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.     

Methods for synthesis of nonabromodiphenyl ethers and a chloro-nonabromodiphenyl ether

Polybrominated diphenyl ethers (PBDEs) have been used extensively as brominated flame retardants (BFRs) in textiles, upholstery and electronics. They are ubiquitous contaminants in wildlife and humans. A low concentration of nonabrominated diphenyl ethers (nonaBDEs) is present in commercial DecaBDE and they are also abiotic and biotic debromination products of decabromodiphenyl ether (BDE-209). The objective of the present work was to develop methods for synthesis of the three nonaBDEs, 2,2',3,3',4,4',5,5',6-nonabromodiphenyl ether (BDE-206), 2,2',3,3',4,4',5,6,6'-nonabromodiphenyl ether (BDE-207) and 2,2',3,3',4,5,5',6,6'-nonabromodiphenyl ether (BDE-208), with the intention of making them available as authentic standards for analytical, toxicological and stability studies, as well as studies regarding physical-chemical properties. Two methods were developed, one based on perbromination of phenoxyanilines and the other via reductive debromination of BDE-209 by sodium borohydride followed by chromatographic separation of the three nonaBDE isomers formed. An additional nonabrominated compound, 4'-chloro-2,2',3,3',4,5,5',6,6'-nonabromodiphenyl ether (Cl-BDE-208), was also synthesized in the present work. Cl-BDE-208, prepared by the perbromination of 4-chlorodiphenyl ether, may be used as an internal standard in analysis of highly brominated diphenyl ethers. BDE-206, BDE-207, BDE-208 and Cl-BDE-208 were characterized by 1H NMR, 13C NMR, electron ionization mass spectra and by their melting points. The structures of all three nonaBDEs have been characterized previously by X-ray crystallography.     

Composite of Natural Polymers and Their Adsorbent Properties on the Dyes and Heavy Metal Ions

Journal of Polymers and the Environment - Natural polymers such as chitosan, chitin, cellulose, alginate, pectin, and lignin produced by renewable organic resources are regarded as greener,...     

Deriving heart rate variability indices from cardiac monitoring—An indicator of driver sleepiness

Objective: Driver fatigue is considered to be a major contributor to road traffic crashes. Cardiac monitoring and heart rate variability (HRV) analysis is a candidate method for early and accurate detection of driver sleepiness. This study has 2 objectives: to evaluate the (1) suitability of different preprocessing strategies for detecting and removing outlier heartbeats and spectral transformation of HRV signals and their impact of driver sleepiness assessment and (2) relation between common HRV indices and subjective sleepiness reported by a large number of drivers in real driving situations, for the first time.

Methods: The study analyzed >3,500 5-min driving epochs from 76 drivers on a public motorway in Sweden. The electrocardiograph (ECG) data were recorded in 3 studies designed to evaluate the physiological differences between awake and sleepy drivers. The drivers reported their perceived level of sleepiness according to the Karolinska Sleepiness Scale (KSS) every 5?min. Two standard methods were used for identifying outlier heartbeats: (1) percentage change (PC), where outliers were defined as interbeat intervals deviating >30% from the mean of the four previous intervals and (2) standard deviation (SD), where outliers were defined as interbeat interval deviating >4 SD from the mean interval duration in the current epoch. Three standard methods were used for spectral transformation, which is needed for deriving HRV indices in the frequency domain: (1) Fourier transform; (2) autoregressive model; and (3) Lomb-Scargle periodogram. Different preprocessing strategies were compared regarding their impact on derivation of common HRV indices and their relation to KSS data distribution, using box plots and statistical tests such as analysis of variance (ANOVA) and Student’s t test.

Results: The ability of HRV indices to discriminate between alert and sleepy drivers does not differ significantly depending on which outlier detection and spectral transformation methods are used. As expected, with increasing sleepiness, the heart rate decreased, whereas heart rate variability overall increased. Furthermore, HRV parameters representing the parasympathetic branch of the autonomous nervous system increased. An unexpected finding was that parameters representing the sympathetic branch of the autonomous nervous system also increased with increasing KSS level. We hypothesize that this increment was due to stress induced by trying to avoid an incident, because the drivers were in real driving situations.

Conclusions: The association of HRV indices to KSS did not depend on the preprocessing strategy. No preprocessing method showed superiority for HRV association to driver sleepiness. This was also true for combinations of methods for frequency domain HRV indices. The results prove clear relationships between HRV indices and perceived sleepiness. Thus, HRV analysis shows promise for driver sleepiness detection.     

What limits predator detection in blue tits (<Emphasis Type="Italic">Parus caeruleus</Emphasis>): posture,task or orientation?

