Phosphorus Limitation, Uptake, and Turnover in Benthic Stream Algae

In this chapter, three different aspects of P utilization by benthic algae will be covered: (1) assessment of P limitation, (2) measurement of P uptake rates, and (3) determination of the release rate of P (expressed as the turnover rate). We note two caveats regarding this chapter. First, we focus exclusively on inorganic P; it is likely that dissolved organic phosphorus plays an important, albeit relatively undefined role, in the nutrient dynamics of freshwater algae (cf. Hwang et al., 1998; Pant et al., 2002), especially in P-limited environments (Karl and Björkman, 2015). However, treatment of this topic is beyond the scope of this chapter (but see Chapter 24). Second, although we use the term benthic algae throughout the chapter, it should be noted that the benthic algae attached to submerged substrata in streams usually exist as part of a complex assemblage variously referred to as periphyton, aufwuchs, or biofilm. This assemblage usually consists of algae, bacteria, fungi, and meiofauna (see Chapters 9e11, 14) that exist within a mucilaginous, poly- saccharide matrix (Lock et al., 1984), and each biotic group has different affinities for P. Indeed, even within a group, P uptake and cycling may be influenced by the abundance of different species and growth forms (cf. Steinman et al., 1992; Davies and Bothwell, 2012)

A superfluid He3 detector for direct dark matter search

MACHe3 (MAtrix of Cells of superfluid He3) is a project of a new detector for direct Dark Matter Search. The idea is to use superfluid He3 as a sensitive medium. The existing device, the superfluid He3 cell, will be briefly introduced. Then a description of the MACHe3 project will be presented, in particular the background rejection and the neutralino event rate that may be achieved with such a device.Comment: 6 pages, 3 figures, Proceedings of the 3rd International Workshop on the Identification of Dark Matter (York, UK, 09/18/2000-09/22/2000

A project of a new detector for direct Dark Matter search: MACHe3

MACHe3 (MAtrix of Cells of superfluid He3) is a project of a new detector for direct Dark Matter (DM) search. A cell of superfluid He3 has been developed and the idea of using a large number of such cells in a high granularity detector is proposed.This contribution presents, after a brief description of the superfluid He3 cell, the simulation of the response of different matrix configurations allowing to define an optimum design as a function of the number of cells and the volume of each cell. The exclusion plot and the predicted interaction cross-section for the neutralino as a photino are presented.Comment: 8 pages, 7 figures, Proceedings of Dark Matter 2000 (Marina Del Rey, Los Angeles, USA, 02/23/2000-02/25/2000

Effects of water level fluctuation and sediment–water nutrient exchange on phosphorus biogeochemistry in two coastal wetlands

Freshwater coastal wetlands are sensitive to drying and wetting events (DWE), associated with water level fluctuations. Prior studies have shown that DWE influence sediment–water nutrient exchange, but the fate of these nutrients has received much less attention. To address this information gap, we investigated how microbial (i.e., bacteria and phytoplankton) structure and function respond to DWE-induced release of nutrients from the sediment of mesotrophic and eutrophic coastal wetlands. Our approach was three-fold: (1) sediment from each wetland was subjected to a desiccation (2 month) and re-wetting (2 days) period in the laboratory; (2) the overlying water column from the DWE was measured for nutrient concentration; and (3) we conducted a microbial response experiment using the water from either DWE experiment (containing the nutrients released during the DWE) or with added nutrients obtained from commercially available sources (but at similar concentrations to those released from the sediments), to test the effect of nutrient concentration and source on microbes inhabiting the water column and their role in the fate of the released nutrients, in particular phosphate (Pi). In both coastal wetlands, the microbial community structure in the water column (absolute and relative abundance of different microbial groups) was modified by bacteria resuspended from the sediment after re-wetting. However, results from the microbial response experiments showed that the microbial communities did not grow following inputs of commercially available nutrients or release of sediment nutrients. In Pi amended treatments, Pi uptake rates increased but not enough to reduce Pi turnover time to values measured in the control. As a result, Pi concentration and turnover remained high. In coastal wetlands with limited hydrologic exchange, these elevated nutrients will accumulate and lead to further eutrophication. However, in open coastal wetlands, with substantial hydrologic exchange, advected nutrients may influence biological activity in the nearshore zone

