Disabling de novo DNA methylation in embryonic stem cells allows an illegitimate fate trajectory.

Genome remethylation is essential for mammalian development but specific reasons are unclear. Here we examined embryonic stem (ES) cell fate in the absence of de novo DNA methyltransferases. We observed that ES cells deficient for both Dnmt3a and Dnmt3b are rapidly eliminated from chimeras. On further investigation we found that in vivo and in vitro the formative pluripotency transition is derailed toward production of trophoblast. This aberrant trajectory is associated with failure to suppress activation of Ascl2Ascl2 encodes a bHLH transcription factor expressed in the placenta. Misexpression of Ascl2 in ES cells provokes transdifferentiation to trophoblast-like cells. Conversely, Ascl2 deletion rescues formative transition of Dnmt3a/b mutants and improves contribution to chimeric epiblast. Thus, de novo DNA methylation safeguards against ectopic activation of Ascl2 However, Dnmt3a/b-deficient cells remain defective in ongoing embryogenesis. We surmise that multiple developmental transitions may be secured by DNA methylation silencing potentially disruptive genes.

Genome-wide methylation data improves dissection of the effect of smoking on body mass index.

Variation in obesity-related traits has a genetic basis with heritabilities between 40 and 70%. While the global obesity pandemic is usually associated with environmental changes related to lifestyle and socioeconomic changes, most genetic studies do not include all relevant environmental covariates, so the genetic contribution to variation in obesity-related traits cannot be accurately assessed. Some studies have described interactions between a few individual genes linked to obesity and environmental variables but there is no agreement on their total contribution to differences between individuals. Here we compared self-reported smoking data and a methylation-based proxy to explore the effect of smoking and genome-by-smoking interactions on obesity related traits from a genome-wide perspective to estimate the amount of variance they explain. Our results indicate that exploiting omic measures can improve models for complex traits such as obesity and can be used as a substitute for, or jointly with, environmental records to better understand causes of disease.

The Nucleosome Remodelling and Deacetylation complex suppresses transcriptional noise during lineage commitment.

Multiprotein chromatin remodelling complexes show remarkable conservation of function amongst metazoans, even though components present in invertebrates are often found as multiple paralogous proteins in vertebrate complexes. In some cases, these paralogues specify distinct biochemical and/or functional activities in vertebrate cells. Here, we set out to define the biochemical and functional diversity encoded by one such group of proteins within the mammalian Nucleosome Remodelling and Deacetylation (NuRD) complex: Mta1, Mta2 and Mta3. We find that, in contrast to what has been described in somatic cells, MTA proteins are not mutually exclusive within embryonic stem (ES) cell NuRD and, despite subtle differences in chromatin binding and biochemical interactions, serve largely redundant functions. ES cells lacking all three MTA proteins exhibit complete NuRD loss of function and are viable, allowing us to identify a previously unreported function for NuRD in reducing transcriptional noise, which is essential for maintaining a proper differentiation trajectory during early stages of lineage commitment.

Stringent Response of Cyanobacteria and Other Bacterioplankton during Different Stages of a Harmful Cyanobacterial Bloom.

