Uncertainty associated with river health assessment in a varying environment: The case of a predictive fish-based index in France

Sensitive biological measures of river ecosystem quality are needed to assess, maintain or restore ecological conditions of water bodies. Since our understanding of these complex systems is imperfect, decision-making requires recognizing uncertainty. In this study, a new predictive multi-metric index based on fish functional traits was developed to assess French rivers. Information on fish assemblage structure, local environment and human-induced disturbances of 1654 French river sites was compiled. A Bayesian framework was used to predict theoretical metric values in absence of human pressure and to estimate the uncertainty associated with these predictions. The uncertainty associated with the index score gives the confidence associated with the evaluation of site ecological conditions.Among the 228 potential metrics tested, only 11 were retained for the index computation. The final index is independent from natural variability and sensitive to human-induced disturbances. In particular, it is affected by the accumulation of different degradations and specific degradations including hydrological perturbations. Predictive uncertainty is globally lower for IPR+ than for underlying metrics.This new methodology seems appropriate to develop bio-indication tools accounting for uncertainty related to reference condition definition and could be extended to other biological groups and areas. Our results support the use of multi-metric indexes to assess rivers and strengthen the idea that examination of uncertainty could contribute greatly to the improvement of the assessment power of bio-indicators.     

Transfer and expression of the tetracyclin resistance transposon Tn925 in Acetobacterium woodii

The Enterococcus faecalis conjugative plasmid pCF10 was used to introduce Tn925 into Acetobacterium woodii by filter mating. Tetracycline resistance was transferred at frequencies of about 10(-6) per donor, but no plasmid DNA was found in the transconjugants. DNA hybridization analyses of HindIII-digested chromosomal DNA demonstrated the insertion of Tn925 at a variety of locations, whereas wild type DNA showed no hybridization at all. The transconjugants were used as donor in mating experiments with tetracycline-sensitive Bacillus subtilis. Transfer of tetracycline resistance was observed at frequencies of 10(-8) per recipient.     

Synthesis and anti-proliferative activities of new derivatives of embelin

Embelin (1), a benzoquinone isolated from Embelia ribes is known to possess variety of biological activities. Despite of several promising biological activities, preclinical efforts on embelin were hampered because of its poor aqueous solubility. In order to address the solubility issue, herein, we have synthesized a series of Mannich products of embelin by treating it with various secondary amines. The synthesized compounds were screened for antiproliferative and antimicrobial activities. In cytotoxicity screening, the benzyl-piperidine linked derivative 8m was found to possess better antiproliferative activity compared to parent natural product embelin against a panel of cell lines including HCT-116, MCF-7, MIAPaCa-2 and PC-3 with IC₅₀ values of 30, 41, 34 and 36 μM, respectively. The mechanistic study of compound 8m revealed that it exhibits cytotoxicity via induction of apoptosis and mitochondrial membrane potential loss. Further, the compounds were tested for antimicrobial activity where dimethylamino- 8a and piperidine linked derivative 8b displayed antibacterial activity against Staphylococcus aureus with MIC values of 8 and 16 μg/mL, respectively. Mannich derivatives did now show improved aqueous solubility, however their hydrochloride salts 8a·HCl, 8b·HCl and 8m·HCl showed significantly improved aqueous solubility without affecting biological activities of parent Mannich derivatives.     

Key factors controlling the size, biomass, and sprouting of Japanese alder swamp forest in Kushiro Mire, Hokkaido, Japan

The interrelation among size, biomass, and sprouting of alder trees was studied to extract the most important hydrochemistrical factors controlling the growth of alder forest in Kushiro Mire, northern Japan. The gradient was mostly explained by chemical variables such as pH, ash content, and P₂O₅, which showed strong positive correlation with each other, and secondarily by fluctuation of the water table (WL, i.e., water level range). These variables are more important than other hydrochemical ones, because neutral and turbid flood water replaces acidic mire water and conveys fine sediment with adsorbed phosphorus, which in turn could regulate the pH and amount of phosphorus. Also, the number of sprouts showed negative correlation mainly with tree size and redox potential (Eh), which suggested a flooded environment. Because of this, the size of alder was suppressed by hydrochemical variables; however, alder individuals producing new sprouts were maintained. We conclude that variation in size, biomass, and sprouting of alder was mainly controlled by acidity and phosphorus availability, and was significantly influenced by water fluctuation.     

Histidine Residue Mediates Radical-induced Hinge Cleavage of Human IgG1

Hydroxyl radicals induce hinge cleavage in a human IgG1 molecule via initial radical formation at the first hinge Cys²³¹ followed by electron transfer to the upper hinge residues. To enable engineering of a stable monoclonal antibody hinge, we investigated the role of the hinge His²²⁹ residue using structure modeling and site-directed mutagenesis. Direct involvement of His²²⁹ in the reaction mechanism is suggested by a 75–85% reduction of the hinge cleavage for variants in which His²²⁹ was substituted with either Gln, Ser, or Ala. In contrast, mutation of Lys²²⁷ to Gln, Ser, or Ala increased hinge cleavage. However, the H229S/K227S double mutant shows hinge cleavage levels similar to that of the single H229S variant, further revealing the importance of His²²⁹. Examination of the hinge structure shows that His²²⁹ is capable of forming hydrogen bonds with surrounding residues. These observations led us to hypothesize that the imidazole ring of His²²⁹ may function to facilitate the cleavage by forming a transient radical center that is capable of extracting a proton from neighboring residues. The work presented here suggests the feasibility of engineering a new generation of monoclonal antibodies capable of resisting hinge cleavage to improve product stability and efficacy.     

