Biochemical evidence for the tyrosine involvement in cationic intermediate stabilization in mouse β-carotene 15, 15'-monooxygenase

<p>Abstract</p> <p>Background</p> <p>β-carotene 15,15'-monooxygenase (BCMO1) catalyzes the crucial first step in vitamin A biosynthesis in animals. We wished to explore the possibility that a carbocation intermediate is formed during the cleavage reaction of BCMO1, as is seen for many isoprenoid biosynthesis enzymes, and to determine which residues in the substrate binding cleft are necessary for catalytic and substrate binding activity. To test this hypothesis, we replaced substrate cleft aromatic and acidic residues by site-directed mutagenesis. Enzymatic activity was measured <it>in vitro </it>using His-tag purified proteins and <it>in vivo </it>in a β-carotene-accumulating <it>E. coli </it>system.</p> <p>Results</p> <p>Our assays show that mutation of either Y235 or Y326 to leucine (no cation-π stabilization) significantly impairs the catalytic activity of the enzyme. Moreover, mutation of Y326 to glutamine (predicted to destabilize a putative carbocation) almost eliminates activity (9.3% of wt activity). However, replacement of these same tyrosines with phenylalanine or tryptophan does not significantly impair activity, indicating that aromaticity at these residues is crucial. Mutations of two other aromatic residues in the binding cleft of BCMO1, F51 and W454, to either another aromatic residue or to leucine do not influence the catalytic activity of the enzyme. Our <it>ab initio </it>model of BCMO1 with β-carotene mounted supports a mechanism involving cation-π stabilization by Y235 and Y326.</p> <p>Conclusions</p> <p>Our data are consistent with the formation of a substrate carbocation intermediate and cation-π stabilization of this intermediate by two aromatic residues in the substrate-binding cleft of BCMO1.</p

Episodic Occurrence of Favourable Weather Constrains Recovery of a Cold Desert Shrubland After Fire

Key to the long-term resilience of dryland ecosystems is the recovery of foundation plant species following disturbance. In ecosystems with high interannual weather variability, understanding the influence of short-term environmental conditions on establishment of foundation species is essential for identifying vulnerable landscapes and developing restoration strategies. We asked how annual environmental conditions affect post-fire establishment of Artemisia tridentata, a shrub species that dominates landscapes across much of the western United States, and evaluated the influence of episodic establishment on population recovery. We collected A. tridentata stem samples from 33 plots in 12 prescribed fire sites that burned 8–11 years before sampling. We determined individual establishment years using annual growth rings. We measured seasonal soil environmental conditions at the study sites and asked if these conditions predicted annual establishment density. We then evaluated whether establishment patterns could be predicted by site-level climate or dominant subspecies. Finally, we tested the effect of the magnitude and frequency of post-fire establishment episodes on long-term population recovery. Annual post-fire recruitment of A. tridentata was driven by the episodic availability of spring soil moisture. Annual establishment was highest with wetter spring soils (relative influence [RI] = 19.4%) and later seasonal dry-down (RI = 11.8%) in the year of establishment. Establishment density declined greatly 4 to 5 years after fire (RI = 17.1%). Post-fire establishment patterns were poorly predicted by site-level mean climate (marginal R2 ≤ 0.18) and dominant subspecies (marginal R2 ≤ 0.43). Population recovery reflected the magnitude, but not the frequency, of early post-fire establishment pulses. Post-fire A. tridentata density and cover (measured 8–11 years after fire) were more strongly related to the magnitude of the largest establishment pulse than to establishment frequency, suggesting that population recovery may occur with a single favourable establishment year. Synthesis and applications. This study demonstrates the importance of episodic periods of favourable weather for long-term plant population recovery following disturbance. Management strategies that increase opportunities for seed availability to coincide with favourable weather conditions, such as retaining unburned patches or repeated seeding treatments, can improve restoration outcomes in high-priority areas

