Vegetation control effects on untreated wood,crude cellulose and holocellulose δ<Superscript>13</Superscript>C of early and latewood in 3- to 5-year-old rings of Douglas-fir

The stable carbon (C) composition of tree rings expressed as δ¹³C, is a measure of intrinsic water-use efficiency and can indicate the occurrence of past water shortages for tree growth. We examined δ¹³C in 3- to 5-year-old rings of Douglas-fir (Pseudotsuga menziesii (Mirb) Franco) trees to elucidate if decreased water supply or uptake was a critical factor in the observed growth reduction of trees competing with understory herb and shrub vegetation compared to those growing without competition. We hypothesized that there would be no differences in δ¹³C of earlywood in trees growing in plots with competing vegetation and those in plots receiving complete vegetation control during 5 years because earlywood formed early in the growing season when soil water was ample. We also hypothesized that δ¹³C in latewood which was formed during the later half of the growing season when precipitation was low, would be greater (less negative) in trees in plots without vegetation control. We then separated early and latewood from rings for three consecutive years and analyzed their δ¹³C composition. No significant differences in earlywood δ¹³C in years 3–5 were observed for trees in the two vegetation control treatments. δ¹³C of untreated latewood separated from wood cores was greater in 4- and 5-year-old rings of trees growing with competing vegetation compared to trees growing without vegetation competition (i.e., −25.5 vs. −26.3‰ for year 4, and −26.1 vs. −26.8‰ for year 5). Results suggest that water shortages occurred in Douglas-fir trees on this coastal Washington site in the latewood-forming portion of the growing season of years 4 and 5 in the no-vegetation control treatment. We also compared δ¹³C from untreated wood, crude cellulose extracted with the Diglyme–HCl method, and holocellulose extracted with toluene–ethanol to see if the extraction method would increase the sensitivity of the analysis. δ¹³C values from the two extraction methods were highly correlated with those from untreated samples (r ² = 0.97, 0.98, respectively). Therefore, using untreated wood would be as effective as using crude cellulose or holocellulose to investigate δ¹³C patterns in young Douglas-fir.     

Novel 2,2'-[1,2-ethanediylbis(nitriloethylidyne)]-bis-hydroquinone double-wall carbon nanotube paste electrode for simultaneous determination of epinephrine, uric acid and folic acid

The electro-oxidation of epinephrine (EP), uric acid (UA), folic acid (FA), and their mixture has been studied by modified carbon nanotube paste electrode of 2,2'-[1,2-ethanediylbis(nitriloethylidyne)]-bis-hydroquinone using cyclic voltammetry, chronoamperometry and differential pulse voltammetry. This modified electrode exhibited potent and persistent electron mediating behavior followed by well-separated oxidation peaks towards EP, UA and FA with activation overpotential. For the ternary mixture containing EP, UA and FA the three compounds can be well separated from each other at the scan rate of 20mVs(-1). The obtained catalytic peak current, was linearly dependent on the EP, UA and FA concentrations in the range of 0.7-1200muM, 25-750muM and 15-800muM and the detection limits for EP, UA and FA were 0.216+/-0.004, 8.8+/-0.2 and 11.0+/-0.3muM, respectively. The diffusion coefficient (D), and the kinetic parameters such as electron transfer coefficient, (alpha) and heterogeneous rate constant, (k') for EP were also determined using electrochemical approaches. The modified electrode showed good sensitivity, selectivity and stability, and was employed for the determination of EP, UA and FA in the real samples.     

