The effects of temperature and exercise training on swimming performance in juvenile qingbo (Spinibarbus sinensis)

To investigate the effects of temperature and exercise training on swimming performance in juvenile qingbo (Spinibarbus sinensis), we measured the following: (1) the resting oxygen consumption rate $ \left( {{\dot{\text{M}}\text{O}}_{{ 2 {\text{rest}}}} } \right) $ , critical swimming speed (U _crit) and active oxygen consumption rate $ \left( {{\dot{\text{M}}\text{O}}_{{ 2 {\text{active}}}} } \right) $ of fish at acclimation temperatures of 10, 15, 20, 25 and 30 °C and (2) the $ \dot{M}{\text{O}}_{{ 2 {\text{rest}}}} $ , U _crit and $ \dot{M}{\text{O}}_{{ 2 {\text{active}}}} $ of both exercise-trained (exhaustive chasing training for 14 days) and control fish at both low and high acclimation temperatures (15 and 25 °C). The relationship between U _crit and temperature (T) approximately followed a bell-shaped curve as temperature increased: U _crit = 8.21/{1 + [(T ? 27.2)/17.0]²} (R ² = 0.915, P < 0.001, N = 40). The optimal temperature for maximal U _crit (8.21 BL s^?1) in juvenile qingbo was 27.2 °C. Both the $ \dot{M}{\text{O}}_{{ 2 {\text{active}}}} $ and the metabolic scope (MS, $ \dot{M}{\text{O}}_{{ 2 {\text{active}}}} - \dot{M}{\text{O}}_{{ 2 {\text{rest}}}} $ ) of qingbo increased with temperature from 10 to 25 °C (P < 0.05), but there were no significant differences between fish acclimated to 25 and 30 °C. The relationships between $ \dot{M}{\text{O}}_{{ 2 {\text{active}}}} $ or MS and temperature were described as $ {\dot{\text{M}}\text{O}}_{{ 2 {\text{active}}}} = 1,214.29/\left\{ {1 + \left[ {\left( {T - 28.8} \right)/10.6} \right]^{2} } \right\}\;\left( {R^{2} = 0.911,\;P < 0.001,\;N = 40} \right) $ and MS = 972.67/{1 + [(T ? 28.0)/9.34]²} (R ² = 0.878, P < 0.001, N = 40). The optimal temperatures for $ \dot{M}{\text{O}}_{{ 2 {\text{active}}}} $ and MS in juvenile qingbo were 28.8 and 28.0 °C, respectively. Exercise training resulted in significant increases in both U _crit and $ \dot{M}{\text{O}}_{{ 2 {\text{active}}}} $ at a low temperature (P < 0.05), but training exhibited no significant effect on either U _crit or $ \dot{M}{\text{O}}_{{ 2 {\text{active}}}} $ at a high temperature. These results suggest that exercise training had different effects on swimming performance at different temperatures. These differences may be related to changes in aerobic metabolic capability, arterial oxygen delivery, available dissolved oxygen, imbalances in ion fluxes and stimuli to remodel tissues with changes in temperature.     

Measurements of CO2 and CH4 evasion from UK peatland headwater streams

Peatland headwater streams are consistently supersaturated with respect to gaseous C and are known to degas CO₂ and CH₄ directly to the atmosphere. Using a combination of injection of a purposeful gas tracer (propane) and a soluble tracer (NaCl) we carried out 49 measurements of the gas transfer coefficient on 12 representative stream reaches to quantify the gas transfer rates of CO₂ and CH₄ in headwater (1st–3rd order) streams draining six UK peatlands. These were compared to measured stream reach physical variables, such as discharge and water travel time. Whilst we found that evasion rates were highly variable in space and time, $ {\text{K}}_{{{\text{CO}}_{2} }} $ (gas transfer coefficient of CO₂) was positively related to discharge. Individual study sites showed a high degree of variability in gas transfer rates; at all 49 sites median/mean values for $ {\text{K}}_{{{\text{CO}}_{2} }} $ were 0.087/0.157 and $ {\text{K}}_{{{\text{CH}}_{4} }} $ 0.092/0.176 min^?1. Median/mean instantaneous CO₂ and CH₄ evasion rates were 133/367 and 0.22/1.45 μg C m^?2 s^?1, respectively. Methane evasion rates were therefore more than two orders of magnitude lower than CO₂, with CH₄ invasion (rather than evasion) measured on 37 % of occasions. Our gas flux measurements from peatland headwater streams are higher than values previously used to estimate landscape scale fluxes and emphasise the importance of the evasion flux term in the overall carbon balance.     

