Change in turnover capacity of crude recombinant dye-decolorizing peroxidase (rDyP) in batch and fed-batch decolorization of Remazol Brilliant Blue R

Decolorization of the representative anthraquinone dye, Remazol Brilliant Blue R (RBBR) was assessed to determine the practical potential of crude recombinant dye-decolorizing peroxidase generated by Aspergillus oryzae (rDyP) in term of turnover capacity of rDyP. The turnover capacity, defined as the milligram of RBBR decolorized per unit of rDyP inactivated over the catalytic life time of rDyP, was quantified under condition by H₂O₂ -mediated rDyP inactivation. In batch culture, equimolar batch addition of H₂O₂ and RBBR yielded complete decolorization of RBBR by rDyP, with a turnover capacity of 4.75. In stepwise fed-batch addition of H₂O₂, the turnover capacity increased to 5.76, and by increasing dye concentration, it reached 14.3. When H₂O₂ was added in continuous fed-batch to minimize rDyP inactivation and 1.6 mM dye was added in stepwise fed-batch mode, the turnover capacity increased to 20.4. At this turnover capacity, 1 l of crude rDyP solution containing 5,000 U could decolorize up to 102 g RBBR in 650 min.     

Influence of day-night and tidal cycles on phenol content and antioxidant capacity in three temperate intertidal brown seaweeds

The daily variations of phenol contents and antioxidant capacities were surveyed in a 32-h field experiment in three temperate brown seaweeds belonging to Fucales - Pelvetia canaliculata, Ascophyllum nodosum and Bifurcaria bifurcata - living at different intertidal levels — high-, mid- and low-tide level, respectively. Phenolic compounds of brown seaweeds are secondary metabolites involved in many different protection mechanisms, as for example against grazer and pathogen attack as well as UV damage. This study was thus aimed at understanding the influence of both day/night and tidal cycles on the brown seaweed phenol pool with respect to their bathymetric level on the shore. These cycles affect the quantity and quality of light received by intertidal seaweeds (protection via the water layer during immersion), and the photoprotective role of phlorotannins was thus evaluated. Phenol levels and antioxidant capacities were monitored every hour during a tidal cycle and a half at the equinox spring tide, in March 2003. The three species contained rather high phenol levels, i.e. about 3, 6 and 4% DW in P. canaliculata, A. nodosum and B. bifurcata, respectively. Antioxidant capacities globally paralleled phenol contents in the three species under study. Moreover, the measured antioxidant capacities and the phenol contents of the extracts were significantly and positively correlated in the three species. Significant effect of neither cycles nor their interaction was observed in any species despite a trend to follow day/night alternation for P. canaliculata, and emersion/immersion cycle for A. nodosum. No trend was observed for the third species. However, significant correlations between phenol levels of P. canaliculata and A. nodosum and measured air-temperature were found. Our results suggested an effect of aerial, aquatic and both conditions on the phenolic pool of P. canaliculata, B. bifurcata and A. nodosum, respectively, and a fast evolution of the phenolic pool on a day time scale.     

Energetic Carrying Capacity of Actively and Passively Managed Wetlands for Migrating Ducks in Ohio

ABSTRACT Habitat conservation strategies of the North American Waterfowl Management Plan (NAWMP) are guided by current understanding of factors that limit growth of waterfowl populations. The 1998 implementation plan of the Upper Mississippi River and Great Lakes Region Joint Venture (UMR and GLRJV) assumed that availability of foraging resources during autumn in wetlands actively managed for waterfowl was the primary limiting factor for duck populations during the nonbreeding season. We used multistage sampling during autumn and spring 2001–2004 to estimate energetic carrying capacity (ECC) of actively and passively managed wetlands in Ohio, USA, and examine this assumption. Energetic carrying capacity during autumn was similar between actively and passively managed wetlands each year. Averaged across years, energetic carrying capacity was 3,446 and 2,047 duck energy-days (DED)/ha for actively and passively managed wetlands, respectively. These estimates exceeded the UMR and GLRJV assumption that 1,236 DED/ha were provided by managed wetland habitats. Energetic carrying capacity declined each year by >80% between autumn and spring migration. Consequently, ECC of actively and passively managed wetlands was low during spring ( = 66–242 DED/ha). These results suggested that duck foraging resources in actively and passively managed wetland habitats are abundant during autumn, but overwinter declines may create food-limiting environments during spring.     

