Transcriptional activation by the sexual pheromone and wounding: a new gene family from Volvox encoding modular proteins with (hydroxy)proline-rich and metalloproteinase homology domains

The green alga Volvox represents the simplest kind of multicellular organism: it is composed of only two cell types, somatic and reproductive, making it suitable as a model system. The sexual development of males and females of Volvox carteri is triggered by a sex-inducing pheromone at a concentration of < 10-16 M. Early biochemical responses to the pheromone involve structural modifications within the extracellular matrix (ECM). By differential screenings of cDNA libraries made from mRNAs of pheromone-treated Volvox, four novel genes were identified that encode four closely related Volvox metalloproteinases that we use to define a new protein family, the VMPs. The existence of several features common to matrix glycoproteins, such as signal peptides, a (hydroxy)proline content of 12-25%, and Ser(Pro)2-4 repeats, suggest an extracellular localization of the VMPs within the ECM. Synthesis of VMP cDNAs is triggered not only by the sex-inducing pheromone, but also by wounding, and is restricted to the somatic cell type. Sequence comparisons suggest that the VMPs are members of the MB clan of zinc-dependent matrix metalloproteinases, although the putative zinc binding site of all VMPs is QEXXHXXGXXH rather than HEXXHXXGXXH. The presence of glutamine instead of histidine in the zinc binding motif suggests a novel family, or even clan, of peptidases. Like the matrixin family of human collagenases, Volvox VMPs exhibit a modular structure: they possess a metalloproteinase homology domain and a (hydroxy)proline-rich domain, and one of them, VMP4, also has two additional domains. Metalloproteinases seem to be crucial for biochemical modifications of the ECM during development or after wounding in the lower eukaryote Volvox with only two cell types, just as in higher organisms.     

An extracellular matrix-localized metalloproteinase with an exceptional QEXXH metal binding site prefers copper for catalytic activity

The extracellular matrix (ECM) of the simple multicellular organism Volvox contains many region-specific morphological elements and mediates a variety of developmental and physiological responses by modification of its components. The fact that >95% of the mature organism is ECM makes Volvox suitable as a model system for ECM investigations. VMPs are a family of Volvox genes that are homologous to zinc-dependent matrix metalloproteinases (MMPs). Here we describe the identification and purification of the first VMP protein, VMP3. The 470-kDa VMP3 glycoprotein is localized within the ECM, and its biosynthesis is induced by the sex pheromone. The metal binding motif of VMP3 is QEXXH, not HEXXH as known for approximately 1300 other metalloproteinases. VMP3 shows proteinase activity and is inhibited by EDTA or the MMP inhibitor GM 6001, but in contrast to all known proteinases, VMP3 clearly prefers copper for activity rather than zinc. The exchange from Q to H within the QEXXH motif abolishes its copper preference. The unique properties of VMP3 suggest a novel type of metalloproteinase.     

Comparison of Four Species Sensitivity Distribution Methods to Calculate Predicted No Effect Concentrations for Bisphenol A

Bisphenol A (BPA, CAS RN 80-05-7) is a high production volume chemical used as an intermediate in the production of polycarbonate plastic and epoxy resins. During its manufacture and use, some emissions to surface waters are anticipated. Chronic predicted no effect concentrations (PNECs) for aquatic systems are used to support the assessment of potential risks to aquatic organisms in receiving waters. PNECs for a compound are considered protective of populations, communities, and ecosystems. Traditionally, PNECs are derived by taking the lowest no-observed effect concentration (NOEC) from a set of toxicity studies and dividing by an assessment factor (e.g., 10 to 1000). This traditional approach is appropriate for substances with few data, but may not be necessary for substances with many valid studies. For well-studied substances, statistical approaches (i.e., development of Species Sensitivity Distribution or SSD methods) can be used to calculate a PNEC that makes use of the full distribution of available NOEC values. Bisphenol A has an extensive set of aquatic toxicity studies covering diverse taxa including algae, hydra, rotifers, mollusks, crustaceans (both benthic and pelagic), insects, annelids, fish, and amphibians. The full chronic data set was used to calculate PNEC values using four SSD methods: (1) the Hazard Concentration (HC5) approach developed by The Netherlands National Institute of Public Health and the Environment (RIVM), (2) the U.S. Environmental Protection Agency's water quality criteria procedure, (3) SigmaPlot (Systat 2000) commercial software that calculates percentile values, and (4) a distributional method consistent with that used by Environment Canada. Using these approaches, PNEC values for BPA range from 11 to 71 μ g/L. Literature studies suggest that application of an additional assessment factor is unwarranted if an SSD-based PNEC is based on chronic data. SSD-derived PNEC values and the traditionally derived PNEC value of 1.6 μ g/L are then compared to concentrations of BPA that have been measured in North American and European surface waters. Adverse risks to aquatic organisms are not anticipated from measured concentrations of BPA in North American and European surface waters.     

