Experience with the Use of Substance Flow Analysis in Denmark

Substance flow analysis (SFA) has been used by the Danish Environmental Protection Agency for more than two decades to identify sources of hazardous substance releases to the environment and to waste streams. More than 35 SFAs have been undertaken using a unified methodology. This article discusses key elements of the methodology and application of the results of the SFAs in Denmark, illustrated by examples. The use of repeated SFAs in the evolution of substance regulation is demonstrated for lead. A cadmium SFA illustrates the use of SFAs to monitor changes in consumption and release of a hazardous substance. Estimates of sources are presented for a number of toxic and resource-intensive metals in solid waste incineration and sewage treatment. Most of the SFAs concern hazardous substances, but aluminum represents a case in which the resource perspective is an important element in a national SFA. The Danish SFA on brominated flame retardants illustrates some methodological problems with conducting national SFAs in a global economy in which the substances are mainly imported in finished products.     

A quick and precise technique for identifying ectomycorrhizas by PCR

A rapid procedure was developed to amplify ITS fragments directly from Tuber ectomycorrhizas either synthesized in a greenhouse or collected from the field. The addition of Bovine Serum Albumin (BSA) to the reaction mixtures overcame the presence of reaction inhibitors present in fungal and root cells, and enabled the amplification of the ITS regions directly from ectomycorrhizal tissues. This method is cheaper and less time-consuming than conventional procedures, and reduces the time required from 1-4 h to a few minutes. It is also much more sensitive, allowing the identification of just a small fragment of a mycorrhizal root tip. Because of this it is possible to select only the target fungal tissue and hence minimise the risk of contamination by saprobic or other mycorrhizal species. The method also avoids the use of toxic or hazardous substances. This method could have a wider application in other areas of applied mycology.     

International Cooperation for Metal Recycling From Waste Electrical and Electronic Equipment

This article addresses a market‐based management concept for waste electrical and electronic equipment (WEEE) known as the “best‐of‐two‐worlds” approach. The concept is based on the idea that recyclers in developing countries and emerging economies can cooperate with technologically advanced refineries in industrialized countries to facilitate efficient recovery of valuable metals, such as gold and palladium, from e‐waste. The article provides an overview of technical and environmental concerns underlying the concept and sheds light on the political framework, the waste‐related trade issues, and the resource economics that need to be considered for further decision making. Building on this synthesis, I conduct a qualitative assessment of sustainability impacts of the proposed concept by analyzing two scenarios and their associated risks. The analysis suggests that, under certain preconditions, the best‐of‐two‐worlds concept could yield significant improvements in terms of management of hazardous substances, resource efficiency, greenhouse gas emissions, income generation, and investments into social and environmental standards. Generally, two potential implementation scenarios were identified: Whereas under Scenario 1 only WEEE generated within developing countries and emerging economies is managed through the best‐of‐two‐worlds approach, Scenario 2 additionally incorporates WEEE imported from industrialized countries. Although both scenarios can yield a variety of benefits, Scenario 2 might cause a net flow of hazardous substances from industrialized countries into developing countries and emerging economies, thus leading to less beneficial sustainability impacts.     

Hazard Reduction in Nanotechnology

The release of hazardous substances is a matter of concern for nanotechnology. This may include some nanoparticles, reactants, by‐products, and solvents. The use of low‐hazard solvents may reduce the hazards from nanoparticle production and nanomaterial processing. The hazards of inorganic nanoparticles may be reduced by modifying their chemical composition, surface characteristics, or structure. In nanomedicine, optimizing the balance between persistence and excretion and preventing the release of toxic degradation products may reduce hazard. In applications of fixed inorganic nanoparticles, the focus should be on preventing the release of such particles and of hazardous compounds during the product life cycle. When, after exhaustion of known hazard reduction options, significant hazard remains, other approaches merit consideration.     

