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1.
J Exp Bot ; 72(8): 3219-3234, 2021 04 02.
Article in English | MEDLINE | ID: mdl-33475728

ABSTRACT

Successful plant defence against microbial pathogens is based on early recognition and fast activation of inducible responses. Key mechanisms include detection of microbe-associated molecular patterns by membrane-localized pattern recognition receptors that induce a basal resistance response. A well-described model of such responses to pathogens involves the interactions between Solanaceae plants and proteinaceous elicitors secreted by oomycetes, called elicitins. It has been hypothesized that the formation of oligomeric structures by elicitins could be involved in their recognition and activation of defensive transduction cascades. In this study, we tested this hypothesis using several approaches, and we observed differences in tobacco plant responses induced by the elicitin ß-cryptogein (ß-CRY) and its homodimer, ß-CRYDIM. We also found that the C-terminal domain of elicitins of other ELI (true-elicitin) clades plays a significant role in stabilization of their oligomeric structure and restraint in the cell wall. In addition, covalently cross-linking ß-CRYDIM impaired the formation of signalling complexes, thereby reducing its capacity to elicit the hypersensitive response and resistance in the host plant, with no significant changes in pathogenesis-related protein expression. By revealing the details of the effects of ß-CRY dimerization on recognition and defence responses in tobacco, our results shed light on the poorly understood role of elicitins' oligomeric structures in the interactions between oomycetes and plants.


Subject(s)
Nicotiana , Oomycetes/pathogenicity , Plant Diseases , Amino Acid Sequence , Nicotiana/metabolism
3.
Eur Spine J ; 28(4): 783-791, 2019 04.
Article in English | MEDLINE | ID: mdl-30506486

ABSTRACT

PURPOSE: The presence of Propionibacterium acnes in a substantial component of resected disc specimens obtained from patients undergoing discectomy or microdiscectomy has led to the suggestion that this prominent human skin and oral commensal may exacerbate the pathology of degenerative disc disease. This hypothesis, therefore, raises the exciting possibility that antibiotics could play an important role in treating this debilitating condition. To date, however, little information about antibiotic penetration into the intervertebral disc is available. METHODS: Intervertebral disc tissue obtained from 54 microdiscectomy patients given prophylactic cefazolin (n = 25), clindamycin (n = 17) or vancomycin (n = 12) was assayed by high-performance liquid chromatography, with cefaclor as an internal standard, to determine the concentration of antibiotic penetrating into the disc tissue. RESULTS: Intervertebral disc tissues from patients receiving the positively charged antibiotic clindamycin contained a significantly greater percentage of the antibacterial dose than the tissue from patients receiving negatively charged cefazolin (P < 0.0001) and vancomycin, which has a slight positive charge (P < 0.0001). CONCLUSION: Positively charged antibiotics appear more appropriate for future studies investigating potential options for the treatment of low-virulence disc infections. These slides can be retrieved under Electronic Supplementary Material.


Subject(s)
Anti-Bacterial Agents/pharmacokinetics , Cefazolin/pharmacokinetics , Clindamycin/pharmacokinetics , Gram-Positive Bacterial Infections/prevention & control , Intervertebral Disc Degeneration/surgery , Intervertebral Disc Displacement/surgery , Intervertebral Disc/metabolism , Propionibacterium acnes , Vancomycin/pharmacokinetics , Adult , Anti-Bacterial Agents/therapeutic use , Cefazolin/therapeutic use , Clindamycin/therapeutic use , Humans , Vancomycin/therapeutic use
4.
J Plant Physiol ; 210: 51-57, 2017 Mar.
Article in English | MEDLINE | ID: mdl-28056387

ABSTRACT

Plant plasma membrane associated proteins play significant roles in Microbe-Associated Molecular Pattern (MAMP) mediated defence responses including signal transduction, membrane transport or energetic metabolism. To elucidate the dynamics of proteins associated with plasma membrane in response to cryptogein, a well-known MAMP of defence reaction secreted by the oomycete Phytophthora cryptogea, 2D-Blue Native/SDS gel electrophoresis of plasma membrane fractions was employed. This approach revealed 21 up- or down-regulated protein spots of which 15 were successfully identified as proteins related to transport through plasma membrane, vesicle trafficking, and metabolic enzymes including cytosolic NADP-malic enzyme and glutamine synthetase. Observed changes in proteins were also confirmed on transcriptional level by qRT-PCR analysis. In addition, a significantly decreased accumulation of transcripts observed after employment of a mutant variant of cryptogein Leu41Phe, exhibiting a conspicuous defect in induction of resistance, sustains the contribution of identified proteins in cryptogein-triggered cellular responses. Our data provide further evidence for dynamic MAMP-induced changes in plasma membrane associated proteins.


Subject(s)
Nicotiana/genetics , Nicotiana/microbiology , Phytophthora/physiology , Cell Membrane/metabolism , Chromatography, Liquid , Electrophoresis, Polyacrylamide Gel , Fungal Proteins/genetics , Fungal Proteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Pathogen-Associated Molecular Pattern Molecules/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Tandem Mass Spectrometry , Nicotiana/metabolism , Trypsin/chemistry
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