We conducted a field study to investigate the role of stringent response in cyanobacteria and coexisting bacterioplankton during nutrient-deprived periods at various stages of bloom in a freshwater lake (Utah Lake) for the first time. Using metagenomics and metatranscriptomics analyses, we examined the cyanobacterial ecology and expression of important functional genes related to stringent response, N and P metabolism, and regulation. Our findings mark a significant advancement in understanding the mechanisms by which toxic cyanobacteria survive and proliferate during nitrogen (N) and phosphorus (P) limitations. We successfully identified and analyzed the metagenome-assembled genomes (MAGs) of the dominant bloom-forming cyanobacteria, namely, Dolichospermum circinale, Aphanizomenon flos-aquae UKL13-PB, Planktothrix agardhii, and Microcystis aeruginosa. By mapping RNA-seq data to the coding sequences of the MAGs, we observed that these four prevalent cyanobacteria species activated multiple functions to adapt to the depletion of inorganic nutrients. During and after the blooms, the four dominant cyanobacteria species expressed high levels of transcripts related to toxin production, such as microcystins (mcy), anatoxins (ana), and cylindrospermopsins (cyr). Additionally, genes associated with polyphosphate (poly-P) storage and the stringent response alarmone (p)ppGpp synthesis/hydrolysis, including ppk, relA, and spoT, were highly activated in both cyanobacteria and bacterioplankton. Under N deficiency, the main N pathways shifted from denitrification and dissimilatory nitrate reduction in bacterioplankton toward N2-fixing and assimilatory nitrate reduction in certain cyanobacteria with a corresponding shift in the community composition. P deprivation triggered a stringent response mediated by spoT-dependent (p)ppGpp accumulation and activation of the Pho regulon in both cyanobacteria and bacterioplankton, facilitating inorganic and organic P uptake. The dominant cyanobacterial MAGs exhibited the presence of multiple alkaline phosphatase (APase) transcripts (e.g., phoA in Dolichospermum, phoX in Planktothrix, and Microcystis), suggesting their ability to synthesize and release APase enzymes to convert ambient organic P into bioavailable forms. Conversely, transcripts associated with bacterioplankton-dominated pathways like denitrification were low and did not align with the occurrence of intense cyanoHABs. The strong correlations observed among N, P, stringent response metabolisms and the succession of blooms caused by dominant cyanobacterial species provide evidence that the stringent response, induced by nutrient limitation, may activate unique N and P functions in toxin-producing cyanobacteria, thereby sustaining cyanoHABs.

Determination of an optimally sensitive and specific chemical exchange saturation transfer MRI quantification metric in relevant biological phantoms.

The purpose of this study was to develop realistic phantom models of the intracellular environment of metastatic breast tumour and naïve brain, and using these models determine an analysis metric for quantification of CEST MRI data that is sensitive to only labile proton exchange rate and concentration. The ability of the optimal metric to quantify pH differences in the phantoms was also evaluated. Novel phantom models were produced, by adding perchloric acid extracts of either metastatic mouse breast carcinoma cells or healthy mouse brain to bovine serum albumin. The phantom model was validated using 1 H NMR spectroscopy, then utilized to determine the sensitivity of CEST MRI to changes in pH, labile proton concentration, T1 time and T2 time; six different CEST MRI analysis metrics (MTRasym , APT*, MTRRex , AREX and CESTR* with and without T1 /T2 compensation) were compared. The new phantom models were highly representative of the in vivo intracellular environment of both tumour and brain tissue. Of the analysis methods compared, CESTR* with T1 and T2 time compensation was optimally specific to changes in the CEST effect (i.e. minimal contamination from T1 or T2 variation). In phantoms with identical protein concentrations, pH differences between phantoms could be quantified with a mean accuracy of 0.6 pH units. We propose that CESTR* with T1 and T2 time compensation is the optimal analysis method for these phantoms. Analysis of CEST MRI data with T1 /T2 time compensated CESTR* is reproducible between phantoms, and its application in vivo may resolve the intracellular alkalosis associated with breast cancer brain metastases without the need for exogenous contrast agents.

Metabolic flux prediction in cancer cells with altered substrate uptake.

Proliferating cells, such as cancer cells, are known to have an unusual metabolism, characterized by an increased rate of glycolysis and amino acid metabolism. Our understanding of this phenomenon is limited but could potentially be used in order to develop new therapies. Computational modelling techniques, such as flux balance analysis (FBA), have been used to predict fluxes in various cell types, but remain of limited use to explain the unusual metabolic shifts and altered substrate uptake in human cancer cells. We implemented a new flux prediction method based on elementary modes (EMs) and structural flux (StruF) analysis and tested them against experimentally measured flux data obtained from (13)C-labelling in a cancer cell line. We assessed the quality of predictions using different objective functions along with different techniques in normalizing a metabolic network with more than one substrate input. Results show a good correlation between predicted and experimental values and indicate that the choice of cellular objective critically affects the quality of predictions. In particular, lactate gives an excellent correlation and correctly predicts the high flux through glycolysis, matching the observed characteristics of cancer cells. In contrast with FBA, which requires a priori definition of all uptake rates, often hard to measure, atomic StruFs (aStruFs) are able to predict uptake rates of multiple substrates.