Can the positive aromatic ring be as ��-electron donor in ��-halogen bond? A MP2 theoretical investigation on the unusual ��-halogen bond interaction between three-membered ring \left( {\hbox{BNN}} \right)_3^{+} and X1X2 (X1, X2?=?F, Cl, Br)

The unusual ??-halogen bond interactions are investigated between $ \left( {\hbox{BNN}} \right)_3^{+} $ and X1X2 (X1, X2?=?F, Cl, Br) employing MP2 at 6-311?+?G(2d) and aug-cc-pVDZ levels according to the ??CP (counterpoise) corrected potential energy surface (PES)?? method. The order of the ??-halogen bond interactions and stabilities of the complexes are obtained to be $ \left( {\hbox{BNN}} \right)_3^{+} \ldots {{\hbox{F}}_2} < \left( {\hbox{BNN}} \right)_3^{+} \ldots {\hbox{ClF < }}\left( {\hbox{BNN}} \right)_3^{+} \ldots {\hbox{C}}{{\hbox{l}}_2} < \left( {\hbox{BNN}} \right)_3^{+} \ldots {\hbox{BrCl}}\quad { < }\quad \left( {\hbox{BNN}} \right)_3^{+} \ldots {\hbox{B}}{{\hbox{r}}_2}\quad { < }\quad \left( {\hbox{BNN}} \right)_3^{+} \ldots {\hbox{BrF}}{.} $ at MP2/aug-cc-pVDZ level. The analyses of the Mulliken charge transfer, natural bond orbital (NBO), atoms in molecules (AIM) theory and electron density shifts reveal that the nature of the ??-halogen bond interaction in the complexes of ClF, BrF and BrCl might partly be charge transfer from the delocalized ??-HOMO orbital of $ \left( {\hbox{BNN}} \right)_3^{+} $ to X1X2. This result suggests that the positive aromatic ring $ \left( {\hbox{BNN}} \right)_3^{+} $ might act as a ??-electron donor to form the ??-halogen bond.

Effects of negative air ions on oxygen uptake kinetics,recovery and performance in exercise: a randomized,double-blinded study

Limited research has suggested that acute exposure to negatively charged ions may enhance cardio-respiratory function, aerobic metabolism and recovery following exercise. To test the physiological effects of negatively charged air ions, 14 trained males (age: 32?±?7 years; \( \overset{\cdotp }{V}{\mathrm{O}}_{2 \max } \) : 57?±?7 mL min^?1 kg^?1) were exposed for 20 min to either a high-concentration of air ions (ION: 220?±?30?×?10³ ions cm^?3) or normal room conditions (PLA: 0.1?±?0.06?×?10³ ions cm^?3) in an ionization chamber in a double-blinded, randomized order, prior to performing: (1) a bout of severe-intensity cycling exercise for determining the time constant of the phase II \( \overset{\cdotp }{V}{\mathrm{O}}_2 \) response (τ) and the magnitude of the \( \overset{\cdotp }{V}{\mathrm{O}}_2 \) slow component (SC); and (2) a 30-s Wingate test that was preceded by three 30-s Wingate tests to measure plasma [adrenaline] (ADR), [nor-adrenaline] (N-ADR) and blood [lactate] (B_Lac) over 20 min during recovery in the ionization chamber. There was no difference between ION and PLA for the phase II \( \overset{\cdotp }{V}{\mathrm{O}}_2 \) τ (32?±?14 s vs. 32?±?14 s; P?=?0.7) or \( \overset{\cdotp }{V}{\mathrm{O}}_2 \) SC (404?±?214 mL vs 482?±?217 mL; P?=?0.17). No differences between ION and PLA were observed at any time-point for ADR, N-ADR and B_Lac as well as on peak and mean power output during the Wingate tests (all P?>?0.05). A high-concentration of negatively charged air ions had no effect on aerobic metabolism during severe-intensity exercise or on performance or the recovery of the adrenergic and metabolic responses after repeated-sprint exercise in trained athletes.     

