How does cell-based non-invasive prenatal test (NIPT) perform against chorionic villus sampling and cell-free NIPT in detecting trisomies and copy number variations? A clinical study from Denmark

Objectives

We aimed to compare cell-based NIPT (cbNIPT) to chorionic villus sampling (CVS) and to examine the test characteristics of cbNIPT in the first clinical validation study of cbNIPT compared to cell-free NIPT (cfNIPT).

Material and Methods

Study 1: Women (N = 92) who accepted CVS were recruited for cbNIPT (53 normal and 39 abnormal). Samples were analyzed with chromosomal microarray (CMA). Study 2: Women (N = 282) who accepted cfNIPT were recruited for cbNIPT. cfNIPT was analyzed using sequencing and cbNIPT by CMA.

Results

Study 1: cbNIPT detected all aberrations (32/32) found in CVS: trisomies 13, 18 and 21 (23/23), pathogenic copy number variations (CNVs) (6/6) and sex chromosome aberrations (3/3). cbNIPT detected 3/8 cases of mosaicism in the placenta. Study 2: cbNIPT detected all trisomies found with cfNIPT (6/6) and had no false positive (0/246). One of the three CNVs called by cbNIPT was confirmed by CVS but was undetected by cfNIPT, two were false positives. cbNIPT detected mosaicism in five samples, of which two were not detected by cfNIPT. cbNIPT failed in 7.8% compared to 2.8% in cfNIPT.

Conclusion

Circulating trophoblasts in the maternal circulation provide the potential of screening for aneuploidies and pathogenic CNVs covering the entire fetal genome.     

Nonthermal GSM RF and ELF EMF effects upon rat BBB permeability

Since the late 1980s, our group has examined the effects of radiofrequency electromagnetic fields (RF-EMF), including pulse-modulated waves of the type emitted by mobile phones, upon the blood?Cbrain barrier. In more than 2,000 rats, we have repeatedly demonstrated a passage of the rats?? own albumin from the blood through the brain capillaries into the surrounding brain parenchyma at SAR values down to 0.1mW/kg. In most of these experiments, the animals were exposed in TEM-cells, ventilated by an external electrical fan at 50 Hz. In the present study, we examined whether the extremely low frequency (ELF) magnetic fields from the fan (50 Hz, 0.3?C1.5 ??T) might add to the RF effect. Sixty-four rats were divided into 4 groups: RF only, ELF only and RF + ELF exposure plus a sham group. The GSM-900 MHz RF exposure was at the very low, nonthermal, average whole-body SAR level 0.4 mW/kg. Demonstration of the normally occurring albumin extravasation in the basal hypothalamus is our inbuilt control proving that the staining is reliable. Two full series of staining of the whole material gave negative results for hypothalamus. Not until we changed to avidin, biotin, and antibodies from a third supplier, we received an acceptable staining. Twenty-five percent of the RF animals had a pathological albumin leakage, while the ELF and RF + ELF groups with three and two pathological findings, respectively, were not significantly different from the control group. We conclude that the use of external fans has had no major influence upon the result.     

Occurrence of organochlorine pesticides in indoor dust

Organochlorine pesticides are present in the environment and suspected of causing serious health effects. Diet has been the main exposure source, but indoor source release is gaining focus. Within a monitoring study of polychlorinated biphenyls of Danish buildings built during the 1960s and 1970s, we coincidently determined extreme levels of dichlorodiphenyltrichloroethane (DDT) levels in two of ten random samples. This raises concern and further large scale investigations are warranted to confirm this.     

