On the other hand, small-molecules also stereoselectively take their biological actions

compared to the AD-A LCLs demonstrating that the AD-N LCLs had a significantly greater ability to adapt to inhibition of the UCP2 protein by increasing proton leak across the inner mitochondrial membrane. Overall, the LCLs exposed to genipin exhibited higher maximal respiratory capacity than the LCLs not exposed to genipin . Maximal respiratory capacity was also higher for AD-A LCLs as compared to AD-N LCLs and for the LCLs not exposed to DMNQ as compared to pretreatment with 10 mM DMNQ. There was a DMNQ by genipin interaction such that the decrease in maximal respiratory capacity with DMNQ was significantly greater for the genipin treated LCLs as compared to the genipin unexposed cells. Overall, reserve capacity was greater for the LCLs exposed to genipin as compared to the unexposed LCLs . Reserve capacity was also greater in the ADA LCLs as compared to the AD-N LCLs and for LCLs not exposed to DMNQ as compared to pretreatment with 10 mM DMNQ. There was a DMNQ by subgroup interaction such that reserve capacity decreased more significantly with DMNQ for the AD-A group than the AD-N group. There was also a DMNQ by genipin interaction such that the decrease in reserve capacity with DMNQ was significantly greater for the genipin treated LCLs as compared 9 Mitochondrial Dysfunction in Autism Cell Lines to the genipin unexposed cells. Lastly, and most interestingly, there was a genipin by LCL subgroup interaction such that reserve capacity changed less for the AD-N LCLs exposed to genipin as compared to the AD-A LCLs exposed to genipin, indicating that the abnormal elevation in reserve capacity seen in the AD-A LCLs was further exacerbated when exposed to genipin. Uncoupling Protein 2 Content Uncoupling Protein 2 is one of the key regulators of proton leak respiration. Since proton leak respiration is one of the major differences in respiratory parameters between the two AD LCL subgroups, we measured UCP2 content by western blots in a subset of LCLs from both the AD-A and AD-N 221244-14-0 subgroups to determine whether UCP2 protein content differed between the two AD subgroups at baseline. As shown in Mitochondrial DNA Copy Number To determine whether the number of mitochondria per cell could account for the differences in the respiratory parameters between the two AD LCL subgroups, we measured mtDNA copy number by calculating the ratio of 3 mitochondria genes, including ND1, ND4 and Cyt B, to the nuclear gene, 6099352 PK. As shown in Intracellular Redox Metabolites and Oxidants in LCLs To verify that ROS production by DMNQ affects glutathione redox balance, glutathione concentrations were measured in 3 control and 5 AD LCLs at DMNQ concentrations of 0, 1, 5, 10, 12.5 and 15 mM. DMNQ significantly decreased GSH and GSH/GSSG and increased GSSG in a linear fashion. These changes were not different across groups. We compared the redox status in the AD and control LCLs by measuring three separate redox couples. AD LCLs demonstrated decreased intracellular GSH and GSH/GSSG ratio but no difference in GSSG as compared to control LCLs. The ratio of reduced cysteine to oxidized cystine was lower in the AD LCLs as compared to the control LCLs although intracellular cysteine and cystine concentrations were not significantly different. The ratio of reduced NADH to oxidized NAD+ was 10355733 also significantly lower in the AD LCLs as compared to control LCLs although intracellular NADH and NAD+ concentrations were not significantly different. 3-nitrotyrosine, a marker of protein oxcompared to the AD-A LCLs demonstrating that the AD-N LCLs had a significantly greater ability to adapt to inhibition of the UCP2 protein by increasing proton leak across the inner mitochondrial membrane. Overall, the LCLs exposed to genipin exhibited higher maximal respiratory capacity than the LCLs not exposed to genipin . Maximal respiratory capacity was also higher for AD-A LCLs as compared to AD-N LCLs and for the LCLs not exposed to DMNQ as compared to pretreatment with 10 mM DMNQ. There was a DMNQ by genipin interaction such that the decrease in maximal respiratory capacity with DMNQ was significantly greater for the genipin treated LCLs as compared to the genipin unexposed cells. Overall, reserve capacity was greater for the LCLs exposed to genipin as compared to the unexposed LCLs . Reserve capacity was also greater in the ADA LCLs as compared to the AD-N LCLs and for LCLs not exposed to DMNQ as compared to pretreatment with 10 mM DMNQ. There was a DMNQ by subgroup interaction such that reserve capacity decreased more significantly with DMNQ for the AD-A group than the AD-N group. There was also a DMNQ by genipin interaction such that the decrease in reserve capacity with DMNQ was significantly greater for the genipin treated LCLs as compared 9 Mitochondrial Dysfunction in Autism Cell Lines to the genipin unexposed cells. Lastly, and most interestingly, there was a genipin by LCL subgroup interaction such that reserve capacity changed less for the AD-N LCLs exposed to genipin as compared to the 7190624 AD-A LCLs exposed to genipin, indicating that the abnormal elevation in reserve capacity seen in the AD-A LCLs was further exacerbated when exposed to genipin. Uncoupling Protein 2 Content Uncoupling Protein 2 is one of the key regulators of proton leak respiration. Since proton leak respiration is one of the major differences in respiratory parameters between the two AD LCL subgroups, we measured UCP2 content by western blots in a subset of LCLs from both the AD-A and AD-N subgroups to determine whether UCP2 protein content differed between the two AD subgroups at baseline. As shown in Mitochondrial DNA Copy Number To determine whether the number of mitochondria per cell could account for the differences in the respiratory parameters between the two AD LCL subgroups, we measured mtDNA copy number by calculating the ratio of 3 mitochondria genes, including ND1, ND4 and Cyt B, to the nuclear gene, PK. As shown in Intracellular Redox Metabolites and Oxidants in LCLs To verify that ROS production by DMNQ affects glutathione redox balance, glutathione concentrations were measured in 3 control and 5 AD LCLs at DMNQ concentrations 15130089 of 0, 1, 5, 10, 12.5 and 15 mM. DMNQ significantly decreased GSH and GSH/GSSG and increased GSSG in a linear fashion. These changes were not different across groups. We compared the redox status in the AD and control LCLs by measuring three separate redox couples. AD LCLs demonstrated decreased intracellular GSH and GSH/GSSG ratio but no difference in GSSG as compared to control LCLs. The ratio of reduced cysteine to oxidized cystine was lower in the AD LCLs as compared to the control LCLs although intracellular cysteine and cystine concentrations were not significantly different. The ratio of reduced NADH to oxidized NAD+ was also significantly lower in the AD LCLs as compared to control LCLs although intracellular NADH and NAD+ concentrations were not significantly different. 3-nitrotyrosine, a marker of protein ox