Influence of Circadian Typology on Drug Consumption,Hazardous Alcohol use,and Hangover Symptoms

Few studies have focused on the influence of circadian typology on drug use, and none has considered the use of illegal drugs and hazardous alcohol consumption. This study analyzes the influence of circadian typology on several types of drug consumption (habitual or sporadic), hangover symptoms (past 12 mos), and, more specifically, hazardous alcohol consumption of young adults. Five hundred seventeen university students (173 males), between 17 and 30 yrs of age, answered the Composite Scale of Morningness (CSM), the Alcohol Use Disorders Identification Test (AUDIT), and a self-referred questionnaire on drug consumption during the previous month and on the prevalence of different hangover symptoms during the previous year. Our results confirm a higher prevalence of consumption of addictive substances, both legal (nicotine and cola drinks) and illegal (cannabis and ecstasy), in evening- compared to morning- and neither-type subjects (p?<?.001 in all cases). Evening-type subjects also obtained a higher total score on the AUDIT (p?<?.001) and showed a greater prevalence in the subscales of potential alcohol problems (p?<?.02), as well as more frequent different hangover symptoms (learning difficulties, thirst, tiredness, headaches, sensorial hypersensitivity, anxiety, and irritability; p?<?.04 in all cases) compared with morning- and neither-type subjects, except for sensorial hypersensitivity and anxiety, for which the evening-type did not differ from the neither-type. Our results provide substantial evidence that the evening circadian typology is a risk factor for the development of drug consumption and that it should be taken into account both in preventive and treatment approaches. Moreover, the data regarding hazardous alcohol use and hangover symptoms emphasize the need to include circadian typology in future studies on the pattern of heavy episodic drinking. (Author correspondence: gprat@ub.edu)     

Invited contribution: Acute toxicity testing,public responsibility and scientific challenges

Knowledge of the acute toxic effects of chemicals is important for the protection of exposed humans. Since sufficient information in humans is often lacking, experiments on laboratory animals must be performed. The LD₅₀ test, which requires large numbers of animals, has become the preferred procedure. It is now widely criticized on scientific and ethical grounds. This paper reviews the possibilities of using fewer animals to obtain relevant information on the acute hazards of chemical substances, but it also identifies the reasons why the traditional testing approaches cannot be changed immediately. An important problem is the practice of basing legal decisions on classification of chemicals in official lists of hazardous substances and for labeling purposes on LD₅₀ values. Proposals are presented on how pain and suffering of the animals included in acute toxicity tests can be reduced. The use of in vitro systems for the evaluation of the hazardous properties of chemicals is discussed.Abbreviations LD₅₀ lethal dose 50%, median lethal dose     

Considering human toxicity as an impact category in life cycle assessment

Characterization of toxic chemicals with relevance to human exposure does normally not belong to Life Cycle Assessments (LCA) and is still a topic of research. The concept of hazard potential classes proposed in this paper is primarily based on threshold limit values that are considered to be a measure of the severity of potential effects. In the absence of threshold limit values the R-phrases of the ordinance of dangerous substances are used. Substances are assigned to five hazard potential classes (A to E). Potentially dangerous chemicals are identified and substances of low toxicological relevance are excluded from further evaluation. The location where a probable exposure might occur (indoor versus outdoor) and inter-media transport of substances is considered. The product comparison is based both on the results of the proposed “semi-quantitative screening method” and on toxicological expert knowledge.     

Identification of a novel multiple environmental factor‐responsive 1‐aminocyclopropane‐1‐carboxylate synthase gene,NT‐ACS2, from tobacco

Many abiotic environmental factors elicit the production of stress‐ethylene in higher plants. To elucidate the molecular mechanisms underlying the regulation of stress‐ethylene production and the physiological roles played by stress‐ethylene in stress responses of plants, we studied the gene expression of ACC synthase in tobacco plants that had been subjected to environmental stresses. Four new tobacco ACC synthase cDNA fragments, NT‐ACS2, NT‐ACS3, NT‐ACS4 and NT‐ACS5, were identified and sequenced. It was found that NT‐ACS2 could be induced by wounding, cold temperature and, especially, sunlight. NT‐ACS4 was induced at a faster kinetics by wounding. The multiple environmental stress‐responsive (MESR) NT‐ACS2 gene was found to contain three introns and four exons and encode a polypeptide of 484 amino acids, 54·6 kDa and pI 6·87. Computer analysis of the 3·4 kb 5 ′ flanking region upstream of the ACS coding region revealed the existence of a group of putative cis‐acting regulatory elements potentially conferring wounding, chilling, and UV light inducibility. Phylogenetic analysis of ACC synthase genes of different plant origins indicated that the chill‐inducible NT‐ACS2 gene is closely related to a chilling‐inducible citrus ACS gene.     

