Kinetic and fluorescence studies of the interaction of p21(ras) with guanine nucleotides and the GTPase activating proteins, p120-GAP and Nfi-GAP

The protein products of the ras proto-oncogenes, p21ras, are 21 KDa guanine nucleotide binding proteins which possess a slow intrinsic GTPase activity. These proteins are thought to be involved in the regulation of cell growth and differentiation, and some single point mutations in the ras genes lead to cell transformation. The biological signal mediated by p21ras is determined by the in vivo concentration of the p21ras.GTP complex since only when bound to GTP are these proteins thought to be biologically active. Nucleotide exchange of bound GDP for cytoplasmic GTP leads to the reformation of the p21ras.GTP complex. Two proteins (GAPs) have been identified which accelerate the rate of GTP hydrolysis by wild p21ras but not by oncogenic p21ras mutants. The kinetic mechanism of the p21ras.GTPase in the presence of the catalytic-domains of two GAPs, p120-GAP and neurofibromin, has been investigated. The studies are based primarily on the use of a fluorescent GTP analogue, 2'(3')-O-(N-methylanthraniloyl)-GTP (mantGTP), which shows changes in fluorescence intensity during several elementary steps in the GTPase mechanism. The experimental results are compatible with a mechanism where the intrinsic hydrolysis of GTP by p21ras is preceded, and controlled, by a protein conformational change in the p21ras.GTP complex. p120-GAP accelerates the overall rate of GTP cleavage by promoting this rate limiting conformational change. The binding of p120-GAP to p21ras.GTP is a rapidly reversible reaction (equilibrium dissociation constant, Kd = 20 μM at l ≈ 20 mM). p120-GAP accelerates the rate of the subsequent conformational change in p21ras.GTP by a factor of 105 compared to the rate of the equivalent reaction in the absence of p120-GAP. A similar extent of activation was observed with neurofibromin although the affinity for p21N-ras.GTP was 20-fold higher than that of p120-GAP. In contrast to p120-GAP, the binding reaction with neurofibromin may not be rapidly reversible. For both proteins, increasing ionic strength lead to a marked increase in the Kd; the rate constant of the conformational change being essentially unaltered. The binding of p120-GAP and neurofibromin to the p21ras.mantGTP complex is associated with an increase in fluorescence intensity, anisotropy and energy transfer (from tryptophan residues in the GAP proteins). These fluorescence signals can be used to determine the equilibrium dissociation constants for the binding of GAP proteins to p21ras. The intrinsic GTPase activity of the Gly 12→ Pro mutant of p21ras is only weakly accelerated by either p120-GAP or neurofibromin although both GAPs bind with an affinity similar to that with the wild type p21ras protein. An explanation for the weakly transforming phenotype of this mutant compared to other Gly 12 mutants is offered. One consequence of this interpretation is that the in vivo rate of GAP-activated GTP hydrolysis and nucleotide exchange may be slower than generally assumed. Finally, the hydrodynamic properties of p21ras proteins have been investigated using time resolved fluorescence techniques combined with a range of fluorescent guanine nucleotide analogues. In all cases, the rotational correlation time of p21ras is consistent with a dimeric structure for these proteins in solution

Syndemics and the PrEP Cascade: Results from a Sample of Young Latino Men Who Have Sex with Men.

Young Latino men who have sex with men (MSM) are a highly vulnerable population for HIV infection. Pre-exposure prophylaxis (PrEP) is a novel biomedical HIV prevention tool that may aid in reducing the disparity in HIV incidence among Latino MSM. However, PrEP use is disproportionally low among Latino MSM and, therefore, identifying barriers along the PrEP continuum of care (the "PrEP cascade") would provide insight into how to best deploy PrEP interventions. Syndemics theory is a prominent framework employed in HIV prevention; however, to date, no known studies have applied this theory to PrEP. Thus, the aim of the current study was to explore the association between syndemics and the PrEP cascade, including the degree to which psychosocial and structural syndemic constructs are related to the PrEP cascade. Participants were 151 young Latino MSM (M age = 24 years; SD = 3) residing in San Diego, California, who completed a battery of online self-report measures. Results indicated high levels of syndemic indicators and varying levels of engagement across the PrEP cascade. As syndemic indicators increased, the odds of engagement across the PrEP cascade were significantly lowered. Psychosocial and structural syndemic factors accounted for unique variance in the PrEP cascade. Results highlight the need for combination interventions that address both psychosocial and structural barriers to PrEP use and persistence among young Latino MSM