Evaluation of Ecotoxicity Effect Indicators for Use in LCIA (10+4 pp)

Goal, Scope and Background The paper describes different ecotoxicity effect indicator methods/approaches. The approaches cover three main groups, viz. PNEC approaches, PAF approaches and damage approaches. Ecotoxicity effect indicators used in life cycle impact assessment (LCIA) are typically modelled to the level of impact, indicating the potential impact on 'ecosystem health'. The few existing indicators, which are modelled all the way to damage, are poorly developed, and even though relevant alternatives from risk assessment exist (e.g. recovery time and mean extinction time), these are unfortunately at a very early stage of development, and only few attempts have been made to include them in LCIA. Methods The approaches are described and evaluated against a set of assessment criteria comprising compatibility with the methodological requirements of LCIA, environmental relevance, reproducibility, data demand, data availability, quantification of uncertainty, transparency and spatial differentiation. Results and Discussion The results of the evaluation of the two impact approaches (i.e. PNEC and PAF) show both pros and cons for each of them. The assessment factor-based PNEC approaches have a low data demand and use only the lowest data (e.g. lowest NOEC value). Because it is developed in tiered risk assessment, and hence makes use of conservative assessment factors, it is not optimal, in its present form, to use in the comparative framework of LCIA, where best estimates are sought. The PAF approaches have a higher data demand but use all data and can be based on effect data (PNEC is no-effect-based), thus making these approaches non-conservative and more suitable for LCIA. However, indiscriminate use of ecotoxicity data tends to make the PAF-approaches no more environmentally relevant than the assessment factor-based PNEC approaches. The PAF approaches, however, can at least in theory be linked to damage modelling. All the approaches for damage modelling which are included here have a high environmental relevance but very low data availability, apart from the 'media recovery-approach', which depends directly on the fate model. They are all at a very early stage of development. Conclusion Recommendations and Outlook. An analysis of the different PAF approaches shows that the crucial point is according to which principles and based on which data the hazardous concentration to 50% of the included species (i.e. HC50) is estimated. The ability to calculate many characterisation factors for ecotoxicity is important for this impact category to be included in LCIA in a proper way. However, the access to effect data for the relevant chemicals is typically limited. So, besides the coupling to damage modelling, the main challenge within the further development and improvement of ecotoxicity effect indicators is to find an optimal method to estimate HC50 based on little data.     

Exposure Assessment of Pharmaceuticals and Their Metabolites in the Aquatic Environment: Application to the French Situation and Preliminary Prioritization

Low levels of pharmaceuticals have been detected in many countries in surface waters. As a wide range of pharmaceuticals can reach aquatic environments, a selection of molecules to survey is the first step before implementing a monitoring program. We used a simple equation to calculate Predicted Environmental Concentrations (PECs), adapted from the European Medicine Agency model used for the Environmental Risk Assessment (ERA) of human pharmaceutical. Excretion fractions for pharmaceuticals were determined for 76 compounds. Using year 2004 French drug consumption data, we determined aquatic PECs for 112 parent molecules and several metabolites. Considering excretion fractions of pharmaceuticals can lead to drastically reduce predicted concentrations reaching the aquatic environment and help to target environmentally relevant pharmaceuticals and metabolites. Calculated PECs using the described methodology are consistent with French field measurements. The simple model for calculating PECs can be used as a valuable estimation of the exposure. Risk quotient ratios were also calculated. Due to the lack of ecotoxicological data, the use of PEC/PNEC ratios is not enough informative to prioritize pharmaceuticals likely to pose a risk for surface waters. Alternative ways to prioritize risk to pharmaceuticals, combining PEC, pharmacological, and ecotoxicological data available from the literature, should be implemented.     

