A Sustained Dietary Change Increases Epigenetic Variation in Isogenic Mice

Epigenetic changes can be induced by adverse environmental exposures, such as nutritional imbalance, but little is known about the nature or extent of these changes. Here we have explored the epigenomic effects of a sustained nutritional change, excess dietary methyl donors, by assessing genomic CpG methylation patterns in isogenic mice exposed for one or six generations. We find stochastic variation in methylation levels at many loci; exposure to methyl donors increases the magnitude of this variation and the number of variable loci. Several gene ontology categories are significantly overrepresented in genes proximal to these methylation-variable loci, suggesting that certain pathways are susceptible to environmental influence on their epigenetic states. Long-term exposure to the diet (six generations) results in a larger number of loci exhibiting epigenetic variability, suggesting that some of the induced changes are heritable. This finding presents the possibility that epigenetic variation within populations can be induced by environmental change, providing a vehicle for disease predisposition and possibly a substrate for natural selection.This work was supported by the Australian Research Council (DP0771859) and the National Health and Medical Research Council (#459412, #635510)

Representation in Westminster in the 1990s : The ghost of Edmund Burke

Why are 'trustee' notions of representation still invoked in the UK House of Commons in the 1990s? In answering this question this article analyses the premises of Burkean theory and the arguments that these premises are of little relevance in the late twentieth century. Despite these dismissals of trusteeship, Burkean ideas are still articulated in the Commons some 200 years after they were first voiced. The idea of trusteeship can prove extremely useful to justify the actions of representatives when those actions conflict with constituency 'opinion', party policy or the wishes of interest groups. Examples of the occasions when Burkean notions have been invoked in the 1990s are provided

Synthetic lethal screening in the mammalian central nervous system identifies Gpx6 as a modulator of Huntington’s disease

Huntington’s disease, the most common inherited neurodegenerative disease, is characterized by a dramatic loss of deep-layer cortical and striatal neurons, as well as morbidity in midlife. Human genetic studies led to the identification of the causative gene, huntingtin. Recent genomic advances have also led to the identification of hundreds of potential interacting partners for huntingtin protein and many hypotheses as to the molecular mechanisms whereby mutant huntingtin leads to cellular dysfunction and death. However, the multitude of possible interacting partners and cellular pathways affected by mutant huntingtin has complicated efforts to understand the etiology of this disease, and to date no curative therapeutic exists. To address the general problem of identifying the disease-phenotype contributing genes from a large number of correlative studies, here we develop a synthetic lethal screening methodology for the mammalian central nervous system, called SLIC, for synthetic lethal in the central nervous system. Applying SLIC to the study of Huntington’s disease, we identify the age-regulated glutathione peroxidase 6 (Gpx6) gene as a modulator of mutant huntingtin toxicity and show that overexpression of Gpx6 can dramatically alleviate both behavioral and molecular phenotypes associated with a mouse model of Huntington’s disease. SLIC can, in principle, be used in the study of any neurodegenerative disease for which a mouse model exists, promising to reveal modulators of neurodegenerative disease in an unbiased fashion, akin to screens in simpler model organisms.National Institute of Neurological Disorders and Stroke (U.S.) (Award R01NS085880)William N. and Bernice E. Bumpus Foundation (Early Career Investigator Innovation Award)JPB FoundationEuropean Molecular Biology Organization (Long-term Fellowship

Efectos de la salinidad sobre la gravedad específica y la viabilidad de huevos de una carpa norteamericana (Ciprinidae)