Age structure, growth and mortality estimates of an endemic Ptychobarbus dipogon (Regan, 1905) (Cyprinidae: Schizothoracinae) in the Lhasa River, Tibet

Ptychobarbus dipogon is an endemic fish in the Yarlung Tsangpo River, but its biology is poorly known. We sampled 582 specimens (total length, TL, between 70.6 and 593.0 mm) from April 2004 to August 2006 in the Lhasa River, Tibet. We estimated ages based on the counts of alternating opaque and translucent zones (annuli) in thin transverse sections of lapilli otoliths. Ages ranged from 1⁺ to 23⁺ years for males and 1⁺ to 44⁺ for females. The observed 44⁺ years was the oldest reported for schizothoracine fishes. Females attained a larger size than males. The TL–weight relationship was W?=?7.12?×?10^?6 TL ^3.006 for combined sexes. The growth parameters fitted von Bertalanffy growth functions were $L_\infty = 598.66\,{\text{mm}}$ , k?=?0.0898 year^?1, t ₀?=??0.7261 year and $W_\infty = 1585.38\;{\text{g}}$ for females and $L_\infty = 494.23{\text{ mm}}$ , k?=?0.1197 year^?1, t ₀?=??0.7296 year and $W_\infty = 904.88{\text{ g}}$ for males. The longevities of 32.7 year for females and 24.3 year for males were similar to the observed ages. Using an empirical model we estimated the instantaneous rate of total mortality (Z) at 0.28 per year in the lower reaches. Z in the upper and middle stocks was close to the M because of unexploited or lightly exploited stock. Protracted longevity, slow growth, low natural mortality and large body size were typical characteristics of P. dipogon. The current declining trend of P. dipogon could be prevented by altering fishing regulations.     

Longitudinal distribution of nitrate δ15N and δ18O in two contrasting tropical rivers: implications for instream nitrogen cycling

The longitudinal variations in the nitrogen (δ¹⁵N) and oxygen (δ¹⁸O) isotopic compositions of nitrate (NO₃ ^?), the carbon isotopic composition (δ¹³C) of dissolved inorganic carbon (DIC) and the δ¹³C and δ¹⁵N of particulate organic matter were determined in two Southeast Asian rivers contrasting in the watershed geology and land use to understand internal nitrogen cycling processes. The $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ became higher longitudinally in the freshwater reach of both rivers. The $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ also increased longitudinally in the river with a relatively steeper longitudinal gradient and a less cultivated watershed, while the $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ gradually decreased in the other river. A simple model for the $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ and the $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ that accounts for simultaneous input and removal of NO₃ ^? suggested that the dynamics of NO₃ ^? in the former river were controlled by the internal production by nitrification and the removal by denitrification, whereas that in the latter river was significantly affected by the anthropogenic NO₃ ^? loading in addition to the denitrification and/or assimilation. In the freshwater-brackish transition zone, heterotrophic activities in the river water were apparently elevated as indicated by minimal dissolved oxygen, minimal δ¹³C_DIC and maximal pCO₂. The δ¹⁵N of suspended particulate nitrogen (PN) varied in parallel to the $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ there, suggesting that the biochemical recycling processes (remineralization of PN coupled to nitrification, and assimilation of NO₃ ^?-N back to PN) played dominant roles in the instream nitrogen transformation. In the brackish zone of both rivers, the $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ displayed a declining trend while the $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ increased sharply. The redox cycling of NO₃ ^?/NO₂ ^? and/or deposition of atmospheric nitrogen oxides may have been the major controlling factor for the estuarine $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ and $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ , however, the exact mechanism behind the observed trends is currently unresolved.     

Hyperfine interactions and electron distribution in FeIIFeI and FeIFeI models for the active site of the [FeFe] hydrogenases: Mössbauer spectroscopy studies of low-spin FeI