China's landscape in oncology drug research: perspectives from research collaboration networks.

OBJECTIVE: Better understanding of China's landscape in oncology drug research is of great significance for discovering anti-cancer drugs in future. This article differs from previous studies by focusing on Chinese oncology drug research communities in co-publication networks at the institutional level. Moreover, this research aims to explore structures and behaviors of relevant research units by thematic community analysis and to address policy recommendations. METHODS: This research used social network analysis to define an institutions network and to identify a community network which is characterized by thematic content. RESULTS: A total of 675 sample articles from 2008 through 2012 were retrieved from the Science Citation Index Expanded (SCIE) database of Web of Science, and top institutions and institutional pairs are highlighted for further discussion. Meanwhile, this study revealed that institutions based in the Chinese mainland are located in a relatively central position, Taiwan's institutions are closely assembled on the side, and Hong Kong's units located in the middle of the Chinese mainland's and Taiwan's. Spatial division and institutional hierarchy are still critical barriers to research collaboration in the field of anti-cancer drugs in China. In addition, the communities focusing on hot research areas show the higher nodal degree, whereas communities giving more attention to rare research subjects are relatively marginalized to the periphery of network. CONCLUSIONS: This paper offers policy recommendations to accelerate cross-regional cooperation, such as through developing information technology and increasing investment. The brokers should focus more on outreach to other institutions. Finally, participation in topics of common interest is conducive to improved efficiency in research and development (R&D) resource allocation.

EUPRO - A reference database on project-based R&D collaboration networks.

The EUPRO database enables the analysis of participation patterns of organisations in and across different European R&D funding initiatives and the investigation of resulting collaborative R&D network structures and dynamics. The perimeter of EUPRO is currently more than 600,000 R&D projects funded by European (EU, transnational or national) research funding organisations, comprising systematic information about contents of the R&D projects, their participating organizations (including organisation type and location), and a number of additional characteristics (e.g. underlying policy instrument and programme). This scientific data descriptor serves as illustrative information source for users, both from science as well as from policy. It discusses the conceptual background and derives respective analytical opportunities for different actual, highly relevant debates in innovation studies and related fields. Moreover, the data collection process is described in a compact manner, as well as how the collected data are harmonized and aggregated into a suitable data model for analytical purposes. Finally, we put forward issues of technical validation, data quality and enrichment, and usage notes on how to access EUPRO.

400 million voting records show profound racial and geographic disparities in voter turnout in the United States.

One of the core tenets of a well-functioning representative democracy is that the people who vote to elect government officials are representative of the public. Here we reinforce the idea that reality is far from this lofty ideal. We document the extent and nature of inequities in voter participation in the United States with a level of granularity and precision that previous research has not afforded. To do so, we use a unique nationwide dataset of approximately 400 million validated voting records across multiple election cycles. With this novel dataset, we document large and persistent gaps in voter turnout by race, age, and political affiliation. Minority citizens, young people, and those who support the Democratic Party are much less likely to vote than whites, older citizens, and Republican Party supporters. Minorities, youth, and democrats are also much more likely to live in local communities where fewer individuals vote-areas that we term turnout deserts. Turnout deserts are especially pernicious given that they are self-reinforcing-bolstered by the social dynamics that fundamentally shape citizens' voting patterns. Our results show just how glaring inequities in political participation are in the US. These patterns threaten the very fabric of our democracy and fundamentally shift the balance of political power in the halls of government towards the interests of whites, older citizens, and republicans. They illustrate that participation in the United States is strikingly unequal-far from the ideals that this country has long aspired to.

Interpretation and reporting of mixed DNA profiles by seven forensic laboratories in the UK and Ireland.

