Animals were randomized, and half had their spleens removed by electrocauterization (BVI Medical, Waltham, MA, USA). In sham‐operated mice, the abdominal wall was opened and then closed after identifying the spleen. All surgical procedures were done under inhalation anesthesia (1% isoflurane) on a heat pad. Body temperature was monitored. Pain was mitigated pre‐ and post‐operatively. A total of 10 mg/kg Carprofen was provided in drinking water 2 d before surgery and continued until 2 d after surgery. In addition, 1 h prior to surgery 5 mg/kg Carprofen was injected s.c. To allow for the inflammation derived from the surgery to resolve, mice were housed for further 70 d without intervention. β‐glucan from barley (Sigma‐Aldrich, St. Louis, MO, USA) was administered i.p. to 40 mice at day 0 and at day 3. PBS was used as a vehicle control. Animals were euthanized at day 7 or at day 7 plus 3 hours after i.p. injection of 10 μg Escherichia coli LPS (E. coli LPS; serotype 055:B5, Sigma‐Aldrich, St. Louis, MO, USA). Sacrifice involved deep inhalation anesthesia (5% isoflurane) followed by cervical dislocation. Two independent experiments were performed.
β glucan from barley
Manufactured by Merck Group
Sourced in United States
β‐glucan from barley is a naturally occurring polysaccharide extracted from the cell walls of barley. It is a soluble dietary fiber that has been shown to have various functional properties. The core function of β‐glucan is to provide structural support and act as a thickening agent in various applications.
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2 protocols using β glucan from barley
1
Splenectomy and β-Glucan Effects on LPS Response
2
Heterologous Expression of Xylanase from Chicken Cecal Metagenome
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The Escherichia coli EPI300™-T1R clone harboring fosmid pCC1FOS carrying a chicken cecal metagenomic DNA fragment containing a xylanase gene was a gift from Dr Kenneth van Driel. All enzymes and dNTPs in this study were purchased from New England BioLabs Inc., USA, and Promega, USA. Plasmid DNA extraction and purification kit was purchased from GE Healthcare, UK. TALON Superflow Metal Affinity Resin (Clonetech) was purchased from TaKaRa (Otsu, Japan). The expression vector pET-32a (Novagen) was used for cloning and expressing the xylanase. E. coli Tuner (DE3)pLysS was used as expression host and was cultivated on Luria–Bertani medium (Difco). The enzyme substrates used were xylan from oat-spelt (Fluka), xylan from beechwood (Megazyme), α-cellulose (Sigma), carboxymethyl cellulose (Sigma), starch (Sigma), β-glucan from barley (Sigma), 4-nitrophenyl-β-D-xylopyranoside (Megazyme), 4-nitrophenyl-β-D-cellobioside (Sigma), and 4-nitrophenyl-α-D-galactopyranoside (Fluka). Molecular weight standard mix containing xylose, xylobiose, xylotriose, xylotetraose, xylopentaose, and xylohexaose (Megazyme) was the gift from Professor Khanok Ratanakhanokchai, KMUTT, Thailand. All other chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA).
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