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6) (14 pts.) While treating his fields with dinitro-ortho-cresol (DNOC)) an insecticide that is now banned by the EPA. a farmer became very ill. He was rushed to the hospital with an elevated body temperature of 39.3°C. He was sweating profusely and had an elevated respiratory rate of 60/mín. Despite attempts to reduce his body temperature he died in a coma shortly after admission to the hospital. Autopsy findings showed a striking absence of subcutaneous and abdominal fat.) In view of the clinical findings. a sample of the pesticide was tested on samples of rat mitochondria incubated in oxygenated buffer and either succinate (Fig 1a), or, malate and glutamate (Fig 1b). Figures la and 1b show the oxygen concentrations measured in the samples as various compounds were added to the incubation mixture. Figure 1. Effects of adding DNOC to rat liver mitochondria. Succinate (5 nMI Glutamate + malate {each 5 mM} ADP (1 mM) ADP M} Admici's Amonimate DNOC (1 mM) [O2] DNOC (1 mM} [O2] Oligomyein (5 mM) KCN 1: mM) Time Time (b) (a) A) Explain why the rate of oxygen consumption in both Figures la and 1b, increases In dramatically Fig 1b, malate oxidation supplies the electrons to the electron transport chain. Why when ADP is added to the suspension. B) is it to also supply glutamate to the suspension? C) than the rate of respiration in the presence of glutamate, malate, and Why necessary is the rate of respiration in the presence of succinate and ADP ADP? (Fig. la) higher E) D) Under Explain normal conditions (No DNOC), what would happen after the addition of the result of adding KCN to the suspension (Fig. la). oligomycin? What do the data allow you to conclude regarding the metabolic effect of DNOC? Provide a sketch of how Fig. 1b would be different. F) (Explain how your conclusion is consistent with the data shown in Fig la and 1b) G) Explain why: i) the victim's body temperature was elevated H) ii) (Bonus) Propose an explanation for why there was a striking absence of body fat found, the victim's respiration was elevated during the autopsy

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a) Why is the rate of respiration in the presence of succinate and ADP (fig 1a) higher than the rate of respiration in the presence of glutamate, malate and ADP?
Respiration rate depends on the mitochondrial respiration, substrate supply and redox ration. Oxygen consumption in state 3 of oxygen respiration depends on the used substrate. The lowest is for pyruvate/malate (NAD related substrates, protons pumped by complexes I, III and IV of the respiratory chain), almost twice higher with succinate (FAD-related substrate, complex I omitted) and several times higher with external reduced cytochrome c or ascorbate +TMPD. Superoxide radical associated oxygen consumption with glutamate/malate is low since glutamate can be converted to succinate by mitochondrial transaminase. Succinate has higher hydrogen supply system which results...
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