Mössbauer studies of [{μ-S(CH₂C(CH₃)₂CH₂S}(μ-CO)Fe^IIFe^I(PMe₃)₂(CO)₃]PF₆ (1 _OX ), a model complex for the oxidized state of the [FeFe] hydrogenases, and the parent Fe^IFe^I derivative are reported. The paramagnetic 1 _OX is part of a series featuring a dimethylpropanedithiolate bridge, introducing steric hindrance with profound impact on the electronic structure of the diiron complex. Well-resolved spectra of 1 _OX allow determination of the magnetic hyperfine couplings for the low-spin distal Fe^I ( $ {\text{Fe}}^{\text{I}} _{\text{ D}} $ Fe D I ) site, A _x,y,z  = [?24 (6), ?12 (2), 20 (2)] MHz, and the detection of significant internal fields (approximately 2.3 T) at the low-spin ferrous site, confirmed by density functional theory (DFT) calculations. Mössbauer spectra of 1 _OX show nonequivalent sites and no evidence of delocalization up to 200 K. Insight from the experimental hyperfine tensors of the Fe^I site is used in correlation with DFT to reveal the spatial distribution of metal orbitals. The Fe–Fe bond in [Fe₂{μ-S(CH₂C(CH₃)₂CH₂S}(PMe₃)₂(CO)₄] (1) involving two $ d_{{z^{2} }} $ d z 2 -type orbitals is crucial in keeping the structure intact in the presence of strain. On oxidation, the distal iron site is not restricted by the Fe–Fe bond, and thus the more stable isomer results from inversion of the square pyramid, rotating the $ d_{{z^{2} }} $ d z 2 orbital of $ {\text{Fe}}^{\text{I}} _{\text{ D}} $ Fe D I . DFT calculations imply that the Mössbauer properties can be traced to this $ d_{{z^{2} }} $ d z 2 orbital. The structure of the magnetic hyperfine coupling tensor, A, of the low-spin Fe^I in 1 _OX is discussed in the context of the known A tensors for the oxidized states of the [FeFe] hydrogenases.     

Theoretical study of the reaction of CH2XO (X = F, Cl, Br) radicals with the NO radical

In this paper, we focus on the multiple-channel reactions of CH₂XO (X = F, Cl, Br) radicals with the NO radical by means of direct dynamic methods. All structures of the stationary points were obtained at the MP2/6-311+G(d,p) level and vibrational frequency analysis was also performed at this level of theory. The minimum energy path (MEP) was obtained via the intrinsic reaction coordinate (IRC) theory at the MP2/6-311+G(d,p) level, and higher-level energetic information was refined by the MC-QCISD method. The rate constants for the three hydrogen abstraction reaction channels over the temperature range 200–1,500 K were calculated by the improved canonical variational transition state theory (ICVT) with a correction for small-curvature tunneling (SCT). The rate constants calculated in this manner were in good agreement with the available experimental data, and the three-parameter rate–temperature formulae for the temperature range 200–1,500 K were $ {k_{1{\text{a}} }}(T)=0.32\times {10^{-18 }}{T^{1.83 }}\exp \left( {1748.54/T} \right) $ , $ {k_{2{\text{a}} }}(T)=0.22\times {10^{-19 }}{T^{2.19 }}\exp \left( {1770.19/T} \right) $ , $ {k_{3{\text{a}} }}(T)=0.88\times {10^{-20 }}{T^{2.20 }}\exp \left( {1513.82/T} \right) $ (in units of cm³ molecule^?1?s^?1).     

Relative CO2/NH3 Permeabilities of Human RhAG, RhBG and RhCG

Mammalian glycosylated rhesus (Rh) proteins include the erythroid RhAG and the nonerythroid RhBG and RhCG. RhBG and RhCG are expressed in multiple tissues, including hepatocytes and the collecting duct (CD) of the kidney. Here, we expressed human RhAG, RhBG and RhCG in Xenopus oocytes (vs. H₂O-injected control oocytes) and used microelectrodes to monitor the maximum transient change in surface pH (ΔpH_S) caused by exposing the same oocyte to 5 % CO₂/33 mM HCO₃ ^? (an increase) or 0.5 mM NH₃/NH₄ ⁺ (a decrease). Subtracting the respective values for day-matched, H₂O-injected control oocytes yielded channel-specific values (*). $({\Updelta {\text{pH}}_{\text{S}}^{*} })_{{{\text{CO}}_{ 2} }}$ and $({ - \Updelta {\text{pH}}_{\text{S}}^{*} })_{{{\text{NH}}_{ 3} }}$ were each significantly >0 for all channels, indicating that RhBG and RhCG—like RhAG—can carry CO₂ and NH₃. We also investigated the role of a conserved aspartate residue, which was reported to inhibit NH₃ transport. However, surface biotinylation experiments indicate the mutants RhBG_D178N and RhCG_D177N have at most a very low abundance in the oocyte plasma membrane. We demonstrate for the first time that RhBG and RhCG—like RhAG—have significant CO₂ permeability, and we confirm that RhAG, RhBG and RhCG all have significant NH₃ permeability. However, as evidenced by $({\Updelta {\text{pH}}_{\text{S}}^{*} })_{{{\text{CO}}_{ 2} }} /({ - \Updelta {\text{pH}}_{\text{S}}^{*} })_{{{\text{NH}}_{ 3} }}$ values, we could not distinguish among the CO₂/NH₃ permeability ratios for RhAG, RhBG and RhCG. Finally, we propose a mechanism whereby RhBG and RhCG contribute to acid secretion in the CD by enhancing the transport of not only NH₃ but also CO₂ across the membranes of CD cells.     