New insights into polyurethane biodegradation and realistic prospects for the development of a sustainable waste recycling process

Polyurethanes are polymeric plastics that were first used as substitutes for traditional polymers suspected to release volatile organic hazardous substances. The limitless conformations and formulations of polyurethanes enabled their use in a wide variety of applications. Because approximately 10 Mt of polyurethanes is produced each year, environmental concern over their considerable contribution to landfill waste accumulation appeared in the 1990s. To date, no recycling processes allow for the efficient reuse of polyurethane waste due to their high resistance to (a)biotic disturbances. To find alternatives to systematic accumulation or incineration of polyurethanes, a bibliographic analysis was performed on major scientific advances in the polyurethane (bio)degradation field to identify opportunities for the development of new technologies to recondition this material. Until polymers exhibiting oxo- or hydro-biodegradative traits are generated, conventional polyurethanes that are known to be only slightly biodegradable are of great concern. The research focused on polyurethane biodegradation highlights recent attempts to reprocess conventional industrial polyurethanes via microbial or enzymatic degradation. This review describes several wonderful opportunities for the establishment of new processes for polyurethane recycling. Meeting these new challenges could lead to the development of sustainable management processes involving polymer recycling or reuse as environmentally safe options for industries. The ability to upgrade polyurethane wastes to chemical compounds with a higher added value would be especially attractive.     

Medicinal and pharmaceutical uses of seaweed natural products: A review

In the last three decades the discovery of metabolites with biological activities from macroalgae has increased significantly. However, despite the intense research effort by academic and corporate institutions, very few products with real potential have been identified or developed. Based on Silverplatter MEDLINE and Aquatic Biology, Aquaculture & Fisheries Resources databases, the literature was searched for natural products from marine macroalgae in the Rhodophyta, Phaeophyta and Chlorophyta with biological and pharmacological activity. Substances that currently receive most attention from pharmaceutical companies for use in drug development, or from researchers in the field of medicine-related research include: sulphated polysaccharides as antiviral substances, halogenated furanones from Delisea pulchra as antifouling compounds, and kahalalide F from a species of Bryopsis as a possible treatment of lung cancer, tumours and AIDS. Other substances such as macroalgal lectins, fucoidans, kainoids and aplysiatoxins are routinely used in biomedical research and a multitude of other substances have known biological activities. The potential pharmaceutical, medicinal and research applications of these compounds are discussed.     

Nitrate tolerance aggravates postischemic myocardial apoptosis and impairs cardiac functional recovery after ischemia

Objectives: This study examined the effects of nitrate tolerance (NT) on myocardial ischemia reperfusion (MI/R) injury and elucidated the potential mechanisms involved. Furthermore, the effects of GSH on postischemic myocardial apoptosis in NT rats were investigated. Methods and results: Male Sprague–Dawley rats were randomized to receive nitroglycerin (60 μg/kg/h) or saline for 12 h followed by 40 min of MI and 4 h of reperfusion. Myocardial apoptosis, infarct size, nitrotyrosine formation, plasma CK and LDH activity, and cardiac function were determined. MI/R resulted in significant apoptotic cell death, which was further increased in animals with NT. In addition, NT further increased plasma CK and LDH activity, enlarged infarct size, and impaired cardiac functional recovery after ischemia. Myocardial nitrotyrosine, a footprint for cytotoxic reactive nitrogen species formation, was further enhanced in the NT heart after MI/R. Treatment of NT animals with exogenous GSH inhibited nitrotyrosine formation, reduced apoptosis, decreased infarct size, and improved cardiac functional recovery. Conclusion: Our results demonstrate that nitrate tolerance markedly enhances MI/R injury and that increased peroxynitrite formation likely plays a role in this pathologic process. In addition, our results suggest that GSH could decrease peroxynitrite formation and reduce MI/R injury in nitrate tolerant hearts.     