Forest and woodland replacement patterns following drought-related mortality

Forest vulnerability to drought is expected to increase under anthropogenic climate change, and drought-induced mortality and community dynamics following drought have major ecological and societal impacts. Here, we show that tree mortality concomitant with drought has led to short-term (mean 5 y, range 1 to 23 y after mortality) vegetation-type conversion in multiple biomes across the world (131 sites). Self-replacement of the dominant tree species was only prevalent in 21% of the examined cases and forests and woodlands shifted to nonwoody vegetation in 10% of them. The ultimate temporal persistence of such changes remains unknown but, given the key role of biological legacies in long-term ecological succession, this emerging picture of postdrought ecological trajectories highlights the potential for major ecosystem reorganization in the coming decades. Community changes were less pronounced under wetter postmortality conditions. Replacement was also influenced by management intensity, and postdrought shrub dominance was higher when pathogens acted as codrivers of tree mortality. Early change in community composition indicates that forests dominated by mesic species generally shifted toward more xeric communities, with replacing tree and shrub species exhibiting drier bioclimatic optima and distribution ranges. However, shifts toward more mesic communities also occurred and multiple pathways of forest replacement were observed for some species. Drought characteristics, species-specific environmental preferences, plant traits, and ecosystem legacies govern post drought species turnover and subsequent ecological trajectories, with potential far-reaching implications for forest biodiversity and ecosystem services.Peer reviewe

Lancet

BACKGROUND: In 2015, the second cycle of the CONCORD programme established global surveillance of cancer survival as a metric of the effectiveness of health systems and to inform global policy on cancer control. CONCORD-3 updates the worldwide surveillance of cancer survival to 2014. METHODS: CONCORD-3 includes individual records for 37.5 million patients diagnosed with cancer during the 15-year period 2000-14. Data were provided by 322 population-based cancer registries in 71 countries and territories, 47 of which provided data with 100% population coverage. The study includes 18 cancers or groups of cancers: oesophagus, stomach, colon, rectum, liver, pancreas, lung, breast (women), cervix, ovary, prostate, and melanoma of the skin in adults, and brain tumours, leukaemias, and lymphomas in both adults and children. Standardised quality control procedures were applied; errors were rectified by the registry concerned. We estimated 5-year net survival. Estimates were age-standardised with the International Cancer Survival Standard weights. FINDINGS: For most cancers, 5-year net survival remains among the highest in the world in the USA and Canada, in Australia and New Zealand, and in Finland, Iceland, Norway, and Sweden. For many cancers, Denmark is closing the survival gap with the other Nordic countries. Survival trends are generally increasing, even for some of the more lethal cancers: in some countries, survival has increased by up to 5% for cancers of the liver, pancreas, and lung. For women diagnosed during 2010-14, 5-year survival for breast cancer is now 89.5% in Australia and 90.2% in the USA, but international differences remain very wide, with levels as low as 66.1% in India. For gastrointestinal cancers, the highest levels of 5-year survival are seen in southeast Asia: in South Korea for cancers of the stomach (68.9%), colon (71.8%), and rectum (71.1%); in Japan for oesophageal cancer (36.0%); and in Taiwan for liver cancer (27.9%). By contrast, in the same world region, survival is generally lower than elsewhere for melanoma of the skin (59.9% in South Korea, 52.1% in Taiwan, and 49.6% in China), and for both lymphoid malignancies (52.5%, 50.5%, and 38.3%) and myeloid malignancies (45.9%, 33.4%, and 24.8%). For children diagnosed during 2010-14, 5-year survival for acute lymphoblastic leukaemia ranged from 49.8% in Ecuador to 95.2% in Finland. 5-year survival from brain tumours in children is higher than for adults but the global range is very wide (from 28.9% in Brazil to nearly 80% in Sweden and Denmark). INTERPRETATION: The CONCORD programme enables timely comparisons of the overall effectiveness of health systems in providing care for 18 cancers that collectively represent 75% of all cancers diagnosed worldwide every year. It contributes to the evidence base for global policy on cancer control. Since 2017, the Organisation for Economic Co-operation and Development has used findings from the CONCORD programme as the official benchmark of cancer survival, among their indicators of the quality of health care in 48 countries worldwide. Governments must recognise population-based cancer registries as key policy tools that can be used to evaluate both the impact of cancer prevention strategies and the effectiveness of health systems for all patients diagnosed with cancer. FUNDING: American Cancer Society; Centers for Disease Control and Prevention; Swiss Re; Swiss Cancer Research foundation; Swiss Cancer League; Institut National du Cancer; La Ligue Contre le Cancer; Rossy Family Foundation; US National Cancer Institute; and the Susan G Komen Foundation