Environmental Risk of Pesticides: Applying the DelAzulPestRisk Model to Freshwaters of an Agricultural Area of Argentina

DelAzulPestRisk is a risk-based chemical ranking model based on human and local biota toxicities that estimates the integrated risk of pesticides in water from their extensive (concentration, risk) and intensive (persistence, bioaccumulation) chemical properties. The model is built on two modules: human health risk factor (estimated based on the probabilistic cancer and non-cancer health risk by using U.S. Environmental Protection Agency models applied to a bathing exposure scenario) and biota health risk factor (quantified on the basis of the probabilistic toxicity exposure ratio –PEC/PNEC– for three local representatives of water biota multiplied by an amplification factor supported by the persistence and bioaccumulation potential). The model was applied to shallow creeks of Tres Arroyos County, Argentina, which flow across wheat and soybean agricultural lands, and in whose waters were detected many organochlorine pesticides (α, γ, y, δ-HCH, aldrin, heptachlor, γ-chlordane, endosulfan, endosulfan sulphate, dieldrin, and DDD). Dieldrin, aldrin, and heptachlor generated the worst potential effects—due mainly to the cancer and non-cancer dermal health risk—although this was not a significant environmental threat. DelAzulPestRisk is a screening assessment tool for water management purposes that become useful in countries lacking efficient water quality control systems.     

Environmental Fate of Neonicotinoids and Classification of Their Potential Risks to Hypogean,Epygean, and Surface Water Ecosystems in Brazil

Due to their reported high toxicity to honey bees, ecotoxicological studies into the side-effects of neonicotinoid insecticides have focused almost exclusively on these organisms. The fate of neonicotinoids and potential toxic side-effects on other (especially non-standard) organisms have received considerably less attention. In the present study, the environmental distribution and leaching potential of neonicotinoids registered for agricultural use in Brazil (acetamiprid, clothianidin, imidacloprid, thiacloprid, and thiamethoxam) were studied by applying several environmental fate models and indices. Potential risks to various environmental compartments were evaluated by applying ranking indices to the maximum application rates recommended in Brazil. Although bees were indeed found to be the most sensitive organism, the neonicotinoids also indicated potential environmental risks to other organism groups. Due to the greater maximum application rates recommended in Brazil as compared to other parts of the world, environmental risk and resistance potential for at least imidacloprid appears especially high in Brazil. Attention should thus also be allocated to organisms other than bees and to resistance potential when performing an environmental risk assessment of neonicotinoids if they are used at relatively high application rates.     

Relationships between Aquatic Hazard Quotients and Probabilistic Risk Estimates: What Is the Significance of a Hazard Quotient >1?

This study examined relationships between hazard quotients (HQ) and probabilistic estimates of aquatic ecological risk. Questions addressed included the magnitude at which an HQ equates to significant risk, and the factors influencing the HQ-risk relationship. The analysis was based upon predicted exposure concentrations (PEC) for copper, hypothetical predicted no effect concentrations (PNEC) distributions, and measured PNEC data for aquatic species acutely and chronically exposed to copper, ammonia, cadmium, cyanide, dieldrin, DDT, phenanthrene, silver. and zinc. The cumulative PNEC and PEC distributions differed in slopes and magnitudes. The relationship between HQ and probabilistic risk, both of which were computed using conventional techniques, depended on the slopes of the PNEC and PEC distributions. Hazard quotients equaling 1.0 affected ～ 5% of the species because they were based on PNECs intended to protect 5% of the species. Hazard quotients greater than 1.0 depended on PNEC slope. For example, HQs for toxicants with the steeper PNEC distributions affected a large percentage of species (18 to 49%, depending on slope) at HQ=2 to 3. Other factors (e.g., variability in both PEC and PNEC data, and use of arithmetic or geometric means or their confidence limits) had variable influences on the HQ-risk relationship.     

