Fork rotation and DNA precatenation are restricted during DNA replication to prevent chromosomal instability

Faithful genome duplication and inheritance require the complete resolution of all intertwines within the parental DNA duplex. This is achieved by topoisomerase action ahead of the replication fork or by fork rotation and subsequent resolution of the DNA precatenation formed. Although fork rotation predominates at replication termination, in vitro studies have suggested that it also occurs frequently during elongation. However, the factors that influence fork rotation and how rotation and precatenation may influence other replication-associated processes are unknown. Here we analyze the causes and consequences of fork rotation in budding yeast. We find that fork rotation and precatenation preferentially occur in contexts that inhibit topoisomerase action ahead of the fork, including stable protein–DNA fragile sites and termination. However, generally, fork rotation and precatenation are actively inhibited by Timeless/Tof1 and Tipin/Csm3. In the absence of Tof1/Timeless, excessive fork rotation and precatenation cause extensive DNA damage following DNA replication. With Tof1, damage related to precatenation is focused on the fragile protein–DNA sites where fork rotation is induced. We conclude that although fork rotation and precatenation facilitate unwinding in hard-to-replicate contexts, they intrinsically disrupt normal chromosome duplication and are therefore restricted by Timeless/Tipin

Learning from the implementation of Universal Free School Meals in Scotland using Normalisation Process Theory:Lessons for policymakers to engage multiple stakeholders

In 2014/15, Universal Free School Meals (UFSM) were introduced in Scotland and England for children in their first three years of primary school. This study examined the implementation of UFSM in Scotland using Noramlisation Process Theory (NPT), a middle-range theory of implementation, to identify areas of learning for policymakers wishing to introduce or extend similar policies. NPT is predominantly used to evaluate interventions or new technologies in healthcare settings. Qualitative data were collected across Scotland using a case study approach shortly after implementation (n = 29 school-level stakeholders) and in the following school year (n = 18 school-level stakeholders and n = 19 local authority-level stakeholders). Observations of lunchtime in each school were conducted at both timepoints. Data were analysed using a thematic framework approach using NPT constructs and sub-constructs. Results suggested education and catering stakeholders experiences of implementation diverged most around the NPT concepts of coherence, cognitive participation, and reflexive monitoring. Lack of coherence around the purpose and long-term benefits of UFSM appeared to reduce education stakeholders’ willingness to engage with the policy beyond operational issues. In contrast, catering stakeholders identified a direct benefit to their everyday work and described receiving additional resources to deliver the policy. Overall, participants described an absence of monitoring data around the areas of greatest salience for education stakeholders. This study successfully used NPT to identify policy learning around school meals. Policymakers must increase the salience of such intersectoral policies for all relevant stakeholders involved before policy implementation, and plan adequate monitoring to evaluate potential long-term benefits

Hypothalamic Vitamin D Improves Glucose Homeostasis and Reduces Weight

Despite clear associations between vitamin D deficiency and obesity and/or type 2 diabetes, a causal relationship is not established. Vitamin D receptors (VDRs) are found within multiple tissues, including the brain. Given the importance of the brain in controlling both glucose levels and body weight, we hypothesized that activation of central VDR links vitamin D to the regulation of glucose and energy homeostasis. Indeed, we found that small doses of active vitamin D, 1α,25-dihydroxyvitamin D3 (1,25D3) (calcitriol), into the third ventricle of the brain improved glucose tolerance and markedly increased hepatic insulin sensitivity, an effect that is dependent upon VDR within the paraventricular nucleus of the hypothalamus. In addition, chronic central administration of 1,25D3 dramatically decreased body weight by lowering food intake in obese rodents. Our data indicate that 1,25D3-mediated changes in food intake occur through action within the arcuate nucleus. We found that VDR colocalized with and activated key appetite-regulating neurons in the arcuate, namely proopiomelanocortin neurons. Together, these findings define a novel pathway for vitamin D regulation of metabolism with unique and divergent roles for central nervous system VDR signaling. Specifically, our data suggest that vitamin D regulates glucose homeostasis via the paraventricular nuclei and energy homeostasis via the arcuate nuclei

Plant Functional Groups and Species Contribute to Ecological Resilience a Decade After Woodland Expansion Treatments

Woody plant expansions are altering ecosystem structure and function, as well as fire regimes, around the globe. Tree-reduction treatments are widely implemented in expanding woodlands to reduce fuel loads, increase ecological resilience, and improve habitat, but few studies have measured treatment outcomes over long timescales or large geographic areas. The Sagebrush Treatment Evaluation Project (SageSTEP) evaluated the ecological effects of prescribed fire and cut-and-leave treatments in sagebrush communities experiencing tree expansion in North American cold desert shrublands. We used 10 yr of data from the SageSTEP network to test how treatments interacted with pre-treatment tree dominance, soil climate, and time since treatment to affect plant functional groups and dominant species. Non-sprouting shrub (Artemisia spp.), sprouting shrub, perennial graminoid, and annual grass responses depended on tree dominance and soil climate, and responses were related to the dominant species\u27 life-history traits. Sites with warm and dry soils showed increased perennial graminoid but reduced Artemisia shrub cover across the tree dominance gradient after prescribed burning, while sites with cool and moist soils showed favorable post-burn responses for both functional types, particularly at low to moderate tree dominance. Cut-and-leave treatments sustained or increased native perennial plant functional groups and experienced smaller increases in exotic annual plants in both soil climates across the tree dominance gradient. Both treatments reduced biocrust cover. Selecting appropriate tree-reduction treatments to achieve desired long-term outcomes requires consideration of dominant species, site environmental conditions, and the degree of woodland expansion. Careful selection of management treatments will reduce the likelihood of undesirable consequences to the ecosystem

Would Functional Agricultural Foods Improve Human Health?