Multicentury changes in ocean and land contributions to the climate-carbon feedback

Author Posting. © American Geophysical Union, 2015. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Global Biogeochemical Cycles 29 (2015): 744–759, doi:10.1002/2014GB005079.Improved constraints on carbon cycle responses to climate change are needed to inform mitigation policy, yet our understanding of how these responses may evolve after 2100 remains highly uncertain. Using the Community Earth System Model (v1.0), we quantified climate-carbon feedbacks from 1850 to 2300 for the Representative Concentration Pathway 8.5 and its extension. In three simulations, land and ocean biogeochemical processes experienced the same trajectory of increasing atmospheric CO2. Each simulation had a different degree of radiative coupling for CO2 and other greenhouse gases and aerosols, enabling diagnosis of feedbacks. In a fully coupled simulation, global mean surface air temperature increased by 9.3 K from 1850 to 2300, with 4.4 K of this warming occurring after 2100. Excluding CO2, warming from other greenhouse gases and aerosols was 1.6 K by 2300, near a 2 K target needed to avoid dangerous anthropogenic interference with the climate system. Ocean contributions to the climate-carbon feedback increased considerably over time and exceeded contributions from land after 2100. The sensitivity of ocean carbon to climate change was found to be proportional to changes in ocean heat content, as a consequence of this heat modifying transport pathways for anthropogenic CO2 inflow and solubility of dissolved inorganic carbon. By 2300, climate change reduced cumulative ocean uptake by 330 Pg C, from 1410 Pg C to 1080 Pg C. Land fluxes similarly diverged over time, with climate change reducing stocks by 232 Pg C. Regional influence of climate change on carbon stocks was largest in the North Atlantic Ocean and tropical forests of South America. Our analysis suggests that after 2100, oceans may become as important as terrestrial ecosystems in regulating the magnitude of the climate-carbon feedback.We are grateful for support from the U.S. Department of Energy Office of Science and the National Science Foundation (NSF). J.T.R. and F.H. received support from the Regional and Global Climate Modeling Program in the Climate and Environmental Sciences Division of the Biological and Environmental Research (BER) Program in the U.S. Department of Energy Office of Science. J.T.R., K.L., E.M., W.F., J.K.M., S.C.D., and N.N.M. received funding from the NSF project “Collaborative Research: Improved Regional and Decadal Predictions of the Carbon Cycle“ (AGS-1048827, AGS-1021776, and AGS-1048890). The Community Earth System Modeling project receives support from both NSF and BER.2015-12-0

Projected 21st century decrease in marine productivity : a multi-model analysis

© Authors, 2010. This work is distributed under the Creative Commons Attribution 3.0 License. The definitive version was published in Biogeosciences 7 (2010): 979-1005, doi: 10.5194/bg-7-979-2010Changes in marine net primary productivity (PP) and export of particulate organic carbon (EP) are projected over the 21st century with four global coupled carbon cycle-climate models. These include representations of marine ecosystems and the carbon cycle of different structure and complexity. All four models show a decrease in global mean PP and EP between 2 and 20% by 2100 relative to preindustrial conditions, for the SRES A2 emission scenario. Two different regimes for productivity changes are consistently identified in all models. The first chain of mechanisms is dominant in the low- and mid-latitude ocean and in the North Atlantic: reduced input of macro-nutrients into the euphotic zone related to enhanced stratification, reduced mixed layer depth, and slowed circulation causes a decrease in macro-nutrient concentrations and in PP and EP. The second regime is projected for parts of the Southern Ocean: an alleviation of light and/or temperature limitation leads to an increase in PP and EP as productivity is fueled by a sustained nutrient input. A region of disagreement among the models is the Arctic, where three models project an increase in PP while one model projects a decrease. Projected changes in seasonal and interannual variability are modest in most regions. Regional model skill metrics are proposed to generate multi-model mean fields that show an improved skill in representing observation-based estimates compared to a simple multi-model average. Model results are compared to recent productivity projections with three different algorithms, usually applied to infer net primary production from satellite observations.This work was funded by the European Union projects CARBOOCEAN (511176-2) and EUROCEANS (511106-2) and is a contribution to the “European Project on Ocean Acidification” (EPOCA) which received funding from the European Community’s Seventh Framework Programme (FP7/2007–2013) under grant agreement no. 211384. Additional support was received from the Swiss National Science Foundation. SCD acknowledges support from the NASA Ocean Biology and Biogeochemistry Program (NNX07AL80G). LB aknowledges support from the EU Project MEECE (Marine Ecosystem Evolution in a Changing Environnement, grant agreement 212085)

