PubMed Health⌕ Search

Biomedical subjects

P A Castric

Publications and source records attributed to P A Castric.

16 recordsLinked to original sources
PubMed1997

Peptide epitope mapping in vaccine development: introduction.

Protection from infectious disease by the host immune response requires specific molecular recognition of unique antigenic determinants of a given pathogen. An epitope is an antigenic determinant which: 1) specifically stimulates the immune response (either B or T cell mediated); and 2) is acted upon by the products of these protective mechanisms. In B cell immunity, antibodies produced from stimulation by specific epitopes recognize and bind to these same antigenic structures. Identification of protective epitopes is extremely valuable to successful vaccine development. In order to be protective these antibodies must, in addition to recognition and binding, interfere with some vital step in pathogenesis such as adherence or toxin action. Protein B cell epitopes are frequently composed of the side chains (R-groups) of the amino acids found at solvent-exposed surfaces. These epitopes are classified as continuous (also linear or sequential) if composed of a single antibody-recognizing element located at a single locus of the primary structure. They are discontinuous (or assembled) if more than one physically separated entity is involved. T cell epitopes are peptides on the surface of antigen-presenting cells (macrophages, dendritic cells, and B cells) that are bound to major histocompatibility proteins; the T cell recognizes this peptide-MHC complex.

Animals↗
PubMed1994

Differentiation of Pseudomonas aeruginosa pili based on sequence and B-cell epitope analyses.

The nucleotide sequences of three previously undescribed Pseudomonas aeruginosa pilin structural genes are presented. Comparisons of deduced pilin primary structure and flanking DNA sequence allowed placement of these and six previously published sequences into one of two groups. Epitope mapping, using overlapping immobilized peptides representing the pilin primary structure, with antipilin monoclonal antibodies revealed several B-cell determinants grouped near the carboxyl terminus of P. aeruginosa 1244 pilin. One determinant was found to reside near the pilin constant region. These determinants were found associated with the pili of 31 of 95 P. aeruginosa clinical isolates.

Amino Acid Sequence↗
PubMed1989

Cloning and sequencing of the Pseudomonas aeruginosa 1244 pilin structural gene.

The pilin structural gene of Pseudomonas aeruginosa 1244 was cloned in both cosmids and lambda. Expression of the cloned gene was detected in P. aeruginosa strains PAO2003, PA103, and 653A by an immunoblot reaction utilizing monoclonal antibodies. Western blot analysis showed that pilin expressed from the cloned gene was slightly larger than native 1244 pilin when produced in strains PAO2003 and 653A, but distinctly smaller in PA103. Bacteriophages specific for the 1244 pilus did not lyse strain PAO2003 containing the cloned 1244 pilin gene, indicating that functional 1244 pili were not assembled in this recombinant strain. Nucleotide sequencing revealed a coding region which when translated would produce a 15,615 dalton peptide. The amino-terminal region of this peptide is identical with published pilin sequences. While the rest of the peptides are generally dissimilar, common residues are seen within potentially antigenic regions.

Amino Acid Sequence↗
PubMed1983

Method for rapid detection of cyanogenic bacteria.

An agar plate method is described in which the production of hydrogen cyanide by as many as 50 microbial isolates per plate may be detected. Cyanide produced by the organisms reacts with copper(II) ethylacetoacetate and 4,4'-methylenebis-(N,N-dimethylaniline) in a paper disk suspended above the microbial colonies. Cell growth occurs in depressions in the agar surface, which allows separation of colonies and enhances sensitivity of hydrogen cyanide detection.

Journal Article↗
PubMed1983

Hydrogen cyanide production by Pseudomonas aeruginosa at reduced oxygen levels.

Hydrogen cyanide production by Pseudomonas aeruginosa growing in a synthetic medium required aerobosis but operated efficiently at low dissolved oxygen concentration. Half maximum levels of cyanogenesis occurred at 0.015 microM oxygen; maximum cyanogenesis occurred over a wide range, 0.1-180 microM, of oxygen concentrations. These cells lost the ability to produce cyanide upon aerobic incubation in the absence of both the carbon energy source (L-glutamate) and the metabolic precursor of hydrogen cyanide (glycine). This loss of cyanogenesis was dependent on oxygen concentration; 1.0 microM oxygen produced no detectable loss, whereas 180 microM oxygen caused a rapid decline in cyanogenic ability. The endogenous cyanide production rate of cells in the presence of carbon energy source was not significantly influenced by oxygen concentration. During the batch culture cycle, the acquisition of the ability to produce HCN was preceded by oxygen reduction to growth-limiting levels. Cells which had lost the ability to produce hydrogen cyanide by oxygen treatment required protein synthesis before they could again become cyanogenic.

Glycine↗
PubMed1980

In vitro susceptibility of Pseudomonas aeruginosa to carbenicillin, glycine, and ethylenediaminetetraacetic acid combinations.

Striking bacterial activity against Pseudomonas aeruginosa 9D-2 was achieved by glycine-carbenicillin, ethylenediaminetetraacetic acid-carbenicillin, and glycine-ethylenediaminetetraacetic acid combinations, whereas none of the agents used alone was capable of the same degree of bactericidal activity. Studies using a microtiter modification of the checkerboard technique were performed to evaluate the comparative activity of these antimicrobial combinations. Isobolograms showed synergistic effects with carbenicillin-glycine, carbenicillin-ethylene-diaminetetraacetic acid, and glycine-ethylenediaminetetraacetic acid combinations. Bacterial growth inhibitory curves with subinhibitory concentrations of these agents in combination confirmed these findings.