Measurements of substrate oxidation using 13CO2-breath testing reveals shifts in fuel mix during starvation

Most fasting animals are believed to sequentially switch from predominantly utilizing one metabolic substrate to another from carbohydrates, to lipids, then to proteins. The timing of these physiological transitions has been estimated using measures of substrate oxidation including changes in respiratory exchange ratios, blood metabolites, nitrogen excretion, or enzyme activities in tissues. Here, we demonstrate how ¹³CO₂-breath testing can be used to partition among the oxidation of distinct nutrient pools in the body (i.e., carbohydrates, lipids, and proteins) that have become artificially enriched in ¹³C. Seventy-two Swiss Webster mice were raised to adulthood on diets supplemented with ¹³C-1-l-leucine, ¹³C-1-palmitic acid, ¹³C-1-d-glucose, or no tracer. Mice were then fasted for 72 h during which $ \dot{V}{\text{O}}_{2} $ , $ \dot{V}{\text{CO}}_{2} $ , δ¹³C of exhaled CO₂, body temperature, body mass, and blood metabolites (i.e., glucose, ketone bodies, and triacylglycerols) were measured. The fasting mice exhibited reductions in body mass (29 %), body temperature (3.3 °C), minimum observed metabolic rates (24 %), and respiratory exchange ratio (0.18), as well as significant changes in blood metabolites; but these responses were not particularly indicative of changes in oxidative fuel mixture. Measurements of endogenous nutrient oxidation by way of ¹³CO₂-breath testing revealed a decrease in the rate of oxidation of carbohydrates from 61 to 10 % of the total energy expenditure during the first 6 h without food. This response was mirrored by a coincidental increase in rate of endogenous lipid oxidation from 18 to 64 %. A transient peak in carbohydrate oxidation occurred between 8 and 14 h, presumably during increased glycogen mobilization. A well-defined period of protein sparing between 8 and 12 h was observed where endogenous protein oxidation accounted for as little as 8 % of the total energy expenditure. Thereafter, protein oxidation continually increased accounting for as much as 24 % of the total energy expenditure by 72 h. This study demonstrates that ¹³CO₂-breath testing may provide a complementary approach to characterizing the timing and magnitude of sequential changes in substrate oxidation that occur during prolonged fasting and starvation.     

Continuous cultivation of Lactobacillus brevis NCL912 for production of gamma-aminobutyric acid

A continuous cultivation method for Lactobacillus brevis NCL912 to synthesize gamma-aminobutyric acid was developed in this work. Different dilution rates were evaluated for obtaining steady state in continuous cultivation. The results showed that steady state could be achieved at dilution rates from 0.08 to 0.12 h^?1. The highest gamma-aminobutyric acid productivity (5.11 g L^?1?h^?1) was obtained at dilution rate of 0.09 h^?1. The kinetic models were established for continuous gamma-aminobutyric acid production by using the Monod equation for microbial growth, and the Luedeking–Piret equation for product formation. The microbial growth and product formation can be described by equations $ \mu = {{{0.1234{C_S}}} \left/ {{\left( {0.9338+{C_S}} \right)}} \right.} $ and $ {Q_P}=6.86\,\mathrm{g}\,{{\mathrm{g}}^{-1 }}\mathrm{cell}\,{{\mathrm{h}}^{-1 }} $ , respectively. The production of gamma-aminobutyric acid by L. brevis NCL912 was non-growth-associated.     