Probabilistic Environmental Risk Assessment for Dibutylphthalate (DBP)

In the risk assessment methods for new and existing chemicals in the European Union (EU), environmental “risk” is characterized by the deterministic quotient of exposure and effects (PEC/PNEC). From a scientific viewpoint, the uncertainty in the risk quotient should be accounted for explicitly in the decision making, which can be done in a probabilistic risk framework. To demonstrate the feasibility and benefits of such a framework, a sample risk assessment for an existing chemical (dibutylphthalate, DBP) is presented in this paper. The example shows a probabilistic framework to be feasible with relatively little extra effort; such a framework also provides more relevant information. The deterministic risk quotients turned out to be worst cases at generally higher than the 95th percentile of the probability distributions. Sensitivity analysis proves to be a powerful tool in identifying the main sources of uncertainty and thus will be effective for efficient further testing. The distributions assigned to the assess ment factors (derivation of the PNEC) dominate the total uncertainty in the risk assessment; uncertainties in the release estimates come second. Large uncertainties are an inherent part of risk assessment that we have to deal with quantitatively. However, the most appropriate way to characterise effects and risks requires further attention. Recommendations for further study are identified.     

Exposure Based Waiving in Environmental Risk Assessment: A Tiered Approach

The REACH legislation describes the “Exposure Based Waiving” (EBW) methodology, that allows reduction of animal tests and that facilitates the Chemical Safety Assessments. We introduced a tiered EBW decision strategy for the aquatic environment. Tier 0 in this strategy justifies waiving if it can be demonstrated that exposure of organisms to a chemical will be practically zero (i.e., if no environmental emissions during the full life-cycle of a chemical are to be expected. Tier 1 uses tabular data, derived from physical–chemical properties of a chemical and emission data, to identify whether the environmental exposure levels are less than the conservative toxicological reference value “Threshold of Toxicological Concern” (TTC). To this end, systematic exposure calculations have been carried out with a hypothetical chemical dataset, covering the full chemical spectrum for neutral organic substances. In tier 2, PEC and PNEC are to be assessed probabilistically in order to address the probability of effects. Following this strategy, risk can be shown to be sufficiently low, using minimal exposure and hazard data. Our tiered EBW concept is illustrated by a case study for the substance dibutylphthalate with the endpoint of fresh water aquatic risks.     

BmVMP90, a large vitelline membrane protein of the domesticated silkmoth Bombyx mori, is an essential component of the developing ovarian follicle

We present the characterization of BmVMP90, a vitelline membrane protein (VMP) of the silkmoth Bombyx mori bearing similarities with dipteran VMPs whose existence had recently been suggested by an in silico analysis of the silkmoth genome and follicular cell RNA expression analyses. Using a specific antibody, we determine the presence of BmVMP90 protein in ovarian follicular cell extracts at the end of vitellogenesis and in vitelline membrane extracts but not in the chorion of fractionated eggshells isolated from ovulated follicles. Whole mount follicle immunofluorescence studies reveal a pattern of BmVMP90 deposition matching the ?imprinted? pattern of follicular cells on the vitelline membrane surface. Antisense DNA-directed inhibition BmVMP90 expression in ex?vivo cultures of early vitellogenic follicles produced a phenotype of kidney- or bean-shaped follicles with detached follicular epithelia, suggestive of the importance of BmVMP90 for the integrity of developing follicles and normal deposition of the chorion structure that follows vitelline membrane formation but no adverse effects on the execution of the follicular cell-imprinted program of choriogenesis per se.     