Membrane binding and oligomer formation by the calcium-dependent lipopeptide antibiotic A54145: a quantitative study with pyrene excimer fluorescence

The final publication is available at Elsevier via http://doi.org/10.1016/j.bpj.2016.07.018 © 2016. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/A54145 is a lipopeptide antibiotic related to daptomycin that permeabilizes bacterial cell membranes. Its action requires both calcium and phosphatidylglycerol in the target membrane, and it is accompanied by the formation of membrane-associated oligomers. We here probed the interaction of A54145 with model membranes composed of dimyristoylphosphatidylcholine and dimyristoylphosphatidylglycerol, using the steady-state and time-resolved fluorescence of a pyrene-labeled derivative (Py-A54145). In solution, the labeled peptide was found to exist as a monomer. Its membrane interaction occurred in two stages that could be clearly distinguished by varying the calcium concentration. In the first stage, which was observed between 0.15 and 1 mM calcium, Py-A54145 bound to the membrane, as indicated by a strong increase in pyrene monomer emission. At the same calcium concentration, excimer emission increased also, suggesting that Py-A54145 had oligomerized. A global analysis of the time-resolved pyrene monomer and excimer fluorescence confirmed that Py-A54145 forms oligomers quantitatively and concomitantly with membrane binding. When calcium was raised beyond 1 mM, a distinct second transition was observed that may correspond to a doubling of the number of oligomer subunits. The collective findings confirm and extend our understanding of the action mode of A54145 and daptomycin.This work was supported by the Natural Sciences and Engineering Research Council of Canada operating grants to J.D. (201603-2013), M.P. (250265-2013), and S.T. (155283-2012)

ROLE OF SECOND MESSENGER SIGNALING PATHWAYS IN THE REGULATION OF SARCOPLASMIC RETICULUM CALCIUM-HANDLING PROPERTIES IN THE LEFT VENTRICLE AND SKELETAL MUSCLES OF DIFFERENT FIBRE TYPE COMPOSITION