Estimation of viscosity from passage time of liquids flowing through a microchannel array

Recently, a microchannel flow analyzer (MC-FAN) has been used to study the flow properties of blood. However, the correlation between blood passage time measured by use of the MC-FAN and hemorheology has not been clarified. In this study, a simple model is proposed for estimation of liquid viscosity from the passage time t _p of liquids. The t _p data for physiological saline were well represented by the model. According to the model, the viscosity of Newtonian fluids was estimated reasonably well from the t _p data. For blood samples, although the viscosity $ \eta_{\text{mc}} $ estimated from t _p was shown to be smaller than the viscosity $ \eta_{{450{\text{s}}^{ - 1} }} $ measured by use of a rotatory viscometer at a shear rate of 450 s^?1, $ \eta_{\text{mc}} $ was correlated with $ \eta_{{450{\text{s}}^{ - 1} }} $ . An empirical equation for estimation of $ \eta_{{450{\text{s}}^{ - 1} }} $ from $ \eta_{\text{mc}} $ of blood samples is proposed.     

Release of non-exchangeable {}^{{{\text{15}}}}{\text{NH}}^{{\text{ + }}}_{{\text{4}}} from subgrade, decomposed granite substrates and uptake by non-mycorrhizal and mycorrhizal California native annual grass, Vulpia microstachys

Release rates of recently fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ from non-exchangeable interlayer sites in 2:1 silicate minerals were determined for decomposed granite (DG) saprolites from three locations in California, USA. Recently-fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ release from the DG substrate was quantified by extracting diffused $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ with H-resin, as well as a native, annual grass Vulpia microstachys. The $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ release data varied with via the method of extraction, which included H-resin pre-treatments (Na⁺ or H⁺) and V. microstachys uptake (mycorrhizal inoculated or uninoculated). After 6 weeks (1008 h), more $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ was recovered from fixed interlayer positions by the H-resins as compared to uptake by V. microstachys. The H⁺ treated H-resins recovered more released $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ (≈94 mg ${\text{NH}}^{{\text{ + }}}_{{\text{4}}} - {\text{N}}\;{\text{kg}}^{1} $ or (12%) of total fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ ) in two of the three DG samples as compared to the Na⁺ treated resins, (which recovered ≈70–78 mg ${\text{NH}}^{{\text{ + }}}_{{\text{4}}} - {\text{N}}\;{\text{kg}}^{{{\text{ - 1}}}} $ (or 9–10%) of the total fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ ). The V. microstachys assimilated 8–9% of the total fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ with mycorrhizal inoculum as compared to only 2% without a mycorrhizal inoculum, over the same time period. The fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ release kinetics from the H-resin experiments were most accurately described by first order and power function models, and can be characterized as biphasic using a heterogeneous diffusion model. Uptake of both the ¹⁵N and ambient, unlabelled N from the soils was closely related to plant biomass. There was no significant difference in percent of N per unit of biomass between the control and mycorrhizal treatments. The findings presented here indicate that observed, long-term $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ release rates from DG in studies utilizing resins, may overestimate the levels of fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ made available to plants and microorganisms. Additionally, the study suggested that mycorrhizae facilitate the acquisition and plant uptake of fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ , resulting in markedly increased plant biomass production.     

Stress fermentation strategies for the production of hyperthermostable superoxide dismutase from Thermus thermophilus HB27: effects of ions

In this study, we explored how ammonium and metal ion stresses affected the production of recombinant hyperthermostable manganese superoxide dismutase (Mn-SOD). To improve Mn-SOD production, fed-batch culture in shake flasks and bioreactor fermentation were undertaken to examine the effects of $ {\text{NH}}_{ 4}^{{^{ + } }} $ and Mn²⁺ feeding. Under the optimized feeding time and concentrations of $ {\text{NH}}_{ 4}^{{^{ + } }} $ and Mn²⁺, the maximal SOD activity obtained from bioreactor fermentation reached some 480 U/ml, over 4 times higher than that in batch cultivation (113 U/ml), indicating a major enhancement of the concentration of Mn-SOD in the scale-up of hyperthermostable Mn-SOD production. In contrast, when the fed-batch culture with appropriate $ {\text{NH}}_{ 4}^{{^{ + } }} $ and Mn²⁺ feeding was carried out in the same 5-L stirred tank bioreactor, a maximal SOD concentration of some 450 U/ml was obtained, again indicating substantial increase in SOD activity as a result of $ {\text{NH}}_{ 4}^{{^{ + } }} $ and Mn²⁺ feeding. The isoelectric point (pI) of the sample was found to be 6.2. It was highly stable at 90 °C and circular dichroism measurements indicated a high α-helical content of 70 % as well, consistent with known SOD properties. This study indicates that $ {\text{NH}}_{ 4}^{{^{ + } }} $ and Mn²⁺ play important roles in Mn-SOD expression. Stress fermentation strategies established in this study are useful for large-scale efficient production of hyperthermostable Mn-SOD and may also be valuable for the scale-up of other extremozymes.     

