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  • Biochemistry and Biotechnology  (10)
  • Candida albicans  (4)
  • 1995-1999  (14)
  • 1960-1964
  • 1
    ISSN: 1573-0832
    Keywords: Candida albicans ; cell wall ; protein and mannoprotein antigens ; Zymolyase ; β-mercaptoethanol ; polyclonal antibodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In order to investigate the antigenic properties of the proteins and mannoproteins present in the cell surface of Candida albicans, and to identify individual antigenic moieties and their distribution, a number of polyclonal antisera were obtained by immunizing rabbits with chemical and enzymatic cell wall extracts obtained from intact cells from both growth forms (yeast and mycelium) of the fungus. Prior to injection, wall moieties present in the extracts were subjected to different treatments and/or purification procedures such as adsorption onto polystyrenelatex microbeads or electrophoretic separation. When used as probes in indirect immunofluorescence assays, the different antisera gave rise to different fluorescence patterns varying in intensity and topological localization of the reactivity in C. albicans cells. When the different antisera were used as probes in Western blots of wall proteinaceous materials solubilized from both blastospores and germ tubes, differences in reactivity and specificity were readily discernible, allowing to identify a number of common and form-specific cell wall components. Of special interest was the fact that one of the antisera raised, after adsorption onto heat-killed blastospores, specifically recognized medium to low molecular weight moieties present only in the cell wall extracts from germ tubes. When this antiserum was used as probe in immunofluorescence assays, reactivity was confined to the hyphal extensions. Together, these observations seem to indicate that the different antibody preparations described in this report could represent important tools in the study of different aspects of the cell wall biology in C. albicans, including the identification and study of the distribution of common and form-specific cell wall-bound antigens.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Mycopathologia 144 (1998), S. 125-129 
    ISSN: 1573-0832
    Keywords: biofilm ; Candida albicans ; Candida tropicalis ; saliva ; serum ; thigmotropism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The biofilm formation of the oral fungal pathogen Candida on denture acrylic strips coated with saliva or serum was examined in relation to the ability to induce hyphae by thigmotropic reaction, using C. albicans (4 isolates), C. glabrata (3 isolates) and C. tropicalis (3 isolates). Both the degree of biofilm formation and the amount of hyphae exhibiting thigmotropism varied depending upon both the species and strains of Candida. Although there was no significant correlation between the amount of hyphae induced by thigmotropic reaction of fungal isolates and biofilm formation on uncoated control specimens (r = 0.577; p 〈 0.05), the ability of hyphae induced by thigmotropic reaction significantly correlated with the amount of both saliva- and serum-admixed biofilms (r = 0.734; p 〈 0.05 and r = 0.793; p 〈 0.01, respectively). Taken together our in vitro data suggested that the hyphal induction by thigmotropic reaction is of importance in candidal biofilm formation on saliva- or serum-coated acrylic surfaces.
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  • 3
    ISSN: 1572-9699
    Keywords: Candida albicans ; cell wall ; culture filtrates ; mannoproteins ; proteins ; surface antigens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Culture filtrate concentrates were obtained fromCandida albicans yeast and mycelial cells grown in the presence of14C-protein hydrolysate for radioactive labeling of cellular polypeptides. Both growth forms released to the medium minor but significant amounts of proteinaceous materials. The analysis of culture filtrate concentrates by means of SDS-polyacrylamide gel electrophoresis and fluorography revealed a similar and complex electrophoretic pattern, though some qualitative and quantitative differences between samples obtained from yeast and mycelial cells were observed. Materials released, mostly composed of mannoproteins as shown by their affinity towards concanavalin A, presented (i) cross-reactivity (by Western immunoblotting and ELISA) against polyclonal antisera to genuine cell wall components (among them the 58-kilodalton fibrinogen-binding mannoprotein) and (ii) high affinity for polystyrene-latex microbeads. Results presented suggest a possible common identity for the molecules shed to the medium and the cell wall protein and mannoprotein constituents.
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  • 4
    ISSN: 0006-3592
    Keywords: metabolic engineering ; carbon metabolism ; Escherichia coli mutants ; microbial growth ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Escherichia coli strains devoid of one or both of the two pyruvate kinase isoenzymes (PKA and PKF), were grown on minimal media in batch fermentations. The strain lacking both PKs showed a 28% decrease on its specific growth rate when compared to the wild type. However, protein and CO2 yields did not change. Using radioactive 1-C14 glucose and collecting the CO2 produced by the cultures, it was found that the mutant lacking both pyruvate kinases, metabolized glucose mainly through the pentose pathway (PP). The increased participation of the PP in glucose metabolism in this strain, was also reflected on the levels of the glucose-6-phosphate and 6-phosphogluconate dehydrogenases.© 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:292-295, 1998.