A unifying theory for methods of systematic analysis

A unifying theory for systematic analysis states that a number of methods should be used jointly to cope with various kinds of data; also that groups should be as consistent as possible, be made with least information loss, and where needed, be polythetic. A test of relationship, homogeneity, can use various kinds of data. It can take account of the internal variation of aggregate items such as genera. It can give due emphasis to smaller clusters that have likely important contexts of external items. It helps in analysing trends, cores and hazes in dendrograms. A proposed detector for formal groups can be based on measures of isolation, identifiability and inclusiveness. Non-mathematical, inter-item reaction tests such as hybridization and serology can also be used in grouping. All relationship data are used polythetically to reveal natural groups. This leads to a unified informational concept for taxa. This is more useful than the biological species concept that is restricted to inter-breeding data. All the methods appear to be analogues of the powerful human grouping instinct. The resulting compatibility is important as precise methods are needed mainly when the data are too complex for the mind to use reliably. Cladograms can be made by self-graded deweighting of homogeneity and agglomerative clustering. Unlike classical cladistics this can reveal any polythetic group. Finding the derived states for making cladograms is often much too hypothetical for a fully cladistic approach to be properly precise. Instead, where the evidence is weak, a milder strength of graded deweighting is used for the cladistic properties, which help to show relationships along with the others. Axiomatic failures of other classes of grouping methods are discussed. Unavoidable remnants of instinctive processing lower the precision of all the methods. The Uniter computer program, based on the theory, is tested with finely graded values of artificially ‘evolved’ items and with coarsely coded cladistic data. The results show that with natural data, the program should act as a fairly sensitive probe of past evolutionary branching. Another test shows how specimens from species complexes can be grouped and how distinctions between groups are analysed.     

Efficient isotopic tryptophan labeling of membrane proteins by an indole controlled process conduct

A protocol for the efficient isotopic labeling of large G protein‐coupled receptors with tryptophan in Escherichia coli as expression host was developed that sufficiently suppressed the naturally occurring L‐tryptophan indole lyase, which cleaves tryptophan into indole, pyruvate, and ammonia resulting in scrambling of the isotopic label in the protein. Indole produced by the tryptophanase is naturally used as messenger for cell–cell communication. Detailed analysis of different process conducts led to the optimal expression strategy, which mimicked cell–cell communication by the addition of indole during expression. Discrete concentrations of indole and ¹⁵N₂‐L‐tryptophan at dedicated time points in the fermentation drastically increased the isotopic labeling efficiency. Isotope scrambling was only observed in glutamine, asparagine, and arginine side chains but not in the backbone. This strategy allows producing specifically tryptophan labeled membrane proteins at high concentrations avoiding the disadvantages of the often low yields of auxotrophic E. coli strains. In the fermentation process carried out according to this protocol, we produced ～15 mg of tryptophan labeled neuropeptide Y receptor type 2 per liter medium. Biotechnol. Bioeng. 2013; 110: 1681–1690. © 2013 Wiley Periodicals, Inc.     

Using stable isotopes to investigate the dietary trophic level of fledgling songbirds

ABSTRACT Use of early successional habitat by mature forest birds during the postfledging period is well documented, but reasons for this habitat shift remain elusive. Although forest‐breeding songbirds are primarily insectivorous during the nestling and early fledgling periods due to high protein requirements, older fledglings may adopt a heavily frugivorous diet. Our objectives were to use stable isotopes to examine the dietary trophic level of juveniles of three species of mature forest songbirds to determine if juvenile songbirds heavily consume fruit resources during the postfledging period and to evaluate a possible link between diet and energetic condition. We collected the outer right rectrix and several body feathers from 34 Wood Thrushes (Hylocichla mustelina), 34 Ovenbirds (Seiurus aurocapilla), and 35 Scarlet Tanagers (Piranga olivacea) captured in regenerating clearcuts in southeastern Ohio in 2005 and 2006. We also collected fruit and arthropod samples from each clearcut. Isotopic values of body feathers were significantly higher (more enriched) than those of rectrices in all cases except values of δ¹³C for Ovenbirds where we found no difference between body and rectrix feathers. These results suggest that juvenile songbirds did not undergo a strong shift to frugivory during the postfledging period, and arthropods were the primary source of protein during the period when rectrix and body feathers were growing. In addition, the energetic condition of birds was not related to the isotopic signature of feathers. Although our results are inconsistent with the hypothesis that juveniles move into regenerating clearcuts enabling them to shift to a primarily frugivorous diet during the postfledging period, they may consume fruit for nonprotein requirements, such as lipids and carbohydrates.     

