Polymerase chain reaction analysis of the cystic fibrosis ΔF508 mutation in human blastomeres following oocyte injection of a single sperm from a carrier

The efficiency of the polymerase chain reaction (PCR) in detecting the cystic fibrosis (CF) ΔF508 mutation (which is the most common mutation of CF) was assessed in single human blastomeres. Twenty-one human immature oocytes (germinal-vesicle-stage oocytes) that had been donated for research were matured in vitro and a single spermatozoon from a carrier of the CF ΔF508 mutation was injected into the ooplasm. Fourteen embryos were obtained after intracytoplasmic sperm injection (ICSI). PCR analysis was carried out on 70 single blastomeres isolated from these 14 embryos. The results showed that the efficiency of DNA amplification by PCR in single nucleate blastomeres was 94 per cent (59/63). There were no false-positive results since none of the blank samples or the blastomeres without a nucleus showed an amplified signal. We found that nine embryos were homozygous for the unaffected genotype and that four embryos were heterozygous since they contained both the unaffected and the ΔF508 genotype. In a four-cell embryo, we observed the homozygous unaffected genotype in one blastomere and a heterozygous ΔF508/unaffected genotype in the other three blastomeres.     

Biocatalytic dechlorination of hexachlorocyclohexane by immobilized bio-Pd in a pilot scale fluidized bed reactor

Lindane (??-hexachlorocyclohexane, ??-HCH) is a recalcitrant and toxic organochlorine insecticide. Due to its non-selective production process and widespread use, HCH isomers and their degradation products have been detected frequently in soils and groundwater. An innovative technology using microbial produced Pd(0) nanoparticles, i.e. bio-Pd, was developed to treat groundwater containing a mixture of HCHs and chlorobenzenes. In a first step, the groundwater was de-ironized and most of the chlorobenzenes were removed in a biological trickling filter. The ??g?L^?1 levels of HCHs and chlorobenzenes were removed in a second step by the bio-Pd-based technology. Therefore, a 200-L pilot scale reactor was developed with 100?mg?L^?1 bio-Pd encapsulated in alginate beads. Hydrogen gas was bubbled at the bottom of the reactor and served to charge the bio-Pd catalyst. The reactor influent contained 5.2???g?L^?1 HCHs and 51.1???g?L^?1 chlorobenzenes. During a test period of 10?days, 29% of the HCH isomers and 63% of the chlorobenzenes were removed applying a nominal hydraulic residence time of 4?h. These removal percentages could be increased to 75 and 68% by doubling the nominal hydraulic residence time to 8?h. This study demonstrated that biologically produced nanoparticles of Pd can be applied for the large-scale remediation of groundwater contaminated with HCHs.     

Sources of PCDD/Fs, non-ortho PCBs and PAHs in sediments of high and low impacted transboundary rivers (Belgium-France)

PAHs, PCDD/Fs and non-ortho PCBs have been assessed in Yser and Upper-Scheldt river sediments. Higher contamination levels were observed in the Upper-Scheldt sediments: maximum concentrations for the 16 US-EPA PAHs, PCDD/Fs and non-ortho PCBs respectively amount to 8.9 mg kg⁻¹, 12 ng TEQ kg⁻¹ and 5.1 ng TEQ kg⁻¹. Diagnostic PAH ratios in sediments and atmospheric samples suggest that the PAH compounds are from pyrolytic origin, more specifically combustion processes. The huge consumption of coal in cokes-ovens and smelters and its use for house-heating in Northern France, although decreasing during the last decades, are in support of that suggestion. PCDD/F fingerprints in sediments and deposition material indicate that OCDD is the dominant congener. In addition use of pentachlorophenol (PCP) in the past led to a minor contribution of PCDD/Fs in our sediment samples. Non-ortho PCBs form a substantial fraction of the total TEQ concentrations observed in the sediments. Since the 1980s and 1990s a substantial reduction of the PCDD/F sediment concentrations is observed, but this is not the case for the PAHs.     