Limits to reproduction and seed size-number trade-offs that shape forest dominance and future recovery

International audienceThe relationships that control seed production in trees are fundamental to understanding the evolution of forest species and their capacity to recover from increasing losses to drought, fire, and harvest. A synthesis of fecundity data from 714 species worldwide allowed us to examine hypotheses that are central to quantifying reproduction, a foundation for assessing fitness in forest trees. Four major findings emerged. First, seed production is not constrained by a strict trade-off between seed size and numbers. Instead, seed numbers vary over ten orders of magnitude, with species that invest in large seeds producing more seeds than expected from the 1:1 trade-off. Second, gymnosperms have lower seed production than angiosperms, potentially due to their extra investments in protective woody cones. Third, nutrient-demanding species, indicated by high foliar phosphorus concentrations, have low seed production. Finally, sensitivity of individual species to soil fertility varies widely, limiting the response of community seed production to fertility gradients. In combination, these findings can inform models of forest response that need to incorporate reproductive potential

Limits to reproduction and seed size-number tradeoffs that shape forest dominance and future recovery

The relationships that control seed production in trees are fundamental to understanding the evolution of forest species and their capacity to recover from increasing losses to drought, fire, and harvest. A synthesis of fecundity data from 714 species worldwide allowed us to examine hypotheses that are central to quantifying reproduction, a foundation for assessing fitness in forest trees. Four major findings emerged. First, seed production is not constrained by a strict trade-off between seed size and numbers. Instead, seed numbers vary over ten orders of magnitude, with species that invest in large seeds producing more seeds than expected from the 1:1 trade-off. Second, gymnosperms have lower seed production than angiosperms, potentially due to their extra investments in protective woody cones. Third, nutrient-demanding species, indicated by high foliar phosphorus concentrations, have low seed production. Finally, sensitivity of individual species to soil fertility varies widely, limiting the response of community seed production to fertility gradients. In combination, these findings can inform models of forest response that need to incorporate reproductive potential

Evaluation of appendicitis risk prediction models in adults with suspected appendicitis

Background Appendicitis is the most common general surgical emergency worldwide, but its diagnosis remains challenging. The aim of this study was to determine whether existing risk prediction models can reliably identify patients presenting to hospital in the UK with acute right iliac fossa (RIF) pain who are at low risk of appendicitis. Methods A systematic search was completed to identify all existing appendicitis risk prediction models. Models were validated using UK data from an international prospective cohort study that captured consecutive patients aged 16–45 years presenting to hospital with acute RIF in March to June 2017. The main outcome was best achievable model specificity (proportion of patients who did not have appendicitis correctly classified as low risk) whilst maintaining a failure rate below 5 per cent (proportion of patients identified as low risk who actually had appendicitis). Results Some 5345 patients across 154 UK hospitals were identified, of which two‐thirds (3613 of 5345, 67·6 per cent) were women. Women were more than twice as likely to undergo surgery with removal of a histologically normal appendix (272 of 964, 28·2 per cent) than men (120 of 993, 12·1 per cent) (relative risk 2·33, 95 per cent c.i. 1·92 to 2·84; P < 0·001). Of 15 validated risk prediction models, the Adult Appendicitis Score performed best (cut‐off score 8 or less, specificity 63·1 per cent, failure rate 3·7 per cent). The Appendicitis Inflammatory Response Score performed best for men (cut‐off score 2 or less, specificity 24·7 per cent, failure rate 2·4 per cent). Conclusion Women in the UK had a disproportionate risk of admission without surgical intervention and had high rates of normal appendicectomy. Risk prediction models to support shared decision‐making by identifying adults in the UK at low risk of appendicitis were identified

Impacts of Climate and Distrubances on Vegetation Dynamics in Piñon-Juniper Woodlands