In 2014, the UK Forensic Science Regulator (FSR) commissioned a collaborative trial to assess the methods used by forensic service providers (FSPs) in the UK and Ireland for analysis, interpretation and reporting of mixed DNA profiles. Five different mixed samples of varying complexity with supporting mock case circumstances were tested using SGMPlus™ and the newly introduced DNA-17(+) multiplexes and reported by participating laboratories. The results demonstrated a high degree of consistency in analytical methods and allele designations, but some variation in the statistical evaluation and reporting of results. Some of the differences noted were attributable to the major technology change to 17(+)-STR systems which had recently been implemented across the UK at that time. The FSR made recommendations based on the trial outcomes which were intended to produce a more consistent approach to mixtures analysis, interpretation and reporting. Four years later, the Association of Forensic Science Providers (AFSP) repeated the trial, with all major UK and Ireland FSPs (both public sector and private companies) again participating. This second trial used the same mixture set as the 2014 trial but was focussed on the methods for interpretation and evaluation. Since 2014, all UK and Ireland FSPs have implemented probabilistic statistical software using continuous models enabling statistical evaluation of more complex mixtures than was possible in 2014. The trial was therefore aimed at investigating the value of these improved capabilities and also to investigate if there appeared to be marked differences between the different software tools in use in the UK. The results demonstrate a high degree of concordance within and between FSPs and across different evaluation models, and will provide important support for the use of such models in evaluation of mixed DNA profiles.

Capture of Mouse and Human Stem Cells with Features of Formative Pluripotency.

Pluripotent cells emerge as a naive founder population in the blastocyst, acquire capacity for germline and soma formation, and then undergo lineage priming. Mouse embryonic stem cells (ESCs) and epiblast-derived stem cells (EpiSCs) represent the initial naive and final primed phases of pluripotency, respectively. Here, we investigate the intermediate formative stage. Using minimal exposure to specification cues, we derive stem cells from formative mouse epiblast. Unlike ESCs or EpiSCs, formative stem (FS) cells respond directly to germ cell induction. They colonize somatic tissues and germline in chimeras. Whole-transcriptome analyses show similarity to pre-gastrulation formative epiblast. Signal responsiveness and chromatin accessibility features reflect lineage capacitation. Furthermore, FS cells show distinct transcription factor dependencies, relying critically on Otx2. Finally, FS cell culture conditions applied to human naive cells or embryos support expansion of similar stem cells, consistent with a conserved staging post on the trajectory of mammalian pluripotency.

The participatory and partisan impacts of mandatory vote-by-mail.

Recently, mandatory vote-by-mail has received a great deal of attention as a means of administering elections in the United States. However, policy-makers disagree on the merits of this approach. Many of these debates hinge on whether mandatory vote-by-mail advantages one political party over the other. Using a unique pairing of historical county-level data that covers the past three decades and more than 40 million voting records from the two states that have conducted a staggered rollout of mandatory vote-by-mail (Washington and Utah), we use several methods for causal inference to show that mandatory vote-by-mail slightly increases voter turnout but has no effect on election outcomes at various levels of government. Our results find meaning given contemporary debates about the merits of mandatory vote-by-mail. Mandatory vote-by-mail ensures that citizens are given a safe means of casting their ballot while simultaneously not advantaging one political party over the other.

Cyanotoxin impact on microbial-mediated nitrogen transformations at the interface of sediment-water column in surface water bodies.

Harmful cyanobacterial blooms produce lethal toxins in many aquatic ecosystems experiencing eutrophication. This manuscript presents results on the effects of cyanotoxins on the aerobic microbial communities residing at the interface of sediments and water columns with the ammonia-oxidizing bacteria (AOB) as the model microbial community. Microcystin-LR (MC-LR), a heavily researched cyanotoxin variant, was used as the model cyanotoxin. To measure cyanotoxin influence on the activity of nitrifying microbial communities, an enriched culture of AOBs collected from an ongoing partial nitrification-nitritation reactor was examined for its exposure to 1, 5 and 10 µg/L of MC-LR. The nitritation kinetics experiment demonstrated MC-LR's ability at 1, 5, and 10 µg/L concentrations to prevent ammonium oxidation with statistically significant differences in nitritation rates between the blanks and spiked samples (One-way ANOVA, p < 0.05). Significantly decreased dissolved oxygen (DO) consumption during oxygen update batch tests demonstrated toxin's influence on AOB's oxidizing capabilities when exposed to even lower concentrations of 0.75, 0.5, and 0.25 µg/L of MC-LR in a separate set of experiments. Based on competitive kinetics, the MC-LR inhibition coefficient-the concentration needed to produce half-maximum inhibition of the mixed community AOBs was determined to be 0.083 µg/L. The stress tests proved the recovery of nitritation to some extent at lower MC-LR concentrations (1 and 5 µg/L), but significant irreversible inhibition was recorded when the AOB population was exposed to 10 µg/L MC-LR. The comparisons of amoA gene expressions corresponded well with nitrifying kinetics. All concentrations of MC-LR spiking were determined to produce a discernible impact on the AOB nitritation rate by either destroying the bacterial cell or immediately inhibiting the amoA gene expression.