The relation between the estimated dietary intake of PCDD/Fs and levels in blood in a Flemish population (50-65 years)

Dioxin-like activity was measured in the serum of 1425 Flemish men and women via the CALUX assay. The adults, aged between 50 and 65 years, participated in a large biomonitoring program, executed by the Flemish Center of Expertise for Environment and Health between 2002 and 2006. Within the context of this biomonitoring program also dietary intake of dioxin-like contaminants was assessed through a food frequency questionnaire. The relation between the estimated dietary intake and the dioxin-like activity in serum was evaluated using multivariate analyses: a logistic model was performed on the total population, while a linear regression analysis was done on the subsample with quantifiable dioxin activity levels in serum. Region, gender, age, BMI, smoking status, as well as dietary habits were entered in the model, with dioxin level as an outcome estimate. Both the logistic and linear model confirmed the contribution of dietary intake to the dioxin activity measured in serum. Also BMI and region were found to be associated with dioxin activity levels.     

Use of monitoring data to support conservation management and policy decisions in Micronesia

Adaptive management implies a continuous knowledge‐based decision‐making process in conservation. Yet, the coupling of scientific monitoring and management frameworks remains rare in practice because formal and informal communication pathways are lacking. We examined 4 cases in Micronesia where conservation practitioners are using new knowledge in the form of monitoring data to advance marine conservation. These cases were drawn from projects in Micronesia Challenge jurisdictions that received funding for coupled monitoring‐to‐management frameworks and encompassed all segments of adaptive management. Monitoring in Helen Reef, Republic of Palau, was catalyzed by coral bleaching and revealed evidence of overfishing that led to increased enforcement and outreach. In Nimpal Channel, Yap, Federated States of Micronesia (FSM), monitoring the recovery of marine food resources after customary restrictions were put in place led to new, more effective enforcement approaches. Monitoring in Laolao Bay, Saipan, Commonwealth of the Northern Mariana Islands, was catalyzed by observable sediment loads from poor land‐use practices and resulted in actions that reduced land‐based threats, particularly littering and illegal burning, and revealed additional threats from overfishing. Pohnpei (FSM) began monitoring after observed declines in grouper spawning aggregations. This data led to adjusting marine conservation area boundaries and implementing market‐based size class restrictions. Two themes emerged from these cases. First, in each case monitoring was conducted in a manner relevant to the social and ecological systems and integrated into the decision‐making process. Second, conservation practitioners and scientists in these cases integrated culturally appropriate stakeholder engagement throughout all phases of the adaptive management cycle. More broadly, our study suggests, when describing adaptive management, providing more details on how monitoring and management activities are linked at similar spatial scales and across similar time frames can enhance the application of knowledge.     

Polycyclic aromatic hydrocarbon release from a soil matrix in the in vitro gastrointestinal tract

Soil ingestion is an important exposure route by which immobile soil contaminants enter the human body. We assessed polycyclic aromatic hydrocarbon (PAH) release from a contaminated soil, containing 49 mg PAH kg(-1), using a SHIME (Simulator of the Human Intestinal Microbial Ecosystem) reactor comprising the stomach, duodenal, and colon compartments. Polycyclic aromatic hydrocarbon release was defined as that fraction remaining in the digest supernatant after centrifugation for 5 min at 1500 x g. The PAH release in the stomach digest was only 0.44% of the total PAH present in soil, resulting in PAH concentrations of 23 micrograms PAH L(-1) chyme. The lower PAH releases in duodenum (0.13%) and colon (0.30%) digests, compared with the stomach digest, were thought to be attributed to combined complexation and precipitation with bile salts, dissolved organic matter, or colon microbiota. We studied these complexation processes in an intestinal suspension more in depth by preparing mixtures of 9-anthracenepropionic acid, a Bacillus subtilis culture, and cholin as model compounds for PAHs, organic matter, and bile salts, respectively. Bile salts or organic matter in the aqueous phase initially enhance PAH desorption from soil. However, desorbed PAHs may form large aggregates with bile and organic matter, lowering the freely dissolved PAH fraction in the supernatant. Using the model compounds, mathematical equations were developed and validated to predict PAH complexation processes in the gastrointestinal tract. Contaminant release and subsequent complexation in the gut is an important prerequisite to intestinal absorption and thus bioavailability of that contaminant. The data from this research may help in understanding the processes to which PAHs are subjected in the gastrointestinal tract, before intestinal absorption.