The overall objective of this thesis was to examine mechanisms involved in the acute regulation of sarcoplasmic reticulum (SR) Ca2+-handling properties by second messenger signaling pathways in skeletal and cardiac muscle. The aim of the first study (Chapter Two) was to characterize changes in the kinetic properties of sarco(endo)-plasmic reticulum Ca2+-ATPase (SERCA) proteins in cardiac and skeletal muscles in response to b-adrenergic, Ca2+-dependent calmodulin kinase II (CaMKII) and protein kinase C (PKC) signaling. The aim of the second study (Chapter Three) was to determine if insulin signaling could acutely regulate SERCA kinetic properties in cardiac and skeletal muscle. The aim of the final study (Chapter Four) was to determine if alterations in plasma glucose, epinephrine and insulin concentrations during exercise are able to influence SR Ca2+-handling properties in contracting human skeletal muscle. Data collected in Chapter Two and Chapter Three were obtained using tissue prepared from a group of 28 male Sprague-Dawley rats (9 weeks of age; mass = 280 ± 4 g: X ± S.E). Crude muscle homogenates (11:1 dilution) were prepared from selected hind limb muscles (soleus, SOL; extensor digitorum longus, EDL; the red portion of gastrocnemius, RG; and the white portion of gastrocnemius, WG) and the left ventricle (LV). Enriched SR membrane fractions, prepared from WG and LV, were also analyzed. A spectrophotometric assay was used to measure kinetic properties of SERCA, namely, maximal SERCA activity (Vmax), and Ca2+-sensitivity was characterized by both the Ca50, which is defined as the free Ca2+-concentration needed to elicit 50% Vmax, and the Hill coefficient (nH), which is defined as the relationship between SERCA activity and Ca2+f for 10 to 90% Vmax. The observations made in Chapter Two indicated that b-adrenergic signaling, activated by epinephrine, increased (P<0.05) Ca2+-sensitivity, as shown by a left-shift in Ca50 (i.e. reduced Ca50), without altering Vmax in LV and SOL but had no effect (P<0.05) on EDL, RG, or WG. Further analysis using a combination of cAMP, the PKA activator forskolin, and/or the PKA inhibitor KT5270 indicated that the reduced Ca50 in LV was activated by cAMP- and PKA-signaling mechanisms. However, although the reduced Ca50 in SOL was cAMP-dependent, it was not influenced by a PKA-dependent mechanism. In contrast to the effects of b-adrenergic signaling, CaMKII activation increased SERCA Ca2+-sensitivity, as shown by a left-shift in Ca50 and increased nh, without altering SERCA Vmax in LV but was without effect in any of the skeletal muscles examined. The PKC activator PMA significantly reduced SERCA Ca2+-sensitivity, by inducing a right-shift in Ca50 and decreased nH in the LV and all skeletal muscles examined. PKC activation also reduced Vmax in the fast-twitch skeletal muscles (i.e. EDL, RG and WG), but did not alter Vmax in LV or SOL. The results of Chapter Three indicated that insulin signaling increased SERCA Ca2+-sensitivity, as shown by a left-shift in Ca50 (i.e. reduced Ca50) and an increased nH, without altering SERCA Vmax in crude muscle homogenates prepared from LV, SOL, EDL, RG, and WG. An increase in SERCA Ca2+-sensitivity was also observed in enriched SERCA1a and SERCA2a vesicles when an activated form of the insulin receptor (A-INS-R) was included during biochemical analyses. Co-immunoprecipitation experiments were conducted and indicated that IRS-1 and IRS-2 proteins bind SERCA1a and SERCA2a in an insulin-dependent manner. However, the binding of IRS proteins with SERCA does not appear to alter the structural integrity of the SERCA Ca2+-binding site since no changes in NCD-4 fluorescence were observed in response to insulin or A-INS-R. Moreover, the increase in SERCA Ca2+-sensitivity due to insulin signaling was not associated with changes in the phosphorylation status of phospholamban (PLN) since Ser16 or Thr17 phosphorylation was not altered by insulin or A-INS-R in LV tissue. The data described in Chapter Four was collected from 15 untrained human participants (peak O2 consumption, VO2peak= 3.45 ± 0.17 L/min) who completed a standardized cycle test (~60% VO2peak) on two occasions during which they were provided either an artificially sweetened placebo (PLAC) or a 6% glucose (GLUC) beverage (~1.00 g CHO per kg body mass). Muscle biopsies were collected from the vastus lateralis at rest, after 30 min and 90 min of exercise and at fatigue in both conditions to allow assessment of metabolic and SR data. Glucose supplementation increased exercise ride time by ~19% (137 ± 7 min) compared to PLAC (115 ± 6 min). This performance increase was associated with elevated plasma glucose and insulin concentrations and reduced catecholamine concentrations during GLUC compared to PLAC. Prolonged exercise reduced (p<0.05) SR Ca2+-uptake, Vmax, Phase 1 and Phase 2 Ca2+-release rates during both PLAC and GLUC. However, no differences in SR Ca2+-handling properties were observed between conditions when direct comparisons were made at matched time points between PLAC and GLUC. In summary, the results of the first study (Chapter Two) indicate that b-adrenergic and CaMKII signaling increases SERCA Ca2+-sensitivity in the LV and SOL; while PKC signaling reduces SERCA Ca2+-sensitivity in all tissues. PKC activation also reduces Vmax in the fast-twitch skeletal muscles (i.e. EDL, RG, and WG) but has no effect on Vmax in the LV and SOL. The results of the second study (Chapter Three) indicate that insulin signaling acutely increases the Ca2+-sensitivity of SERCA1a and SERCA2a in all tissues examined, without altering the Vmax. Based on our observations, it appears that the increase in SERCA Ca2+-sensitivity may be regulated, in part, through the interaction of IRS proteins with SERCA1a and SERCA2a. The results of the final study (Chapter Four) indicate that alterations in plasma glucose, epinephrine and insulin concentrations associated with glucose supplementation during exercise, do not alter the time course or magnitude of reductions in SERCA or Ca2+-release channel (CRC) function in working human skeletal muscle. Although glucose supplementation did increase exercise ride time to fatigue in this study, our data does not reveal an association with SR Ca2+-cycling measured in vitro. It is possible that the strength of exercise signal overrides the hormonal influences observed in resting muscles. Additionally, these data do not rule out the possibility that glucose supplementation may influence E-C coupling processes or SR Ca2+-cycling properties in vivo

