Ecological effects of diffuse mixed pollution are site-specific and require higher-tier risk assessment to improve site management decisions: a discussion paper

Many Dutch ecosystems, whether terrestrial, aquatic or sediment-based, are diffusely polluted by mixtures of contaminants, whose concentrations often exceed regulatory Safe Values or other generic quality criteria. This situation has unclear consequences, especially when local authorities are confronted with such pollution. Water managers are frequently in doubt whether their water systems satisfy the criteria for ‘Good Ecological Status’ as defined in the EU's Water Framework Directive. In case of soils, soil users may wonder whether the soil is ‘fit for use’. In case of nature conservation, the problem is that protected species might suffer from toxic stress. Official regulations in these cases call for appropriate action, but it is unclear whether the diffuse exposure causes adverse effects, and what the action should be. This paper proposes and discusses a site-oriented approach in the risk assessment of diffusely contaminated sites that can be used in addition to the compound-oriented policies from which the abovementioned generic quality criteria were derived. The site-oriented approach can be of help in reducing site-specific risks of diffuse contamination.Reflecting on the results of a large Dutch research effort in systems-oriented ecotoxicological effects, the conclusion is drawn that exposure and effects of diffuse pollution are site-specific in kind and magnitude, determined by the local combination of source-pathway-receptor issues, and often not clearly detectable (though often present). To assist in risk management, higher-tier methods can address various aspects, like addressing local mixture composition, bioavailability, and sensitivity of local species groups. Higher-tier risk assessment methods have as yet been developed mainly for cases of serious contamination, like for pesticide management and Risk-Based Land Management. For diffuse pollution, site-specific information can also be used to obtain site-specific exposure and impact information, while practical and ecology-based approaches can be introduced to obtain an integrated overview of the meaning of site contamination and to derive options for managing and reducing the local risks. These issues are discussed against the background of current major policy shifts, in The Netherlands and elsewhere, from a pollutant-oriented assessment to an additional ecological and site-oriented assessment. The latter is most clearly represented in the Good Ecological Status aim of the EU-Water Framework Directive. The paper assesses, integrates and discusses the results of the Dutch research effort in this policy context.     

项目和大型项目计划管理审计——如何提高项目管理和大型项目计划管理质量（下）

2．4项目审计标准取决于项目管理方法 审计标准取决于审计中所依据的项目管理方法。在项目型组织中，项目管理指导原则和标准可作为参考基线。学习效果受审计标准所限。如果项目管理审计以传统项目管理方法(例如：项目管理知识体系指南PMBOK介绍的方法)为基础，则审计标准将会局限于传统项目管理方法(包括范围、进度和成本管理方法)范围内。例如，只有在使用PRINCE2 (OGC，2002)的情况下，     

An activity, stability and selectivity comparison of propioin synthesis by thiamine diphosphate-dependent enzymes in a solid/gas bioreactor

Enzymatic carboligation in a solid/gas bioreactor represents a new challenge in biotechnology. In this paper, the continuous gas-phase production of propioin from two propanal molecules by using thiamine diphosphate-dependent enzymes was studied. Two enzymes were used, namely benzaldehyde lyase (BAL) from Pseudomonas fluorescens and benzoylformate decarboxylase (BFD) from Pseudomonas putida. The enzymes are homologous and catalyze carboligase and carbolyase reactions in which no external cofactor regeneration is needed. The influence of water and substrate activity on the initial reaction rate and biocatalyst stability was investigated. An increase in water activity raised the initial reaction rates to the maximal values of 250 and 80 U g(-1) for BAL and BFD, respectively. The half-life showed the same trend with maximal values of 50 and 78 min for BAL and BFD, respectively. The increase in the half-life by increasing water activity was unexpected. It was also observed that BFD is more stable than BAL in the presence of the substrate propanal. Both enzymes showed substrate inhibition in the kinetic studies, and BAL was also deactivated during the reaction. Unexpectedly, the stereoselectivity of both enzymes (ee of 19 % for BAL and racemic mixture for BFD) was significantly impaired in the gas phase compared to the liquid phase.     