Carbenicillin↗
PubMed1977

The effect of inorganic phosphate on cyanogenesis by Pseudomonas aeruginosa.

The biosynthesis of hydrogen cyanide (HCN) by a strain of Pseudomonas aeruginosa is found to be significantly influenced by inorganic phosphate. Optimum HCN production occurs when the phosphate concentration is between 1 and 10 mM. Above and below this concentration the amount of HCN produced decreases sharply and at 0.1 and 100 mM phosphate low HCN production occurs. If a culture growing at 0.1 mM phosphate and producing low HCN is shifted to 10 mM phosphate, HCN biosynthesis resumes. Experiments with chloramphenicol indicate that de novo-protein synthesis is required for the process.

Bacterial Proteins↗
PubMed1977

Purification and properties of the urea amidolyase from Candida utilis.

Urea amidolyase was purified to homogeneity from extracts of Candida utilis. The purification involves protamine sulfate precipitation, ammonium sulfate precipitation, polyethylene glycol precipitation, Sepharose 6B gel filtration, DEAE-cellulose column chromatography, and hydroxylapatite column chromatography. The final preparation is pure as judged by disc-gel electrophoresis. The molecular weight of urea amidolyase, as determined by gel filtration and disc-gel electrophoresis, is between 500,000 and 520,000. Treatment with sodium dodecyl sulfate results in two peptides with molecular weights of 70,000 and 170,000. The urea carboxylase and allophanate hydrolase activities of urea amidolyase may be distinguished from one another on the basis of (a) the effect of the stabilizers, urea and glycerol, (b) the effect of storage pH on activity, and (c) selective inhibition by sulfhydryl reagents.

Candida↗
PubMed1977

Glycine metabolism by Pseudomonas aeruginosa: hydrogen cyanide biosynthesis.

Hydrogen cyanide (HCN) production by Pseudomonas aeruginosa in a synthetic medium is stimulated by the presence of glycine. Methionine enhances this stimulation but will not substitute for glycine as a stimulator of cyanogenesis. Threonine and phenylalanine are effective substitutes for glycine in the stimulation of HCN production. Glycine, threonine, and serine are good radioisotope precursors of HCN, but methionine and phenylalanine are not. Cell extracts of P. aeruginosa convert [14C]threonine to [14C]glycine. H14CN is produced with low dilution of label from either [1-14C]glycine or [2-14C]glycine, indicating a randomization of label either in the primary or secondary metabolism of glycine. When whole cells were fed [1,2-14C]glycine, cyanide and bicarbonate were the only radioactive extracellular products observed.

Carbon Radioisotopes↗
PubMed1976

Urea amidolyase of Candida utilis. Characterization of the urea cleavage reactions.

Evidence is presented that the enzymes catalyzing the three reactions involved in urea cleavage in Candida utilis, biotin carboxylation, urea carboxylation, and allophanate hydrolysis occur as a complex of enzymes. The allophanate-hydrolyzing activity could not be separated from the urea-cleaving activity using common methods of protein purification. Further, urea cleavage and allophanate hydrolysis activities are induced coordinately in cells grown on various nitrogen sources. The reactions involved in urea cleavage can be distinguished from one another on the basis of their sensitivities to (a) heat, (b) pH, and (c) chemical inhibitors. Evidence is presented for the product of the first reaction in urea cleavage, biotin carboxylation. Production of carboxylated enzyme is ATP dependent and avidin sensitive. Carboxylated enzyme is not observed in the presence of 1 mM urea.

Acetamides↗
PubMed1975

Hydrogen cyanide, a secondary metabolite of Pseudomonas aeruginosa.

Seventy-four of 110 strains of Pseudomonas aeruginosa tested produced detectable amounts of HCN from growth in 2% peptone or nutrient agar. Of the 25 species of12 bacterial and fungal genera tested, other than P. aeruginosa, only P. fluorescens and P. polycolor gave positive HCN tests. Cyanide is produced after cessation of active growth. Iron was stimulatory to cyanogenesis in concentration above 1 muM, while copper, zinc, cobalt, and manganese at concentrations of 20 muM had no effect. Cyanogenesis id dependent on the temperature of incubation within ranges which allow complete growth. Inorganic phosphate in concentrations between 90 and 300 mM allows growth but inhibits HCN production. Growth of cells anaerobically, using nitrate as the electron acceptor, results in low cyanide yields, which can be partially reversed by subsequent aerobic incubation. These results indicate that HCN is a secondary metabolite of P. aeruginosa.

Ferric Compounds↗
PubMed1971

Formation of -cyanoalanine by O-acetylserine sulfhydrylase.

Cell-free extracts of Bacillus megaterium form beta-cyanoalanine (beta-CNA)-(14)C from Na(14)CN and l-cysteine, O-acetyl-l-serine or, to a lesser extent, l-serine. However, the presence of cyanide in the growth medium does not increase the capacity of cell extracts to catalyze the formation of beta-CNA from cysteine and cyanide. The formation of beta-CNA is readily detected in extracts of cells grown in synthetic media with sulfate or l-djenkolic acid as sulfur sources; such cells also exhibit an increased ability to form cysteine when compared with cells grown on cysteine as the sulfur source. beta-CNA formation could not be detected in extracts of cells grown on cysteine as the sulfur source. A 40-fold purification of the O-acetyl-serine sulfhydrylase resulted in the co-purification of the beta-CNA-forming activity. The sulfhydrylase and the beta-CNA-forming activity co-chromatographed on diethyl-aminoethyl cellulose and Sephadex G-100.

Acetates↗