**10. Liver tissue bioenergetics in concanavalin A hepatitis in mice**

Concanavalin A (Con A) is a plant lectin from the seeds of Canavalia ensiformis (jack bean). This toxin serves as a polyclonal T-cell mitogen. It produces fulminant hepatitis in mice, a disease that mimics human infection with hepatitis B virus (Tiegs et al., 1992 & 1997). The hepatic injury is typically noted within 3 hr of intravenous injection of > 1.5 mg/kg of Con A and progresses with time (Tiegs et al., 1992). Activation and recruitment of Natural Killer (NK) T-cells and other cells of the innate immune system are early events, which lead to increased secretion of various inflammatory cytokines (e.g., TNF-, IL-2, IL-10, IL-12 and IFN-) (Takeda et al., 2000; Margalit et al., 2005; Chen et al., 2010; Sass et al., 2002). This immune response targets multiple organs including the liver. Its outcome is irreversible hepatotoxicity, which includes inflammatory infiltrates and necrosis (Leist et al., 1996).

The above described *in vitro* system is employed to assess liver tissue respiration in Con A treated C57BL/6 mice. The purpose of the work was to estimate hepatocyte bioenergetics in this well-studied hepatitis model. The mice were injected intravenously with 12 mg/kg Con A or PBS. Specimens (20 to 30 mg each) were cut from the liver of anesthetized (urethane, 100 L per 10 g body weight, using 25% solution, w/v, in 0.9% NaCl) mice using a sharp scissor (Moria Vannas Wolg Spring, cat. # ST15024-10) (Al Samri et al., 2011). The specimens were immediately immersed in ice-cold Krebs-Henseleit buffer (115 mM NaCl, 25 mM NaHCO3, 1.23 mM NaH2PO4, 1.2 mM Na2SO4, 5.9 mM KCl, 1.25 mM CaCl2, 1.18 mM MgCl2 and 6 mM glucose, pH ~7.2), gassed with 95% O2:5% CO2. Pieces were then weighed and placed in 1-ml Pd phosphor solution (Krebes-Henseleit buffer containing 0.5% albumin and 3 M Pd phosphor) for O2 measurement. The results are summarized in Table 3.

