Glucose oxidase immobilized electrode for potentiometric estimation of glucose

Glucose oxidase has been immobilized onto a thin platinum strip, by co-crosslinking with bovine serum albumin and glutaraldehyde. The retention of redox characteristics of glucose oxidase has been verified by cyclic voltammetry. The activity of the immobilized enzyme reduces to a quarter of its value when the enzyme is in solution but improves when coimmobilized with 1 urea. The potentiometric response builds up and remains stable after 100 s. It is sensitive to the thickness of the immobilizing matrix, pH and temperature. An improvement in the performance of the electrode has been achieved by coimmobilizing 2 urea and metal ions such as Mg²⁺ and Mn²⁺. The presence of Cu has been proved to be detrimental. The electrode has been calibrated in the 0.1–5.0 mM glucose concentration range. It gives a stable response for more than 50 independent assays and can be stored for 60 days without significant loss of function.     

Whole-lake experimentation as a tool to assess ecosystem health,response to stress and recovery: the Experimental Lakes Area experience

The rationale and methods for and value of whole-lake experimentation are described using the Experimental Lakes Area (ELA), northwestern Ontario, as the example. The ELA consists of 46 lakes (< 100 ha in surface area), their watersheds, and several streams protected for research purposes in near-pristine boreal forest on the Precambrian Shield near Kenora, Ontario. Over more than 20 y, whole-lake experimentation has provided unique information on the effects on lakes of nutrient additions, acidification, Cd addition, and biomanipulation. Experiments are planned to study the effects of PCB addition and flooding. Recovery, mitigation, and remediation have been explored in some experiments. As well, the fate of radioactive metals in a lake and the effects of acidification on a poor fen and an upland watershed are studied. Comparison between the experimental systems and unmanipulated reference systems has proven to be essential. These reference systems also have a role in defining (absolute) aquatic ecosystem health for small, pristine Precambrian Shield lakes. The ELA experimental data base is available, as well, for calibrating indices of relative aquatic ecosystem health, i.e., environmental degradation, using the dose-responses of lakes to eutrophication, acidification, Cd addition, and other stressors.     

The pH-independent effect of aluminum on cultures of phytoplankton from an acidic Wisconsin lake

Negative correlations between aluminum and planktonic algal abundance have been reported in acidic lakes. Natural assemblages of phytoplankton from a low-pH, low-Al lake (Franklin Lake, WI) were grown in semi-continuous cultures consisting of four treatments at pH 5.7 with 0.0, 50, 100, and 200 μg Al L⁻¹ and one treatment at pH 4.7 with no Al added. Asterionella ralfsii var. americana (a common diatom plankter in acidic lakes) grew well at both pH 4.7 and 5.7 when no Al was added but declined in all other treatments and so may be useful as an indicator of acidic, low monomeric-Al conditions. Other common plankters that showed this pattern included: Arthrodesmus indentatus, Ar. octocornus, Ar. quiriferus, Staurastrum arachne var. curvatum, S. longipes var. contractum, and S. pentacerum. Common taxa showing no toxic effects of Al were Dinobryon bavaricum, Peridinium limbatum, Stenokalyx monilifera, Elaktothrix sp. and Oedogonium sp. We hypothesize that metal toxicity as a pulse at spring snowmelt could dramatically change algal succession in moderately acidic lakes. The experimental results agreed well with field observations. These types of experiments are useful for predicting the responses of natural phytoplankton communities to increases in Al concentration.     