Characterization of a recombinant l-fucose isomerase from Caldicellulosiruptor saccharolyticus that isomerizes l-fucose, d-arabinose, d-altrose, and l-galactose

A recombinant l-fucose isomerase from Caldicellulosiruptor saccharolyticus was purified as a single 68 kDa band with an activity of 76 U mg^?1. The molecular mass of the native enzyme was 204 kDa as a trimer. The maximum activity for l-fucose isomerization was at pH 7 and 75°C in the presence of 1 mM Mn²⁺. Its half-life at 70°C was 6.1 h. For aldose substrates, the enzyme displayed activity in decreasing order for l-fucose, with a k _cat of 11,910 min^?1 and a K _m of 140 mM, d-arabinose, d-altrose, and l-galactose. These aldoses were converted to the ketoses l-fuculose, d-ribulose, d-psicose, and l-tagatose, respectively, with 24, 24, 85, 55% conversion yields after 3 h.     

Net ecosystem carbon exchange and the greenhouse gas balance of tidal marshes along an estuarine salinity gradient

Tidal wetlands are productive ecosystems with the capacity to sequester large amounts of carbon (C), but we know relatively little about the impact of climate change on wetland C cycling in lower salinity (oligohaline and tidal freshwater) coastal marshes. In this study we assessed plant production, C cycling and sequestration, and microbial organic matter mineralization at tidal freshwater, oligohaline, and salt marsh sites along the salinity gradient in the Delaware River Estuary over four years. We measured aboveground plant biomass, carbon dioxide (CO₂) and methane (CH₄) exchange between the marsh and atmosphere, microbial sulfate reduction and methanogenesis in marsh soils, soil biogeochemistry, and C sequestration with radiodating of soils. A simple model was constructed to estimate monthly and annually integrated rates of gross ecosystem production (GEP), ecosystem respiration (ER) to carbon dioxide ( \( {\text{ER}}_{{{\text{CO}}_{2} }} \) ) or methane ( \( {\text{ER}}_{{{\text{CH}}_{4} }} \) ), net ecosystem production (NEP), the contribution of sulfate reduction and methanogenesis to ER, and the greenhouse gas (GHG) source or sink status of the wetland for 2 years (2007 and 2008). All three marsh types were highly productive but evidenced different patterns of C sequestration and GHG source/sink status. The contribution of sulfate reduction to total ER increased along the salinity gradient from tidal freshwater to salt marsh. The Spartina alterniflora dominated salt marsh was a C sink as indicated by both NEP (~140 g C m^?2 year^?1) and ²¹⁰Pb radiodating (336 g C m^?2 year^?1), a minor sink for atmospheric CH₄, and a GHG sink (~620 g CO_2-eq m^?2 year^?1). The tidal freshwater marsh was a source of CH₄ to the atmosphere (~22 g C–CH₄ m^?2 year^?1). There were large interannual differences in plant production and therefore C and GHG source/sink status at the tidal freshwater marsh, though ²¹⁰Pb radiodating indicated modest C accretion (110 g C m^?2 year^?1). The oligohaline marsh site experienced seasonal saltwater intrusion in the late summer and fall (up to 10 mS cm^?1) and the Zizania aquatica monoculture at this site responded with sharp declines in biomass and GEP in late summer. Salinity intrusion was also linked to large effluxes of CH₄ at the oligohaline site (>80 g C–CH₄ m^?2 year^?1), making this site a significant GHG source (>2,000 g CO_2-eq m^?2 year^?1). The oligohaline site did not accumulate C over the 2 year study period, though ²¹⁰Pb dating indicated long term C accumulation (250 g C m^?2 year^?1), suggesting seasonal salt-water intrusion can significantly alter C cycling and GHG exchange dynamics in tidal marsh ecosystems.     

