Metal levels in feathers of cormorants, flamingos and gulls from the coast of Namibia in southern Africa

Arsenic, cadmium, chromium, lead, manganese, mercury, selemium,and tin concentrations were measured in the feathers of Capecormorant (Phalacrocorax capensis), Hartlaubs gull(Larus hartlaubii), kelp gull (Larus dominicanus), andlesser flamingo (Phoeniconaias minor) from the coast ofNamibia in southern Africa. Metal concentrations in feathers represent the concentrations in the blood supply at the time offeather formation. Cape Cormorants are piscivores; kelp gullsare primarily piscivores; Hartlaubs gull is an omnivore; and lesser flamingos eat primarily blue-green algae and invertebrates filtered from the water and sediment ofhypersaline lagoons. We predicted that metal concentrationswould reflect these trophic level differences. There weresignificant species differences in the concentrations of allmetals, with flamingos having the lowest levels, and cormorantshaving the highest levels of 4 metals but not mercury. The gulls hadthe highest levels of mercury, perhaps reflecting their morescavenging behavior.     

PGD for monogenic disorders: aspects of molecular biology

Preimplantation genetic diagnosis (PGD) for monogenic diseases has known a considerable evolution since its first application in the early 1990s. Especially the technical aspects of the genetic diagnosis itself, the single-cell genetic analysis, has constantly evolved to reach levels of accuracy and efficiency nearing those of genetic diagnosis on regular DNA samples. In this review, we will focus on the molecular biological techniques that are currently in use in the most advanced centers for PGD for monogenic disorders, including multiplex polymerase chain reaction (PCR) and post-PCR diagnostic methods, whole genome amplification (WGA) and multiple displacement amplification (MDA). As it becomes more and more clear that when it comes to ethically difficult indications, PGD goes further than prenatal diagnosis (PND), we will also briefly discuss ethical issues. Copyright © 2008 John Wiley & Sons, Ltd.     

Detonation transition criteria from the interaction of supersonic shock-flame complexes with different shaped obstacles

The present study is an experimental investigation of the last stages of the deflagration-to-detonation transition. A fast flame following a lead shock was generated by passing a detonation wave through a perforated plate. The shock flame complex then interacts with an obstacle of different shape. We study the influence of the obstacle shape on the transition mechanism to a detonation. The obstacles studied are a single round or square obstacle, a flat plate, a C-shaped and an H-shaped obstacle. The experiments were performed in a thin transparent channel permitting high speed schlieren visualization. Stoichiometric propane-oxygen was investigated at sub-atmospheric conditions. For each obstacle configuration, the initial pressure was changed to modify the flame burning velocity and the Mach number of the leading shock. The burning velocity prior to the interaction was measured experimentally from the displacement velocity of the flame in the videos. This required estimating the speed of the gas ahead of the flame. A linear correction to the speed immediately behind the lead shock was applied using the shock change equations and the measured pressure gradient behind the lead shock in order to account for the non-steadiness of the lead shock and viscous losses to the walls. Three main findings were that the obstacle shape had a minimal influence on the critical flame strength required for transition, although obstacles with a forward facing cavity were able to suppress the transition by isolating the re-initiation event inside the cavity. The main transition mechanism for all geometries was the enhancement of the flame burning velocity through the flame interaction with the shock reflected on the obstacle leading to Richtmyer-Meshkov instability. Finally, it was found that the flame burning velocity of the initial flame required for transition was closely approximated by the Chapman-Jouguet burning velocity. Consistent with the visual observations, this supports the view that transition is favored when the flame is in phase with the acoustic waves, and strong internal pressure waves can be amplified.     

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.     

Whole genome amplification from a single cell: a new era for preimplantation genetic diagnosis

Preimplantation genetic diagnosis (PGD) is a technique used for determining the genetic status of a single cell biopsied from embryos or oocytes. Genetic analysis from a single cell is both rewarding and challenging, especially in PGD. The starting material is very limited and not replaceable, and the diagnosis has to be made in a very short time. Different whole genome amplification (WGA) techniques have been developed to specifically increase the DNA quantities originating from clinical samples with limited DNA contents. In this review, currently available WGA techniques are introduced and, among them, multiple displacement amplification (MDA) is discussed in detail. MDA generates abundant assay-ready DNA to perform broad panels of genetic assays through its ability to rapidly amplify genomes from single cells. The utilization of MDA for single-cell molecular analysis is expanding at a high rate, and MDA is expected to soon become an integral part of PGD. Copyright © 2007 John Wiley & Sons, Ltd.     

