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Adventures in Restorative Listening

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Taking place two years after the events of the first game, our now more matured main character finds himself reunited with his estranged sister, Mia, when he accepts an invitation from his aunt to spend his Summer break from college with them.


Prenatal inflammation and infection influence the postnatal immuneresponse, which includes susceptibility to infection, hypersensitivitydisorders, and vaccination response. Several studies have shown thatprenatal inflammation can programme changes in mature immune celloutput, but the drivers of long-term changes in offspring immunity areunknown [6]. In early development, "sterile" inflammatory signallingis required for HSC emergence in the developing embryo. However,beyond emergence, little is known about whether developing HSCsrespond to maternal inflammation in utero, how they respond, and theimplications for postnatal hematopoietic and immune development.The foetal HSC response to prenatal inflammation, we hypothesise,causes long-term changes in hematopoietic output in offspring,shaping the postnatal immune response [7].


RAW 264.7 cells, a type of mouse mononuclear macrophageleukaemia cell, were obtained from the Advanced Research Center atCentral South University's cell repository. RAW 264.7 cells were grownin Dulbecco's modified Eagle's medium (DMEM), which contained 10%FBS and 1% penicillin-streptomycin. Primary bone marrow-derivedmacrophages (BMDMs) were isolated from C57BL/6 mice femurs andtibias, as previously described. Using ACK lysis buffer, erythrocyteswere extracted from bone marrow. To re-suspend bone marrow cells,RPMI 1640 culture medium containing 10% foetal bovine serum, 1%penicillin-streptomycin, and 40 ng/ml macrophage colony-stimulatingfactor was used. The cells were cultured in a 75 cm2 cell culture flaskat 37 C and 5% CO2. Adherent cells differentiated into maturemacrophages after 7 days of culture (M0). The M1 macrophages werethen stimulated for 24 hours with LPS (50 ng/ml) and IFN- (20 ng/ml)(M1). M1 macrophages were stimulated for 24 hours with either QCT(10uM) or Fer-1 (10uM) in separate experiments. The M2 macrophageswere then stimulated for 24 hours with IL-4 (10 ng/ml) and IL-13 (10ng/ml) (M1). M2 macrophages were stimulated for 24 hours witheither QCT (10uM) or Fer-1 (10uM) in separate experiments. 041b061a72


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