Estrogen-induced alterations in amh and dmrt1 expression signal for disruption in male sexual development in the zebrafish

Dmrt1 and amh are genes involved in vertebrate sex differentiation. In this study, we cloned dmrt1 and amh cDNAs in zebrafish (Danio rerio) and investigated the effects of exposure to 17a-ethinylestradiol (EE2), during early life on their patterns of expression and impact on the subsequent gonadal phenotype. Expression of both amh and dmrt1 in embryos was detected as early as at 1 day post fertilization (dpf) and enhanced expression of amh from 25 dpf was associated with the period of early gonadal differentiation. Sex-dependent differences in enhanced green fluorescent protein transgene expression driven by the promoter of the germ cell-specific vas gene were exploited to show that at 28dpf and 56dpf both amh and dmrt1 mRNA were overexpressed in males compared with females. Exposure during early life to environmentally relevant concentrations of EE2 had a suppressive effect on the expression of both amh and dmrt1 mRNAs and this was associated with a cessation/retardation in male gonadal sex development. Our findings indicate that estrogen-induced suppression in expression of dmrt1 and amh during early life correlate with subsequent disruptive effects on the sexual phenotype in males.     

Novel Simple Conjugation Chemistries for Decoration of GMMA with Heterologous Antigens

Outer Membrane Vesicles (OMV) constitute a promising platform for the development of efficient vaccines. OMV can be decorated with heterologous antigens (proteins or polysaccharides), becoming attractive novel carriers for the development of multicomponent vaccines. Chemical conjugation represents a tool for linking antigens, also from phylogenetically distant pathogens, to OMV. Here we develop two simple and widely applicable conjugation chemistries targeting proteins or lipopolysaccharides on the surface of Generalized Modules for Membrane Antigens (GMMA), OMV spontaneously released from Gram-negative bacteria mutated to increase vesicle yield and reduce potential reactogenicity. A Design of Experiment approach was used to identify optimal conditions for GMMA activation before conjugation, resulting in consistent processes and ensuring conjugation efficiency. Conjugates produced by both chemistries induced strong humoral response against the heterologous antigen and GMMA. Additionally, the use of the two orthogonal chemistries allowed to control the linkage of two different antigens on the same GMMA particle. This work supports the further advancement of this novel platform with great potential for the design of effective vaccines.     

Multilevel converters for single-phase grid connected photovoltaic systems: an overview

Multilevel voltage source inverters offer several advantages compared to their conventional counterparts. By synthesising the AC output terminal voltage from several levels of DC voltages, staircase waveforms can be produced, which approach the sinusoidal waveform with low harmonic distortion, thus reducing filter requirements. The need of several sources on the DC side of the converter makes multilevel technology attractive for photovoltaic applications. This paper provides an overview on different multilevel topologies and investigates their suitability for single-phase grid connected photovoltaic systems. Several transformerless photovoltaic systems incorporating multilevel converters are compared regarding issues such as component count and stress, system power rating and the influence of the photovoltaic array earth capacitance.     

Mild Muscle Mitochondrial Fusion Distress Extends Drosophila Lifespan through an Early and Systemic Metabolome Reorganization

In a global aging population, it is important to understand the factors affecting systemic aging and lifespan. Mitohormesis, an adaptive response caused by different insults affecting the mitochondrial network, triggers a response from the nuclear genome inducing several pathways that promote longevity and metabolic health. Understanding the role of mitochondrial function during the aging process could help biomarker identification and the development of novel strategies for healthy aging. Herein, we interfered the muscle expression of the Drosophila genes Marf and Opa1, two genes that encode for proteins promoting mitochondrial fusion, orthologues of human MFN2 and OPA1. Silencing of Marf and Opa1 in muscle increases lifespan, improves locomotor capacities in the long term, and maintains muscular integrity. A metabolomic analysis revealed that muscle down-regulation of Marf and Opa1 promotes a non-autonomous systemic metabolome reorganization, mainly affecting metabolites involved in the energetic homeostasis: carbohydrates, lipids and aminoacids. Interestingly, the differences are consistently more evident in younger flies, implying that there may exist an anticipative adaptation mediating the protective changes at the older age. We demonstrate that mild mitochondrial muscle disturbance plays an important role in Drosophila fitness and reveals metabolic connections between tissues. This study opens new avenues to explore the link of mitochondrial dynamics and inter-organ communication, as well as their relationship with muscle-related pathologies, or in which muscle aging is a risk factor for their appearance. Our results suggest that early intervention in muscle may prevent sarcopenia and promote healthy aging.     

