Allele-specific amplification for preimplantation genetic diagnosis (PGD) of spinal muscular atrophy

We have developed a new allele-specific amplification method for the preimplantation genetic diagnosis (PGD) of spinal muscular atrophy (SMA; Werdnig-Hoffmann disease) from a single cell. This method is based on the detection of the deletion of exon 7 of the telomeric copy of the survival motor neurone (SMN^t) gene. An oligonucleotide was designed to be specific to the SMN^t nucleotidic sequence with exonic mismatch G (for SMN^t)→A (for SMN^c) at its 3′ end. This test produces reliable PCR products in 95% of single lymphoblasts (85/88) tested as well as in 16/16 blastomeres from normal controls. Specificity analysis showed that we were able to detect homozygous deletion of the SMN^t gene in 99% of single lymphoblasts (103/104) from a SMA patient. No contamination was detected in 68 blanks tested. Multiple cell and DNA dilution analysis revealed that the test is accurate and specific up to 100 pg DNA and should thus also be suitable for PGD at the blastocyst stage. This rapid procedure requires a single round of fluorescent PCR and no restriction digestion, while previously described single cell methods include nested PCR followed by restriction enzyme digestion. Two PGD cycles for SMA using this procedure were performed in our centre. Copyright © 2001 John Wiley & Sons, Ltd.     

Choosing a sustainable demolition waste management strategy using multicriteria decision analysis

This paper presents an application of the ELECTRE III decision-aid method in the context of choosing a sustainable demolition waste management strategy for a case study in the city of Lyon, France. This choice of waste management strategy takes into consideration the sustainable development objectives, i.e. economic aspects, environmental consequences, and social issues. Nine alternatives for demolition waste management were compared with the aid of eight criteria, taking into account energy consumption, depletion of abiotic resources, global warming, dispersion of dangerous substances in the environment, economic activity, employment, and quality of life of the local population. The case study concerned the demolition of 25 buildings of an old military camp. Each alternative was illustrated with different waste treatments, such as material recovery, recycling, landfilling, and energy recovery. The recommended solution for sustainable demolition waste management for the case study is a selective deconstruction of each building with local material recovery in road engineering of inert wastes, local energy recovery of wood wastes, and specific treatments for hazardous wastes.     

Sorption of polycyclic aromatic hydrocarbons (PAHs) to low and high density polyethylene (PE)

Background, aim, and scope

According to their high sorption capacity polyethylene (PE) passive samplers are often used for the analysis of polycyclic aromatic hydrocarbons (PAHs) in the aquatic environment. PE is also one of the primary synthetic polymers found in oceans, and sorption of PAHs to marine PE debris may determine PAH exposure and therefore hazards in marine ecosystems. Thus, an understanding of the sorption process is of great importance. In the present study, the sorption of several PAHs with different polarities to low density polyethylene (LDPE) and high density polyethylene (HDPE) was studied in order to improve our understanding of the influence of material properties on the Fickian diffusion of PAHs into PE.

Materials and methods

Batch sorption experiments were performed with aqueous solutions containing acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, and LPDE or HDPE pellets. Samples were shaken in the dark at 20?±?1°C for 16 time intervals within one week. Concentrations of PAHs were determined in the aqueous samples using solid-phase microextraction coupled with gas chromatography–mass spectrometry. The distribution coefficients (K _PE) between PE and water were estimated from different models reported in the literature. Kinetic sorption of the PAHs into the plastic pellets was described by a diffusion model based on Fick’s second law in spherical coordinates.

Results and discussion

A comparison between different models describing the equilibrium distribution of PAHs between PE and water revealed that the sorption equilibrium seemed to be driven by parameters other than, or in addition to, organic carbon. For both plastic types, diffusion coefficients decreased while the molecular weight of the PAHs increased which indicates a hindered diffusion through the matrix as a result of a larger molecule size. Higher diffusion coefficients were derived for LPDE than for HDPE indicating a greater sorption velocity for LPDE according to the lower polymer density.

Conclusions

Our results revealed that equilibrium time could be shortened during passive sampling as polymer membranes of lower density are used. In some areas, marine ecosystems may not be in equilibrium with respect to concentrations of organic contaminants and abundance of marine plastic debris. In such cases, different polymer densities should be taken into account in risk assessments.

