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Search results for: Anti Aspergillus Purified Preparations

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#29719541   2018/04/17 To Up

A Reappraisal on the Potential Ability of Human Neutrophils to Express and Produce IL-17 Family Members : Failure to Reproducibly Detect It.

Neutrophils are known to perform a series of effector functions that are crucial for the innate and adaptive responses, including the synthesis and secretion of a variety of cytokines. In light of the controversial data in the literature, the main objective of this study was to more in-depth reevaluate the capacity of human neutrophils to express and produce cytokines of the IL-17 family . By reverse transcription quantitative real-time PCR, protein measurement commercial ELISA, immunohistochemistry (IHC) and immunofluorescence (IF), flow cytometry, immunoblotting, chromatin immunoprecipitation (ChIP), and ChIP-seq experiments, we found that highly pure (>99.7%) populations of human neutrophils do not express/produce IL-17A, IL-17F, IL-17AF, or IL-17B mRNA/protein upon incubation with a variety of agonists. Similar findings were observed by analyzing neutrophils isolated from active psoriatic patients. In contrast with published studies, IL-17A and IL-17F mRNA expression/production was not even found when neutrophils were incubated with extremely high concentrations of IL-6 plus IL-23, regardless of their combination with inactivated hyphae or conidia from . Consistently, no deposition of histone marks for active (H3K27Ac) and poised (H3K4me1) genomic regulatory elements was detected at the IL-17A and IL-17F locus of resting and IL-6 plus IL-23-stimulated neutrophils, indicating a closed chromatin conformation. Concurrent experiments revealed that some commercial anti-IL-17A and anti-IL-17B antibodies (Abs), although staining neutrophils either spotted on cytospin slides or present in inflamed tissue samples by IHC/IF, do not recognize intracellular protein having the molecular weight corresponding to IL-17A or IL-17B, respectively, in immunoblotting experiments of whole neutrophil lysates. By contrast, the same Abs were found to more specifically recognize other intracellular proteins of neutrophils, suggesting that their ability to positively stain neutrophils in cytospin preparations and, eventually, tissue samples derives from IL-17A- or IL-17B-independent detections. In sum, our data confirm and extend, also at epigenetic level, previous findings on the inability of highly purified populations of human neutrophils to express/produce IL-17A, IL-17B, and IL-17F mRNAs/proteins , at least under the experimental conditions herein tested. Data also provide a number of justifications explaining, in part, why it is possible to false positively detect IL-17A-neutrophils.
Nicola Tamassia, Fabio Arruda-Silva, Federica Calzetti, Silvia Lonardi, Sara Gasperini, Elisa Gardiman, Francisco Bianchetto-Aguilera, Luisa Benerini Gatta, Giampiero Girolomoni, Alberto Mantovani, William Vermi, Marco A Cassatella

1362 related Products with: A Reappraisal on the Potential Ability of Human Neutrophils to Express and Produce IL-17 Family Members : Failure to Reproducibly Detect It.

10 1mg25 5 100 UG50 µg100 μg50 100ul0.1 mg 1 G

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#17962084   2007/10/24 To Up

In vitro studies on liposomal amphotericin B obtained by supercritical carbon dioxide-mediated process.

Nanotechnology in drug delivery is a rapidly expanding field. Nanosized liposomal preparations are already in use for efficient drug delivery with better therapeutic indices. Existing methods of liposome preparation are limited by problems of scale-up, difficulty in controlling size, and intercalation efficiency. Here we prepare amphotericin B-intercalated liposomes by a novel process where amphotericin B and purified phosphatidyl choline are solubilized in suitable solvent and precipitated in supercritical fluid carbon dioxide (known as a gas antisolvent technique), to obtain microsized particles that are subsequently introduced into a buffer solution. The morphology of liposomes was characterized through a phase-contrast microscope, and the particle size distribution studied by laser technique showed nanosize with a narrow range of size distribution (between 0.5 and 15 microm) and a higher intercalation efficiency. In vitro studies conducted using Aspergillus fumigatus (MTCC 870) strain proved to be efficient in the retardation of the growth of the organism.
Udaya Sankar Kadimi, Deepan Raja Balasubramanian, Usha Rani Ganni, Manohar Balaraman, Venkateswaran Govindarajulu

1293 related Products with: In vitro studies on liposomal amphotericin B obtained by supercritical carbon dioxide-mediated process.

100.00 ug2ug100ug1 g100ug100 mg0.1mg

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#17933209   // To Up

Development and evaluation of a microemulsion formulation for transdermal delivery of terbinafine.