Desert dust and anthropogenic aerosol interactions in the Community Climate System Model coupled-carbon-climate model

© The Authors, 2011. This article is distributed under the terms of the Creative Commons Attribution 3.0 License. The definitive version was published in Biogeosciences 8 (2011): 387-414, doi:10.5194/bg-8-387-2011.Coupled-carbon-climate simulations are an essential tool for predicting the impact of human activity onto the climate and biogeochemistry. Here we incorporate prognostic desert dust and anthropogenic aerosols into the CCSM3.1 coupled carbon-climate model and explore the resulting interactions with climate and biogeochemical dynamics through a series of transient anthropogenic simulations (20th and 21st centuries) and sensitivity studies. The inclusion of prognostic aerosols into this model has a small net global cooling effect on climate but does not significantly impact the globally averaged carbon cycle; we argue that this is likely to be because the CCSM3.1 model has a small climate feedback onto the carbon cycle. We propose a mechanism for including desert dust and anthropogenic aerosols into a simple carbon-climate feedback analysis to explain the results of our and previous studies. Inclusion of aerosols has statistically significant impacts on regional climate and biogeochemistry, in particular through the effects on the ocean nitrogen cycle and primary productivity of altered iron inputs from desert dust deposition.This work was done under the auspices of NASA NNG06G127G, NSF grants 0748369, 0932946, 0745961 and 0832782. The work of C. J. was supported by the Joint DECC/Defra Met Office Hadley Centre Climate Programme (GA01101)

Symplastic solute transport and avocado fruit development : a decline in cytokinin/ABA ratio is related to appearance of the Hass small fruit variant

Studies on the effect of fruit size on endogenous ABA and isopentenyladenine (iP) in developing avocado (Persea americana Mill. cv. Hass) fruit revealed that ABA content was negatively correlated with fruit size whilst the iP/ABA ratio showed a linear relationship with increasing size of fruit harvested 226 d after full bloom. The effect of this change in hormone balance on the relationship between symplastic solute transport and appearance of the small fruit variant was examined following manipulation of the endogenous cytokinin (CK)/ABA ratio. Application of ABA caused seed coat senescence and retarded fruit growth but these effects were absent in fruit treated with equal amounts of ABA plus iP. Thus, the underlying physiological mechanisms associated with ABA-induced retardation of Hass avocado fruit growth appeared to be inextricably linked to a decline in CK content and included: diminution of mesocarp and seed coat plasmodesmatal branching, gating of mesocarp and seed coat plasmodesmata by deposition of electron dense material in the neck region, abolishment of the electrochemical gradient between mesocarp and seed coat parenchyma, and arrest of cell-to-cell chemical communication

Carbon-nitrogen interactions regulate climate-carbon cycle feedbacks : results from an atmosphere-ocean general circulation model

