Klebsiella pneumoniae

Manufactured by Merck Group

Sourced in China, United States, United Kingdom

Klebsiella pneumoniae is a species of Gram-negative, rod-shaped bacteria. It is a common inhabitant of the human gastrointestinal tract and can be found in the environment. Klebsiella pneumoniae is often used as a model organism in laboratory research and testing.

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7 protocols using klebsiella pneumoniae

Neutrophil Histone Citrullination Assay

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Bone marrow–derived neutrophils were resuspended in RPMI 1640 with Hepes and allowed to adhere at 37°C in 5% CO₂ to glass-bottom plates (previously coated with poly-l-lysine) for 20 min before stimulation with LPS (10 μg/ml) from Klebsiella pneumoniae (Sigma-Aldrich). After 2 hours, cells were fixed in 2% (v/v) paraformaldehyde. Cells were then stained for citrullinated histone 3 as described below.

Bernal A, & Kimbrell D.A. (2000). Drosophila Thor participates in host immune defense and connects a translational regulator with innate immunity. Proceedings of the National Academy of Sciences of the United States of America, 97(11), 6019-6024.

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Bacterial LPS and MIPS-9451 Synthesis

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LPS from the bacterial species, Escherichia coli serotype: 0111:B4, E. coli serotype: 026:B6, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica serotype: abortus equi, S. enterica serotype: enteritidis, S. enterica serotype: Minnesota R595 (rough strain), S. enterica, serotype: Minnesota (smooth strain), S. enterica, serotype: typhimurium, S. typhosa and S. marcescens, all purified by phenol extraction, and 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES buffer) were purchased from Sigma Aldrich (St. Louis, MO). LPS from the bacterial species Campylobacter jejuni, Proteus mirabilis, Proteus vulgaris, all purified by phenol extraction, and Helicobacter pylori and Porphyromonas gingivalis, purified by ultracentrifugation, were purchased from Wako chemicals, Osaka, Japan. Polymyxin B was purchased from BetaPharma (Shanghai, China). The synthesis of MIPS-9451 has been previously described elsewhere.^{14 (link)}

McInerney M., Roberts K., Thompson P., Li J., Nation R., Velkov T, & Nicolazzo J. (2016). Quantitation of polymyxin-lipopolysaccharide interactions using an image-based fluorescent probe. Journal of pharmaceutical sciences, 105(2), 1006-1010.

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Antimicrobial Chitosan Nanocomposite Synthesis

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CS (degree of deacetylation, 85%; M_w, around 600 kDa), silver nitrate, sodium periodate, glycerol, acetic acid, sodium tri-polyphosphate and Sulforhodamine B were purchased from Shanghai branch of Sigma Aldrich Chemical Co., Ltd (Shanghai, China). CNC aqueous suspension (1 mg/mL) was prepared by our group according to a procedure modified from the literature [17 (link)]. All the other chemicals and reagents were of analytical grade and double distilled water was used throughout. Dulbecco's modified eagle medium (DMEM) and beef extract peptone medium used for the cultivation of bacteria were obtained from Sigma Co. Ltd. The clinical strains (C) of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Enterobacter cloacae, Streptococcus pneumoniae and Pseudomonas aeruginosa, Candida albicans, Candida glabrata and Candida krusei were donated by the Second Affiliated Hospital of Qiqihar Medical College (Qiqihar, China). Staphylococcus aureus standard strain (S) (ATCC25923), Escherichia coli (S) (ATCC25922), Pseudomonas aeruginosa (S) (ATCC27853)and NIH3T3 cell lines were obtained from Laboratory of Biochemistry of Qiqihar University (Qiqihar, China).

Dong F, & Li S. (2018). Wound Dressings Based on Chitosan-Dialdehyde Cellulose Nanocrystals-Silver Nanoparticles: Mechanical Strength, Antibacterial Activity and Cytotoxicity. Polymers, 10(6), 673.

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Lipopolysaccharide Detection Protocol