Interaction properties of biosynthesized cadmium sulphide quantum dots with human serum albumin: further investigation of antibacterial activities and sensing applications

The synthesis of small-sized quantum dots (QDs) (1–10 nm) via the green route has garnered great interest regarding their prospective use in many biological applications (diagnosis, drug delivery and in vivo sensing); this is difficult to achieve using chemical synthesis methods, which produce larger size QD particles and also require hazardous reagents. Here, we synthesized biogenic cadmium sulphide (CdS) QDs using green tea extract as the reducing agent to produce particles that were homogeneous and a smaller size of 2–4 nm. We also elucidated the (a) protein binding, (b) antibacterial use and (c) sensing applications of biogenic CdS QDs in this present work. The biosynthesized CdS QDs were found to have extensive antibacterial activity against both Gram-negative Escherichia coli and Gram-positive Enterococcus faecalis bacterial strains. The introduction of QDs in biological medium can lead to the formation of protein–QD complexes; therefore we investigated the binding interaction of CdS QDs with the carrier protein human serum albumin (HSA) in vitro. The synthesized CdS QDs quenched the intrinsic fluorescence of HSA through a static quenching mechanism and the binding constant (K_b) was in the order of 10⁴ M⁻¹. It was also observed that the presence of biogenic CdS QDs affected the HSA–ligand interactions in vitro. The synthesized CdS made highly effective sensors for tetracycline, rifampicin, and bilirubin with limit of detection (LOD) values of 99, 141 and 29 ng/ml, respectively.     

Application of a direct toxicity assessment approach to assess the hazard of potential pesticide exposure at selected sites on the Crocodile and Magalies rivers,South Africa

The potentially hazardous effects of agricultural pesticide usage in the Crocodile (west) Marico catchment were evaluated using the Danio rerio and Daphnia pulex lethality, Selenastrum capricornutum growth inhibition and the Ames mutagenicity plate incorporation assays. Hazard assessment categories are proposed to standardise the output of the different toxicity assessments. Sites were categorised according to the toxicity hazard indicated and were related to the gradients of agricultural land-use intensity at each site. Intensive agricultural sites showed the highest effects to all tested biota. Receiving water at urban sites associated with increased nutrients and lowest pesticide usage showed few adverse effects, while the relatively unimpacted site indicated no hazard to any organism, and only a slight stimulation to algal growth. Weighted hazard scores indicated that the unimpacted sites were least hazardous, falling within a B category, the urban sites were moderately hazardous (C category), and the agricultural sites (D category) had the highest potential impacts on aquatic organisms. This study demonstrated the usefulness of using the hazard assessment approach and the role it could play in assessing site-specific potential toxicity hazards of river water impacted by agrochemicals. It can be used together with other assessment methods, such as biological indices, in a tiered approach.     

The Drive to Lower Lead Levels in Food Chemicals

Abstract

Because chemicals that are intentionally added to food during its production constitute a significant portion of the US diet, reducing lead levels in them is one of the long-term objectives of the Committee on Food Chemicals Codex, a committee of the Food and Nutrition Board within the Institute of Medicine. The Committee recommends limits for lead, as well as for other heavy metals, arsenic, and other potentially hazardous constituents in chemicals used in food production. The Food Chemicals Codex is a compendium of purity specifications that often become legal standards for food chemical purity in the United States and in many countries worldwide. The Committee plans to lower existing lead limits for food chemicals based on their level of consumption or reported use. Data regarding food chemical use is available from the 1987 Poundage and Technical Effects Update of Substances Added to Food. Because of their high level of consumption, sweeteners constitute a group of food chemicals in which lead limits could be lowered. However, the Committee's ability to recommend lower lead limits in these products depends on the availability of a test method capable of measuring lead at lower levels. This Committee intends to make its contribution in the effort to lower dietary intake of lead.     

