potentially compromised by interaction or adsorption of the dye or its metabolic products.

Treatment of middle ear ventilation disorders: sheep as animal model for stenting the human Eustachian tube--a cadaver study.

Eustachian tube disorders can lead to chronic otitis media with consecutive conductive hearing loss. To improve treatment and to develop new types of implants such as stents, an adequate experimental animal model is required. As the middle ear of sheep is known to be comparable to the human middle ear, the dimensions of the Eustachian tube in two strains of sheep were investigated.

Cloning and expression of candidate allergens from Culicoides obsoletus for diagnosis of insect bite hypersensitivity in horses.

Insect bite hypersensitivity (IBH) is an IgE-mediated (Type I) hypersensitivity reaction induced by allergens from biting midges of the Culicoides spp. The aim of the present study was to identify, clone and express recombinant allergens from C. obsoletus, the main species found feeding on horses in the Netherlands, by sequence homology searches on the C.

Culicoides obsoletus extract relevant for diagnostics of insect bite hypersensitivity in horses.

Insect bite hypersensitivity (IBH) is an allergic dermatitis in horses caused by the bites of Culicoides species. The aim of the present study was to evaluate the applicability of whole body extracts of C. obsoletus (the main species found feeding on horses in the Netherlands), C.

Identification, expression and characterisation of a major salivary allergen (Cul s 1) of the biting midge Culicoides sonorensis relevant for summer eczema in horses.

Salivary proteins of Culicoides biting midges are thought to play a key role in summer eczema (SE), a seasonal recurrent allergic dermatitis in horses. The present study describes the identification, expression and clinical relevance of a candidate allergen of the North American midge Culicoides sonorensis. Immunoblot analysis of midge saliva revealed a 66 kDa protein (Cul s 1) that was bound by IgE from several SE-affected (SE+) horses.

Monitoring of basophil sensitization to antigens of the cat flea (Ctenocephalides felis felis): a new tool for the diagnosis of feline flea bite hypersensitivity?

Suitability of blood basophils for in vitro diagnosis of flea allergy dermatitis (FAD) or flea bite hypersensitivity was studied in cats. A functional in vitro test (FIT) for sensitized type I allergic effector cells was used to evaluate the degree and kinetics of in vivo basophil sensitization against flea antigens in cats under long-term flea exposure. FIT results were compared with intradermal (IDT) and serological testing.

Transmission of Chlamydophila pneumoniae from dendritic cells to macrophages does not require cell-to-cell contact in vitro.

Chlamydophila pneumoniae (C. pneumoniae) has been detected in macrophages (Mø) and dendritic cells (DC) in vascular diseases. To understand the importance of these cell types in C.

Comparison of cellular and humoral immunoassays for the assessment of summer eczema in horses.

The objective of this study was to compare and analyze three common diagnostic methods for summer eczema (SE) in horses, an allergic dermatitis caused by bites of Culicoides spp. Nine horses with a medical history of SE and nine control animals were intradermally challenged with whole body extracts (WBE) and the saliva of a native (C. nubeculosus) and exotic (C.

Collection and analysis of salivary proteins from the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae).

Salivary proteins of hematophagous Culicoides spp. are thought to play an important role in pathogen transmission and skin hypersensitivity. Analysis of these proteins, however, has been problematic due to the difficulty in obtaining adequate amounts of secreted Culicoides saliva.

Modulation of carp (Cyprinus carpio) neutrophil functions during an infection with the haemoparasite Trypanoplasma borreli.

Trypanoplasma borreli is an extracellular blood parasite of common carp (Cyprinus carpio) transmitted by fish-biting leeches. The infestation with this parasite in juvenile carp may range between 75% and 100%, especially in fish recovering from the first hibernation period. T.

IgE and IgG antibodies in skin allergy of the horse.

In horses, allergies have been characterized by clinical signs and/or intradermal (i.d.) allergen testing.

Platelet-bound antibodies detected by a flow cytometric assay in cats with thrombocytopenia.

In cats, primary or secondary immune-mediated thrombocytopenia have rarely been described or characterised. The objective of this study was to determine platelet-bound antibodies (PBA) by a flow cytometric assay in both healthy and thrombocytopenic cats. Direct PBA testing was performed in 42 thrombocytopenic cats (platelet counts 6-179 x 10(9)/l, median 56 x 10(9)/l).

Primary immune-mediated hemolytic anemia in 19 cats: diagnosis, therapy, and outcome (1998-2004).

Immune-mediated hemolytic anemia (IMHA) occurs less frequently in cats than in dogs. The value of the Coombs' test (CT) has been questioned, but detailed surveys of its use are lacking. The objective of this study was to describe 19 cats with primary IMHA (pIMHA) and to examine the diagnostic value of the direct CT.