Dredging activity and associated sound have negligible effects on adult Atlantic sturgeon migration to spawning habitat in a large coastal river.

Dredging is considered a major threat/impedance to anadromous fish migrating to spawning habitat. Due to the perceived threat caused by dredging, environmental windows that restrict dredge operations are enforced within many rivers along the east coast. However, it is generally unknown how anadromous fish react to encountering an active dredge during spawning migrations. Atlantic sturgeon (ATS) are an endangered, anadromous species along the Atlantic slope of North America. To determine if and how an active dredge may affect ATS spawning migration, a Vemco Positioning System array was deployed around an active hydraulic-cutterhead dredge that adult ATS must traverse to reach spawning habitat in the James River, VA. Telemetry data showed that all ATS that entered the study area survived. ATS that migrated upstream during dredge operations (N = 103) traversed the dredge area and continued upstream to spawning habitat. Many ATS made multiple trips through the study area during dredge operations. There was no noticeable difference in swim behavior regardless of whether the dredge was absent or working within the study area. We suggest that dredging in the lower James River does not create a barrier for adult ATS migrating to spawning habitat or cause adults to significantly modify swim behavior. This is the first study to utilize fine-scale telemetry data to describe how an organism moves in relation to an active dredge. This methodology could be used to describe dredge-sturgeon interactions on different life stages and in other locations and could be expanded to other aquatic organisms of concern.

Bone Marrow Mesenchymal Stem Cells Support Acute Myeloid Leukemia Bioenergetics and Enhance Antioxidant Defense and Escape from Chemotherapy.

Like normal hematopoietic stem cells, leukemic stem cells depend on their bone marrow (BM) microenvironment for survival, but the underlying mechanisms remain largely unknown. We have studied the contribution of nestin+ BM mesenchymal stem cells (BMSCs) to MLL-AF9-driven acute myeloid leukemia (AML) development and chemoresistance in vivo. Unlike bulk stroma, nestin+ BMSC numbers are not reduced in AML, but their function changes to support AML cells, at the expense of non-mutated hematopoietic stem cells (HSCs). Nestin+ cell depletion delays leukemogenesis in primary AML mice and selectively decreases AML, but not normal, cells in chimeric mice. Nestin+ BMSCs support survival and chemotherapy relapse of AML through increased oxidative phosphorylation, tricarboxylic acid (TCA) cycle activity, and glutathione (GSH)-mediated antioxidant defense. Therefore, AML cells co-opt energy sources and antioxidant defense mechanisms from BMSCs to survive chemotherapy.

Detecting network communities by propagating labels under constraints.

We investigate the recently proposed label-propagation algorithm (LPA) for identifying network communities. We reformulate the LPA as an equivalent optimization problem, giving an objective function whose maxima correspond to community solutions. By considering properties of the objective function, we identify conceptual and practical drawbacks of the label-propagation approach, most importantly the disparity between increasing the value of the objective function and improving the quality of communities found. To address the drawbacks, we modify the objective function in the optimization problem, producing a variety of algorithms that propagate labels subject to constraints; of particular interest is a variant that maximizes the modularity measure of community quality. Performance properties and implementation details of the proposed algorithms are discussed. Bipartite as well as unipartite networks are considered.

High-throughput DNA sequencing reveals the dominance of pico- and other filamentous cyanobacteria in an urban freshwater Lake.