The influence of salinity on survival, specific gravity, and size of eggs of the endangered Hybognathus amarus (Rio Grande silvery minnow) was studied to provide insight into factors affecting their potential dispersal and fate. Under low salinity conditions egg specific gravity declined significantly in the first hour after spawning as the perivitelline space of the egg filled with water. Egg specific gravity achieved a minimum value approximately 1 h post-spawning and remained approximately constant until hatching, which occurred near 48 h post-spawning at 20°C. Specific gravity of the egg depended on the salinity of the water surrounding the egg: hardened eggs changed rapidly in diameter and specific gravity when exposed to water of higher salinity. Size and specific gravity of H. amarus eggs also differed when the eggs were incubated in different groundwater sources. Experiments indicated that calcium chloride saline solution had a greater effect on egg specific gravity and size than did solutions of sodium or potassium chlorides. Survival of H. amarus eggs declined sharply at salinity greater than 3 (practical salinity units, PSU) and was only 5% at a salinity of 8. Habitat restoration to benefit H. amarus should consider the salinity of habitats in which eggs incubate.El efecto de la salinidad sobre la supervivencia, gravedad específica y talla de los huevos del ciprínido puesto en peligro de extinción, Hybognathus amarus, fue estudiado para proporcionar información sobre los factores que afectan la dispersión y el destino de los huevos. Bajo condiciones de salinidad baja la gravedad específica del huevo disminuyó de forma significativa durante la primera hora después de la puesta, a medida que el espacio del perivitelino del huevo se fue llenando de agua. la gravedad específica del huevo alcanzó un valor mínimo 1 hora después de la puesta y quedó aproximadamente constante hasta eclosión, que ocurrió alrededor de 48 horas tras la puesta a 20˚C. la gravedad específica del huevo dependió de la salinidad del agua que le rodeaba. los huevos con corion endurecido cambiaron rápidamente en diámetro y gravedad específica cuando fueron expuestos a agua de salinidad más alta. El tamaño y la gravedad específica de los huevos de H. amarus también se diferenciaron cuando los huevos fueron incubados en diferentes fuentes de agua subterránea. Los experimentos indicaron que la solución salina del cloruro de calcio tuvo un mayor efecto sobre la gravedad específica del huevo que las soluciones de los cloruros del sodio o del potasio. Expuestos a soluciones de salinidad de más de 3 (PsU), la supervivencia de los huevos de H. amarus disminuyó claramente, bajando a sólo el 5% en la solución de 8. la restauración del hábitat con el objetivo de beneficiar a H. amarus debe considerar la salinidad de los hábitats en los cuales el huevo incuba

Mice Expressing Low Levels of CalDAG-GEFI Exhibit Markedly Impaired Platelet Activation With Minor Impact on HemostasisHighlights

OBJECTIVE: The tight regulation of platelet adhesiveness, mediated by the αIIbβ3 integrin, is critical for hemostasis and prevention of thrombosis. We recently demonstrated that integrin affinity in platelets is controlled by the guanine nucleotide exchange factor, CalDAG-GEFI (CD-GEFI), and its target, RAP1. In this study, we investigated whether low-level expression of CD-GEFI leads to protection from thrombosis without pathological bleeding in mice. APPROACH AND RESULTS: Cdg1(low) mice were generated by knockin of human CD-GEFI cDNA into the mouse Cdg1 locus. CD-GEFI expression in platelets from Cdg1(low) mice was reduced by ≈90% when compared with controls. Activation of RAP1 and αIIbβ3 was abolished at low agonist concentrations and partially inhibited at high agonist concentrations in Cdg1(low) platelets. Consistently, the aggregation response of Cdg1(low) platelets was weaker than that of wild-type platelets, but more efficient than that observed in Cdg1(-/-) platelets. Importantly, Cdg1(low) mice were strongly protected from arterial and immune complex-mediated thrombosis, with only minimal impact on primary hemostasis. CONCLUSIONS: Together, our studies suggest the partial inhibition of CD-GEFI function as a powerful new approach to safely prevent thrombotic complications

Structure of the Reduced Copper Active Site in Pre-Processed Galactose Oxidase: Ligand Tuning for One-Electron O2 Activation in Cofactor Biogenesis