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Sodium dodecyl sulfate (SDS), sodium tetraborate, and sodium hydroxide were obtained from Merck (Darmstadt, Germany). Fluorescein isothiocyanate (FITC), (1-3)-β-D-glucans were purchased from Sigma - Aldrich (St. Louis, MO, USA). Lipopolysaccharides from Escherichia coli 0111:B4 (Lot # 024M4019V), Escherichia coli 055:B5(Lot #025M4040V), Escherichia coli 026:B6 (Lot # 053M4060V), Escherichia coli 0127:B8 (Lot # 103M4051V), Salmonella enterica serotype enteritidis (Lot # 064M4035V), Pseudomonas aeruginosa 10 (Lot # 100M4101V), Salmonella enterica serotype typhimurium (Lot # 093M4088V), Escherichia coli J5 (Rc mutant, rough strains) (Lot # 053M4112V), Salmonella typhosa (Lot # 063M4017V), Escherichia coli F583 (Rd mutant, rough strains) (Lot # 055M4005V), Salmonella enterica serotype minnesota (Lot # 064M4015V), Escherichia coli 0128:B12 (Lot # 063M4014V), Klebsiella pneumoniae (Lot # 111M4038V), Escherichia coli K-235 (Lot # 031M4076V), Salmonella enterica serotype abortusequi (Lot # 104M4064V), and Serratia marcescens (Lot # 013M4078V) were purchased from Sigma Aldrich (St. Louis, MO, USA). Buffers were prepared with ultrapure water obtained from Merck Millipore (Millipore, Bedford, MA, USA). All other reagents were of analytical grade. Experiments were carried out using freshly prepared and filtered solutions.

Fung F.M., Su M., Feng H.T, & Li S.F. (2017). Extraction, separation and characterization of endotoxins in water samples using solid phase extraction and capillary electrophoresis-laser induced fluorescence. Scientific Reports, 7, 10774.

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Rodent Model of Acute-on-Chronic Liver Failure

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The rodent model of ACLF utilised in this study was as described previously^{47 (link)}. Eight groups of adult male Sprague–Dawley rats (RRID:RGD_10395233) (n = 12 per group) were studied 4-weeks after either sham-operation or BDL. Animals were treated either with saline or recombinant human alkaline phosphatase (recAP; gifted AMPharma, Netherlands) − 1000 U/kg intraperitoneal on four consecutive days before LPS injection (days 25–28 after BDL). The last recAP injection was performed 3-hours before ACLF induction with LPS derived from Klebsiella pneumoniae (Sigma, UK) (0.3 mg/kg/hr, intravenously) or saline 0.9% (154 mmol sodium chloride (NaCl), vehicle). The animals in each group were terminated by exsanguination under general anaesthesia with isoflurane (Piramal Healthcare, USA) 3 h after LPS administration. The groups were as described in Supplementary Fig. 1. Hemodynamic measurements including PP and RBF was performed.

Engelmann C., Adebayo D., Oria M., De Chiara F., Novelli S., Habtesion A., Davies N., Andreola F, & Jalan R. (2020). Recombinant Alkaline Phosphatase Prevents Acute on Chronic Liver Failure. Scientific Reports, 10, 389.

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Antimicrobial Activities of Natural Products

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Types of Gram-positive bacteria Staphylococcus aureus and Streptococcus pyogenes, Gram-negative bacteria Escherichia coli and Klebsiella pneumoniae, and fungi Aspergillus niger, Alternaria alternate, Curvularia lunata, and Candida albicans were from the National Research Centre, Department of Chemistry of Natural and Microbial Products, Giza, Egypt, and cefotaxime sodium, nystatin, and DMSO were purchased from Sigma-Aldrich.

Abu-Hashem A.A, & Al-Hussain S.A. (2022). The Synthesis, Antimicrobial Activity, and Molecular Docking of New 1, 2, 4-Triazole, 1, 2, 4-Triazepine, Quinoline, and Pyrimidine Scaffolds Condensed to Naturally Occurring Furochromones. Pharmaceuticals, 15(10), 1232.

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Antimicrobial Evaluation of Bacterial Strains

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The bacterial strains: Escherichia coli ATCC 10536, Klebsiella pneumoniae ATCC 33495, Pseudomonas aeruginosa ATCC 9721, Staphylococcus aureus ATCC 25923, and Streptococcus pyogenes ATCC 19615 were acquired from Sigma-Aldrich (Johannesburg, South Africa). Chromobacterium violaceum ATCC 12472 (biomonitor strain) was used for the qualitative and quantitative evaluation of AQS, which was kindly provided by the Centre for Microbial, Ecology and Genomics (CMEG), University of Pretoria, South Africa. Ethics approval for the use of these microorganisms was obtained from the University of Pretoria, Faculty of Natural and Agricultural Sciences Ethics Committee (reference number: NAS158/2021).
The Clinical and Laboratory Standard Institute guidelines were followed for these bacterial growth media and conditions (Clinical and Laboratory Standards Institute, 2020) . Bacterial cultures of E.

Itumeleng T.B., Idowu J.A., Abdullahi A.Y., & Cosa S. (2022). Antibacterial, antiquorum sensing, antibiofilm activities and chemical profiling of selected South African medicinal plants against multi-drug resistant bacteria. Journal of Medicinal Plants Research, 16(2), 52-65.

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