© 2009 The Authors. This article is distributed under the terms of the Creative Commons Attribution 3.0 License. The definitive version was published in Biogeosciences 6 (2009): 2099-2120, doi:10.5194/bg-6-2099-2009Inclusion of fundamental ecological interactions between carbon and nitrogen cycles in the land component of an atmosphere-ocean general circulation model (AOGCM) leads to decreased carbon uptake associated with CO2 fertilization, and increased carbon uptake associated with warming of the climate system. The balance of these two opposing effects is to reduce the fraction of anthropogenic CO2 predicted to be sequestered in land ecosystems. The primary mechanism responsible for increased land carbon storage under radiatively forced climate change is shown to be fertilization of plant growth by increased mineralization of nitrogen directly associated with increased decomposition of soil organic matter under a warming climate, which in this particular model results in a negative gain for the climate-carbon feedback. Estimates for the land and ocean sink fractions of recent anthropogenic emissions are individually within the range of observational estimates, but the combined land plus ocean sink fractions produce an airborne fraction which is too high compared to observations. This bias is likely due in part to an underestimation of the ocean sink fraction. Our results show a significant growth in the airborne fraction of anthropogenic CO2 emissions over the coming century, attributable in part to a steady decline in the ocean sink fraction. Comparison to experimental studies on the fate of radio-labeled nitrogen tracers in temperate forests indicates that the model representation of competition between plants and microbes for new mineral nitrogen resources is reasonable. Our results suggest a weaker dependence of net land carbon flux on soil moisture changes in tropical regions, and a stronger positive growth response to warming in those regions, than predicted by a similar AOGCM implemented without land carbon-nitrogen interactions. We expect that the between-model uncertainty in predictions of future atmospheric CO2 concentration and associated anthropogenic climate change will be reduced as additional climate models introduce carbon-nitrogen cycle interactions in their land components.This work was supported in part by NASA Earth Science Enterprise, Terrestrial Ecology Program, grant #W19,953 to P. E. Thornton. Support was provided by the National Center for Atmospheric Research (NCAR) through the NCAR Community Climate System Modeling program, and through the NCAR Biogeosciences program. Additional support was provided by the US Department of Energy, Office of Science, Office of Biological and Environmental Research. I. Fung, S. Doney, N. Mahowald, and J. Randerson acknowledge support from National Science Foundation, Atmospheric Sciences Division, through the Carbon and Water Initiative

Aerosol trace metal leaching and impacts on marine microorganisms

Metal dissolution from atmospheric aerosol deposition to the oceans is important in enhancing and inhibiting phytoplankton growth rates and modifying plankton community structure, thus impacting marine biogeochemistry. Here we review the current state of knowledge on the causes and effects of the leaching of multiple trace metals from natural and anthropogenic aerosols. Aerosol deposition is considered both on short timescales over which phytoplankton respond directly to aerosol metal inputs, as well as longer timescales over which biogeochemical cycles are affected by aerosols

Wage losses in the year after breast cancer: Extent and determinants among Canadian women

This article is available open access through the publisher’s website at the link below. © The Author 2008.Background - Wage losses after breast cancer may result in considerable financial burden. Their assessment is made more urgent because more women now participate in the workforce and because breast cancer is managed using multiple treatment modalities that could lead to long work absences. We evaluated wage losses, their determinants, and the associations between wage losses and changes for the worse in the family's financial situation among Canadian women over the first 12 months after diagnosis of early breast cancer. Methods - We conducted a prospective cohort study among women with breast cancer from eight hospitals throughout the province of Quebec. Information that permitted the calculation of wage losses and information on potential determinants of wage losses were collected by three pretested telephone interviews conducted over the year following the start of treatment. Information on medical characteristics was obtained from medical records. The main outcome was the proportion of annual wages lost because of breast cancer. Multivariable analysis of variance using the general linear model was used to identify personal, medical, and employment characteristics associated with the proportion of wages lost. All statistical tests were two-sided. Results - Among 962 eligible breast cancer patients, 800 completed all three interviews. Of these, 459 had a paying job during the month before diagnosis. On average, these working women lost 27% of their projected usual annual wages (median = 19%) after compensation received had been taken into account. Multivariable analysis showed that a higher percentage of lost wages was statistically significantly associated with a lower level of education (Ptrend = .0018), living 50 km or more from the hospital where surgery was performed (P = .070), lower social support (P = .012), having invasive disease (P = .086), receipt of chemotherapy (P < .001), self-employment (P < .001), shorter tenure in the job (Ptrend < .001), and part-time work (P < .001). Conclusion - Wage losses and their effects on financial situation constitute an important adverse consequence of breast cancer in Canada.The Canadian Breast Cancer Research Alliance, Canadian Institutes of Health Research, and Fondation de l’Université Laval

