Use of Inhibitors of Scavenger Receptor Class Proteins for the Treatment of Infectious Diseases
a technology of scavenger receptor and inhibitor, which is applied in the direction of amide active ingredients, drug compositions, tetracycline active ingredients, etc., can solve the problem of even more depressing reality, and achieve the effect of inhibiting high density lipoprotein (hdl) uptak
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example 1
Cell Cultivation and Seeding
[0288]Huh human hepatoma cells, were cultivated in RPMI (Gibco / Invitrogen) medium containing 10% FBS, 1% non-essential amino acid solution (Gibco / Invitrogen), 1% penicillin / streptomycin solution (Gibco / Invitrogen), 1% glutamine (Gibco / Invitrogen) and 1% Hepes pH 8 (Gibco / Invitrogen).
[0289]Hepa 1-6 cells were cultured in complete DMEM medium (Gibco / Invitrogen) supplemented with 10% fetal calf serum (Gibco / Invitrogen) and 1% penicillin / streptomycin (Gibco / Invitrogen) incubated at 37° C. and 5% CO2.
[0290]Cells were split twice per week by seeding 106 cells in 15 ml complete medium in 75 ml culture flasks (Nunc). For passaging, cells were detached from the flask by incubation with 3 ml Trypsin solution (Gibco / Invitrogen) for 5 min at 37° C. Trypsin was inactivated by adding 10 ml of complete medium to the flask.
[0291]Cell based experiments were performed in black, optical 96 well plates (Costar / Coming). 4000-6000 cells per well were seeded in a volume of 100 ...
example 2
Isolation of Sporozoites of Plasmodium berghei ANKA or Plasmodium yoelii from Mosquitoes
[0293]Sporozoites were obtained from Anopheles stephensi mosquitoes infected with P. berghei ANKA or P. yoelii. Salivary glands were dissected and collected in RPMI medium (GIBCO) on ice. Collected tissues were gently ground in the medium to release sporozoites. Tissue fragments were removed by centrifugation at 40×g for 3 min, and sporozoites were collected from the supernatant.
example 3
Treatment of Cells with Chemical Compounds Prior to Infection
[0294]Blt-1, Blt-2, and Blt-4 were purchased from ChemBridge Corporation (San Diego, USA). Ezetimibe (for chemical structure see FIG. 6 and structure (XXVI)) was derived from powdered Ezeterol tablets (Essex Pharma, UK). Each compound was dissolved in DMSO at a final concentration of 50 mM. 48 hours after seeding of 6000 Huh-7 cells per well, growth medium was replaced by fresh complete culture medium, containing the compounds in 4 different concentrations, generated by dilution series of the compound stock solutions in complete growth medium: 8 μM; 1.6 μM; 320 nM; 64 nM. Controls media were prepared according to DMSO concentrations in the 4 compound dilutions, i.e. 0.016%; 0.0032%; 0.00064%; 0.000128%. Huh-7 cells were equilibrated for 1 h with compound / DMSO-containing medium at 37° C. before infection with 10,000 sporozoites per well (FIG. 1 to 4).
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