Regulating effects of climate,net primary productivity,and nitrogen on carbon sequestration rates in temperate wetlands,Northeast China

Temperate wetlands in the Northern Hemisphere have high long-term carbon sequestration rates, and play critical roles in mitigating regional and global atmospheric CO₂ increases at the century timescale. We measured soil organic carbon (SOC), total nitrogen (TN), and total phosphorus (TP) from 11 typical freshwater wetlands (Heilongjiang Province) and one saline wetland (Jilin Province) in Northeast China, and estimated carbon sequestration rates using ²¹⁰Pb and ¹³⁷Cs dating technology. Effects of climate, net primary productivity, and nutrient availability on carbon sequestration rates (R_carbon) were also evaluated. Chronological results showed that surface soil within the 0–40 cm depth formed during the past 70–205 years. Soil accretion rates ranged from 2.20 to 5.83 mm yr⁻¹, with an average of 3.84 ± 1.25 mm yr⁻¹ (mean ± SD). R_carbon ranged from 61.60 to 318.5 gC m⁻² yr⁻¹ and was significantly different among wetland types. Average R_carbon was 202.7 gC m⁻² yr⁻¹ in the freshwater wetlands and 61.6 gC m⁻² yr⁻¹ in the saline marsh. About 1.04 × 10⁸ tons of carbon was estimated to be captured by temperate wetland soils annually in Heilongjiang Province (in the scope of 45.381–51.085°N, 125.132–132.324°E). Correlation analysis showed little impact of net primary productivity (NPP) and soil nutrient contents on R_carbon, whereas climate, specifically the combined dynamics of temperature and precipitation, was the predominant factor affecting R_carbon. The negative relationship observed between R_carbon and annual mean temperature (T) indicates that warming in Northeast China could reduce R_carbon. Significant positive relationships were observed between annual precipitation (P), the hydrothermal coefficient (defined as P/AT, where AT was accumulative temperature ≥10 °C), and R_carbon, indicating that a cold, humid climate would enhance R_carbon. Current climate change in Northeast China, characterized by warming and drought, may form positive feedbacks with R_carbon in temperate wetlands and accelerate carbon loss from wetland soils.     

A comparison of soil carbon pools and profiles in wetlands in Costa Rica and Ohio

Wetlands are large carbon pools and play important roles in global carbon cycles as natural carbon sinks. This study analyzes the variation of total soil carbon with depth in two temperate (Ohio) and three tropical (humid and dry) wetlands in Costa Rica and compares their total soil C pool to determine C accumulation in wetland soils. The temperate wetlands had significantly greater (P < 0.01) C pools (17.6 kg C m⁻²) than did the wetlands located in tropical climates (9.7 kg C m⁻²) in the top 24 cm of soil. Carbon profiles showed a rapid decrease of concentrations with soil depth in the tropical sites, whereas in the temperate wetlands they tended to increase with depth, up to a maximum at 18–24 cm, after which they started decreasing. The two wetlands in Ohio had about ten times the mean total C concentration of adjacent upland soils (e.g., 161 g C kg⁻¹ were measured in a central Ohio isolated forested wetland, and 17 g C kg⁻¹ in an adjacent upland site), and their soil C pools were significantly higher (P < 0.01). Among the five wetland study sites, three main wetland types were identified – isolated forested, riverine flow-through, and slow-flow slough. In the top 24 cm of soil, isolated forested wetlands had the greatest pool (10.8 kg C m⁻²), significantly higher (P < 0.05) than the other two types (7.9 kg C m⁻² in the riverine flow-though wetlands and 8.0 kg C m⁻² in a slowly flowing slough), indicating that the type of organic matter entering into the system and the type of wetland may be key factors in defining its soil C pool. A riverine flow-through wetland in Ohio showed a significantly higher C pool (P < 0.05) in the permanently flooded location (18.5 kg C m⁻²) than in the edge location with fluctuating hydrology, where the soil is intermittently flooded (14.6 kg C m⁻²).     