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  • 5
    ISSN: 0884-3996
    Keywords: chemiluminescence ; flow injection ; isoniazid ; solid-phase reactors ; pharmaceutical analysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A flow injection procedure for the indirect chemiluminescent determination of isoniazid is proposed. The method is performed in a flow-injection manifold provided with a solid-phase reactor. The reactor was made from manganese dioxide physically entrapped by polymerization; the redox reaction isoniazid-manganese dioxide released Mn(II) which was monitored through its inhibitory effect on the reaction between luminol and hydrogen peroxide in presence of potassium hexacyanoferrate(III). The procedure resulted in a linear calibration graph over the range 5-15 mg/L of isoniazid with a sample throughput of 43 samples/h. The influence of foreign compounds was studied and the method was applied to determination of the drug in a pharmaceutical formulation. © 1998 John Wiley & Sons, Ltd.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 56 (1997), S. 62-70 
    ISSN: 0006-3592
    Keywords: osmotic shock ; water permeability ; mixing time constant ; mathematical model ; yeast ; leukemia K562 cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Water permeability (Lp), calculated from the volume variations of cells subjected to an osmotic shock, is classically used to characterize cell membrane properties. In this work, we have shown the importance of the kind of mixing reactor used to measure the Lp parameter. A mathematical model including the mixing time constant has been proposed allowing an accurate Lp estimation even though the mixing time constant is higher than the cell time constant obtained in response to a perfect shock. The estimated Lp values of human leukemia K562 cells were found to be the same whatever the mixing time constant. The Lp value of Saccharomyces cerevisiae could not be exactly estimated. However, S. cerevisiae has unexpectedly high water permeability, implying that this yeast may contain water channels in the membrane. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 62-70, 1997.
    Additional Material: 3 Ill.
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  • 7
    ISSN: 1075-2617
    Keywords: β-bend ; 310-helix ; peptide conformational analysis ; spin-labelled amino acid ; TOAC peptides ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A variety of host L-alanine homo-peptides (to the pentamer) containing one or two spin-labelled TOAC (2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid) residues were synthesized by solution methods and fully characterized. The conformational features of the terminally blocked, doubly spin-labelled-TOAC-(Ala)2-TOAC-Ala- pentapeptide were examined in the crystal state by X-ray diffraction and in solution using a combination of techniques (Fourier transform infrared, circular dichroism, cyclic voltammetry and electron spin resonance) in comparison with singly labelled shorter peptides. The 310-helical structure of the pentapeptide, promoted by the two Cα,α-disubstituted glycines under favourable experimental conditions, allows an interaction to take place between the two nitroxide TOAC side chains spaced by one turn of the helix. Taken together, these results suggest that TOAC is an excellent probe for exploring bends and helices in doubly labelled peptides.
    Additional Material: 8 Ill.
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  • 8
    ISSN: 1075-2617
    Keywords: β-amino-alcohol ; statine ; UNCAs ; vicinal tricarbonyl compounds ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: N-Urethane-protected N-carboxyanhydrides (UNCAs) are very reactives. They have been successfully used in peptide synthesis, in both solution and solid phase. We have demonstrated that UNCAs are interesting starting materials for the synthesis of various amino acid derivatives. Chemoselective reduction of UNCAs with sodium borohydride led the corresponding N-protected β amino alcohols. Reaction of UNCAs with Meldrum's acid, followed by cyclisation, yielded enantiomerially pure tetramic acid derivatives. Diastereoselective reduction of tetramic acid derivatives produced (4S,5S)-N-alkoxycarbonyl-4-hydroxy-5-alkylpyrrolidin-2-ones derived from amino acids, which after hydrolysis yielded statine and statine analogues. Tetramic acid derivatives could also be obtained by reaction of UNCAs with benzyl ethyl followed by hydrogenolytic deprotection and decarboxylation. UNCAs also reacted with phosphoranes to produce the ketophosphorane in excellent yields. Subsequent oxidation with oxone or with [bis(acetoxy)-iodol]-benzene produced vicinal tricarbonyl derivatives. These reactions usually proceeded smoothly and with high yields.
    Additional Material: 4 Ill.
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  • 9
    ISSN: 0173-0835
    Keywords: Two-dimensional polyacrylamide gel electrophoresis ; Silver staining ; Mass spectrometry ; Proteome ; Mitochondria ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Owing to the complexity of higher eukaryotic cells, characterization of a complete proteome is likely to be difficult to achieve. However, advantage can be taken of the cell compartmentalization to build organelle proteomes, which can moreover be viewed as specialized tools to study specifically the biology and “physiology” of the target organelle. Within this frame, we report here the construction of the human mitochondrial proteome, using placenta as the source tissue. Protein identification was carried out mainly by peptide mass fingerprinting, but other methods were also used (N-terminal microsequencing, blotting). The optimization steps in two-dimensional (2-D) electrophoresis needed for proteome research are discussed. However, the relative paucity of data concerning mitochondrial proteins is still the major limiting factor in building the corresponding proteome, which should be a useful tool for researchers working on human mitochondria and their deficienciesThis work can be found on the Internet at the following address: http://www-dsv.cea.fr/MitoPick/Default.html.
    Additional Material: 4 Ill.
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  • 10
    ISSN: 0749-503X
    Keywords: Candida albicans ; heat-shock (stress) proteins ; nucleotide sequence ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have cloned and sequenced a Candida albicans gene (SSB1) encoding a potential member of the heat-shock protein seventy (hsp70) family. The protein encoded by this gene contains 613 amino acids and shows a high degree (85%) of sequence identity to the ssb subfamily (ssb1 and ssb2) of the Saccharomyces cerevisiae hsp70 family. The transcribed mRNA (2·1 kb) is present in similar amounts both in yeast and germ tube cells of C. albicans. The sequence data has been deposited in the GenBank data library under the Accession Number X97723. © John Wiley & Sons, Ltd.
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