Noninvasive Digital Detection of Fetal DNA in Plasma of 4-Week-Pregnant Women following In Vitro Fertilization and Embryo Transfer

The discovery of cell-free fetal DNA (cfDNA) circulating in the maternal blood has provided new opportunities for noninvasive prenatal diagnosis (NIPD). However, the extremely low levels of cfDNA within a high background of the maternal DNA in maternal circulation necessitate highly sensitive molecular techniques for its reliable use in NIPD. In this proof of principle study, we evaluated the earliest possible detection of cfDNA in the maternal plasma by a bead-based emulsion PCR technology known as BEAMing (beads, emulsion, amplification, magnetics). Blood samples were collected from in vitro fertilization (IVF) patients at 2 to 6 weeks following embryo transfer (i.e., 4 to 8 week pregnancies) and plasma DNA was extracted. The genomic regions of both X and Y chromosome-specific sequences (AMELX and AMELY) were concurrently amplified in two sequential PCRs; first by conventional PCR then by BEAMing. The positive beads either for AMELX or AMELY gene sequences were counted by a flow cytometer. Our results showed that the pregnancies yielding boys had significantly higher plasma AMELY gene fractions (0.512 ± 0.221) than the ones yielding girls (0.028 ± 0.003) or non-pregnant women (0.020 ± 0.005, P= 0.0059). Here, we clearly demonstrated that the BEAMing technique is capable of reliably detecting cfDNA in the blood circulation of 4-week-pregnant women, which is only two weeks after the embryo transfer. BEAMing technique can also be used to early detect fetal DNA alterations in other pregnancy-associated disorders.     

Influence of prey species and stages on predatory efficiency and development of two phytoseiid mites

The effect of prey species and the different stages of prey on the predatory efficiency and biology of the phytoseiid mites,Amblyseius gossipi Elbadry andTyphlodromus mangiferus sp. n. was studied. It was found that feeding either predator onTetranychus cucurbitacearum (Sayed) promoted faster development and a higher rate of oviposition than rearing on the twospotted spider mite,T. urticae (Koch). Different stages of both prey species also produced different responses in the biological activities of these predaceous mites.

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Résumé On a étudié l'effet de l'espèce de la proie et de ses différents stades sur l'efficacité et la biologie des acariens phytoseiides,Amblyseius gossipi Elbadry etTyphlodromus mangiferus sp. n. Il a été constaté que l'alimentation des 2 prédateurs avecTetranychus cucurbitacearus (Sayed) assure un dévelopment plus rapide et une fécondité plus élevée que leur élevage surT. urticae (Koch). Les différents stades de ces 2 proies produisent des réponses différentes dans les activités biologiques des 2 acariens prédateurs.

     

Detoxification of aflatoxin B1 by acidogenous yoghurt

Serial concentrations of aflatoxin B₁ ranged from 200 to 1000 p.p.b. were assayed for detoxification by acidogenous yoghurt. Thin-layer chromatography analysis revealed a complete transformation of 800 p.p.b. of aflatoxin B₁ to a new fluorescing compound corresponding to aflatoxin B_2a which is referred as hydroxydihydroaflatoxin B₁. Partial conversion was present in yoghurt sample containing 1000 p.p.b. Toxicity test on chickens, confirmed Ciegler findings.     

Setaria equina: In vivo effect of diethylcarbamazine citrate on microfilariae in albino rats

Although diethylcarbamazine citrate (DEC) is successful drug in eliminating human filariasis, yet, its mode of action is still debatable. Herein, the effect of DEC to treat albino rats infected with the animal filarial parasite Setaria equina was tested. Microfilarial (mf) counts and sections from liver, lung, kidney as well as spleen were investigated at different time points after treatment by light microscopy. After 45 and 300 min of treatment, a significant decrease in blood mf was observed accompanied by adherence of degenerated mf to both kupffer cells and leukocyte in liver sections. In lung sections, loss of sheath was observed at 45 min, while degeneration was observed at later time points. In kidney sections, more mf counts and less matrix were observed in the glomeruli at all time points after treatment. Degenerated mf were observed in spleen sections only at, late time point, 480 min after treatment. In conclusion, one of the possible mechanisms by which DEC reduces blood microfilarial count is trapping larvae in organs and killing them through cellular adherence.     