Equine Chorionic Gonadotropin as an Effective FSH Replacement for In Vitro Ovine Follicle and Oocyte Development

The use of assisted reproductive technologies (ART) still requires strategies through which to maximize individual fertility chances. In vitro folliculogenesis (ivF) may represent a valid option to convey the large source of immature oocytes in ART. Several efforts have been made to set up ivF cultural protocols in medium-sized mammals, starting with the identification of the most suitable gonadotropic stimulus. In this study, Equine Chorionic Gonadotropin (eCG) is proposed as an alternative to Follicle Stimulating Hormone (FSH) based on its long superovulation use, trans-species validation, long half-life, and low costs. The use of 3D ivF on single-ovine preantral (PA) follicles allowed us to compare the hormonal effects and to validate their influence under two different cultural conditions. The use of eCG helped to stimulate the in vitro growth of ovine PA follicles by maximizing its influence under FBS-free medium. Higher performance of follicular growth, antrum formation, steroidogenic activity and gap junction marker expression were recorded. In addition, eCG, promoted a positive effect on the germinal compartment, leading to a higher incidence of meiotic competent oocytes. These findings should help to widen the use of eCG to ivF as a valid and largely available hormonal support enabling a synchronized in vitro follicle and oocyte development.     

Isolation,Culture and Comprehensive Characterization of Biological Properties of Human Urine-Derived Stem Cells

Mesenchymal stem cells (MSCs) represent an attractive source within the field of tissue engineering. However, their harvesting often requires invasive medical procedures. Urine-derived stem cells (UDSCs) display similar properties to MSCs, and their obtention and further processing is non-invasive for the donors as well as low cost. Here, we offer a comprehensive analysis of their biological properties. The goal of this study was to analyze their morphology, stemness, differentiation potential and cytokine profile. We have successfully isolated UDSCs from 25 urine samples. First colonies emerged up to 9 days after the initial seeding. Cell doubling time was 45 ± 0.24 SD, and when seeded at the density of 100 cells/cm², they formed 42 ± 6.5 SD colonies within 10 days. Morphological analyzes revealed that two different types of the cell populations have been present. The first type had a rice-grain shape and the second one was characterized by a polyhedral shape. In several cell cultures, dome-shaped cells were observed as well. All examined UDSCs expressed typical MSC-like surface markers, CD73, CD90 and CD105. Moreover, conditioned media from UDSCs were harvested, and cytokine profile has been evaluated showing a significantly higher secretory rate of IL-8, IL-6 and chemokines MCP-1 and GM-CSF. We have also successfully induced human UDSCs into chondrogenic, osteogenic and myogenic cell lineages. Our findings indicate that UDSCs might have immense potential in the regeneration of the damaged tissues.     

Konformation und Rotationsbarrieren substituierter Glyoxylsäureamide

Conformation and Rotation Barriers of Substituted Glyoxylic Acid Amides Semiempirical calculations predict an orthogonal orientation of the carbonyl groups in tertiary glyoxylic acid amides, which is in good agreement with an X-ray structure analysis of 5 . Due to the influence of the α-carbonyl group, the rotation barrier in the substituted glyoxylic acid amides 2a–d, 3a, 3b , and 4–6 (ΔG^#_c = 84–92 kJ mol⁻¹) is about 10 kJ/mol higher than in simple acid amides, as was found by dynamic NMR line shape analysis.