Concern over diet-health relationships has moved to the forefront of public health concerns in the UK and much of the developed world. It has been estimated, for example, that obesity costs the UK National Health Service up to £6b per year (Rayner and Scarborough, 2005), but if all consumers were to follow recommended healthy eating guidelines there would be major implications for food consumption, land use and international trade (Srinivasan et al, 2006). This is unlikely to happen, at least in the short term, but it is realistic to anticipate some dietary adjustment toward the recommendations, resulting in an improvement in diet quality (Mazzocchi et al, 2007). Although consumers are reluctant to make major changes to their diets, they may be prepared to substitute existing foods for healthier alternatives. Three of the most prominent nutritional recommendations are to consume more fruit and vegetables, which contain phytochemicals beneficial to health, reduce consumption of saturated fatty acids (SFA) and increase intake of long-chain n-3 fatty acids (FA). In the first case, consumption of fruit and vegetables has been stable at around three 80 g portions per person per day according to the Health Survey for England. It is estimated that 42,200 deaths per year could be avoided in England and 411,000 Quality Adjusted Life Years (QALYs) could be saved if fruit and vegetable consumption were increased to the recommended 5 portions per day (Ofcom 2006). As well as continuing to encourage people to eat more, it could be desirable to ‘intensify’ the beneficial phytochemical content of existing fruit and vegetables.Agribusiness, Agricultural and Food Policy, Farm Management, Food Consumption/Nutrition/Food Safety, Industrial Organization,

Hubble Space Telescope Ultraviolet Spectroscopy of Fourteen Low-Redshift Quasars

We present low-resolution ultraviolet spectra of 14 low redshift (z<0.8) quasars observed with HST/STIS as part of a Snap project to understand the relationship between quasar outflows and luminosity. By design, all observations cover the CIV emission line. Nine of the quasars are from the Hamburg-ESO catalog, three are from the Palomar-Green catalog, and one is from the Parkes catalog. The sample contains a few interesting quasars including two broad absorption line (BAL) quasars (HE0143-3535, HE0436-2614), one quasar with a mini-BAL (HE1105-0746), and one quasar with associated narrow absorption (HE0409-5004). These BAL quasars are among the brightest known (though not the most luminous) since they lie at z<0.8. We compare the properties of these BAL quasars to the z1.4 Large Bright Quasar samples. By design, our objects sample luminosities in between these two surveys, and our four absorbed objects are consistent with the v ~ L^0.62 relation derived by Laor & Brandt (2002). Another quasar, HE0441-2826, contains extremely weak emission lines and our spectrum is consistent with a simple power-law continuum. The quasar is radio-loud, but has a steep spectral index and a lobe-dominated morphology, which argues against it being a blazar. The unusual spectrum of this quasar resembles the spectra of the quasars PG1407+265, SDSSJ1136+0242, and PKS1004+13 for which several possible explanations have been entertained.Comment: Uses aastex.cls, 21 pages in preprint mode, including 6 figures and 2 tables; accepted for publication in The Astronomical Journal (projected vol 133

A Target-Based High Throughput Screen Yields Trypanosoma brucei Hexokinase Small Molecule Inhibitors with Antiparasitic Activity

African sleeping sickness is a disease found in sub-Saharan Africa that is caused by the single-celled parasite Trypanosoma brucei. The drugs used widely now to treat infections are 50 years old and notable for their toxicity, emphasizing the need for development of new therapeutics. In the search for potential drug targets, researchers typically focus on enzymes or proteins that are essential to the survival of the infectious agent while being distinct enough from the host to avoid accidental targeting of the host enzyme. This work describes our research on one such trypanosome enzyme, hexokinase, which is a protein that the parasite requires to make energy. Here we describe the results of our search for inhibitors of the parasite enzyme. By screening 220,223 compounds for anti-hexokinase activity, we have identified new inhibitors of the parasite enzyme. Some of these are toxic to trypanosomes while having no effect on mammalian cells, suggesting that they may hold promise for the development of new anti-parasitic compounds

A Full-Genomic Sequence-Verified Protein-Coding Gene Collection for Francisella tularensis

The rapid development of new technologies for the high throughput (HT) study of proteins has increased the demand for comprehensive plasmid clone resources that support protein expression. These clones must be full-length, sequence-verified and in a flexible format. The generation of these resources requires automated pipelines supported by software management systems. Although the availability of clone resources is growing, current collections are either not complete or not fully sequence-verified. We report an automated pipeline, supported by several software applications that enabled the construction of the first comprehensive sequence-verified plasmid clone resource for more than 96% of protein coding sequences of the genome of F. tularensis, a highly virulent human pathogen and the causative agent of tularemia. This clone resource was applied to a HT protein purification pipeline successfully producing recombinant proteins for 72% of the genes. These methods and resources represent significant technological steps towards exploiting the genomic information of F. tularensis in discovery applications