To detect threats and reduce predation risk prey animals need to be alert. Early predator detection and rapid anti-predatory action increase the likelihood of survival. We investigated how foraging affects predator detection and time to take-off in blue tits (Parus caeruleus) by subjecting them to a simulated raptor attack. To investigate the impact of body posture we compared birds feeding head-down with birds feeding head-up, but could not find any effect of posture on either time to detection or time to take-off. To investigate the impact of orientation we compared birds having their side towards the attacking predator with birds having their back towards it. Predator detection, but not time to take-off, was delayed when the back was oriented towards the predator. We also investigated the impact of foraging task by comparing birds that were either not foraging, foraging on chopped mealworms, or foraging on whole ones. Foraging on chopped mealworms did not delay detection compared to nonforaging showing that foraging does not always restrict vigilance. However, detection was delayed more than 150% when the birds were foraging on whole, live mealworms, which apparently demanded much attention and handling skill. Time to take-off was affected by foraging task in the same way as detection was. We show that when studying foraging and vigilance one must include the difficulty of the foraging task and prey orientation.Communicated by P.A. Bednekoff     

Aluminium metabolism in chronic renat failure: Environmental influences and regional differences in Norway

Many aspects of Al metabolism in chronic renal failure are poorly understood. A longitudinal study of serum Al concentrations in predialysis patients and healthy control subjects revealed very high values during the autumn of 1984 and 1985. Renal Al clearance was low during the autumnal spike in serum Al but increased substantially when the serum Al concentration declined. A second study confirmed that by using citric acid as a chelator, the gastrointestinal absorption of Al from Al(OH)₃ may be considerably augmented as reflected by increases in both serum Al concentrations and renal Al clearance. The individual differences in Al absorption in this study were large.The first study suggests the existence of an unidentified environmental factor, possibly water borne, with profound effects on Al absorption and excretion. The citric acid/Al(OH)₃ experiment suggests that the existence of such a factor is likely. The implications of these results are not known.A histomorphometric study of bone biopsies from 138 hemodialysis and 66 predialysis patients without clinical evidence of Al related disease, revealed Al deposits after staining with aurin tricarboxylic acid in 78% of the biopsies from the former and 24% of the latter patients. Serum Al concentrations did not differ between predialysis and hemodialysis patients with Al positive biopsies. Stratification of the hemodialysis patients, who came from all parts of Norway, revealed that patients living in regions with slightly Al contaminated drinking water (Al <30 g/L) had lower serum Al concentrations than patients from regions with highly contaminated water (Al >100 g/L). The prevalence of Al-positive biopsies was the same in both regions. Patients with Al-positive biopsies did not differ in serum Al level from those with Al-negative biopsies within the same region. Predialysis patients with Al-positive biopsies had significantly higher serum Al levels than predialysis patients with Al negative biopsies.Stainable Al deposits are commonly found in the bone of patients with chronic renal failure. Cross-sectionally obtained serum Al concentrations do not reflect the prevalence of stainable bone Al in renal patients but are related to the degree of Al contamination of water of the region.     

On plankton production in Kungsbacka Fjord,an estuary on the Swedish west coast

Environmental conditions, primary production, and zooplankton populations were studied from May, 1969 to November, 1970 at one station in Kungsbacka Fjord, Sweden. The fjord, with an arca of 53 km², is a moderately polluted estuary, with a small tidal range. Data for primary production and environmental parameters were correlated using Spearman's rank correlation coefficient. The annual rate of primary production in 1970 was about 100 gC·m^-2. Carbon fixation was about 80 g·m^-2 in May–November in1969 and 1970. The average monthly rate was highest in June, 1970, with 25 gC·m^-2; about 15 gC·m^-2 was recorded in August–October of both years. Carbon fixation by the phytoplankton was estimated to be about 2,800 tons in the whole fjord in 1970. The average fresh-water inflow to the fjord, amounting to about 13 m³·sec^-1, added about 380 tons of organic carbon, 45 tons of nitrogen, and 4.5 tons of phosphorus per month. Primary production displayed strong correlation with temperature at different depths (P<0.05 to 0.001), indicating the sediments to be the most important nutrient source. A total of 19 holoplanktonic zooplankton species was identified, copepods being the dominant group. The highest zooplankton biomass, 800 to 900 mg·m^-3, was recorded in June of both years. The production of copepods in May–October was about 1 gC·m^-2 in both years. The total secondary production of the zooplankton was calculated as only 1.8 gC·m^-2 in 1970.     

The indirect effects of eutrophication on habitat choice and survival of fish larvae in the Baltic Sea

The structure of the habitat is usually crucial for growth and survival of young life stages. Presently, some nursery areas of fish larvae are changing due to eutrophication, e.g. due to enhanced growth of ephemeral filamentous algae at the expense of perennial species. We studied the influence of two habitats, one with filamentous algae (Cladophora glomerata) and the other with bladder wrack (Fucus vesiculosus), on habitat choice of pike larvae (Esox lucius) in the absence/presence of a predator or a competitor. We further tested whether the habitat choice is adaptive in increasing survival under predation threat. In contrast to expectations, pike larvae preferred the habitat with ephemeral filamentous algae to the bladder wrack, thriving in clean waters, independent of the presence/absence of both predator/competitor. In addition, the survival of the larvae was higher in the filamentous algae in the presence of predators, which suggested that the habitat preference of the larvae was adaptive. The structure of the bladderwrack habitat was probably too open for newly hatched larvae, which implies that F. vesiculosus and other large brown algae are not as important refuges for young larvae as previously thought.