The aim of the present study is to develop and evaluate microemulsion formulations for Terbinafine (TB) with a view to enhance its permeability through the skin and provide release for 24 h. Various o/w microemulsions were prepared by the spontaneous emulsification method. Oleic acid was chosen as the oil phase, Caprylo caproyl macrogol-8- glyceride (Labrasol S) and purified diethylene glycol monoethyl ether (Transcutol P) were used as surfactant and cosurfactant, respectively, on the basis of solubility studies. Pseudoternary phase diagrams were constructed to obtain the concentration range of oil, surfactant, cosurfactant, and water for microemulsion formulation. The optimized microemulsion consisted of 2% w/w TB, 8% w/w oleic acid, 31% w/w labrasol S, 31% w/w transcutol P, and 30% w/w distilled water. Permeability parameters like Jss and Kp were found to be significantly higher for formulation F4 as compared to other formulations (P < 0.05). Microbiological studies of TB in microemulsion showed better anti-fungal activity against Candida albicans and Aspergillus flavus as compared to marketed product (P < 0.05).
S Baboota, A Al-Azaki, K Kohli, J Ali, N Dixit, F Shakeel

1857 related Products with: Development and evaluation of a microemulsion formulation for transdermal delivery of terbinafine.

500 G100ul300 units200ul2.5 mg25 mg10 mg1000 tests1000 TESTS/0.65ml10 mg

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#16714088   2006/04/07 To Up

Esterification of ferulic acid with polyols using a ferulic acid esterase from Aspergillus niger.

Commercially available enzyme preparations were screened for enzymes that have a high ability to catalyze direct ester-synthesis of ferulic acid with glycerol. Only a preparation, Pectinase PL "Amano" produced by Aspergillus niger, feruloylated glycerol under the experimental conditions. The enzyme responsible for the esterification was purified and characterized. This enzyme, called FAE-PL, was found to be quite similar to an A. niger ferulic acid esterase (FAE-III) in terms of molecular mass, pH and temperature optima, substrate specificity on synthetic substrates, and the N-terminal amino acid sequence. FAE-PL highly catalyzed direct esterification of ferulic acid and sinapinic acid with glycerol. FAE-PL could feruloylate monomeric sugars including arabinose, fructose, galactose, glucose, and xylose. We determined the suitable conditions for direct esterification of ferulic acid with glycerol to be as follows: 1% ferulic acid in the presence of 85% glycerol and 5% dimethyl sulfoxide at pH 4.0 and 50 degrees C. Under these conditions, 81% of ferulic acid could be converted to 1-glyceryl ferulate, which was identified by (1)H-NMR. The ability of 1-glyceryl ferulate to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals was higher than that of the anti-oxidant butyl hydroxytoluene.
Moriyasu Tsuchiyama, Tatsuji Sakamoto, Tomoyuki Fujita, Shuichi Murata, Haruhiko Kawasaki

2049 related Products with: Esterification of ferulic acid with polyols using a ferulic acid esterase from Aspergillus niger.

1 G 25 G500 mg5 mg500 mg100 assays 5 G 25 G 5 G 100 G10 ML

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#10361723   // To Up

Glucose 1- and 2-oxidases from fungal strains: isolation and production of monoclonal antibodies.

Monoclonal antibodies (Mabs) against purified glucose 2-oxidase (EC 1.1.3.10) from Coriolus versicolor were raised by hybridoma technology using Sp2/0 myeloma cells as a fusion partner. Hybrid growth was observed in 42% of culture wells and 30% of these (i.e. 30 culture wells) contained anti-glucose 2-oxidase activity. Three positive wells containing hybrid cell lines were selected and cloned twice by the limiting dilution method and two hybridoma clones (E1A5 and E1A6) secreting Mabs were selected at random for purification and characterisation purposes. Both cell lines secreted Mabs of IgM class which were purified by gel filtration chromatography on a Sephacryl S-200 column with a final recovery of 80% and a purification factor of 16. The purified preparations were apparently homogeneous on native PAGE running with a M(r) of 950 kDa. Mabs were highly specific for glucose 2-oxidase as determined by Western blotting. These Mabs also crossreacted with glucose 1- and 2-oxidases from other fungal sources (Phanerochaeta chrysosporium, Penicillium amagasakiense and Aspergillus niger) as determined by Western blotting and by ELISA. Both glucose 1- and 2-oxidases from C. versicolor, P. chrysosporium, P. amagasakiense and A. niger were purified by hydrophobic interaction chromatography on Sepharose 4B-triazine dye with a recovery of enzyme activity in the range 85-92%. Purified preparations of glucose oxidases from fungal strains were apparently homogeneous on native PAGE. Glucose 2-oxidases were more hydrophobic than glucose 1-oxidases as determined by their chomatographic behaviour on Sepharose 4B-Cibacron Red G-E which could be used to study their roles in lignin biodegradation.
A Karmali, P Oliveira

2035 related Products with: Glucose 1- and 2-oxidases from fungal strains: isolation and production of monoclonal antibodies.

