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.     

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.     

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.     

Severity models of cross-median and rollover crashes on rural divided highways in Pennsylvania

Introduction

Crossover and rollover crashes in earth-divided, traversable medians on rural divided highways can lead to severe injury outcomes. This study estimated severity models of these two crash types. Vehicle, driver, roadway, and median cross-section design data were factors considered in the models. A unique aspect of the data used to estimate the models were the availability of median cross-slope data, which are not commonly included in roadway inventory data files.

Methods

A binary logit model of cross-median crash severity and a multinomial logit model of rollover crash severity were estimated using five years of data from rural divided highways in Pennsylvania.

Results

The highest probability of a fatal or major injury in cross-median and rollover crashes was found to occur in cases when a driver was not wearing a seatbelt. While flatter cross-slopes and narrower medians were associated with more severe cross-median crash outcomes, steeper cross-slopes and narrower medians significantly increased rollover crash severity outcomes. The presence of horizontal curves was associated with increased probabilities of high-severity outcomes in a median rollover crash.

Impact on Industry

Modeling results in this study confirmed that cross-median and median rollover crash severity outcomes are associated with median cross-section design characteristics. Based on the estimated models, it appears that flatter and narrower medians lead to more severe injury outcomes in cross-median crashes. Steeper median cross-slopes and narrower medians were associated with higher probabilities of more severe outcomes in median rollover crashes. The results presented in this study suggest that there is a trade-off between median cross-section design and cross-median and rollover crashes in earth-divided, traversable medians on rural divided highways. While the severity models can be included in a framework to develop design guidance in relation to this trade-off, models of crash frequency should also be considered.     

Overview of motorcycling in the United States: a national telephone survey

Objectives

Motorcycle registrations have risen in recent years. Although motorcyclist crash fatalities in 2009 were 16% lower than in 2008, they were double the number of deaths in 1997. The present study examined current motorcyclists’ travel patterns and views of motorcycle helmets and other safety topics.

Methods

Motorcycle drivers were interviewed in a national telephone survey conducted in 2009. A weighted sample of 1,606 motorcyclists resulted from adjusting for the oversampling of those younger than 40 and those in the three states without a motorcycle helmet use law (Illinois, Iowa, New Hampshire). All analyses were based on the weighted sample, which was intended to result in a nationally representative sample of motorcyclists.

Results

About one-quarter of respondents said they did not always wear helmets. Of these respondents, 57% said a law requiring helmet use would persuade them to do so, and 27% said nothing would. Ninety-four percent of respondents in states with universal helmet laws said they always ride helmeted, compared with about half of respondents in other states. About half of all respondents favored these laws. About three-quarters said they believe helmets keep riders safer, including two-thirds of respondents who oppose universal laws and almost half of drivers who rarely/never wear helmets. Drivers ages 18–29 and drivers of sport/unclad sport, sport touring, and super sport motorcycles were more likely to always wear helmets, support universal helmet laws, and believe helmets keep riders safer. About half of respondents said antilock braking systems (ABS) enhance safety and that they would get ABS on their next motorcycle. Less than one-quarter thought an airbag would protect a motorcyclist in a crash, and even fewer would consider getting one on their next motorcycle. Forty-three percent of motorcyclists said they had crashed at least once; 62% of the most recent crashes involved no vehicles besides the motorcycle. Respondents reported riding their motorcycles about 5,400 miles, on average, during the past year. Drivers ages 18–29 reported riding fewer miles, on average, than older drivers and more often rode at night and to/from work or school. Drivers of touring and sport touring motorcycles traveled more miles and took more long trips.

Conclusions

Motorcyclists’ travel patterns and views vary widely, but there are distinct patterns by driver age and motorcycle type. Drivers who believe helmets keep riders safer are more likely to always wear them, but this belief appears insufficient to motivate some drivers to wear them. However, universal helmet laws appear effective in increasing helmet use. Many drivers are receptive to purchasing ABS on their next motorcycle.

