dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression.

Distinct epigenomic profiles of histone marks have been associated with gene expression, but questions regarding the causal relationship remain. Here we investigated the activity of a broad collection of genomically targeted epigenetic regulators that could write epigenetic marks associated with a repressed chromatin state (G9A, SUV39H1, Krüppel-associated box (KRAB), DNMT3A as well as the first targetable versions of Ezh2 and Friend of GATA-1 (FOG1)). dCas9 fusions produced target gene repression over a range of 0- to 10-fold that varied by locus and cell type. dCpf1 fusions were unable to repress gene expression. The most persistent gene repression required the action of several effector domains; however, KRAB-dCas9 did not contribute to persistence in contrast to previous reports. A 'direct tethering' strategy attaching the Ezh2 methyltransferase enzyme to dCas9, as well as a 'recruitment' strategy attaching the N-terminal 45 residues of FOG1 to dCas9 to recruit the endogenous nucleosome remodeling and deacetylase complex, were both successful in targeted deposition of H3K27me3. Surprisingly, however, repression was not correlated with deposition of either H3K9me3 or H3K27me3. Our results suggest that so-called repressive histone modifications are not sufficient for gene repression. The easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation

The rotation and coma profiles of comet C/2004 Q2 (Machholz)

Aims. Rotation periods of cometary nuclei are scarce, though important when studying the nature and origin of these objects. Our aim is to derive a rotation period for the nucleus of comet C/2004 Q2 (Machholz). Methods. C/2004 Q2 (Machholz) was monitored using the Merope CCD camera on the Mercator telescope at La Palma, Spain, in January 2005, during its closest approach to Earth, implying a high spatial resolution (50km per pixel). One hundred seventy images were recorded in three different photometric broadband filters, two blue ones (Geneva U and B) and one red (Cousins I). Magnitudes for the comet's optocentre were derived with very small apertures to isolate the contribution of the nucleus to the bright coma, including correction for the seeing. Our CCD photometry also permitted us to study the coma profile of the inner coma in the different bands. Results. A rotation period for the nucleus of P = 9.1 +/- 0.2 h was derived. The period is on the short side compared to published periods of other comets, but still shorter periods are known. Nevertheless, comparing our results with images obtained in the narrowband CN filter, the possibility that our method sampled P/2 instead of P cannot be excluded. Coma profiles are also presented, and a terminal ejection velocity of the grains v_gr = 1609 +/- 48 m/s is found from the continuum profile in the I band.Comment: 11 pages, 9 figures, accepted by A&

Is the Forest Service Keeping Up With the Times?

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75358/1/j.1541-0072.1995.tb01747.x.pd

The role of DNA methylation in directing the functional organization of the cancer epigenome

The holistic role of DNA methylation in the organization of the cancer epigenome is not well understood. Here we perform a comprehensive, high-resolution analysis of chromatin structure to compare the landscapes of HCT116 colon cancer cells and a DNA methylation-deficient derivative. The NOMe-seq accessibility assay unexpectedly revealed symmetrical and transcription-independent nucleosomal phasing across active, poised, and inactive genomic elements. DNA methylation abolished this phasing primarily at enhancers and CpG island (CGI) promoters, with little effect on insulators and non-CGI promoters. Abolishment of DNA methylation led to the context-specific reestablishment of the poised and active states of normal colon cells, which were marked in methylation-deficient cells by distinct H3K27 modifications and the presence of either well-phased nucleosomes or nucleosome-depleted regions, respectively. At higher-order genomic scales, we found that long, H3K9me3-marked domains had lower accessibility, consistent with a more compact chromatin structure. Taken together, our results demonstrate the nuanced and context-dependent role of DNA methylation in the functional, multiscale organization of cancer epigenomes.Charles Heidelberger Memorial Fellowshi

Changes in household wealth in communities living in proximity to a large-scale copper mine in Zambia

