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The spleens were manually dissociated, and the cells were cleaned by passage through a 70 m strainer followed by a 2-minute incubation in ACK medium (37 mM NH4Cl, 10 mM KHCO3, 0.1 M EDTA) at room temperature

Following Fourier transformation with 0.3 Hz line broadening and a zero-filling to 64 K, ACD Lab Processor Academic Edition (Advanced Chemistry Development, 12.01, 2010) was used to perform phase adjustment and baseline correction of 1 H-NMR spectra as well as to set spectral chemical shift referencing on the TSP CH 3 signal at 0.00 ppm

The global burden of disease attributable to alcohol and drug use in 195 countries and territories, 19902016: a systematic analysis for the Global Burden of Disease Study 2016

Insonation was applied if needed (2 MHz, 2.0 W/cm 2 , 20% duty cycle, 5 min)

doi:10.1212/01.wnl.0000078887.16593.12 Connor JR, Ponnuru P, Wang XS, et al