食源性致病菌风险排名方法

综述食品风险排名的相关概念及其在风险评估中的重要性,探讨发达国家进行风险分析的方法与经验,其中致病菌风险排名的研究因病原菌的动态性和宿主特异性而呈现出独特的挑战性。重点详细探讨美国对食源性致病菌排名的创新方法——专家组启发法,此法作为弥补数据不足和不确定性的手段,结合传统的＂自上而下＂和＂自下而上＂方法,可以为风险管理者提供了更全面更有效的食源性致病菌风险排名。     

Pyrithiones as antifoulants: Environmental chemistry and preliminary risk assessment

Pyrithiones are widely used as bactericides, fungicides, or algicides in a variety of products such as shampoos, metal working fluids, adhesives, sealants, and coatings. This broad antimicrobial activity, along with low water solubility and favorable environmental chemistry, makes zinc pyrithione and copper pyrithione potentially ideal replacements for TBT in marine antifouling paints. Several studies on the toxicity and environ- ? mental fate of these pyrithiones have been conducted in freshwater and saltwater systems. Environmental fate studies show that pyrithiones rapidly degrade in the water column to less toxic compounds. Sediment accumulation is also prevented by the facile reduction of a critical functional group under anaerobic conditions. Modeling programs were used to calculate the predicted environmental concentration (PEC) for pyrithione. Comparison of PECs calculated for more persistent antifoulants with actual measured concentrations provided a measure of the bias inherent to the models. The results indicate a pyrithione risk quotient (PEC/PNEC) < 1. The findings are consistent with the absence of ecological effects during the long history of the use of zinc pyrithione as an antidandruff agent.     

In search of relevant predictors for marine species distribution modelling using the MarineSPEED benchmark dataset

Aim

Ideally, datasets for species distribution modelling (SDM) contain evenly sampled records covering the entire distribution of the species, confirmed absences and auxiliary ecophysiological data allowing informed decisions on relevant predictors. Unfortunately, these criteria are rarely met for marine organisms for which distributions are too often only scantly characterized and absences generally not recorded. Here, we investigate predictor relevance as a function of modelling algorithms and settings for a global dataset of marine species.

Location

Global marine.

Methods

We selected well‐studied and identifiable species from all major marine taxonomic groups. Distribution records were compiled from public sources (e.g., OBIS, GBIF, Reef Life Survey) and linked to environmental data from Bio‐ORACLE and MARSPEC. Using this dataset, predictor relevance was analysed under different variations of modelling algorithms, numbers of predictor variables, cross‐validation strategies, sampling bias mitigation methods, evaluation methods and ranking methods. SDMs for all combinations of predictors from eight correlation groups were fitted and ranked, from which the top five predictors were selected as the most relevant.

Results

We collected two million distribution records from 514 species across 18 phyla. Mean sea surface temperature and calcite are, respectively, the most relevant and irrelevant predictors. A less clear pattern was derived from the other predictors. The biggest differences in predictor relevance were induced by varying the number of predictors, the modelling algorithm and the sample selection bias correction. The distribution data and associated environmental data are made available through the R package marinespeed and at http://marinespeed.org .

Main conclusions

While temperature is a relevant predictor of global marine species distributions, considerable variation in predictor relevance is linked to the SDM set‐up. We promote the usage of a standardized benchmark dataset (MarineSPEED) for methodological SDM studies.

     

Uncertainties in the Environmental Risk Assessment of Metals

As life has evolved in the presence of metals, the assessment of the potential adverse effects of metals on ecosystems requires a different approach than those presently used for man-made organic substances. This article provides a brief review of applications and limitations of current techniques and presents, based on recent research results, suggestions for improving the scientific relevance and accuracy of environmental risk assessments of metals. The importance of the following factors responsible for major uncertainties in current environmental risk assessments of metals are discussed: factors affecting metal bioavailability and toxicity, the potential importance of deficiency effects (for essential metals), and field extrapolation of laboratory toxicity data. Possible (regulatory) consequences of inaccurately assessing the natural background concentrations of metals and acclimatization/adaptation potential of laboratory organisms and resident communities are illustrated using examples of recent research, hypothesis development, and a probabilistic environmental risk assessment.     

