Duplex,triplex and quadruplex PCR for the preimplantation genetic diagnosis (PGD) of cystic fibrosis (CF), an exhaustive approach

Most of cystic fibrosis (CF) pre-implantation genetic diagnosis (PGD) cases described to date are limited to the detection of ΔF508. Beside this predominant mutation, over 1000 mutations have been identified, rendering the development of a mutation-based PGD protocol impracticable. This is the reason why we, as well as the others, have developed PGD strategies on the basis of the identification of the pathogenic haplotype instead of the mutation(s). In a previous article, we reported the conditions for the co-amplification of two intragenic polymorphic markers and the F508 locus. Here we describe an improved protocol allowing the additional amplification of two new intragenic markers, intron 1 CA repeat (I1CA) and IVS17bTA. This new protocol should, theoretically, allow us to provide a diagnosis to all couples requiring PGD for CF. Using single lymphoblasts, we have tested four different PCR configurations, including one duplex, two triplexes and one quadruplex PCR. All of them gave results compatible with a clinical application. The number of single lymphoblasts tested in each series varied from 89 to 155. PCR efficiency ranged from 95.4 to 100%. A complete haplotype was achieved for 83.2 to 90.7% of the tested cells, with an allele drop out (ADO) rate comprised between 6.0 and 11.6%. We present here three cases that we performed either with the former test (one case using the triplex PCR combining F508, IVS8CA and IVS17bCA) or with the new one (one case using the triplex combining F508, I1CA and IVS17bTA and one case using a quadruplex test). We obtained two single pregnancies. Copyright © 2004 John Wiley & Sons, Ltd.     

Soil organic horizons as a major source for radiocesium biorecycling in forest ecosystems

Here we review some of the main processes and key parameters affecting the mobility of radiocesium in soils of semi-natural areas. We further illustrate them in a collection of soil surface horizons which largely differ in their organic matter contents. In soils, specific retention of radiocesium occurs in a very small number of sorbing sites, which are the frayed edge sites (FES) born out of weathered micaceous minerals. The FES abundance directly governs the mobility of trace Cs in the rhizosphere and thus its transfer from soil to plant. Here, we show that the accumulation of organic matter in topsoils can exert a dilution of FES-bearing minerals in the thick humus of some forest soils. Consequently, such accumulation significantly contributes to increasing 137Cs soil-to-plant transfer. Potassium depletion and extensive exploration of the organic horizons by plant roots can further enhance the contamination hazard. As humus thickness depends on both ecological conditions and forest management. our observations support the following ideas: (1) forest ecosystems can be classified according to their sensitivity to radiocesium bio-recycling, (2) specific forest management could be searched to decrease such bio-recycling.     

Use of commercial plant species in a hydroponic system to treat domestic wastewaters

The objectives in this work were to investigate a conceptual layout for an inexpensive and simple system that would treat primary municipal wastewater to discharge standards. A commercial hydroponic system was adapted for this study and the wastewater was used to irrigate wooly digitalis (Digitalis lanata Ehrh.) and foxglove (Digitalis purpurea L.). These plants are medicinal and produce cardenolide compounds. Influent and effluent samples were collected once a month for six months and analyzed to determine the various parameters relating to water quality. The legal discharge levels for total suspended solids (SS), biochemical oxygen demand (BOD5), and chemical oxygen demand (COD) were reached for the two tested plants after 48 h of wastewater treatment; the removal was 82, 93, and 79%, respectively, for wooly digitalis and 92, 92, and 84%, respectively, for foxglove. Similar results were obtained during a 6-mo period although the sewage composition varied widely. The system tended to be unable to remove N and P to concentrations below regulated levels. Compared with the nutrient solution composition, the wastewater was more concentrated in Na+ and Cl- and less in N, K+, and Ca2+. These variations can lead to the decline of wooly digitalis plants. Foxglove developed a significant root system to increase mineral absorption wastewater being used as the unique nutritive source. After 10 wk all the wooly digitalis seedlings were dead. Despite this fact, however, the root system remained in place for a significant time (< 4 mo), thus continuing to filter wastewater and to be used as a bacterial support thus making it possible to have a security period to replace the dead plants.     