Protein instability during HIC: describing the effects of mobile phase conditions on instability and chromatographic retention

Hydrophobic interaction chromatography (HIC) is known to be potentially denaturing to proteins, but the effects of mobile phase conditions on chromatographic behavior are not well understood. In this study, we apply a model describing the effects of secondary protein unfolding equilibrium on chromatographic behavior, including the effects of salt concentration on both stability and adsorption. We use alpha-lactalbumin as a model protein that in the presence and absence of calcium, allows evaluation of adsorption parameters for folded and unfolded species independently. The HIC adsorption equilibrium under linear binding conditions and solution phase protein stability have been obtained from a combination of literature and new experiments. The effect of salt concentration on protein stability and the rate constant for unfolding on the chromatographic surface have been determined by fitting the model to isocratic chromatography data under marginally stable conditions. The model successfully describes the effects of added calcium and ammonium sulfate. The results demonstrate the importance of considering the effects on stability of mobile phase modifiers when applying HIC to marginally stable     

Water management practices and climate in ancient agriculture: inferences from the stable isotope composition of archaeobotanical remains

Carbon isotope discrimination (Δ¹³C) in charred grains from archaeological sites provides reliable information about water availability of ancient crops. However, as cereals are cultivated plants, they may reflect not only climatic fluctuations, but also the effect on water status of certain agronomic practices, such as sowing in naturally wet soils or irrigation. In this work, we propose a methodological approach to combine Δ¹³C data from different plant species, in order to discriminate between climate-derived and anthropogenic effects on ancient crops. We updated previous models for estimating water inputs from Δ¹³C of cereal grains of Hordeum vulgare and Triticum aestivum/durum, and we applied them to published data from several archaeological sites, including samples from the Neolithic to the present day in northeast and southeast Spain, as well as from the Neolithic site of Tell Halula (northwest Syria). We found an important decrease in water availability from the Neolithic to the present time in the three areas of study, especially clear for the two driest areas (southeast Spain and northwest Syria). Potential differences in water management practices between wheat and barley, as well as between cereal and legume crops (Vicia faba and Lens culinaris), are also discussed on the basis of the comparison of Δ¹³C values across several archaeological sites.     

Association between tree-ring and needle δ<Superscript>13</Superscript>C and leaf gas exchange in <Emphasis Type="Italic">Pinus halepensis</Emphasis> under semi-arid conditions

Associations between δ¹³C values and leaf gas exchanges and tree-ring or needle growth, used in ecophysiological compositions, can be complex depending on the relative timing of CO₂ uptake and subsequent redistribution and allocation of carbon to needle and stem components. For palaeoenvironmental and dendroecological studies it is often interpreted in terms of a simple model of δ¹³C fractionation in C₃ plants. However, in spite of potential complicating factors, few studies have actually examined these relationships in mature trees over inter- and intra-annual time-scales. Here, we present results from a 4 years study that investigated the links between variations in leaf gas-exchange properties, growth, and dated δ¹³C values along the needles and across tree rings of Aleppo pine trees growing in a semi-arid region under natural conditions or with supplemental summer irrigation. Sub-sections of tissue across annual rings and along needles, for which time of formation was resolved from growth rate analyses, showed rapid growth and δ¹³C responses to changing environmental conditions. Seasonal cycles of growth and δ¹³C (up to ~4‰) significantly correlated (P<0.01) with photosynthetically active radiation, vapour pressure deficit, air temperature, and soil water content. The irrigation significantly increased leaf net assimilation, stomatal conductance and needle and tree-ring growth rate, and markedly decreased needle and tree-ring δ¹³C values and its sensitivity to environmental parameters. The δ¹³C estimates derived from gas-exchange parameters, and weighted by assimilation, compared closely with seasonal and inter-annual δ¹³C values of needle- and tree-ring tissue. Higher stomatal conductances of the irrigated trees (0.22 vs. 0.08 mol m⁻² s⁻¹ on average) corresponded with ~2.0‰ lower average δ¹³C values, both measured and derived. Derived and measured δ¹³C values also indicated that needle growth, which occurs throughout the stressful summer was supported by carbon from concurrent, low rate assimilation. For Aleppo pine under semi-arid and irrigated conditions, the δ¹³C of tree-ring and needle material proved, in general, to be a reasonable indicator of integrated leaf gas-exchange properties.