Atmospheric carbon dioxide variability in the Community Earth System Model : evaluation and transient dynamics during the twentieth and twenty-first centuries

Author Posting. © American Meteorological Society, 2013. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Climate 26 (2013): 4447–4475, doi:10.1175/JCLI-D-12-00589.1.Changes in atmospheric CO2 variability during the twenty-first century may provide insight about ecosystem responses to climate change and have implications for the design of carbon monitoring programs. This paper describes changes in the three-dimensional structure of atmospheric CO2 for several representative concentration pathways (RCPs 4.5 and 8.5) using the Community Earth System Model–Biogeochemistry (CESM1-BGC). CO2 simulated for the historical period was first compared to surface, aircraft, and column observations. In a second step, the evolution of spatial and temporal gradients during the twenty-first century was examined. The mean annual cycle in atmospheric CO2 was underestimated for the historical period throughout the Northern Hemisphere, suggesting that the growing season net flux in the Community Land Model (the land component of CESM) was too weak. Consistent with weak summer drawdown in Northern Hemisphere high latitudes, simulated CO2 showed correspondingly weak north–south and vertical gradients during the summer. In the simulations of the twenty-first century, CESM predicted increases in the mean annual cycle of atmospheric CO2 and larger horizontal gradients. Not only did the mean north–south gradient increase due to fossil fuel emissions, but east–west contrasts in CO2 also strengthened because of changing patterns in fossil fuel emissions and terrestrial carbon exchange. In the RCP8.5 simulation, where CO2 increased to 1150 ppm by 2100, the CESM predicted increases in interannual variability in the Northern Hemisphere midlatitudes of up to 60% relative to present variability for time series filtered with a 2–10-yr bandpass. Such an increase in variability may impact detection of changing surface fluxes from atmospheric observations.The CESM project is supported by the National Science Foundation and the Office of Science (BER) of the U.S. Department of Energy. Computing resources were provided by the Climate Simulation Laboratory at NCAR’s Computational and Information Systems Laboratory (CISL), sponsored by the National Science Foundation and other agencies. G.K.A. acknowledges support of a NOAA Climate and Global Change postdoctoral fellowship. J.T.R., N.M.M., S.C.D., K.L., and J.K.M. acknowledge support of Collaborative Research: Improved Regional and Decadal Predictions of the Carbon Cycle (NSF AGS-1048827, AGS-1021776,AGS-1048890). TheHIPPO Programwas supported byNSF GrantsATM-0628575,ATM-0628519, and ATM-0628388 to Harvard University, University of California (San Diego), and by University Corporation for Atmospheric Research, University of Colorado/ CIRES, by the NCAR and by the NOAAEarth System Research Laboratory. Sunyoung Park, Greg Santoni, Eric Kort, and Jasna Pittman collected data during HIPPO. The ACME project was supported by the Office of Biological and Environmental Research of the U.S. Department of Energy under Contract DE-AC02- 05CH11231 as part of the Atmospheric Radiation Measurement Program (ARM), the ARM Aerial Facility, and the Terrestrial EcosystemScience Program. TCCON measurements at Eureka were made by the Canadian Network for Detection of Atmospheric Composition Change (CANDAC) with additional support from the Canadian Space Agency. The Lauder TCCON program was funded by the New Zealand Foundation for Research Science and Technology contracts CO1X0204, CO1X0703, and CO1X0406. Measurements at Darwin andWollongong were supported by Australian Research Council Grants DP0879468 and DP110103118 and were undertaken by David Griffith, Nicholas Deutscher, and Ronald Macatangay. We thank Pauli Heikkinen, Petteri Ahonen, and Esko Kyr€o of the Finnish Meteorological Institute for contributing the Sodankyl€a TCCON data. Measurements at Park Falls, Lamont, and Pasadena were supported byNASAGrant NNX11AG01G and the NASA Orbiting Carbon Observatory Program. Data at these sites were obtained by Geoff Toon, Jean- Francois Blavier, Coleen Roehl, and Debra Wunch.2014-01-0