Water current velocity as an environmental factor regulating the distribution of amphipod species in Gamo lagoon, Japan

The role of current velocity as an environmental factor affecting the distribution of amphipod species was investigated in the brackish Gamo lagoon, Japan. Two gammaridean amphipod species, Eogammarus possjeticus and Melita setiflagella, were found inhabiting stations of different water current with same substrates. E. possjeticus and M. setiflagella selected substrates at low and high water current, respectively. The lifestyles and anatomical features of these two free-living amphipods contributed to their adaptation to habitats with different water currents. The results of field and experimental observations suggest the existence of some type of interspecific competition between the two species. The presence of E. possjeticus caused a displacement of the fundamental niche of M. setiflagella. Received: October 10, 2000 / Accepted: August 17, 2001     

The method matters: A guide for indicator aggregation in ecological assessments

Ecological assessment requires the integration of many physical, chemical, and/or biological quality elements. The choice of the aggregation method of such partial assessments into an overall assessment can considerably affect the assessment outcome – an issue that has been controversially discussed within the scientific community for the last decade. Current practice often considers only two different aggregation methods, the weighted arithmetic mean (additive aggregation) and the one-out, all-out method (minimum aggregation). However, both have important drawbacks. Additive aggregation compensates a bad status of one quality element by a number of elements featuring good status. Minimum aggregation can lead to overly pessimistic assessment results, since only the quality element in the worst status is considered. Here, we introduce a toolbox containing current and new aggregation methods, demonstrate and discuss their properties with simple, didactical examples, and suggest in which situations best to use them. Then, we illustrate the consequences of selected aggregation schemes for ecological river assessment with the case study of the Swiss Modular Concept of stream assessment (SMC), which we apply to ten river reaches in the Mönchaltdorfer Aa catchment in Switzerland. To be able to do so, we used multi-criteria decision analysis, i.e., multi-attribute value theory, to arrange the SMC quality elements into an objectives hierarchy, and to translate their individual assessments into value functions. Our case study revealed that choosing the most appropriate aggregation method particularly matters, if objectives with significantly different qualities are aggregated. We argue that redundant objectives (i.e., quality elements), often found at the lower levels of the objectives hierarchy, should best be aggregated additively allowing for compensation to increase the statistical significance of the results. Further, we suggest that complementary sub-objectives that often occur at higher levels may be optimally aggregated with a mixture of additive and minimum aggregation. Such a mixed method will allow some compensation, but nevertheless penalize for very bad states. Since here we compare commonly used aggregation methods with some which we believe have never been discussed in an assessment context before, our study concurrently informs ecological assessment in theory and in practice.     

Responses of aquatic macrophyte assemblages to nutrient enrichment in a lowland river basin of western Greece

Abstract

In Greece, as in many other Mediterranean countries, there is a lack of information enabling the use of aquatic macrophytes as biological indicators of the water trophic status. This research aimed to investigate the trophic preferences of the aquatic macrophyte assemblages encountered in selected standing water bodies of a lowland river basin of western Greece. It also aimed to assess the response of the macrophyte species that occur in the studied water bodies to different nutrient concentrations. A total of 56 macrophyte species were identified. TWINSPAN analysis distinguished four main vegetation groups potentially related to three trophic types. The Trophic Ranking Score developed in the UK was applied in order to investigate the trend in aquatic macrophytes in response to the different trophic conditions. The results indicated that TRS_UK was not in accordance to the water trophic indices. After the addition of 14 new species and the re-scoring of the plants from the UK list, the re-calculation of TRS enabled a better classification of the trophic status convergent with the information from the water trophic indices. The current study constitutes a reference document providing basic information, and must be improved by updating the plant list with new data from other sites.     

Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells

Luminescence Resonance Energy Transfer, or LRET, is a powerful technique used to measure distances between two sites in proteins within the distance range of 10-100 Å. By measuring the distances under various ligated conditions, conformational changes of the protein can be easily assessed. With LRET, a lanthanide, most often chelated terbium, is used as the donor fluorophore, affording advantages such as a longer donor-only emission lifetime, the flexibility to use multiple acceptor fluorophores, and the opportunity to detect sensitized acceptor emission as an easy way to measure energy transfer without the risk of also detecting donor-only signal. Here, we describe a method to use LRET on membrane proteins expressed and assayed on the surface of intact mammalian cells. We introduce a protease cleavage site between the LRET fluorophore pair. After obtaining the original LRET signal, cleavage at that site removes the specific LRET signal from the protein of interest allowing us to quantitatively subtract the background signal that remains after cleavage. This method allows for more physiologically relevant measurements to be made without the need for purification of protein.