On the Social Behaviour of Cells

Polystyrene Petri dishes are in use in hundreds of thousands of laboratories world wide. Cell culture experiments performed in them provide fundamental information in a wide range of applications, including but not limited to testing novel biomaterials and pharmaceuticals, and stem cell research. These experiments cost billions of dollars per year. In this study we report on a potential deficiency of polystyrene Petri dishes, possibly caused by an increase in interracial pH under relevant culture condi-tions and affecting cell performance. We conclude that cell performance on Petri dishes could be improved by improving the Petri dishes. As a spin-off of our study we postulate the concept that cancer cells and stem cells are social. It is impossible to validate this concept on the basis of the model established in this paper. However. the coherence of our insights may encourage further study and lead to the development of a qualitative improvement of cell culture devices, including Petri dishes and culture flasks, to the identification of potential strategies for chemotherapy and chemoprevention that could suppress progression of metastasis, and to the establishment of improved settings for tissue engineering and stem cell research. An immediate recom-mendation of our study is to use chemically and biologically inert substrates for important cell culture experiments, for example, nanocrystalline diamond.     

Effect of hypophysectomy, adrenalectomy, pituitary hormone secretion and gastric acid secretion on neurotensin induced gastric protection against stress gastric lesions

In previous studies we have established that intracisternal (i.c.) but not peripheral (intravenous) administration of neurotensin (NT), a brain and gastrointestinal tridecapeptide, totally prevents the development of gastric lesions produced by cold-restraint stress (CRS) with food-deprived rats. In this investigation, removal of the pituitary and adrenal gland, anterior pituitary hormone secretion and gastric acid secretion were evaluated independently as potential intermediates for NT's protective effect. NT (30 micrograms) produced a significant reduction of gastric lesions incidence and severity in intact and sham-operated controls. Adrenalectomy, but not hypophysectomy totally blocked the protective effect of i.c. NT. In addition, replacement therapy with s.c. prednisone (1 mg/kg) for 5 days following adrenalectomy did not restore the protective activity of central (i.c.) NT in adrenalectomized rats. A significant reduction of serum levels of TSH, PRL and GH following i.c. NT (30 micrograms) was observed after 2h of CRS. The gastrosecretory studies revealed that i.c. NT (30 micrograms) did not affect gastric acid secretion in pylorus ligated rats. However, blockade of peripheral (gut) cholinergic (muscarinic) receptors with i.p. atropine methylbromide (1 mg/kg) significantly raised gastric pH and reduced gastric acid concentration and output. In conclusion, the results of this study indicate that the acute protective effect of brain NT appears to be mediated, at least in part, by the sympathoadrenomedullary axis, and not by the pituitary gland or substances derived from the pituitary or by inhibition of gastric acid secretion.     

Methane and nitrous oxide emissions under no‐till farming in China: a meta‐analysis

No‐till (NT) practices are among promising options toward adaptation and mitigation of climate change. However, the mitigation effectiveness of NT depends not only on its carbon sequestration potential but also on soil‐derived CH₄ and N₂O emissions. A meta‐analysis was conducted, using a dataset involving 136 comparisons from 39 studies in China, to identify site‐specific factors which influence CH₄ emission, CH₄ uptake, and N₂O emission under NT. Comparative treatments involved NT without residue retention (NT0), NT with residue retention (NTR), compared to plow tillage (PT) with residue removed (PT0). Overall, NT0 significantly decreased CH₄ emission by ~30% (P < 0.05) compared to PT0 with an average emission 218.8 kg ha⁻¹ for rice paddies. However, the increase in N₂O emission could partly offset the benefits of the decrease in CH₄ emission under NT compared to PT0. NTR significantly enhanced N₂O emission by 82.1%, 25.5%, and 20.8% (P < 0.05) compared to PT0 for rice paddies, acid soils, and the first 5 years of the experiments, respectively. The results from categorical meta‐analysis indicated that the higher N₂O emission could be mitigated by adopting NT within alkaline soils, for long‐term duration, and with less N fertilization input when compared to PT0. In addition, the natural log (lnR) of response ratio of CH₄ and N₂O emissions under NT correlated positively (enhancing emission) with climate factors (temperature and precipitation) and negatively (reducing emission) with experimental duration, suggesting that avoiding excess soil wetness and using NT for a long term could enhance the benefits of NT. Therefore, a thorough understanding of the conditions favoring greenhouse gas(es) reductions is essential to achieving climate change mitigation and advancing food security in China.     

