Screen shots of serious game.

<p>A) Each case included a 2-D rendering of the patient, a chief complaint, vital signs, a history, and a written description of the physical exam. Physicians had 42 minutes (a simulated 8 hour shift) to complete the ten cases. A clock at the top right of the screen helped track the passage of time. B) Physicians could manage patients by selecting from a pre-specified list of 250 medications, studies, and procedures. C) We included audio-visual distractors, including nursing requests for help with disruptive patients to increase the verisimilitude of the experience.</p

Sampling frame for study.

<p>There was a 79% response rate and an 86% completion rate.</p

Case Descriptions.

<p>NR – non-representative; R – representative.</p

AICAR improves renal function in mice during CLP.

<p>Serum cystatin and blood urea nitrogen (BUN) concentrations after CLP or SHAM operation in adult mice receiving either AICAR or diluent control (n = 6 mice per group per timepoint). Data are means ± s.e.m.; rank sum test.</p

Inhibition of VPS34 attenuates autophagy and inhibits recovery of renal function in young mice during endotoxemia.

<p>Immunofluorescent microscopy (20X) of renal cortex harvested at 48 hours after LPS in young mice from 4 experimental groups: NT siRNA and saline control, VPS34 siRNA and saline control, NT siRNA and LPS, and VPS34 siRNA and LPS. (a) VPS34 (Cy3, red), actin (Alexa488, green), nucleus (Hoechst, blue). Representative images of n = 4 experiments (4 mice per experiment). (b) LC3 (Cy3, red), actin (Alexa488, green), nucleus (Hoechst, blue). Representative images of n = 4 experiments (4 mice per experiment). (c) Immunoblot analysis of renal cortex lysate detecting LC3b (16KD) in untreated mice (lanes 1 and 2) and mice treated with Non-target siRNA and LPS (lanes 3 and 4) or VPS34 siRNA and LPS (lanes 5 and 6). Representative blot is shown at 48 h timepoint after LPS from n = 2 experiments (6 mice per experiment).</p

Temsirolimus improves renal function and increases renal autophagy in adult mice during endotoxemia.

<p>(a) Time course plots for serum cystatin and blood urea nitrogen (BUN) concentrations after LPS in adult mice receiving either temsirolimus or diluent control (n = 8–11 mice per group per timepoint). (b) Immunoblot analysis of renal cortex lysate detecting LC3b (16KD) and p-Ser²⁴⁴⁸ mTOR (289KD) in mice treated with temsirolimus (lanes 1 and 2), diluent control and LPS (lanes 3–5), or temsirolimus and LPS (lanes 6–8). Representative blot is shown at 48 h timepoint after LPS from n = 4 experiments (8 mice per experiment). Graphs summarizing corresponding density of LC3b and p-Ser²⁴⁴⁸ mTOR for each experimental condition. (c) Immunofluorescent microscopy (20X) of renal cortex harvested at 48 hours after LPS in 4 experimental groups: diluent control, temsirolimus, diluent control and LPS, and temsirolimus and LPS. LC3 (Cy3, red), actin (Alexa488, green), nucleus (Hoechst, blue). Representative images of n = 4 experiments (8 mice per experiment). (d) Transmission electron microscopy (10⁵X) of renal cortex harvested at 48 hours after LPS in 4 experimental groups. Inset (5×10⁵X) Representative images of n = 4 experiments (8 mice per experiment). arrowheads, autophagosomes; arrows, autophagolysosomes. (e) Serum cystatin and blood urea nitrogen (BUN) concentrations in adult mice receiving either temsirolimus or diluent control after endotoxemia (n = 8–11 mice per group per timepoint). Abbreviations: TEM, temsirolimus. Data are means ± s.e.m.; rank sum test.</p

Arterial blood gas measurements in mice bled to and maintained at a MAP of 20 mm Hg for 30 minutes with and without inhaled CO (250 ppm).

<p>*P<0.05 compared to sham mice.</p><p>^§P<0.05 compared to shock mice.</p><p>Sham mice underwent anesthesia and surgical manipulation without hemorrhage. Mice were bled to a pressure of 20 mm Hg over 15 minutes. CO therapy was started once a pressure of 20 mm Hg was reached.</p

CO protects against organ injury and inflammation in a dose dependent fashion in murine model of hemorrhagic shock and resuscitation.

<p>Lung myeloperoxidase activity (MPO; <b>A.</b>) and serum ALT (<b>B.</b>) at 4 hours after resuscitation in mice demonstrates lung and liver injury, respectively. CO limits this injury in a dose-dependent fashion when treated for 30 minutes (25–500 ppm) starting 90 minutes into hypotension. C. Serum TNF-alpha and IL-6 levels were also increased by hemorrhagic shock and resuscitation at a 4 hour time point, and CO therapy limited these markers of inflammation in a dose dependent fashion. Results are mean±SEM for 8 mice per group. *P<0.05 compared to sham and #P<0.05 compared to shock. ANOVA was utilized for above comparisons.</p

CO therapy protects against mortality in a model of murine hemorrhagic shock and resuscitation.

<p>A severe hemorrhagic shock and resuscitation model resulted in 80% mortality by 36 hours. CO therapy (250 ppm) limited mortality to 20% (P<0.05). Kaplan-Meier method was used to compare survival.</p

CO decreases oxygen consumption and limits the development of cellular hypoxia in hepatocytes in vitro.

<p>A. Oxygen consumption rates of primary murine hepatocytes were demonstrated <i>in vitro</i> in normoxic cells or in hepatocytes immediately following 30 minutes of hypoxia. CO treatment (250ppm) occurred during this normoxic or hypoxic periods. Hypoxia decreased oxygen consumption rates (*P<0.01 compared to normoxic cells) and this was further decreased by CO therapy (#P<0.05 compared to hypoxia alone). Results of four independent experiments, with each condition performed in triplicate. <b>B, C.</b> Representative immunocytochemistry and quantitative mean fluorescence of hypoxyprobe staining in hepatocytes under normoxic, normoxic+CO, hypoxic, or hypoxic +CO conditions for 30 minutes. Increased green staining represents increased cellular hypoxia. ANOVA was utilized for above comparisons.</p