Unfolding energetics and conformational stability of DLC8 monomer

To understand the rules governing the protein folding process it is essential to study the stability and unfolding of small monomeric proteins. Here, I present the pH dependent thermal unfolding energetics and conformational stability analysis of monomeric Dynein light chain protein (DLC8) in the pH range 3.5-2.0. DLC8 is the smallest and the most conserved light chain among the light chains of the dynein motor assembly. Thermal unfolding of DLC8 monomer is much complex with the presence of transient intermediates, which is in contrast to the notion that small proteins unfold via simple two-state process. The unfolding seems to be more cooperative at lower pH and the temperature of highest conformational stability (T(s)) is found to be maximum (295.7 K) at pH 2.76. Stability curves have been simulated to understand the thermodynamic parameters that govern the shapes of the experimentally obtained curves. Further, an effort has been made to correlate the observed differences in the denaturation energetics with the protein sequence in order to throw light on the structure-folding paradigm of the DLC8 monomer.     

The effect of muscularity on thermal responses,muscle performance,and dexterity during whole body exposure to 10 °C

1.

The study evaluated the effects of exposure to cold air (10 °C) on thermal responses, muscle performance and dexterity of muscular subjects and their matched lean counterparts.     

A unique and highly efficient non-viral DNA/siRNA delivery system based on PEI-bisepoxide nanoparticles

Delivery of DNA and siRNA into mammalian cells is a powerful technique in treating various diseases caused by single gene defects. Herein, we report a highly efficient delivery system using 1,4-butanediol diglycidyl ether (bisepoxide) crosslinked polyethylenimine (PEI) nanoparticles (PN). The nanoparticle/DNA complexes (nanoplexes) exibited approximately 2.5- to 5.0-fold gene transfer efficacy and decreased cytotoxicity in cultured cell lines, compared to the native PEI (25 kDa) (gold standard) and commercially available transfection agents such as Lipofectamine 2000 and Fugene. The bisepoxide crosslinking results in change in amine ratio in PEI; however, it retains the net charge on PN unaltered. A series of nanoparticles obtained by varying the degree of crosslinking was found to be in the size range of 69-77 nm and the zeta potential varying from +35 to 40 mV. The proposed system was also found to deliver siRNA efficiently into HEK cells, resulting in approximately 70% suppression of the targetted gene (GFP).     

BPPred: a Web-based computational tool for predicting biophysical parameters of proteins

We exploit the availability of recent experimental data on a variety of proteins to develop a Web-based prediction algorithm (BPPred) to calculate several biophysical parameters commonly used to describe the folding process. These parameters include the equilibrium m-values, the length of proteins, and the changes upon unfolding in the solvent-accessible surface area, in the heat capacity, and in the radius of gyration. We also show that the knowledge of any one of these quantities allows an estimate of the others to be obtained, and describe the confidence limits with which these estimations can be made. Furthermore, we discuss how the kinetic m-values, or the Beta Tanford values, may provide an estimate of the solvent-accessible surface area and the radius of gyration of the transition state for protein folding. Taken together, these results suggest that BPPred should represent a valuable tool for interpreting experimental measurements, as well as the results of molecular dynamics simulations.     

A simulation model for nitrogen retention in a papyrus wetland near Lake Victoria,Uganda (East Africa)

