Klebsiella pneumoniae is a clinically significant, Gram-negative pathogen in which the production of extracellular polysaccharides is a key virulence factor. Extracellular polysaccharides such as the capsule and its mucoviscosity play a significant role in K. pneumoniae infection. In this article, we explain several standard protocols used to characterize the extracellular polysaccharides of K. pneumoniae. Several of these protocols are adapted specifically for K. pneumoniae and describe methods to purify and quantify the extracellular polysaccharide of K. pneumoniae. We also present a standardized protocol to quantify K. pneumoniae mucoviscosity, a unique feature of K. pneumoniae extracellular polysaccharide. These protocols will help create uniformity in standard protocols used in K. pneumoniae extracellular polysaccharide studies. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Extracellular polysaccharide isolation and purification Basic Protocol 2: Large-scale isolation and purification of extracellular polysaccharide Basic Protocol 3: Uronic acid quantification of extracellular polysaccharide Basic Protocol 4: Extracellular polysaccharide visualization by SDS-PAGE Basic Protocol 5: Klebsiella pneumoniae mucoviscosity measurement by sedimentation resistance assay Alternate Protocol 5: 96-well plate-based Klebsiella pneumoniae sedimentation resistance assay Support Protocol 5: Determination of plate to cuvette conversion factor.
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http://dx.doi.org/10.1002/cpz1.937 | DOI Listing |
J Agric Food Chem
January 2025
State Key Laboratory of Green Pesticide, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Center for R&D of Fine Chemicals of Guizhou University, Guiyang 550025, China.
To develop antibacterial agents with a novel mechanism of action, a series of sulfone compounds containing a 1,2,4-triazolo[4,3-]pyridine were designed and synthesized by progressive molecular structure optimization. The antibacterial activities of some derivatives against the four plant pathogens (), (), (), and () were evaluated. Among them, compound demonstrated significant antibacterial activities against , , and , with EC values of 1.
View Article and Find Full Text PDFNPJ Biofilms Microbiomes
January 2025
Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand.
Chronic infections represent a significant global health and economic challenge. Biofilms, which are bacterial communities encased in an extracellular polysaccharide matrix, contribute to approximately 80% of these infections. In particular, pathogens such as Pseudomonas aeruginosa and Staphylococcus aureus are frequently co-isolated from the sputum of patients with cystic fibrosis and are commonly found in chronic wound infections.
View Article and Find Full Text PDFCarbohydr Polym
March 2025
Laboratory of Biochemistry and Biomedical Materials, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, PR China. Electronic address:
Ultraviolet B (UVB) irradiation from sunlight is one of the primary environmental factors that causes photodamage to the skin. The aim of this study was to prepare succinyl-chitosan oligosaccharide (SU-COS) and evaluate its protective effects and related molecular mechanisms against UVB-induced photodamage for the first time. SU-COS (substitution degree: 69.
View Article and Find Full Text PDFFitoterapia
January 2025
Translational Chinese Medicine Key Laboratory of Sichuan Province, Sichuan Academy of Chinese Medicine Sciences, Sichuan Institute for Translational Chinese Medicine,Chengdu, Sichuan 610041, PR China. Electronic address:
Background: Hepatic fibrosis is a wound healing response that leads to excessive deposition of extracellular matrix (ECM) due to sustained liver injury. Hepatic stellate cells (HSCs) are key players in ECM synthesis, with the TGF-β/Smad signaling pathway being central to their activation. Despite advances in understanding the pathogenesis of hepatic fibrosis, effective anti-fibrotic therapies are still lacking.
View Article and Find Full Text PDFFASEB J
January 2025
Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Neutrophils are peripheral blood-circulating leukocytes that play a pivotal role in host defense against bacterial pathogens which upon activation, they release web-like chromatin structures called neutrophil extracellular traps (NETs). Here, we analyzed and compared the importance of myeloid differentiation factor 88 (MYD88), peptidyl arginine deiminase 4 (PAD4), and gasdermin D (GSDMD) for NET formation in vivo following sepsis and neutrophilia challenge. Injection of lipopolysaccharide (LPS)/E.
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