Global climate change and altered disturbance regimes have already and are predicted to continue to cause significant shifts in vegetation distribution. Regional warming has increased tree mortality rates over the past several decades through increasing water deficits and insect outbreaks, which have dramatically changed forest and woodland structure and altered water and energy fluxes and carbon stocks. For the first part of my dissertation, I use field surveys and historical data to examine how changes in climate and recent piñon pine (Pinus edulis) mortality events may affect tree regeneration dynamics in the widely distribution piñon-juniper (Juniperus osteosperma and J. monosperma) woodlands of the southwestern USA. First, I show that piñon seed cone production declined by 40% from the 1974 decade (1969-1978) to the 2008 decade (2003-2012) in revisited stands throughout New Mexico and northwestern Oklahoma. Seed cone production was highly correlated with late summer temperatures at the time of cone initiation. Further, declines in seed cone production were greatest among populations that experienced the greatest increases in growing season temperatures, which were the populations located at the cooler, upper elevations. Second, I examine the effects of increasing temperatures and recent piñon mortality on tree recruitment and growth across the southwestern USA and determine how these effects are moderated by local climate, biotic interactions, and soil properties. In addition to changing climate and recent mortality events, large tracts of land across the western U.S. have been managed over the last century in an effort to increase forage production and timber harvesting yields, reduce the risk of wildland fires, and/or restore ecosystem structure, and these human disturbances can also dramatically alter these woodland ecosystems. The second part of my dissertation research documents the spatial extent and cost of past management treatments done within piñon-juniper woodlands of the Colorado Plateau. In addition, I determine the legacy effects of these past treatments on understory vegetation and woodland structure as well as the efficacy of current management practices at accomplishing management goals. Overall, my findings have provided important insights into how woodlands recover following disturbances under a warmer climate

Data from: Declines in pinyon pine cone production associated with regional warming

Global climate change is expected to produce large shifts in vegetation distribution and has already increased tree mortality, altering forest structure. However, long-term shifts will be partly dependent on the ability of species to reproduce under a novel climate. Few studies have examined the impact of climate change on the reproductive output of long-lived ‘masting' species, or species characterized by episodic reproductive events. Here, we show that seed cone production among pinyon pine (Pinus edulis), a masting species, declined by 40% from the 1974 decade (1969–1978) to the 2008 decade (2003–2012) in revisited stands throughout New Mexico and northwestern Oklahoma. Seed cone production was highly correlated with late summer temperatures at the time of cone initiation. Further, declines in seed cone production were greatest among populations that experienced the greatest increases in growing season temperatures, which were the populations located at the cooler, upper elevations. As growing season temperatures are predicted to increase across this region over the next century, these findings suggest seed cone production may be an increasingly important bottleneck for future pinyon pine regeneration, especially in areas with greater increases in temperature. Declines in seed cone production may not only affect pinyon pine population dynamics but also the various wildlife species that rely on pinyon pine seeds. Because pinyon pine has similar reproductive strategies as other semi-arid pine species, increasing temperature may negatively influence reproductive output of other conifers. Further investigation into the full geographic and taxonomic extent of these seed declines is warranted

Managing for ecological resilience of pinyon–juniper ecosystems during an era of woodland contraction

Abstract Dryland woodland ecosystems worldwide have experienced widespread drought‐ and heat‐related tree mortality events coupled with extreme wildfire behavior. In contrast to other forest types where the emphasis has been on the silvicultural enhancement of ecosystem resilience and restoration of structural heterogeneity, limited frameworks are available for management to improve drought resilience in semiarid woodlands. This challenge is especially acute in pinyon–juniper woodlands, a dominant vegetation type across western North America that has experienced extensive tree die‐off over the past several decades while simultaneously undergoing expansion in portions of its range. In this paper, we describe the critical and urgent need to manage for future drought resilience in these highly vulnerable ecosystems and synthesize the current state of knowledge on how to enhance woodland resilience to drought coupled with high temperatures and associated disturbances. We present a landscape prioritization framework for guiding management goals and practices that requires prioritization of efforts based on the need for action and the probability of a positive outcome. Four guiding factors include historical woodland structure and drivers of long‐term landscape change, current vegetation structure and composition, future climate suitability, and habitat and resource value. In summarizing the strength of evidence supporting our recommendations, we identify critical knowledge gaps and highlight the importance of adaptive management strategies that reflect current uncertainties. This will ultimately allow for improved management of diverse semiarid woodland ecosystems that are undergoing substantial changes due to past and present land use, biological invasions, and climate change