A Retrospective Isotopic Study of Spruce Decline in the Vosges Mountains (France)

The objective of this study was to assess the time variation of mineral and water stress levels across the life of a declining, Mg-deficient, spruce stand, in order to clarify the factors that caused the decline. Since 1985, strong soil acidification linked to a large leaching of nitrate and base cations was measured at the study site. In 1994, 5 trees were felled and tree rings were measured and analysed for Ca, Mg, K, Sr, ¹³C¹²C and ⁸⁷Sr/⁸⁶Sr isotopic ratios. Strontium pools and fluxes as well as root Sr isotope ratio in relation to depth were also measured. Wood chemical concentrations and isotope ratios were strongly related to the dominance status of each tree. On average during the study period, the ⁸⁷Sr/⁸⁶Sr ratio of spruce wood decreased. Using a mechanistic model computing long term variations of ⁸⁷Sr/⁸⁶Sr ratio in trees and soils, we reproduced the observed trend by simulating soil acidification – increasing Sr drainage from the whole profile, and particularly from the organic horizon –, and root uptake becoming more superficial with time. Between 1952 and 1976, tree ring ¹³C decreased strongly and continuously, which, in addition to other factors, might be related to an increase in water stress. Thus, a decrease in rooting depth, possibly related to soil acidification, appeared as a possible cause for the long term increase in water stress. The extreme drought event of 1976 appears to have revealed and triggered the decline.     

Suspended particulate matter determines physical speciation of Fe,Mn, and trace metals in surface waters of Loire watershed

This study investigates the spatiotemporal variability of major and trace elements, dissolved organic carbon (DOC), total dissolved solids (TDS), and suspended particulate matter (SPM) in surface waters of several hydrosystems of the Loire River watershed in France. In particular, this study aims to delineate the impact of the abovementioned water physicochemical parameters on natural iron and manganese physical speciation (homoaggregation/heteroaggregation) among fine colloidal and dissolved (<?10 nm), colloidal (10–450 nm) and particulate (>?450 nm) phases in Loire River watershed. Results show that the chemistry of the Loire River watershed is controlled by two end members: magmatic and metamorphic petrographic context on the upper part of the watershed; and sedimentary rocks for the middle and low part of the Loire. The percentage of particulate Fe and Mn increased downstream concurrent with the increase in SPM and major cations concentration, whereas the percentage of colloidal Fe and Mn decreased downstream. Transmission electron microscopy analyses of the colloidal and particulate fractions (from the non-filtered water sample) revealed that heteroaggregation of Fe and Mn rich natural nanoparticles and natural organic matter to the particulate phase is the dominant mechanism. The heteroaggregation controls the partitioning of Fe and Mn in the different fractions, potentially due to the increase in the ionic strength, and divalent cations concentration downstream, and SPM concentration. These findings imply that SPM concentration plays an important role in controlling the fate and behavior of Fe and Mn in various sized fractions.

Physical speciation by heteroaggregation of (Fe-Mn) compounds: high [SPM]?→?[Fe-Mn] particulate faction; low {SPM]?→?[Fe-Mn] colloid-dissolved fraction.

     

Cloudwater studies at a high elevation site in the Vosges Mountains (France)

Cloud and rainwater samples have been collected at a high elevation site in the Vosges Mountains. An automated collection system has been used to collect bulk cloudwater and small cloudwater droplets. Bulk cloudwater concentrations were up to 10 times more concentrated than rainwater concentrations. Small clouddroplets showed generally higher concentrations than bulk cloudwater. Nevertheless, the enrichment factors depend on the compounds under study and appear to be related to the composition of the cloud condensation nuclei forming small or large clouddroplets. Principal component analysis and factor analysis were applied to the collected datasets and confirmed the influence of the cloud condensation nuclei on the composition difference between small and large cloudwater droplets.     

Europe-wide survey of estrogenicity in wastewater treatment plant effluents: the need for the effect-based monitoring

A pan-European monitoring campaign of the wastewater treatment plant (WWTP) effluents was conducted to obtain a concise picture on a broad range of pollutants including estrogenic compounds. Snapshot samples from 75 WWTP effluents were collected and analysed for concentrations of 150 polar organic and 20 inorganic compounds as well as estrogenicity using the MVLN reporter gene assay. The effect-based assessment determined estrogenicity in 27 of 75 samples tested with the concentrations ranging from 0.53 to 17.9 ng/L of 17-beta-estradiol equivalents (EEQ). Approximately one third of municipal WWTP effluents contained EEQ greater than 0.5 ng/L EEQ, which confirmed the importance of cities as the major contamination source. Beside municipal WWTPs, some treated industrial wastewaters also exhibited detectable EEQ, indicating the importance to investigate phytoestrogens released from plant processing factories. No steroid estrogens were detected in any of the samples by instrumental methods above their limits of quantification of 10 ng/L, and none of the other analysed classes of chemicals showed correlation with detected EEQs. The study demonstrates the need of effect-based monitoring to assess certain classes of contaminants such as estrogens, which are known to occur at low concentrations being of serious toxicological concern for aquatic biota.     

