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Fatima's project - Coggle Diagram
Fatima's project
Cross Epithelial Transport
M cells
in vivo
Fiber Staining in Peyer's patch Tissue Sections
Mouse Pellet stain
M137 and LM-11 indistinguishable from autofluorescence
Xylanase treatment
still not disntinguishable
Psyllium feeding (5 days) Peyer's patch section stain with M72
can't see PSY stained with M72
GP2 Staining
Successful
in vitro
Caco-2 growth
Induce epithelial permeability
in vivo feeding
Galactomannan G
Psyllium
Detect in P.P. with antibodies (ELISA/ Tissue Staining)
Polybiotin
Detected in
liver
17 hours after oral gavage
Microscopy
Not associated with Kupffer cells
Streptavidin Sandwich ELISA
Glycan-Immune Interaction
Glycans
Psyllium
reduces LPS/ATP induced IL-1B in RAW macrophages
does not bind to reduce IL-1B detection
Xylan
reduces LPS/ATP induced IL-1B in RAW macrophages
does not bind to reduce IL-1B detection
Does not stimulate LDH release
Binds to RAW macrophages (biotin-xyl, Christian M)
Receptor mediated?
test candidate receptors (CRISPR)
Internalized?
Degradation?
Time course assay (Christian M)
Mechanism of degradation
ROS Sam M, in progress
Exocytosis Assay
Other cytokines: qPCR
TNF-alpha
IL-6
in vivo
Induce inflammation, IP injection of xylan, measure IL-1B in peritoneum
Induce inflammation by DSS, feed xylan, measure inflammation + disease score
I.P injection in vivo
induces serum antibodies?
YES
Black 6 conventional (FR12)
Swiss Webster conventionalized (FR14)
NO
Swiss Webster Germ Free (FR13)
-Swiss Webster Conventionalized (FR15)
Swiss Webster Germ Free (FR15)
Detected in Peritoneal cell lysate?
YES
Swiss Webster Conventionalized (FR14)
Swiss Webster Germ Free (FR13)
NO
Swiss Webster Conventionalized (FR15)
Swiss Webster Germ Free (FR15)
detected in liver lysate?
YES
Swiss Webster Conventionalized (FR14)
Swiss Webster Germ Free (FR13)
Serum IL-10?
Levan
reduces LPS/ATP induced IL-1B in RAW macrophages
does not bind to reduce IL-1B detection
Alginate
reduces LPS/ATP stimulated IL-1B
Does not stimulate LDH release
Galactomannan G