Extraction of DNA from amniotic fluid cells for the early prenatal diagnosis of genetic disease

Ten-ml samples of amniotic fluid were taken from pregnancies being terminated at 8–14 weeks' gestation. DNA was extracted from the amniotic cells by sequential centrifugation and analysed using the polymerase chain reaction (PCR). Fifteen samples were analysed for evidence of maternal contamination using Mfd5 oligo-nucleotide primers for repeat polymorphisms. Ten amniotic fluid samples were tested for the Delta-F₅₀₈ deletion characteristic of cystic fibrosis to demonstrate a diagnostic application for the technique. In each case, DNA extracted from fetal tissue from the same pregnancy was included in the controls. In 14 of the 15 cases tested with the Mfd5 primers, both the amniotic fluid DNA and the fetal DNA showed no evidence of contaminating DNA. In one case, neither the amniotic fluid cells nor the fetal cells yielded results. In nine of the ten cases tested with the Delta-F₅₀₈ primers, the amniotic fluid cell DNA provided accurate information about the genetic status of the fetus; in the tenth, the fetal DNA failed to amplify. The results indicate that adequate DNA can be extracted from amniotic fluid from 8 weeks' gestation onward and these samples are suitable for prenatal diagnosis using PCR.     

Urban Flood Damage and Greenhouse Scenarios - The Implications for Policy: An Example from Australia

Urban flooding is often used as an illustration of the potentially adverse effects of greenhouse-induced climate change on extreme events. There is however, a paucity of studies that convert climate scenarios into changes in flood damage. This account summarises the use of modelling techniques, for three flood prone urban catchments in south eastern Australia, to assess changes to urban flood losses for the 'most wet' and 'most dry' scenarios for the year 2070. The most wet scenario indicates that annual average flood damage could increase within the range of 2.5 to 10 times, under the most dry scenario flood regimes would be similar to those experienced at present. The socio-economic scenarios based on the changes to flood losses are used to consider policy responses. It is unlikely that many local government authorities will respond because of lack of interest and because of major changes to the climate scenarios proposed over the last decade. Any response is likely to be incremental and accord with the 'no regrets' and the precautionary principle'.     

Superovulation fails to increase human blastocyst yield after uterine lavage

Uterine lavage affords the potential for non-invasive human blastocyst recovery, with obvious potential for preimplantation genetic diagnosis. In an effort to duplicate in women the multiple blastocyst recovery per cycle that can be achieved in several other species, we initiated a programme in which fertile women underwent superovulation, followed by lavage and embryo collection. We superovulated 15 fertile women, aged 21–40, in 29 cycles using one of four regimens. Insemination was by either intercourse or artificial intracervical donor insemination with cryopreserved sperm from men of proven fertility. In 28 of 29 cycles, the uterus was lavaged daily for 1, 2, or 3 days between 5 and 10 days after human chorionic gonadotropin (hCG) administration or luteinizing hormone (LH) surge. Almost total fluid volume was recovered in every lavage. There were no retained pregnancies and no complications. Surprisingly, only two morulae, one blastocyst, and four unfertilized ova were recovered. Thus, alterations in ovulation induction, insemination timing, or lavage techniques must be contemplated in order to increase the blastocyst yield and thus fulfil the potential of uterine lavage for preimplantation diagnosis.     

Early amniocentesis at 11–14 weeks' gestation for the diagnosis of fetal chromosomal abnormality—a clinical evaluation

Early arnniocentesis between 11 and 14 weeks' gestation was offered to 110 women at risk of a chromosomally abnormal fetus due to maternal age. Four were found to be unsuitable for the procedure, and 106 early amniocenteses were performed. In 102 cases, clear amniotic fluid was obtained with a single tap. There were two dry taps and two bloodstained tapis; sampling was repeated in three of these cases before 15 weeks. In the fourth case, placental biopsy was performed at 16 weeks. Thus, we were able to obtain a satisfactory sample in all but three cases(2.8 percent). Karyotyping of cells harvested from the early amniotic fluid samples was successful in all the 105 cases. Cell culture from the initial samples revealed a normal karyotype in 99 cases, two balanced translocations, two tetraploid karyotypes, and two cases of pseudomosaicism. Of the 105 pregnancies successfully sampled, there have been two losses to date (1–8 per cent). Two further patients presented with premature rupture of membranes, both pregnancies having successful outcomes. Sixty-two babies have delivered to date, with four congenital anomalies. There were no respiratory problems. Twenty-nine pregnancies are continuing without known complications, and details are not yet available on the remaining 12. The results indicate that early arnniocentesis may replace the traditional test at 15–17 weeks.     

Wood materials used as a means to reduce greenhouse gases (GHGs): An examination of wooden utility poles

There has been growing concern over the build-up of greenhouse gase(GHGs) in the atmosphere, particularly carbon dioxide (CO₂), as acause of global warming. The IPCC Third Assessment Report (2001) suggests two ways in which the choice of materials could berelevant. First, some materials, particularly wood, have the advantage thatthey continue to hold carbon (C)in their cells even after being convertedto products. The implications of this feature are well researched. Second,an area that is not well researched relates to the different energyrequirements for producing similar products made with different materials. Using the findings of recent research, this paper compares the energyrequirements and C emissions of manufacturing a product using wood withthat of other materials. The case study of utility poles demonstrates thepositive C and global warming consequences of the lower energyrequirements of wood in the U.S., compared to other materials such assteel or concrete. It demonstrates that GHG emissions associated withutility poles are a small but significant percent of total US annual emissions. Wood utility poles are associated with GHG emission reductions of 163Terragrams (Tg) of CO₂ when compared with steel poles. This isabout 2.8 percent of US annual GHG emissions, which are estimated atabout 5.28 Petragrams (Pg) of CO₂ annually. Thus, the use ofwooden utility poles rather than steel results in a small but significantreduction in total US emissions.     

Prenatal diagnosis for dystrophia myotonica using the polymerase chain reaction

The polymerase chain reaction has been used to detect an abundant class of short repeat DNA families of the form (dC-dA)n.(dG-dT)n, known as microsatellites. These units are found throughout the human genome and have been characterized for several loci including APOC2 on chromosome 19ql2-ql3.2. The locus APOC2 is linked to the gene for dystrophia myotonica and a microsatellite within this locus was used to derive polymorphisms in a family to predict the inheritance of the disease. Chorionic villus sampling (CVS) was performed at 151/2 weeks' gestation. Following DNA extraction from the CVS material and parental blood samples, microsatellite analysis was carried out by the polymerase chain reaction.