Oxygen removal from water versus arterial oxygen delivery: calibrating the Fick equation in Pacific salmon

While it is well known that O₂ is directly removed from the water by skin and gill tissues of fish, the mismatch between O₂ removal from water (O₂ uptake; \(\dot{V}{\text{O}}_{ 2}\) ) and the O₂ delivered to tissues by the primary circulation (O₂ consumption; \(\dot{V}{\text{aO}}_{ 2}\) ) has never been measured directly. Using data from four recent studies that simultaneously measured \(\dot{V}{\text{O}}_{ 2}\) and \(\dot{V}{\text{aO}}_{ 2}\) in 2–5 kg Pacific salmon, our analysis revealed that sockeye salmon can remove an additional 12–48 % more O₂ from the water than the primary circulation delivers to the systemic tissues. This percentage did not change significantly during swimming activity, a result that contradicts an earlier prediction that the difference should decrease when \(\dot{V}{\text{O}}_{ 2}\) increases during exercise. In resting Chinook salmon, a similar percentage difference in simultaneously measured \(\dot{V}{\text{O}}_{ 2}\) and \(\dot{V}{\text{O}}_{ 2}\) was observed, yet the difference tended to disappear during acute heat stress to a near lethal temperature. These results emphasize that caution should be exercised when using the Fick equation to estimate cardiac output because the overestimate of cardiac output that results from using the Fick equation in Pacific salmon is not small, may not be fixed and may exist in other teleosts.     

Mercury(II) complex formation with glutathione in alkaline aqueous solution

The structure and speciation of the complexes formed between mercury(II) ions and glutathione (GSH = L-glutamyl-L-cysteinyl-glycine) have been studied for a series of alkaline aqueous solutions (\( C_{{{\text{Hg}}^{{2 + }}}}\,{\sim18\,{\rm{mmol}}\,{\rm{{dm^{-3}}}}}\) and C _GSH = 40–200 mmol dm^?3 at pH ～10.5) by means of extended X-ray absorption fine structure (EXAFS) and ¹⁹⁹Hg NMR spectroscopy at ambient temperature. The dominant complexes are [Hg(GS)₂]^4? and [Hg(GS)₃]^7?, with mean Hg–S bond distances of 2.32(1) and 2.42(2) Å observed in digonal and trigonal Hg–S coordination, respectively. The proportions of the Hg²⁺–glutathione complexes were evaluated by fitting linear combinations of model EXAFS oscillations representing each species to the experimental EXAFS spectra. The [Hg(GS)₄]^10? complex, with four sulfur atoms coordinated at a mean Hg–S bond distance of 2.52(2) Å, is present in minor amounts (<30%) in solutions containing a large excess of glutathione (C _GSH ≥ 160 mmol dm^?3). Comparable alkaline mercury(II) cysteine (H₂Cys) solutions were also investigated and a reduced tendency to form higher complexes was observed, because the deprotonated amino group of Cys^2? allows the stable [Hg(S,N-Cys)₂]^2? chelate to form. The effect of temperature on the distribution of the Hg²⁺–glutathione complexes was studied by comparing the EXAFS spectra at ambient temperature and at 25 K of a series of glycerol/water (33/67, v/v) frozen glasses with \( C_{{{\text{Hg}}^{{2 + }} }} \,{\sim7\,{\rm{mmol}}\,{\rm{{dm^{-3}}}}} \) and C _GSH = 16–81 mmol dm^?3. Complexes with high Hg–S coordination numbers, [Hg(GS)₃]^7? and [Hg(GS)₄]^10?, became strongly favored when just a moderate excess of glutathione (C _GSH ≥28 mmol dm^?3) was used in the glassy samples, as expected for a stepwise exothermic bond formation. Addition of glycerol had no effect on the Hg(II)–glutathione speciation, as shown by the similarity of the EXAFS spectra obtained at room temperature for two parallel series of Hg(II)-glutathione solutions with \( C_{{{\text{Hg}}^{{2 + }} }} \,{\sim7\,{\rm{mmol}}\,{\rm{{dm^{-3}}}}},\) with and without 33% glycerol. Also, the ¹⁹⁹Hg NMR chemical shifts of a series of ～18 mmol dm^?3 mercury(II) glutathione solutions with 33% glycerol were not significantly different from those of the corresponding series in aqueous solution.