Faba bean grain yield, N2 fixation, and weed infestation in a long-term tillage experiment under rainfed Mediterranean conditions

Background and aims

Long-term experiments could provide valuable information to determine the effects of an agronomic practice on agro-ecosystem productivity and stability. This study evaluated the long-term (18-year) impact of different tillage systems on faba bean (Vicia faba L.) productivity, including weed and broomrape incidence, and N₂ fixation.

Methods

The experiment was carried out on a Vertisol under rainfed Mediterranean conditions. It was set up as a strip-plot design. The tillage systems were: conventional tillage (CT) with moldboard plow, reduced tillage (RT) with chisel plow, and no tillage (NT). Nitrogen fixation was estimated over 2?years in the final phase of the experiment using the ¹⁵N isotope dilution technique.

Results

On average, grain yield was 31?% and 23?% higher under NT than under CT and RT, respectively; however, the effect of tillage varied by year. The yield advantage of NT over CT was pronounced when rainfall was scarce. Average broomrape infestation was lower under NT than under CT, but a reduction in tillage intensity resulted in an increase in weed biomass. Tillage had no effect on weed richness, but the use of the NT technique progressively altered the weed composition by determining conditions conducive to the growth of a few weed species that are hard to control. Nitrogen fixation was strongly influenced by tillage, being higher under NT than under CT.

Conclusions

The results suggest that NT is a valuable option in the rainfed cereal–legume rotation systems of Mediterranean environments; however, the success of NT technique depends on the use of effective weed control strategies.     

DNA methylation levels in porcine fetal fibroblasts induced by an inhibitor of methylation, 5-azacytidine

Removal of the somatic DNA methylation pattern from donor cells and remodeling of embryonic status have been suggested as integral processes for successful nuclear transfer (NT) reprogramming. This study has investigated the effects of 5-azacytidine (5-azaC), a DNA methylation inhibitor, on global methylation changes in porcine fetal fibroblasts (PFF); this may improve NT attributable to the potential reprogramming of the methyl groups. PFF in 5th passage cultures were treated with 0, 0.5, 1.0, 2.0, and 3.0 μM 5-azaC for 96 h; 5-azaC inhibited the growth at all tested concentrations. At the higher concentrations of 5-azaC used, cells appeared to exhibit morphological changes and to become apoptotic as observed by TUNEL assay. Thus, cells were negatively affected by 5-azaC. Differences in cellular ploidy were also observed at higher concentrations. Analysis showed no considerable changes in the proportion of cells at the G₁-phase of the cell cycle with 5-azaC concentrations. The fractional part of the methylated DNA of these cells was significantly reduced by 5-azaC treatment. Confocal microscopy confirmed the inhibition of methylation levels in PFF with increased concentrations of 5-azaC. Exposure to 5-azaC altered the expression of genes involved in imprinting (IGF2) or pro-apoptosis (BAX), whereas there was a reduction in the expression of the main enzyme responsible for replicating the DNA methylation pattern (DNMT1) and anti-apoptosis (BCL2L1). Therefore, 5-azaC induces a relative reduction in methylation in PFF, and cells treated with 0.5 μM 5-azaC may have enhanced potential for porcine NT.The financial support of BioGreen 21 (grant no. 100052004002000) and KOSEF (grant no. R05-2004-000-10702-0) in Korea is gratefully acknowledged.