1000 TESTS/0.65ml0.1 mg1 ml100.00 ug100.00 ug100 ug100 ug200.00 ug100.00 ug100.00 ug100 ul100 ug

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#8755864   // To Up

Isolation of a gene involved in 1,3-beta-glucan synthesis in Aspergillus nidulans and purification of the corresponding protein.

Saccharomyces cerevisiae has two highly homologous genes, FKS1 and FKS2, which encode interchangeable putative catalytic subunits of 1,3-beta-glucan synthase (GS), an enzyme that synthesizes an essential polymer of the fungal cell wall. To determine if GS in Aspergillus species is similar, an FKS homolog, fksA, was cloned from Aspergillus nidulans by cross-hybridization, and the corresponding protein was purified. Sequence analysis revealed a 5,716-nucleotide coding region interrupted by two 56-bp introns. The fksA gene encodes a predicted peptide of 229 kDa, FksAp, that shows a remarkable degree of conservation in size, charge, amino acid identity, and predicted membrane topology with the S. cerevisiae FKS proteins (Fksps). FksAp exhibits 64 and 65% identity to Fks1p and Fks2p, respectively, and 79% similarity. Hydropathy analysis of FksAp suggests an integral membrane protein with 16 transmembrane helices that coincide with the transmembrane helices of the Saccharomyces Fksps. The sizes of the nontransmembrane domains are strikingly similar to those of Fks1p. The region of FksAp most homologous to the Saccharomyces FKS polypeptides is a large hydrophilic domain of 578 amino acids that is predicted to be cytoplasmic. This domain is 86% identical to the corresponding region of Fks1p and is a good candidate for the location of the catalytic site. Antibodies raised against a peptide derived from the FksAp sequence recognize a protein of approximately 200 kDa in crude membranes and detergent-solubilized active extracts. This protein is enriched approximately 300-fold in GS purified by product entrapment. Purified anti-FksAp immunoglobulin G immunodepletes nearly all of the GS activity in crude or purified extracts when Staphylococcus aureus cells are used to precipitate the antibodies, although it does not inhibit enzymatic activity when added to extracts. The purified GS is inhibited by echinocandins with a sensitivity equal to that displayed by whole cells. Thus, the product of fksA is important for the activity of highly purified preparations of GS, either as the catalytic subunit itself or as an associated copurifying subunit that mediates susceptibility of enzymatic activity to echinocandin inhibition.
R Kelly, E Register, M J Hsu, M Kurtz, J Nielsen

1263 related Products with: Isolation of a gene involved in 1,3-beta-glucan synthesis in Aspergillus nidulans and purification of the corresponding protein.

5ug100ug Lyophilized100ug Lyophilized1 Set1 Set1 Set1 Set100ug Lyophilized1 Set1 Set1 Set1 Set

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#3097110   // To Up

Enzyme profile and immunochemical characterization of Aspergillus fumigatus antigens.

We have compared the immunochemical characteristics of culture-filtrate antigens (Ag) from Aspergillus fumigatus extracted in our laboratory with commercially available Ags. A total of 20 different preparations were studied for protein and carbohydrate content, presence of endotoxins, mycotoxins, and hemolytic toxins. These extracts were analyzed by two-dimensional electrophoresis for protein components. The immunogenicity of the preparations was determined by rocket electrophoresis with rabbit anti-A. fumigatus sera and by agar gel diffusion with sera from patients with allergic bronchopulmonary aspergillosis, aspergilloma, and normal control subjects. In order to have dependable immunologic results, the Ags must be sufficiently pure and reproducible. Until such time as pure and standardized Ags are available, the crude Ags used should be characterized to the extent that adequate reproducibility between preparations can be ascertained. The enzyme profile of the Ag preparations provides a fair indication of the quality of antigenic components, and together with other immunochemical parameters, it will be of use in determining the suitability of the extracts in immunodiagnosis. Immunochemical results demonstrate that commercial Ags contain less proteins and carbohydrates and fewer enzymes than the homemade antigens. In addition, fewer patients demonstrated specific precipitins against commercial Ags than with homemade Ags. This study once again confirms the need for pure standardized Ags for studying the immunologic response in patients with Aspergillus-induced diseases. Until such preparations are readily available, partially purified or crude Ags with known immunochemical properties and enzyme profile may be the choice for immunodiagnosis.
V P Kurup, A Resnick, G H Scribner, M Gunasekaran, J N Fink

1940 related Products with: Enzyme profile and immunochemical characterization of Aspergillus fumigatus antigens.

100μg1x96 well plate50 ul100 mg 500 ml 100ul100.00 ug1-99 mg/ml/ea price x 210 mg1mg100ug100ug

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