Localised uptake of63nickel into dinoflagellate chromosomes: An autoradiographic study

Summary The uptake of⁶³Ni into cells of the binucleate dinoflagellateGlenodinium foliaceum was investigated using insoluble compound light and electron microscope autoradiography. Cells labelled over a period of 2 hours showed active uptake throughout the whole population, with an increase in mean cell grain count when the labelling period was extended to 4 hours and 24 hours. The mean grain count did not vary with type of fixation (glutaraldehyde, paraformaldehyde or acetic alcohol) suggesting that retention of⁶³ Ni is not a specific fixation-binding artefact. At light microscope level, silver grains were not localised to any major cell component, but with the greater resolution of electron microscope autoradiography, a high degree of localisation was demonstrated in the typical dinoflagellate (dinocaryotic) nucleus-which contained about 83% of the cell label (cytoplasm 16%, supernumerary nucleus 1%). Silver grain distribution within the dinocaryotic nucleus was consistent with some degree of localisation to the condensed chromatin.The autoradiographic results corroborate previous X-ray microanalytical data which demonstrated high levels of transition metals in dinoflagellate nuclei. The distinction between the two types of nucleus inGlenodinium is further emphasised, giving additional support to the concept of a separate phyllogenetic origin of the supernumerary nucleus.     

ZINC ADSORPTION AND TRANSPORT BY CHLAMYDOMONAS VARUIABILIS AND SCENEDESMUS SUBSPICATUS (CHLOROPHYCEAE) GROWN IN SEMICONTINUOUS CULTURE1

The amount of zinc adsorbed onto the cell surface of the unicellular green algae Scenedesmus subspicatus Hodat and Chlamydomonas variabilis Dangeard was operationally defined by extraction with EDTA; it was a function of the concentration of free ionic zinc remaining in the growth medium, rather than that of the total (free plus complexed) zinc concentration, and could be described by Langmuir isotherms. Conditional adsorption equilibrium constants for zinc were 0.123 and 0.039 L ·μmol^?1 for S. subspicatus and C. variabilis, respectively. A portion of the zinc adsorbed onto C. variabilis was released into solution after 1 h of contact with the metal, providing a possible tolerance mechanism for this alga; the division rate of C. variabilis was not altered by up to 12 μmol Zn²⁺· L^?1, although the cell yield obtained during the stationary phase was significantly decreased. The amount of transported or cellular zinc, for both algal species, was operationally defined as the zinc remaining with the cell after EDTA-extraction; it was a linear function of the free ionic zinc concentration remaining in solution, suggesting that the zinc transported into the cell was not derived from the total adsorbed fraction, although the latter may contain some zinc originating from specific sites leading to zinc transport.     

Nickel requirement for chemolithotrophic growth in hydrogen-oxidizing bacteria

In a comparative study the requirement of several strains of autotrophic hydrogen-oxidizing bacteria for nickel was examined. Autotrophic growth was studied both in liquid media, previously freed from trace metals; and on solidified media, using a plate diffusion assay. The latter assay was based on the observation that EDTA causes complete inhibition of autotrophic growth on agar medium as a result of nickel deficiency. Nickel was shown to be required as a trace element in five strains of Alcaligenes eutrophus, in two strains of Xanthobacter autotrophicus, in Pseudomonas flava, in Arthrobacter spec. 11X and in strain 12X. In these bacteria nickel was not replaceable by cobalt, copper, manganese or zinc ions. No significant nickel requirement was detected by these methods, however, for Paracoccus denitrificans and Nocardia opaca 1b.     

Heavy metal composition of polysomal fractions following cadmium challenge

Endogeneous levels of zinc and copper were found to be 1.2±0.1×10⁻² and 0.3±0.1×10⁻² μg/A260 unit, respectively, in polysomal fractions from control animals; cadmium, however, was undetectable. In experimental animals (injected with cadmium) zinc, copper, and cadmium were found in polysomal fractions isolated by two different methods. One hour after a cadmium injection there was a rise in both the zinc and copper content of the polysomal fractions, which then declined steadily to below control levels by 16 h. Neither zinc nor cadmium were dialyzable from these fractions by a TRIS buffer; however, addition of 0.01M EDTA to the buffer resulted in removal of 75% of the zinc and all of the detectable cadmium. The addition of cadmium (CdCl₂) to control supernatants (adjusted to the cadmium concentration present in supernatants 6 h after in vivo exposure) resulted in metal binding to polysomal fractions in levels comparable to those observed after in vivo exposures to the metal. When cadmium was added in the form of cadmium thionein, a smaller fraction of the metal was isolated with the polysomal fraction. Cadmium bound to polysomal fractions in vivo (24 h after exposure) was sensitive to release by protease digestion, but insensitive to release by ribonuclease digestion.     