Galactose oxidase (GO) is a copper-dependent enzyme that accomplishes 2e- substrate oxidation by pairing a single copper with an unusual cysteinylated tyrosine (Cys-Tyr) redox cofactor. Previous studies have demonstrated that the post-translational biogenesis of Cys-Tyr is copper- and O2-dependent, resulting in a self-processing enzyme system. To investigate the mechanism of cofactor biogenesis in GO, the active-site structure of Cu(I)-loaded GO was determined using X-ray absorption near edge structure (XANES) and extended X-ray absorption fine structure (EXAFS) spectroscopy, and density-functional theory (DFT) calculations were performed on this model. Our results show that the active-site tyrosine lowers the Cu potential to enable the thermodynamically unfavorable 1e- reduction of O2, and the resulting Cu(II)-O2¿- is activated toward H atom abstraction from cysteine. The final step of biogenesis is a concerted reaction involving coordinated Tyr ring deprotonation where Cu(II) coordination enables formation of the Cys-Tyr cross-link. These spectroscopic and computational results highlight the role of the Cu(I) in enabling O2 activation by 1e- and the role of the resulting Cu(II) in enabling substrate activation for biogenesis

Six supersoft X-ray binaries: system parameters and twin-jet outflows

A comparison is made between the properties of CAL 83, CAL 87, RX J0513.9-6951, 1E 0035.4-7230 (SMC 13), RX J0019.8+2156, and RX J0925.7-4758, all supersoft X-ray binaries. Spectra with the same resolution and wavelength coverage of these systems are compared and contrasted. Some new photometry is also presented. The equivalent widths of the principal emission lines of H and He II differ by more than an order of magnitude among these sources, although those of the highest ionization lines (e.g. O VI) are very similar. In individual systems, the velocity curves derived from various ions often differ in phasing and amplitude, but those whose phasing is consistent with the light curves (implying the lines are formed near the compact star) give masses of $\sim 1.2M_{\odot}$ and $\sim 0.5M_{\odot}$ for the degenerate and mass-losing stars, respectively. This finding is in conflict with currently prevailing theoretical models for supersoft binaries. The three highest luminosity sources show evidence of "jet" outflows, with velocities of $\sim 1-4 \times10^3 km/s$. In CAL 83 the shape of the He II 4686\AA profile continues to show evidence that these jets may precess with a period of $\sim 69$ days.Comment: 27 pages including 5 tables, plus 6 figures. To appear in Ap

PIK3CA mutant tumors depend on oxoglutarate dehydrogenase

Oncogenic PIK3CA mutations are found in a significant fraction of human cancers, but therapeutic inhibition of PI3K has only shown limited success in clinical trials. To understand how mutant PIK3CA contributes to cancer cell proliferation, we used genome scale loss-of-function screening in a large number of genomically annotated cancer cell lines. As expected, we found that PIK3CA mutant cancer cells require PIK3CA but also require the expression of the TCA cycle enzyme 2-oxoglutarate dehydrogenase (OGDH). To understand the relationship between oncogenic PIK3CA and OGDH function, we interrogated metabolic requirements and found an increased reliance on glucose metabolism to sustain PIK3CA mutant cell proliferation. Functional metabolic studies revealed that OGDH suppression increased levels of the metabolite 2-oxoglutarate (2OG). We found that this increase in 2OG levels, either by OGDH suppression or exogenous 2OG treatment, resulted in aspartate depletion that was specifically manifested as auxotrophy within PIK3CA mutant cells. Reduced levels of aspartate deregulated the malate-aspartate shuttle, which is important for cytoplasmic NAD + regeneration that sustains rapid glucose breakdown through glycolysis. Consequently, because PIK3CA mutant cells exhibit a profound reliance on glucose metabolism, malate-aspartate shuttle deregulation leads to a specific proliferative block due to the inability to maintain NAD + /NADH homeostasis. Together these observations define a precise metabolic vulnerability imposed by a recurrently mutated oncogene. Keyword: PIK3CA; 2OG; OGDH; TCA cycle; glycolysisDamon Runyon Cancer Research Foundation (HHMI Fellowship