21,000 Years of Ethiopian African monsoon variability recorded in sediments of the western Nile deep-sea fan

The Nile delta sedimentation constitutes a continuous high-resolution record of Ethiopian African monsoon (EAM) regime intensity. Multi-proxy analyses performed on hemipelagic sediments deposited on the Nile deep-sea fan allow the quantification of the Saharan aeolian dust and the Blue/White Nile River suspended matter frequency fluctuations during the last 21,000 years. The radiogenic strontium and neodymium isotopes, clay mineralogy, elemental composition and preliminary palynological analyses reveal large changes in source components, oscillating between a dominant aeolian Saharan contribution during the Last Glacial Maximum (LGM) and the late Holocene (~4,000–2,000 years), a dominant Blue/Atbara Nile River contribution during the early Holocene (15,000–8,000 years) and a probable White Nile River contribution during the middle Holocene (8,000–4,000 years). The following main features are highlighted: (1) The rapid shift from the LGM arid conditions to the African Humid Period (AHP) started at about 15,000 years. The AHP extends until 8,000 years, and we suggest that the EAM maximum between 15,000 and 8,000 years is responsible for a larger Blue/Atbara Nile sediment load and freshwater input into the eastern Mediterranean Sea. (2) The transition between the AHP and the arid late Holocene is gradual and occurs in two main phases between 8,400–6,500 years and 6,500–3,200 years. We suggest that the main rain belt shifted southward from 8,000 to ~4,000 years and was responsible for progressively reduced sediment load and freshwater input into the eastern Mediterranean Sea. (3) The aridification along the Nile catchments occurred from ~4,000 to 2,000 years. This dry period, which culminates at 3,200 year, seems to coincide with a re-establishment of increased oceanic primary productivity in the western Mediterranean Sea. Such a pattern imposes a large and rapid northward shift of the rain belt over the Ethiopian highlands (5–15°N) since 15,000 years. Precipitation over Ethiopia increased from 15,000 to 8,000 years. It was followed by a gradual southward shift of the rain belt over the equator from 8,000 to 4,000 years and finally a large shift of the rain belt south the equator between 4,000 and 2,000 years inducing North African aridification. We postulate that the decrease in thermohaline water Mediterranean circulation could be part of a response to huge volumes of freshwater delivered principally by the Nile River from 15,000 to 8,000 years in the eastern Mediterranean.     

Effects of chronic exposure to radiofrequency electromagnetic fields on energy balance in developing rats

The effects of radiofrequency electromagnetic fields (RF-EMF) on the control of body energy balance in developing organisms have not been studied, despite the involvement of energy status in vital physiological functions. We examined the effects of chronic RF-EMF exposure (900 MHz, 1 V?m^?1) on the main functions involved in body energy homeostasis (feeding behaviour, sleep and thermoregulatory processes). Thirteen juvenile male Wistar rats were exposed to continuous RF-EMF for 5 weeks at 24 °C of air temperature (T _a) and compared with 11 non-exposed animals. Hence, at the beginning of the 6th week of exposure, the functions were recorded at T _a of 24 °C and then at 31 °C. We showed that the frequency of rapid eye movement sleep episodes was greater in the RF-EMF-exposed group, independently of T _a (+42.1 % at 24 °C and +31.6 % at 31 °C). The other effects of RF-EMF exposure on several sleep parameters were dependent on T _a. At 31 °C, RF-EMF-exposed animals had a significantly lower subcutaneous tail temperature (?1.21 °C) than controls at all sleep stages; this suggested peripheral vasoconstriction, which was confirmed in an experiment with the vasodilatator prazosin. Exposure to RF-EMF also increased daytime food intake (+0.22 g?h^?1). Most of the observed effects of RF-EMF exposure were dependent on T _a. Exposure to RF-EMF appears to modify the functioning of vasomotor tone by acting peripherally through α-adrenoceptors. The elicited vasoconstriction may restrict body cooling, whereas energy intake increases. Our results show that RF-EMF exposure can induce energy-saving processes without strongly disturbing the overall sleep pattern.