Impact on industry

States should be encouraged to enact universal helmet laws, and motorcycle manufacturers should be encouraged to offer ABS.     

Implementing falls prevention research into policy and practice in Australia: past, present and future

Introduction

Falls in older Australians are a significant public health issue with one in three older people falling one or more times each year.

Method

Many fall prevention randomized controlled trials have been conducted in Australia as well as across the world.

Results

The findings of these studies now constitute a substantial evidence base that can provide direction for health and lifestyle interventions for preventing falls in older people. This research evidence has contributed to health policy in Australia to some extent, but is yet to be widely implemented into practice. This opinion piece overviews previous policy initiatives and describes a new Partnership research program funded by the Australian National Health and Medical Research Council (NHMRC), which seeks to further influence health policy and address the ongoing research-practice gap.     

Expanding multiple marker screening for Down's syndrome to include Edward'S syndrome

Information on maternal age and maternal serum alpha-fetoprotein, unconjugated oestriol (uE₃), and human chorionic gonadotrophin (hCG) levels was used to investigate retrospectively the effect of estimating Edward's syndrome risk in women having multi-marker screening for Down's syndrome. The screened population comprised 15 pregnancies affected by Edward's syndrome, 15 with Down's syndrome and 5472 unaffected pregnancies. The use of all three markers to estimate Edward's syndrome risk would have led to the detection of 10–12 (67–80 per cent) cases with a false-positive rate of 0.3–0.6 per cent depending on the risk cut-off. A further case would have been detected as a result of screening for Down's syndrome alone. Similar results were obtained when the Edward's syndrome risk was based on uE₃ and hCG only. These data suggest that extending Down's syndrome screening to include Edward's syndrome risk will yield a high detection rate with only a small increase in the false-positive rate.     

Serum PAPP-A measurements in first-trimester screening for Down syndrome

Serum PAPP-A measurements taken from 254 women in the first trimester are reported. Eleven chromosomal abnormalities were detected. The mean serum PAPP-A levels in cases of Down syndrome were 0.44 MOM at 9 weeks gestation, 0.15 MOM at 10 weeks, and 0.29 MOM at 11 weeks. The PAPP-A level at 10 weeks was below those of pregnancies which aborted spontaneously. At 11 weeks, the pregnancies with Down syndrome recorded the lowest PAPP-A levels at that gestation. On this small sample, offering chorionic villus sampling to women with singleton pregnancies and a PAPP-A level below 0.3 MOM (approximately 6.5 per cent of this at-risk group) would have detected all the Down syndrome fetuses at 10 weeks and 50 per cent at 11 weeks without selecting those cases destined to abort. This suggests that serum PAPP-A should continue to be investigated as a potential first-trimester screening test for Down syndrome.     

Insect damage influences heat and water stress resistance gene expression in field-grown popcorn: implications in developing crop varieties adapted to climate change

Warming climatic conditions can pose problems for crop production in many parts of the world, but detailed information on the expression of heat and drought stress resistance genes of potentially affected crop plants is lacking. This information is important to have in order to most efficiently guide the breeding of crops that are adapted to new climatic conditions. A maize (Zea mays) gene microarray, a method used worldwide to evaluate the expression of tens of thousands of genes at once, was used to investigate changes in expression of genes involved in resistance to heat and water stress in milk stage popcorn kernels from undamaged and insect-damaged ears. Popcorn is a form of maize that is more susceptible to heat and drought stress due to its smaller root system. In years of heat and drought stress, expression of many heat shock- and senescence-related proteins increased compared to the year when weather was closer to average conditions, but the expression of many genes related to drought stress resistance decreased in years of weather stress. A different complex of heat shock protein and water stress resistance protein genes had higher expression in kernels from undamaged compared to insect-damaged ears in years of heat and drought stress. These results indicate that the interaction of biotic components, such as insects, are important to consider in developing crop lines with adaptation to stress as this will help identify additional genes and their regulatory components involved in heat and drought stress resistance that might otherwise be overlooked, and will likely be an important strategy for the most effective development of climate stress-tolerant crops globally.