Use of leukocytes as treatment for endometritis in mares experimentally infected with Streptococcus equi subsp. zooepidemicus.

This study compared four treatments for bacterial endometritis in mares experimentally infected with Streptococcus zooepidemicus. Twenty-five mares were used, 20 resistant and five susceptible to endometritis. Mares would be in estrus when infected.

Quantitative and qualitative assessment of serum- and inflammatory mediator-induced migration of carp (Cyprinus carpio) head kidney neutrophils in vitro.

A quantitative transmigration system, permitting the harvest of transmigrated cells for further analysis, was used to study carp head kidney (HK) granulocyte migration in vitro. Pooled carp serum and leukotriene B4 (LTB-4), but not recombinant human C-X-C chemokine ligand 8 (rhCXCL8), recombinant human complement component 5a (rhC5a) or N-formyl-methionyl-leucyl-phenylalanine (FMLP) induced strong migration (up to 70%) of carp HK granulocytes. The transmigrated cells were viable (>or=96%) and uniform (purity >or=97%).

Occurrence of IgE in foals: evidence for transfer of maternal IgE by the colostrum and late onset of endogenous IgE production in the horse.

IgE is the key antibody involved in type I allergies. Allergen mediated crosslinking of IgE bound to high affinity Fcepsilon-receptors on mast cells and basophils stimulates cellular degranulation and release of inflammatory mediators and cytokines. In this report, we demonstrate that IgE antibodies can be transferred from the mother to offspring in horses via the colostrum.

Dexamethasone depresses the expression of L-selectin but not the in vivo migration of bovine neutrophils into the uterus.

There are several indications that periparturient depression of functional properties of polymorphonuclear neutrophilic granulocytes (PMN) may be of great importance for the pathogenesis of genital infections after calving. As the periparturient period is characterized by high plasma levels of corticosteroids, the hypothesis to be tested was that high concentrations of glucocorticoids during the periparturient period suppress uterine PMN in number and functionality. An in vivo endometritis model was applied to examine uterine PMN.

Alpha-1-acid glycoprotein inhibits phorbol ester-induced but not Fc-receptor-induced generation of reactive oxygen species in bovine peripheral blood neutrophils.

Alpha-1-acid glycoprotein (AGP) is an acute-phase protein with anti-inflammatory and immunomodulating properties. AGP is described as a potent inhibitor of the production of reactive oxygen species (ROS) in human neutrophils. However, published reports about the mechanism of inhibition are conflicting.

Flow cytometric evaluation of bound IgG on erythrocytes of anaemic dogs.

The detection of red blood cell (RBC)-bound immunoglobulins in case of anaemia with the direct agglutination test (DAT or Coombs test) has been reported to be of low sensitivity. We therefore tested the applicability of flow cytometry for the detection of canine IgG on RBC using two different IgG-specific secondary reagents: goat-anti-dog IgG (GalphaD-IgG) and rabbit-anti-dog IgG (RalphaD-IgG). Membrane staining RBC samples were performed at 4 degrees C.

Effect of hexavalent carcinogenic chromium on carp Cyprinus carpio immune cells.

Chromium is widely used in industrial processes, and is released into aquatic environments by electroplating, tannery and textile industries. Fishes in natural waters or in aquaculture facilities supplied with these waters are exposed to chromium waste and are presumed to be affected by deposits. Herein, we examine the effect of hexavalent chromium on carp Cyprinus carpio derived immune cells.

[Bacterial pathogen associated molecular pattern and superantigens indirectly induce the accelerated death of bovine neutrophilic granulocytes].

Pathogen associated molecular patterns (PAMPs) and bacterial superantigens have many effects on mononuclear cells (MNC) and macrophages. Influences on neutrophilic granulocytes (PMN), especially by non methylated CpG motifs, gained less attention. Here we investigated whether PAMPs and the superantigen SEA have a direct or indirect influence on the survival rate of bovine PMN.

Some immune parameters in carp cyprinus Carpio susceptible and resistant to the haemoflagellate Trypanoplasma borreli.

The present study addresses aspects of the (specific) immune response of carp to the haemoflagellate Trypanoplasma borreli. Sera of resistant carp contained antibodies, which agglutinated the flagellates in vitro. When flagellates were incubated in fish sera from resistant carp, binding of antibodies to flagellates could be demonstrated by flow cytometry, and T.

Measurement of phagocytosis and oxidative burst of canine neutrophils: high variation in healthy dogs.

Quantitative analysis of phagocytosis and oxidative burst in canine polymorphonuclear (PMN) cells was performed by flow cytometry techniques. Different concentrations of phorbol myristate acetate (PMA) were used to modulate PMN phagocytosis. A low concentration of PMA (3 nmol) resulted in increased phagocytic activity of canine PMN, which could not be enhanced by higher dosages.