Generation and Bioenergetic Profiles of Cybrids with East Asian mtDNA Haplogroups

Human mitochondrial DNA (mtDNA) variants and haplogroups may contribute to susceptibility to various diseases and pathological conditions, but the underlying mechanisms are not well understood. To address this issue, we established a cytoplasmic hybrid (cybrid) system to investigate the role of mtDNA haplogroups in human disease; specifically, we examined the effects of East Asian mtDNA genetic backgrounds on oxidative phosphorylation (OxPhos). We found that mtDNA single nucleotide polymorphisms such as m.489T>C, m.10398A>G, m.10400C>T, m.C16223T, and m.T16362C affected mitochondrial function at the level of mtDNA, mtRNA, or the OxPhos complex. Macrohaplogroup M exhibited higher respiratory activity than haplogroup N owing to its higher mtDNA content, mtRNA transcript levels, and complex III abundance. Additionally, haplogroup M had higher reactive oxygen species levels and NAD+/NADH ratios than haplogroup N, suggesting difference in mitonuclear interactions. Notably, subhaplogroups G2, B4, and F1 appeared to contribute significantly to the differences between haplogroups M and N. Thus, our cybrid-based system can provide insight into the mechanistic basis for the role of mtDNA haplogroups in human diseases and the effect of mtDNA variants on mitochondrial OxPhos function. In addition, studies of mitonuclear interaction using this system can reveal predisposition to certain diseases conferred by variations in mtDNA

Mitochondrial complex 1 activity measured by spectrophotometry is reduced across all brain regions in ageing and more specifically in neurodegeneration

Mitochondrial function, in particular complex 1 of the electron transport chain (ETC), has been shown to decrease during normal ageing and in neurodegenerative disease. However, there is some debate concerning which area of the brain has the greatest complex 1 activity. It is important to identify the pattern of activity in order to be able to gauge the effect of age or disease related changes. We determined complex 1 activity spectrophotometrically in the cortex, brainstem and cerebellum of middle aged mice (70–71 weeks), a cerebellar ataxic neurodegeneration model (pcd5J) and young wild type controls. We share our updated protocol on the measurements of complex1 activity and find that mitochondrial fractions isolated from frozen tissues can be measured for robust activity. We show that complex 1 activity is clearly highest in the cortex when compared with brainstem and cerebellum (p<0.003). Cerebellum and brainstem mitochondria exhibit similar levels of complex 1 activity in wild type brains. In the aged brain we see similar levels of complex 1 activity in all three-brain regions. The specific activity of complex 1 measured in the aged cortex is significantly decreased when compared with controls (p<0.0001). Both the cerebellum and brainstem mitochondria also show significantly reduced activity with ageing (p<0.05). The mouse model of ataxia predictably has a lower complex 1 activity in the cerebellum, and although reductions are measured in the cortex and brain stem, the remaining activity is higher than in the aged brains. We present clear evidence that complex 1 activity decreases across the brain with age and much more specifically in the cerebellum of the pcd5j mouse. Mitochondrial impairment can be a region specific phenomenon in disease, but in ageing appears to affect the entire brain, abolishing the pattern of higher activity in cortical regions

A High-Density Genome-Wide Association Screen of Sporadic ALS in US Veterans

Following reports of an increased incidence of amyotrophic lateral sclerosis (ALS) in U.S. veterans, we have conducted a high-density genome-wide association study (GWAS) of ALS outcome and survival time in a sample of U.S. veterans. We tested ∼1.3 million single nucleotide polymorphisms (SNPs) for association with ALS outcome in 442 incident Caucasian veteran cases diagnosed with definite or probable ALS and 348 Caucasian veteran controls. To increase power, we also included genotypes from 5909 publicly-available non-veteran controls in the analysis. In the survival analysis, we tested for association between SNPs and post-diagnosis survival time in 639 Caucasian veteran cases with definite or probable ALS. After this discovery phase, we performed follow-up genotyping of 299 SNPs in an independent replication sample of Caucasian veterans and non-veterans (ALS outcome: 183 cases and 961 controls; survival: 118 cases). Although no SNPs reached genome-wide significance in the discovery phase for either phenotype, three SNPs were statistically significant in the replication analysis of ALS outcome: rs6080539 (177 kb from PCSK2), rs7000234 (4 kb from ZNF704), and rs3113494 (13 kb from LOC100506746). Two SNPs located in genes that were implicated by previous GWA studies of ALS were marginally significant in the pooled analysis of discovery and replication samples: rs17174381 in DPP6 (p = 4.4×10−4) and rs6985069 near ELP3 (p = 4.8×10−4). Our results underscore the difficulty of identifying and convincingly replicating genetic associations with a rare and genetically heterogeneous disorder such as ALS, and suggest that common SNPs are unlikely to account for a substantial proportion of patients affected by this devastating disorder