     

Bacteriolytic Bacillus species isolated from brackish waters of the Southern Baltic Sea

The present study examines antagonistic relationships between different microorganisms inhabiting brackish water and includes a systematic screening for bacteriolytic prokaryotes in the brackish waters of the Southern Baltic Sea (Nordruegenscher Bodden) sampled in July/August 2009. Ten of the 35 marine bacteriolytic isolates belong to the genus Bacillus. Five isolates (B. pumilus, B. subtilis, B. megaterium, B. licheniformis) lysed living microbial cells such as the bacteria Arthrobacter citreus, Arthrobacter protophormiae, Micrococcus luteus, Pseudomonas putida and the yeast Trichosporon mucoides. These and other bacteria (Aeromonas sp., Bacillus subtilis, Chromobacterium violaceum, Citrobacter freundii, Enterobacter aerogenes, Proteus mirabilis, Pseudomonas aeruginosa and Serratia marcescens) were also lysed as autoclaved and pasteurized cells on agar plates. One isolate of Bacillus pumilus showed a distinct bacteriolysis activity against pasteurized cells of A. citreus in liquid culture. Our results suggest that Bacillus species may play a role as opportunistic predators in the marine microbial food web.     

Predictable annual mass release of gametes by the coral reef sponge Neofibularia nolitangere (Porifera: Demospongiae)

Mass release of gametes of the sponge Neofibularia nolitangere (Duch. & Mich., 1864) occurs simultaneosly along the leeward coast of Curaçao over a period of three subsequent days, without any apparent spatial pattern in the sperad of activity. A population of 99 individual sponges was monitored from August through November 1984 for development and subsequent release of gametes. Release started every day of 3 3-d period (12 to 14 October) at about 1400 hrs and lasted until just after sunset (1830 hrs). Ninety percent of the population showed reproductive activity. Exactly one lunar month later (11 to 12 November), a second release of gametes occurred. In the following year the same sequence of events was observed for the original population (2 to 4 October and 1 to 2 November, 1985). In all instances the first gamete release began on the third day after the full moon. These and earlier observations on this phenomenon show a strong correlation between moon phase and the time of gamete release. Histological and field observations show separate development and release of gametes (sex-ratio males: females 1.5:1) with external fertilization. No sex-reversal occurred in the population during release activity the following year, demonstrating a clear gonochoristic and oviparous type of reproduction for N. nolitangere. First development of oocytes and spermatocytes has been observed at 36 and 7 d, respectively, before the date of first release. In both males and females, a major part of the mesohyl of the total sponge was involved in gamete development. Spermatozoa are released through the osculum as a dense white smoke; eggs consisting of oocyte-nurse cell globules, reinforced with spicules, are separately released en masse with the outgoing water stream of the female sponge. Shortly after release the eggs become sticky and show a negative buoyancy. The reproductive strategy of N. nolitangere appears to be directed at maximizing the number of surviving recruits by maintaining a high reproductive output at a short specific time interval.     

The political significance of local agenda 21: The early stages of some European experience

Sustainable development, as it emerged in Agenda 21 from the Rio conference in 1992, will only be meaningful when it touches the lives of ordinary people; then it becomes a reality. Local Agenda 21 (LA21) seeks to achieve that objective. This article assesses the origins of LA21, reviews its social and political significance, and considers its prospects in the light of case study experience emerging from the UK, Germany and Norway, focusing on the role of local government as a major stakeholder in Agenda 21 (A21). The range of response to LA21 has proved to be varied. A successful transformation to a more sustainable world will require visionary political leadership, supportive administrations, networks of experience sharing, alliances with non-governmental organisations and local industry, and effective community mobilisation. All of that, in turn, requires equally supportive economic and social policy backing from national governments. This article will indicate that, not surprisingly, it is the domestic political context, nationally and locally, which in the main determines the speed and nature of response to LA21, now and in the future. By understanding and being aware of these contexts, factors impeding progress towards LA21 may be addressed, whilst at the same time retaining the diversity of response which is an essential part of local sustainability.