Opportunities for web-based indicators in environmental sciences

This paper proposes a set of web-based indicators for quantifying and ranking the relevance of terms related to key-issues in Ecology and Sustainability Science. Search engines that operate in different contexts (e.g. global, social, scientific) are considered as web information carriers (WICs) and are able to analyse; (i) relevance on different levels: global web, individual/personal sphere, on-line news, and culture/science; (ii) time trends of relevance; (iii) relevance of keywords for environmental governance. For the purposes of this study, several indicators and specific indices (relational indices and dynamic indices) were applied to a test-set of 24 keywords. Outputs consistently show that traditional study topics in environmental sciences such as water and air have remained the most quantitatively relevant keywords, while interest in systemic issues (i.e. ecosystem and landscape) has grown over the last 20 years. Nowadays, the relevance of new concepts such as resilience and ecosystem services is increasing, but the actual ability of these concepts to influence environmental governance needs to be further studied and understood. The proposed approach, which is based on intuitive and easily replicable procedures, can support the decision-making processes related to environmental governance.     

Comparison of new Brazilian legislation for the approval of advanced therapy medicinal products with existing systems in the USA,European Union and Japan

Background aimsAdvanced therapy medicinal products (ATMPs) are a class of biological products for human use that are based on genes, cells and tissues. The first ATMP received marketing authorization in Europe in 2009, whereas Brazil granted the first authorization in 2020. The objective of this study was to compare the regulatory models adopted by Brazil, the USA, Japan and the European Union, which comprise the member countries of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use, with regard to the marketing authorization of ATMPs.MethodsThe authors performed a review of the scientific literature and official documents of the regulatory agencies in the aforementioned countries.ResultsThe legislation and regulatory guidelines adopted by the regulatory agencies exhibit similarities and differences. It was not possible to assess whether these differences can be translated into divergent final recommendations by regulatory authorities upon a request for marketing authorization.ConclusionsIn the future, it will be appropriate to start a progressive process of harmonization between these agencies in terms of terminology, legal recommendations and characterization requirements. This is particularly important for emerging countries such as Brazil. In this sense, some measures can be taken to achieve alignment between regulators.     

Using Bioassays and Species Sensitivity Distributions to Assess Herbicide Toxicity towards Benthic Diatoms

Although benthic diatoms are widely used in ecological studies of aquatic systems, there is still a dearth of data concerning species sensitivities towards several contaminants. Within the same community, different species may respond differently depending on their physiological and ecological characteristics. This lack of knowledge makes specific appropriate risk assessment impossible. To find out whether species sensitivity distribution (SSD) could be used to estimate the risk of herbicide toxicity for diatoms, we need to know whether their sensitivity depends on their physiological and ecological characteristics. We carried out single-species bioassays on 11 diatom species exposed to 8 herbicides. Dose-responses relationships were used to extrapolate the Effective Concentration 5 (EC(5)) and the Effective Concentration 50 (EC(50)) for each exposure. These data were used to fit a SSD curve for each herbicide, and to determine the Hazardous concentration 5 (HC(5)) and 50 (HC(50)). Our results revealed a high level of variability of the sensitivity in the set of species tested. For photosystem-II inhibitor (PSII) herbicides, diatoms species displayed a typical grouping of sensitivity levels consistent with their trophic mode and their ecological guild. N-heterotroph and "motile" guild species were more tolerant of PSII inhibitors, while N-autotroph and "low profile" guild species were more sensitive. Comprehensive SSD curves were obtained for 5 herbicides, but not for sulfonylurea herbicides or for dimetachlor, which had toxicity levels that were below the range of concentration tested. The SSD curves provided the following ranking of toxicity: diuron> terbutryn> isoproturon> atrazine> metolachlor. The HC that affected 5% of the species revealed that, even at the usual environmental concentrations of herbicides, diatom assemblages could be affected, especially by isoproturon, terbutryn, and diuron.