Carbon isotope composition and its evolution around the base of the Drumian Stage in Linzhou area, northern Henan

Carbon isotope stratigraphy is an important means to subdivide and correlate the Cambrian strata. Based on the carbon isotope analysis of the carbonate strata of Lütuogou Section, northern Henan Province, it is found the δ13C value of carbonate samples collected from the base of the Drumian Stage at Lütuogou Section, northern Henan, ranges from 3.1‰ to -1.7‰, and the δ18O value ranges from -3.9‰ to -9.4‰. No covariance exists between δ13C and δ18O, which implies that the carbon isotope composition is nearly stable. Value of δ13C around the base of the Drumian Stage consists of a negative excursion, the amplitude of the negative excursion is 3.0‰, and the minimum value of δ13C is -1.7‰, which occurs at the place 15 m below the first occurrence of trilobite protasaphiscus. The position and amplitude of the negative excursion are similar to those of the drum carbon isotope excursion (DICE) tested in the Wangcun Section, western Hunan, South China, and the Drum Mountains section in Utah, USA, which suggests that the DICE can be used as a global indicator for the correlation of the Drumian Stage. Furthermore, the cyan shale with thin limestone at the position of minimum δ13C overlies the oolitic limestone; therefore, DICE occurs in the transgressive succession in the early Miaolingian

Carbon-Isotope Excursions Recorded in the Cambrian System, South China: Implications for Mass Extinctions and Sea-Level Fluctuations

Cambrian carbonates with abundant fossils of agnostoid trilobites deposited on the southern slope (Jiangnan slope belt) of the Yangtze Platform and in the Jiangnan deepwater basin are well exposed in the Wangcun Section of western Hunan, South China, and in the Duibian A Section of western Zhejiang, southeastern China, respectively. To better understand the response of carbon-isotope excursions to depositional environment changes, mass extinctions and eustatic events, we collected 530 carbonate samples in fresh roadcut exposures of the two measured sections for analysis of carbon and oxygen isotopic compositions. Data of δ13C from the Wangcun Section, western Hunan, South China, demonstrate that the Cambrian carbon-isotope profile includes three remarkable positive excursions CPEwc-1, 2, 3 in the Upper Series 2, in the Lower and in the Middle Furongian Series. Three distinctive negative excursions CNEwc-1, 2, 3 were separately tested in the Lower Terreneuvian Series, Lower Series 3 and in the Upper Furongian Series. Similarly, in the corresponding horizons in the Duibian A Section, Zhejiang Province, southeastern China, three positive excursions CPEdb-1, 2, 3 and three negative excursions CNEdb-1, 2, 3 also have been discovered. We interpret these significant carbon-isotope excursions as being associated with enhanced biogenic productivity, mass extinctions and eustatic events

Simultaneous Improvement of Efficiency and Lifetime of Quantum Dot Light-Emitting Diodes with a Bilayer Hole Injection Layer Consisting of PEDOT:PSS and Solution-Processed WO₃

Even though chemically stable metal oxides (MOs), as substitutes for poly­(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS), have been successfully adopted for improving device stability in solution-processed quantum dot light-emitting diodes (QLEDs), the efficiencies of QLEDs are at a relatively low level. In this work, a novel architecture of QLEDs has been introduced, in which inorganic/organic bilayer hole injection layers (HILs) were delicately designed by inserting an amorphous WO₃ interlayer between PEDOT:PSS and the indium tin oxide anode. As a result, the efficiency and operational lifetime of QLEDs were improved simultaneously. The results show that the novel architecture QLEDs relative to conventional PEDOT:PSS-based QLEDs have an enhanced external quantum efficiency by a factor of 50%, increasing from 8.31 to 12.47%, meanwhile exhibit a relatively long operational lifetime (12 551 h) and high maximum brightness (>40 000 cd m^–2) resulting from a better pathway for hole injection with staircase energy-level alignment of the HILs and reduction of surface roughness. Our results demonstrate that the novel architecture QLEDs using bilayer MO/PEDOT:PSS HILs can achieve long operational lifetime without sacrificing efficiency