褐菖鲇肝HSC70基因的环介导等温扩增技术的建立及在石油污染剂量效应研究中的应用

从褐菖鲉肝脏中克隆了热休克蛋白HSC70基因,利用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)建立HSC70基因的定量检测方法.为检测该方法的可行性,将褐菖鲉分别暴露于石油水溶性成分(water-soluble fraction,WSF) 20、60、180 μg·L-11d后,利用real-time PCR及LAMP技术同时测定褐菖鲉肝HSC70 mRNA表达量,两种方法测定结果基本一致,证实LAMP技术可用于褐菖鲉肝HSC70基因的定量.为更细致了解石油WSF影响褐菖鲉肝脏HSC70基因表达的剂量-效应关系,将褐菖鲉分别暴露于25、50、75、100、125、150、175 μg·L-1 WSF中,5d后采样,用LAMP技术定量检测HSC70 mRNA.结果表明,HSC70 mRNA表达量在50 μg· L-1浓度组即被显著诱导,在75μg·L-1浓度下达到最大值,这说明褐菖鲉肝HSC70基因对石油污染较敏感,有潜力作为海洋石油污染的生物标志物.     

16S rRNA基因技术在环境科学领域中的应用

随着分子生物学的发展，16S rRNA基因技术被逐渐应用到环境科学领域中。目前在环境保护和治理中，该技术主要被用于鉴定污染物的生物降解菌和分析环境样品中的微生物群落多样性，由于它不必将微生物培养分离出来，也就避免了在培养过程中可能出现的微生物去失的情况。本文对16S rRNA基因技术及其在环境科学领域中的应用现状和发展作了一简要介绍，并对16S rRNA基因技术存在的不足进行了讨论。     

微波功率放大组件加速贮存数据处理及寿命评估

目的 以微波功率放大组件为研究对象,开展加速贮存试验退化数据的处理、建模、分析,给出组件的贮存期评估值和激活能等贮存特征参数。方法 在已完成微波功率放大组件加速贮存试验的基础上,借鉴已有的数据建模及处理方法,开展加速贮存数据处理及寿命评估,应用多种退化轨迹拟合寻优、基于性能退化模型参数折算建模、多种寿命分布建模及拟合优度检验等技术,对贮存数据进行分析处理。结果 给出了微波功率放大组件退化轨迹模型、寿命分布模型、加速因子、激活能等贮存特征参数和可靠寿命评估值。结论 描述的加速贮存试验数据处理方法,适应性好,具有较高的优良性,可为类似具有退化数据的电子设备提供借鉴。     

Interactive effects of cadmium and hypoxia on metabolic responses and bacterial loads of eastern oysters Crassostrea virginica Gmelin

Pollution by toxic metals including cadmium (Cd) and hypoxia are important stressors in estuaries and coastal waters which may interactively affect sessile benthic organisms, such as oysters. We studied metabolic responses to prolonged hypoxic acclimation (2 weeks at 5% O₂) in control and Cd-exposed (30 d at 50 μg L⁻¹ Cd) oysters Crassostrea virginica, and analyzed the effects of these stressors on abundance of Vibrio spp. in oysters. Hypoxia-acclimated oysters retained normal standard metabolic rates (SMR) at 5% O₂, in contrast to a decline of SMR observed during acute hypoxia. However, oysters spent more time actively ventilating in hypoxia than normoxia resulting in enhanced Cd uptake and 2.7-fold higher tissue Cd burdens in hypoxia. Cd exposure led to a significant decrease in tissue glycogen stores, increase in free glucose levels and elevated activity of glycolytic enzymes (hexokinase and aldolase) indicating a greater dependence on carbohydrate catabolism. A compensatory increase in activities of two key mitochondrial enzymes (citrate synthase and cytochrome c oxidase) was found during prolonged hypoxia in control oysters but suppressed in Cd-exposed ones. Cd exposure also resulted in a significant increase in abundance of Vibrio parahaemolyticus and Vibrio vulnificus levels during normoxia and hypoxia, respectively. Overall, Cd- and hypoxia-induced changes in metabolic profile, Cd accumulation and bacterial flora of oysters indicate that these stressors can synergistically impact energy homeostasis, performance and survival of oysters in polluted estuaries and have significant consequences for transfer of Cd and bacterial pathogens to the higher levels of the food chain.     

Alga-lysing bioreactor and the dominant bacteria strain

Alga-lysing bacteria have been paid much attention to in recent years. In this study, the alga-lysing strain P05 which was isolated from an immobilizing biosystem was immobilized by coke and elastic filler, forming two biological reactors. The removal efficiencies of algae, NH3-N and organic matter using the two reactors were studied. The results showed that strain P05 was an ideal algal-lysing bacteria strain because it was easy to be immobilized by coke and elastic filler which are of cheap, low biodegradability and the simple immobilization procedure. After 7 d filming, the biological film could be formed and the reactors were used to treat the eutrophic water. These two reactors were of stability and high effect with low cost and easy operation. The optimal hydraulic retention time (HRT) of each reactor was 4 h. The algae removal rates were 80.38% and 82.1% (in term of Chl-a) of coke reactor and filler reactor, respectively. And that of NH3-N were 52.3% and 52.7%. The removal rates of CODMn were 39.03% and 39.64%. The strain P05 was identified as Bacillus sp. by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database.