Fast Fourier Optimization: Sparsity Matters

Many interesting and fundamentally practical optimization problems, ranging from optics, to signal processing, to radar and acoustics, involve constraints on the Fourier transform of a function. It is well-known that the {\em fast Fourier transform} (fft) is a recursive algorithm that can dramatically improve the efficiency for computing the discrete Fourier transform. However, because it is recursive, it is difficult to embed into a linear optimization problem. In this paper, we explain the main idea behind the fast Fourier transform and show how to adapt it in such a manner as to make it encodable as constraints in an optimization problem. We demonstrate a real-world problem from the field of high-contrast imaging. On this problem, dramatic improvements are translated to an ability to solve problems with a much finer grid of discretized points. As we shall show, in general, the "fast Fourier" version of the optimization constraints produces a larger but sparser constraint matrix and therefore one can think of the fast Fourier transform as a method of sparsifying the constraints in an optimization problem, which is usually a good thing.Comment: 16 pages, 8 figure

Phosphate availability and the ultimate control of new nitrogen input by nitrogen fixation in the tropical Pacific Ocean

International audienceDue to the low atmospheric input of phosphate into the open ocean, it is one of the key nutrients that could ultimately control primary production and carbon export into the deep ocean. The observed trend over the last 20 years has shown a decrease in the dissolved inorganic phosphate (DIP) pool in the North Pacific gyre, which has been correlated to the increase in di-nitrogen (N2) fixation rates. Following a NW-SE transect, in the Southeast Pacific during the early austral summer (BIOSOPE cruise), we present data on DIP, dissolved organic phosphate (DOP) and particulate phosphate (PP) pools along with DIP turnover times (TDIP) and N2 fixation rates. We observed a decrease in DIP concentration from the edges to the centre of the gyre. Nevertheless the DIP concentrations remained above 100 nmol L-1 and T DIP was more than 6 months in the centre of the gyre; DIP availability remained largely above the level required for phosphate limitation to occur and the absence of Trichodesmium spp and low nitrogen fixation rates were likely to be controlled by other factors such as temperature or iron availability. This contrasts with recent observations in the North Pacific Ocean at the ALOHA station and in the western Pacific Ocean at the same latitude (DIAPALIS cruises) where lower DIP concentrations (-1) and T DIP 2 fixation rates and possibly carbon dioxide sequestration, if the primary ecophysiological controls, temperature and/or iron availability, were alleviated

Fracture of a biopolymer gel as a viscoplastic disentanglement process

We present an extensive experimental study of mode-I, steady, slow crack dynamics in gelatin gels. Taking advantage of the sensitivity of the elastic stiffness to gel composition and history we confirm and extend the model for fracture of physical hydrogels which we proposed in a previous paper (Nature Materials, doi:10.1038/nmat1666 (2006)), which attributes decohesion to the viscoplastic pull-out of the network-constituting chains. So, we propose that, in contrast with chemically cross-linked ones, reversible gels fracture without chain scission