The current study presents findings related to algal blooms in a fresh water lake, which has been experiencing severe cyanobacterial blooms (CyanoHABs). Primarily, picocyanobacteria belonging to the genus Synechococcus and filamentous cyanobacterial group belonging to Aphanizomenon and Dolichospermum dominated top water column during non-bloom and bloom periods respectively. The dominance of Synechococcus in early summer informs that blooming in Utah Lake starts in early summer and then later is taken over by other bloom-forming cyanobacteria, such as species belonging to the genus Aphanizomenon. A strong negative correlation (r = -0.9, p < 0.001) was found between the occurrence of Aphanizomenon and Synechococcus which correlates very well with the fact that the blooms of these two different cyanobacteria never coexisted. The predominance of cyanobacteria in 2017 was attributed more to temperature (r = 0.18, p < 0.001). The Actinobacteria was negatively correlated with primary production and high chlorophyll a concentration. Flavobacterium and Limnohabitans were the main phytoplankton colonizers and predators detected that could secrete extracellular enzymes to degrade algal exudates (such as proteins and polysaccharides). Additionally, cyanotoxins producers Microcystis aeruginosa and Planktothrix accounted for up to 12.43% and 7.04% of total cyanobacteria abundance during blooms. The relative abundance of chloroplast reads was overall lower than the cyanobacteria reads, except for the May 5th sampling in 2017. There was inter-annual variability in the bloom-associated heterotrophic bacterial populations, but these populations were consistent with bloom-associated bacterial populations found in other lakes. Community diversity analysis for both Shannon and Simpson indices indicated lower community diversity during the bloom period. The beta diversity conducted by PCoA and UPGMA trees suggested the significant temporal rather than spatial impacts on shaping the phytoplankton community structures during the summer season.

Modularity and community detection in bipartite networks.

The modularity of a network quantifies the extent, relative to a null model network, to which vertices cluster into community groups. We define a null model appropriate for bipartite networks, and use it to define a bipartite modularity. The bipartite modularity is presented in terms of a modularity matrix B; some key properties of the eigenspectrum of B are identified and used to describe an algorithm for identifying modules in bipartite networks. The algorithm is based on the idea that the modules in the two parts of the network are dependent, with each part mutually being used to induce the vertices for the other part into the modules. We apply the algorithm to real-world network data, showing that the algorithm successfully identifies the modular structure of bipartite networks.

Structure and regulation of acetyl-CoA carboxylase genes of metazoa.

Acetyl-CoA carboxylase (ACC) plays a fundamental role in fatty acid metabolism. The reaction product, malonyl-CoA, is both an intermediate in the de novo synthesis of long-chain fatty acids and also a substrate for distinct fatty acyl-CoA elongation enzymes. In metazoans, which have evolved energy storage tissues to fuel locomotion and to survive periods of starvation, energy charge sensing at the level of the individual cell plays a role in fuel selection and metabolic orchestration between tissues. In mammals, and probably other metazoans, ACC forms a component of an energy sensor with malonyl-CoA, acting as a signal to reciprocally control the mitochondrial transport step of long-chain fatty acid oxidation through the inhibition of carnitine palmitoyltransferase I (CPT I). To reflect this pivotal role in cell function, ACC is subject to complex regulation. Higher metazoan evolution is associated with the duplication of an ancestral ACC gene, and with organismal complexity, there is an increasing diversity of transcripts from the ACC paraloges with the potential for the existence of several isozymes. This review focuses on the structure of ACC genes and the putative individual roles of their gene products in fatty acid metabolism, taking an evolutionary viewpoint provided by data in genome databases.

Multiple thresholds in a model system of noisy ion channels.

Voltage-activated ion channels vary randomly between open and closed states, influenced by the membrane potential and other factors. Signal transduction is enhanced by noise in a simple ion channel model. The enhancement occurs in a finite range of signals; the range can be extended using populations of channels. The range increases more rapidly in multiple-threshold channel populations than in single-threshold populations. The diversity of ion channels may thus be present as a strategy to reduce the metabolic costs of handling a broad class of electrochemical signals.