Large-scale mining can alter the living conditions of surrounding communities in positive and negative ways. A health impact assessment conducted in the context of a newly developed large-scale copper mine in rural Zambia gave us the opportunity to measure changes in health determinants over time. We conducted periodic household surveys at baseline in 2011, during the construction phase in 2015 and during the operational phase in 2019. Data collected included economic indicators that were based on the standardized list of household assets used in the Zambia Demographic and Health Survey, which we subsequently converted into a wealth score using principal component analysis. We compared mean wealth scores in six communities directly impacted by the mine with comparison communities, as well as the rest of the North-Western province of Zambia. A difference-indifferences linear regression model was used to compare changes over time. Mean wealth of the communities near the mine was significantly lower at baseline than that of the North-Western province (? 0.54 points; pvalue< 0.001) in 2011, but surpassed the regional average in 2019 (+1.07 points; p-value <0.001). Mean wealth increased more rapidly in communities directly impacted by mine than in the comparison communities (+0.30 points, p-value <0.001). These results suggest a positive impact on living conditions in communities living near this copper mine. Our findings underscore the potential of the mining sector to contribute to economic development in Zambia

Epigenetic Modulation of miR-122 Facilitates Human Embryonic Stem Cell Self-Renewal and Hepatocellular Carcinoma Proliferation

The self-renewal capacity ascribed to hESCs is paralleled in cancer cell proliferation, suggesting that a common network of genes may facilitate the promotion of these traits. However, the molecular mechanisms that are involved in regulating the silencing of these genes as stem cells differentiate into quiescent cellular lineages remain poorly understood. Here, we show that a differentiated cell specific miR-122 exemplifies this regulatory attribute by suppressing the translation of a gene, Pkm2, which is commonly enriched in hESCs and liver cancer cells (HCCs), and facilitates self-renewal and proliferation. Through a series of gene expression analysis, we show that miR-122 expression is highly elevated in quiescent human primary hepatocytes (hPHs) but lost or attenuated in hESCs and HCCs, while an opposing expression pattern is observed for Pkm2. Depleting hESCs and HCCs of Pkm2, or overexpressing miR-122, leads to a common deficiency in self-renewal and proliferation. Likewise, during the differentiation process of hESCs into hepatocytes, a reciprocal expression pattern is observed between miR-122 and Pkm2. An examination of the genomic region upstream of miR-122 uncovered hyper-methylation in hESCs and HCCs, while the same region is de-methylated and occupied by a transcription initiating protein, RNA polymerase II (RNAPII), in hPHs. These findings indicate that one possible mechanism by which hESC self-renewal is modulated in quiescent hepatic derivatives of hESCs is through the regulatory activity of a differentiated cell-specific miR-122, and that a failure to properly turn “on” this miRNA is observed in uncontrollably proliferating HCCs

The Claims Culture: A Taxonomy of Industry Attitudes

This paper presents an analysis of a familiar aspect of construction industry culture that we have dubbed 'the claims culture'. This is a culture of contract administration that lays a strong emphasis on the planning and management of claims. The principal elements of the analysis are two sets of distinctions. The first comprises economic and occupational orders, referring to two kinds of control that are exercised over the construction process; predicated respectively on economic ownership and occupational competence. The second refers to contrasting attitudes towards relationships and problem solving within these orders: respectively 'distributive' and 'integrative'. The concepts of economic and occupational order entail further sub-categories. The various attitudes associated with these categories and sub-categories are described. They are assessed as to their consequences for change initiatives in the industry

Preventing Perinatal Transmission of HIV Costs and Effectiveness of a Recommended Intervention

In the United States, nearly all new human immunodeficiency virus (HIV) infections in children are acquired through perinatal (mother-to-infant) transmission. Each year, approximately 7000 infants are born to HIV-infected women in the United States.1 Without intervention, an estimated 15-30% of these infants would become infected.2 In 1994, results of the AIDS Clinical Trial Group (ACTG) Protocol 076 showed that treatment of infected pregnant women and their infants with zidovudine (ZDV) reduced the rate of perinatal HIV transmission from 25% to 8%.3,4 Following these findings, the Public Health Service (PHS) issued recommendations for ZDV therapy to prevent perinatal HIV transmission5 and for HIV counseling and voluntary testing of pregnant women.