A distributed modelling system for simulation of monthly runoff and nitrogen sources, loads and sinks for ungauged catchments in Denmark

We validated an existing physically based 3D MIKE SHE groundwater resource model (DK-model) at 175 Danish gauging stations covering different catchment sizes in order to calculate monthly water runoff in the 50% ungauged part of Denmark. Model performance was in most cases good (61% of gauging stations had a Nash-Sutcliffe (NS) coefficient >0.60) but nevertheless showed a large seasonal and georegion specific bias. Therefore, bias correction factors had to be developed before applying the DK-model simulations of runoff in the ungauged areas. Simulated monthly runoff from ungauged areas and the measured monthly runoff from 178 gauging stations were distributed to 2663 smaller Hydrological Units (ca. 15 km(2)) and linked with a new empirical model for flow-weighted monthly total nitrogen (TN) concentrations (R(2) = 0.43; P < 0.0001) developed based on 20 years of observations (1990-2009) in 83 small catchments for calculation of monthly gross diffuse TN-loads from HU's. Nitrogen retention was calculated in streams, lakes and wetlands utilising both lake specific models and rate coefficients to calculate N retention in surface water bodies. The whole model complex was linked in the DK-QN concept for simulation of monthly TN losses from point sources and diffuse sources, TN retention and resulting loadings to Danish coastal waters. The DK-QN model was validated in 118 gauged catchments and the model simulations had for >25% of the observations of monthly discharge weighted TN concentrations a NS larger than 0.26. Catchment specific monthly TN-loadings were modelled with a higher performance as 50% of the catchments had a NS greater than 0.75. The model concept allows calculation of N retention in streams, lakes and wetlands and the average annual model calculated N retention amounted to 21% of the modelled gross riverine TN loadings. The average annual gross TN loading to surface freshwater in Denmark derived from diffuse sources amounted to 97 000 tonnes N (91% of gross TN loadings) which is 54% of the total estimated N-leaching from the root zone on the Danish land area (212 000 tonnes N) during the period 1990-2009.     

Ignition of dust-air atmospheres by ultrasonic waves

This paper shows the results of our investigations on the ignition source ultrasound in dust-air atmospheres. Ultrasound is, on the one hand, considered to be an ignition source according to international safety standards (EN 1127–1 (2011)). On the other hand, though, ultrasound is used for various applications in gases and air, such as level and flow measurement, or in the process industry, but no explosion accidents have yet been reported. Our research now shows that it is indeed possible to ignite dust-air mixtures in ultrasound fields under certain conditions. We conducted our experiments in an ultrasound standing wave field and used maize starch, calcium stearate and sulfur dust. For ignitions, an absorbing target was needed to convert the acoustic energy into heat. From theoretical estimations and experiments critical conditions that provoke ultrasonically triggered explosions are identified.     

Uncertainty modelling in multi-criteria analysis of water safety measures

Water utilities must assess risks and make decisions on safety measures in order to obtain a safe and sustainable drinking water supply. The World Health Organization emphasises preparation of water safety plans, in which risk ranking by means of risk matrices with discretised probability and consequence scales is commonly used. Risk ranking enables prioritisation of risks, but there is currently no common and structured way of performing uncertainty analysis and using risk ranking for evaluating and comparing water safety measures. To enable a proper prioritisation of safety measures and an efficient use of available resources for risk reduction, two alternative models linking risk ranking and multi-criteria decision analysis (MCDA) are presented and evaluated. The two models specifically enable uncertainty modelling in MCDA, and they differ in terms of how uncertainties in risk levels are considered. The need of formal handling of risk and uncertainty in MCDA is emphasised in the literature, and the suggested models provide innovations that are not dependent on the application domain. In the case study application presented here, possible safety measures are evaluated based on the benefit of estimated risk reduction, the cost of implementation and the probability of not achieving an acceptable risk level. Additional criteria such as environmental impact and consumer trust may also be included when applying the models. The case study shows how safety measures can be ranked based on preference scores or cost-effectiveness and how measures not reducing the risk enough can be identified and disqualified. Furthermore, the probability of each safety measure being ranked highest can be calculated. The two models provide a stepwise procedure for prioritising safety measures and enable a formalised handling of uncertainties in input data and results.     