Papyrus wetlands around Lake Victoria, East Africa play an important role in the nutrient flows from the catchment to the lake. A dynamic model for nitrogen cycling was constructed to understand the processes contributing to nitrogen retention in the wetland and to evaluate the effects of papyrus harvesting on the nitrogen absorption capacity of the wetlands. The model had four layers: papyrus mat, water, sludge and sediment. Papyrus growth was modelled as the difference between nitrogen uptake and loss. Nitrogen uptake was modelled with a logistic equation combined with a Monod-type nitrogen limitation. Nitrogen compartments were papyrus plants, organic material in the floating mat; and total ammonia, nitrate and organic nitrogen in the water, sludge and sediment. Apart from the uptake and decay rates of the papyrus, the model included sloughing and settling of mat material into the water, mineralization of organic matter, and nitrification and diffusion of dissolved inorganic nitrogen. Literature data and field measurements were used for parameterization. The model was calibrated with data from Kirinya wetland in Jinja, Uganda which receives effluent from a municipal wastewater treatment plant. The model simulated realistic concentrations of dissolved nitrogen with a stable biomass density of papyrus and predicted accumulation of organic sludge in the wetland. Assuming that this sludge is not washed out of the wetland, the overall nitrogen retention of the wetland over a three-year period was 21.5 g N m⁻² year⁻¹ or about 25% of input. Harvesting 10, 20 and 30% of the papyrus biomass per year increased nitrogen retention capacity of the wetland to 32.3, 36.8 and 38.1 g m⁻² year⁻¹, respectively. Although the nutrient flows estimated by the model are within the ranges found in other papyrus wetlands, the model could be improved with regard to the dynamics of detrital nitrogen. Actual net retention of nitrogen in the sludge is likely to be lower than 21.5 g N m⁻² year⁻¹ because of flushing out of the sludge to the lake during the rainy season.     

The Oxidative Cost of Acoustic Signals: Examining Steroid Versus Aerobic Activity Hypotheses in a Wild Bird

Vertebrate vocalizations are widespread secondary sexual signals used for mate attraction and territory defence, and variation in signal quality is often condition dependent and impacts reproductive outcomes. Although vocal signal performance is known to reflect various aspects of male quality, few studies have examined the underlying mechanisms mediating its costs and hence its honesty. Using a population of Arctic‐breeding snow buntings (Plectrophenax nivalis), we compared the ‘Oxidation Handicap Hypothesis’, which predicts that testosterone‐induced increases in oxidative stress provide a direct mechanistic basis for ensuring the honesty of many secondary sexual signals, to the ‘Aerobic Activity Hypothesis, which predicts that it is the aerobic activity involved with signal production (i.e. vocal performance or defending a large territory) and not testosterone directly that links signal quality and oxidative stress. Males singing at faster rates had higher levels of both reactive oxygen metabolites and non‐enzymatic antioxidant capacity in the plasma (i.e. without an increase in overall oxidative stress), enabling certain males to produce high‐quality signals while also mitigating the costs of an associated increase in oxidative stress. However, these results were completely independent of plasma testosterone levels, supporting the role of aerobic performance in directly affecting oxidative stress. Although song performance was not linked to reproductive parameters in our data set, our research is the first to test these competing hypotheses in a behavioural trait and results suggest that oxidative stress may be an underlying physiological cost preventing low‐quality individuals from producing high‐quality signals.     

Relationship of aerobic and anaerobic parameters with 400 m front crawl swimming performance

The aims of the present study were to investigate the relationship of aerobic and anaerobic parameters with 400 m performance, and establish which variable better explains long distance performance in swimming. Twenty-two swimmers (19.1±1.5 years, height 173.9±10.0 cm, body mass 71.2±10.2 kg; 76.6±5.3% of 400 m world record) underwent a lactate minimum test to determine lactate minimum speed (LMS) (i.e., aerobic capacity index). Moreover, the swimmers performed a 400 m maximal effort to determine mean speed (S400m), peak oxygen uptake (V.O2PEAK) and total anaerobic contribution (C_ANA). The C_ANA was assumed as the sum of alactic and lactic contributions. Physiological parameters of 400 m were determined using the backward extrapolation technique (V.O2PEAK and alactic contributions of C_ANA) and blood lactate concentration analysis (lactic anaerobic contributions of C_ANA). The Pearson correlation test and backward multiple regression analysis were used to verify the possible correlations between the physiological indices (predictor factors) and S400m (independent variable) (p < 0.05). Values are presented as mean ± standard deviation. Significant correlations were observed between S400m (1.4±0.1 m·s^-1) and LMS (1.3±0.1 m·s^-1; r = 0.80), V.O2PEAK (4.5±3.9 L·min^-1; r = 0.72) and C_ANA (4.7±1.5 L·O₂; r= 0.44). The best model constructed using multiple regression analysis demonstrated that LMS and V.O2PEAK explained 85% of the 400 m performance variance. When backward multiple regression analysis was performed, C_ANA lost significance. Thus, the results demonstrated that both aerobic parameters (capacity and power) can be used to predict 400 m swimming performance.