Characterisation of dissolved organic matter in Parisian urban aquatic systems: predominance of hydrophilic and proteinaceous structures

Understanding the nature of organic matter is a necessary first step in assessing contaminant bioavailability and allowing water supply managers to optimise the treatment train in the aim of providing safe and inexpensive drinking water. This study provides further insight into the composition, structure and functional groups of dissolved organic matter (DOM) (both hydrophobic and hydrophilic) from urban aquatic systems by means of various analytical techniques (DAX-8/XAD-4 fractionation, elemental analysis, UV and FTIR spectroscopies, ¹³C and ¹⁵N isotopic analysis, size exclusion chromatography and Pyrolysis-GC-MS). The analytical range chosen for this study constitutes a powerful tool in the characterisation of DOM in urban water. The inclusion of information from one technique to the next might not only serve as a support to each one, but also as a complement. The DOM fraction from treated effluent and, more generally, DOM from urban water (i.e. receiving treated effluent) display a strong hydrophilic characteristic [i.e. low humic substance (HS) content, low SUVA], along with a high distribution in molecular weights observed by SEC and low average molecular weight. Due to the origin of this DOM, proteinaceous structures constitute the main compounds, as observed by FTIR and Py-GC-MS. Such characteristics (i.e. heterogeneity, low average molecular weight and diverse functional groups, which make up a total of N) could explain that DOM from treated effluent displayed a strong reactive potential metals pollutants as previously demonstrated.     

Prey preference of the wolf spider,Pardosa pseudoannulata (Boesenberg et Strand)

Functional responses of the wolf spider, Pardosa pseudoannulata (Boesenberg et Strand) attacking the rice brown planthopper, Nilaparvata lugens (Stål.), and the mirid predator Cyrtorhinus lividipennis Reuter were both those of Holling Type II. The attack rate was higher and handling time lower for C. lividipennis. However, when caged with the two prey, the wolf spider showed a significant preference for N. lugens at a lower prey proportion. Proportions of prey attacked were significantly different from the expected ratios of prey available as well as from the predicted preferences derived from the functional response parameters. As proportions of N. lugens attacked changed from greater to less than expected as the proportions of N. lugens available increased, a “reverse switch” behaviour seems to be evident.     

Aflatoxin and aflatoxicosis: V. The kinetic behaviour of dietary aflatoxins in colostrum drawn from cows postpartum

This work was conducted in order to study the kinetic behaviour of dietary aflatoxins in the colostrum of a pregnant cow exposed to contaminated feeds for a short period.In this study, two pregnant cows received a single dose of dietary aflatoxins in the form of rice powder contained 31.20 ppm aflatoxin B and 19.68 ppm aflatoxin G during the last stage of pregnancy, at about two weeks before parturition.Samples of colostrum were collected from dams and assayed for the presence of toxic metabolites as well as its conjugations by electrophoretic analysis.The results revealed that the intake of aflatoxins appeared in the colostrum pospartum as AFM₁ and also AFB_2a which is a non toxic metabolite. Moreover, it was found that the excreted metabolites including AFB_2a were conjugated to the immunoglobulin protein fraction of the colostrum.The significance of the obtained results to the newborn calf are discussed.     

Activation of potassium channels by hypoxia and reoxygenation in the human lung adenocarcinoma cell line A549

Active oxygen species are generated in cells during pathophysiologic conditions such as illflammation and postischemic reperfusion. If oxygen radical scavengers are added before reperfusion, then the magnitude of injury is reduced. We inves-tigated whether free radicals generated following exposure to hypoxia and reoxygenation activate voltage-dependent K⁺ ion channels in tumor cells in vitro. Using the technique of whole cell voltage clamping, we recorded currents from two families of potassium (K⁺) channels that were activated following reoxygenation. One of these groups possessed the electrophysical characteristics of a tetraethylammonium (TEA)-sensitive delayed rectifier channel and the other possessed characteristics of a Tea-insensitive slow inactivating channel. We present evidence which suggests that K⁺ channels are activated following reoxygenation but not during the hypoxia phase. The K⁺ currents decayed with time following reoxygenation. The decay characteristics of the K⁺ currents depended on the duration and level of hypoxia to which the cells were exposed. To determine whether activation of K⁺ channels by reoxygenation was initiated by free radicals, we pretreated cells with N-Acetyl L-Cysteine (NAC), a free radical scavenger, and found that this pretreatment abolished the currents induced by reoxygenation. We also present evidence that free radicals do not directly act on the channel itself, but activate a protein kinase which, in turn, activates the K⁺ channels. Taken together, these results indicate that one of the early responses to oxidative stress is the activation of K⁺ currents. © 1993 Wiley-Liss, Inc.