Y chromosome detection by Real Time PCR and pyrophosphorolysis-activated polymerisation using free fetal DNA isolated from maternal plasma

Objectives To validate the use of Real Time PCR, a widely used technique that can detect very low levels of Y chromosomal sequence, and to assess the use of a highly sensitive PCR technique, pyrophosphorolysis-activated polymerisation (PAP), for fetal sex determination using free fetal DNA (ffDNA). Methods The fetal sex was determined by Real Time PCR in 58 pregnancies using ffDNA isolated from maternal plasma. In parallel with the Real Time PCR experiments, the presence of Y chromosome sequence was also determined using PAP on 54 isolated ffDNA samples. Results Both techniques detected Y chromosome sequence at very low levels with 98% specificity and 100% sensitivity (Real Time n = 44, PAP n = 54). Furthermore, the PAP technique was shown to be more robust than the Real Time PCR as none of the samples tested failed to meet the acceptance criteria. Combining the two techniques for male fetal sex detection from maternal blood plasma increases the sensitivity and specificity to 100% in this series. Conclusions This study shows that both Real Time PCR and PAP can be used for Y chromosome detection on ffDNA. Furthermore, by using PAP in combination with Real Time PCR more reliable early prenatal sexing can be performed using ffDNA. Copyright © 2007 John Wiley & Sons, Ltd.     

Concurrent Chemical and Histological Analyses: Are They Compatible?

Bivalves are often used as sentinel organisms in monitoring programmes for trace organic contaminants. the animal's physiological state may be important in interpreting trends in contaminant body burden. Simultaneous evaluation of physiological state and organic contaminant concentration in bivalves typically involves removal of a lipid-rich cross-section of the body mass for histopathological and/or gonadal analysis.

In this study, the bias introduced by this technique in the final trace organic concentrations, e.g. of polynuclear aromatic hydrocarbons, chlorinated pesticides and polychlorinated biphenyls, are evaluated on five different size groups of oysters. As a test case, we evaluated the use of this method in the NOAA's Status & Trends Mussel Watch (NS&T) Programme. the average biases introduced by this technique in the final trace organic concentrations in Gulf of Mexico oysters have been increasing since 1986 as a consequence of a continuous decrease in the size of the individuals sampled.     

Winter-spring essential reorganization of cell proliferation in the digestive tract epithelia in the mussel Crenomytilus grayanus

A study of the seasonal dynamics of cell proliferation in the digestive tract epithelia of the mussel Crenomytilus grayanus (Bivalvia) collected in the Sea of Japan between 1971 and 1984 revealed two opposite states so far unknown for marine molluscan tissues: proliferative resting in winter and a following outburst of cell division in spring. In winter and early spring (at water temperatures below or near 0°C), mitotic figures in the lining epithelia of labial palps and intestine strongly reduced in number and later almost disappeared (February-March); DNA-synthesizing cells in these epithelia on thymidine radioautographs became sparse and only a restricted number of new cells entered the cell cycle. A proliferation block was relieved at the beginning of April (at a temperature of 2 to 4°C): an initiation of a many-fold increase in the mitotic index and some increase in the ³H-T-labelled nuclei index and their advance to the maximum in late April to May (up to more than 0.2 and 5 to 6%, accordingly) were found. In late spring (at temperatures of 5 to 10°C), a striking rise in the number of cells entering the cell cycle was observed with a probable acceleration of the cycle proper. Both seasonal states of cell proliferation — a deep depression and outburst, established in the winter-spring period are interpreted as essential events in the life cycle of the mussel studied.     

Nanocomposites of Cellulose Acetate Butyrate Reinforced with Cellulose Nanocrystals

Nanocrystals were prepared by acid hydrolysis of bacterial cellulose microfibrils. These were topochemically trimethylsilylated, in an attempt to reduce their hydrophilicity. Composites were made by dispersing either native or silylated crystals in cellulose acetatebutyrate matrixes and solution casting of the dispersions. Particles were characterized by transmission electron microscopy. The composites were characterized by differential scanning calorimetry and dynamic mechanical analysis. The unmodified cellulose crystals exhibited better reinforcement characteristics than the trimethylsilated crystals.     

Effects of predation by common eiders (Somateria mollissima) in an intertidal rockweed bed relative to an adjacent mussel bed

Marine Biology - Little is known about the effect of commercial rockweed (Ascophyllum nodosum) harvest on intertidal community structure, and on interactions among species. Using predator-exclusion...