Hill coefficient-based stochastic switch-like signal directly governs damage-recovery dynamics in freshwater fish in response to pulse copper

Growing evidence demonstrates that fluctuating metal stressors can have profound impact on the ecophysiological responses in aquatic species. However, how environmental stochasticity affects the complex damage-recovery dynamics in organisms remains difficult to predict. The objective of this paper was to investigate the stochastic behavior in the damage-recovery dynamics in tilapia in response to pulse waterborne copper (Cu). We developed a mathematical framework that allows discrimination between damage and recovery processes in tilapia exposed to designed pulse Cu scenarios. We built deterministic nonlinear models for the damage-recovery dynamics that produce response surfaces describing killing/recovery rate–Cu-pulse interval interactions. Here we showed that the stochastic switching behavior arose from competition among killing, recovery rates, and Cu pulse frequency. This competition resulted in an ultrasensitivity appeared in whole body, gills, muscle, liver, and kidney with Hill coefficients of ≥7, 4, 7, 5, and 5, respectively, at Cu 3 mg L⁻¹, dilution rate 0.05 h⁻¹, and pulse interval 72 h, indicating that a stochastic switch-like response was generated. We argue that the role of gill-associated Hill coefficient as a direct signal of the stochastic switch-like response in the damage-recovery dynamics in response to pulse metal stressor can serve as a sensitive indicator for risk detection in fluctuating environments. Our approach constitutes a general method to identify the stochastic switch-like response for aquatic species exposed to fluctuating metal stressors, which may help to predict and, eventually, expand our understanding of the damage-recovery dynamics. Finally, we implicate that Hill coefficient-based switch-like signal and its damage with hazard response can be linked in an information theoretic framework to handle environmental stochasticity.     

Human Health Risk Assessment of Soils Contaminated with Metal(loid)s by Using DGT Uptake: A Case Study of a Former Korean Metal Refinery Site

The human health risk of soils contaminated with As, Pb, Cu, and Zn was evaluated based on pseudo-total concentrations of metal(loid)s, the physiologically based extraction test (PBET), and diffusive gradients in thin films (DGT). Non-carcinogenic (NCR) and carcinogenic (CR) risks exceeded the U.S. Environmental Protection Agency criteria under both the residential and non-residential scenarios. Human bioavailable concentrations (PBET) were much lower than pseudo-total concentrations. The Hazardous Index of NCR (HI (NCR)) for the PBET in the studied soils was 67% and 94% less than that for pseudo-total concentration, respectively, under the non-residential and residential scenarios. Similarly, CR for the PBET was also 65% and 93% less for the two soils. The concentration of metal(loid)s accumulated in the DGT resin was highly correlated with the PBET-extractable concentration (R² > 0.649). Therefore, for both the CR and HI (NCR), the DGT-calculated risk was linearly related to the PBET-calculated risk for the studied soils under both scenarios. The results suggest that DGT uptake and PBET-extracted concentrations are good surrogates for risk estimation and that both J1 and J2 soils require remediation before their use for residential or non-residential purposes.     

X-Ray Diffractometric Studies on Starches

A linear amylodextrin (average degree of polymerization 12.6) from sweetpotato was crystallized at different temperatures and from different concentrations of solution, and the crystalline type of the recrystallized amylodextrin was examined by X-ray diffraction. It was shown that the crystal type was dependent on the crystallization conditions. The conditions required for the formation of A-, C- and B-type crystals were limited by the following functional relations: 2.5T+C>84, 72<2.5T+C<84, and 2.5T+C<72, respectively, where T is the temperature (°C) at which the amylodextrin was crystallized and C is the concentration (%) of the amylodextrin solution. This relation showed that high temperatures and high concentrations of amylodextrin were favorable for A-type crystal formation and low temperatures and concentrations for B-type crystals, and that the type of crystals formed was more affected by temperature than concentration of the solution.