The importance of experience in the interpretation of conspecific chemical signals

Foraging bumblebees scent mark flowers with hydrocarbon secretions. Several studies have found these scent marks act as a repellent to bee foragers. This was thought to minimize the risk of visiting recently depleted flowers. Some studies, however, have found a reverse, attractive effect of scent marks left on flowers. Do bees mark flowers with different scents, or could the same scent be interpreted differently depending on the bees’ previous experience with reward levels in flowers? We use a simple experimental design to investigate if the scent marks can become attractive when bees forage on artificial flowers that remain rewarding upon the bees’ return after having depleted them. We contrast this with bees trained in the more natural scenario where revisits to recently emptied flowers are unrewarding. The bees’ association between scent mark and reward value was tested with flowers scent marked from the same source. We find that the bees’ experience with the level of reward determines how the scent mark is interpreted: the same scent can act as both an attractant and a repellent. How experience and learning influence the interpretation of the meaning of chemical signals deposited by animals for communication has rarely been investigated.     

The evaluation of an analytical method for polychlorinated terpenes (PCC) in biological samples using an internal standard

The quantitative analysis of PCC (Toxaphene) residues in biological samples using gas chromatography with electron capture detection has been evaluated. Dechlorane 603, a chloroalicyclic compound is used as internal standard and modifications of the injection system for proper evaporation of this high-boiling compound is described. In the cleanup procedure PCB (and 1 – 3 % of the PCC) are removed by adsorption chromatography. Chlordane components and DDT compounds remaining in the PCC fraction constitute 5 – 10 % and 13 – 16 %, respectively, of the peak area in the chromatograms of the samples investigated. The quantitative results are influenced by the number of PCC-peaks chosen for the calculations and have to be specified together with the PCC-levels reported. The recoveries of PCC from spiked samples (20 ug PCC per gram crude fat above “natural” level) are 61 – 98 %, the lower figures obtained with samples containing low levels of PCC-residues. The practical limit of detection is about 10 ng PCC per g extracted fat.     

Characterisation, dissemination and persistence of gentamicin resistant Escherichia coli from a Danish university hospital to the waste water environment

The aim of the study was to investigate the potential spread of gentamicin resistant (GEN(R)) Escherichia coli isolates or GEN(R) determinants from a Danish university hospital to the waste water environment. Waste water samples were collected monthly from the outlets of the hospital bed wards and the inlet of the related waste water treatment plant (WWTP) from October 2002 to August 2003. Waste water samples were also collected monthly from a residential area in the same period to be able to compare the prevalence of GEN(R)E. coli isolates from hospital related and residential waste water. The waste water isolates were compared to GEN(R)E. coli isolates obtained consecutively from September 2002 to September 2003 from patients mainly with urinary tract infections at the hospital with respect to Pulsed Field Gel Electrophoresis (PFGE) profiles. All isolates were investigated for GEN(R) mechanisms (aac(3)-II, aac(3)-IV, ant(2')-I, armA), phenotypic resistance pattern, and virulence genes (hlyA, chuA, sfaS, fogG, malX, traT, iutA, fyuA, iroN, cnf1) to investigate if the hospital and waste water could be reservoirs of antimicrobial resistance and virulence. The ability for GEN(R) determinants to transfer horizontally was investigated by mating experiments. A total of 38, 15, 21, and two GEN(R)E. coli were isolated from patients, the hospital outlets, the inlet of the WWTP, and the residential area, respectively. GEN(R)E. coli were more prevalent in waste water from the hospital and the WWTP than in waste water from the residential area. PFGE profiling revealed no spread of specific patient isolates to the waste water. The aac(3)-II gene was detected both in patient and waste water isolates. Furthermore horizontal transfer of the aac(3)-II gene of patient origin to a recipient was shown in vitro, indicating a potential spread of the gene from patient isolates to waste water isolates. Regardless of origin, most isolates exhibited multi-resistance and contained several virulence genes. In conclusion, our study showed a possible spread of aac(3)-II from the hospital to the waste water. Most of the GEN(R)E. coli isolates from both patients and waste water had a multi-resistant phenotype and contained virulence genes and should therefore be considered reservoirs of antimicrobial resistance and virulence genes.