Denaturing high-performance liquid chromatography (DHPLC)-based prenatal diagnosis for tuberous sclerosis

Tuberous sclerosis (TSC) is a frequent autosomal-dominant condition (affecting 1 in 6000 individuals) caused by various mutations in either the hamartin (TSC1) or the tuberin gene (TSC2). This allelic and non-allelic heterogeneity makes genetic counseling and prenatal diagnosis difficult, especially as a significant proportion of TSC cases are due to de novo mutations. For this reason the identification of the disease causing mutation is mandatory for accurate counseling, yet current mutation detection methods such as single-strand conformation polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE) are labor intensive with limited detection efficiency. Denaturing high-performance liquid chromatography (DHPLC) is a high-throughput, semi-automated mutation detection system with a reported mutation detection rate close to 100% for PCR fragments of up to 800 bp. We used a recently described DHPLC assay allowing the efficient detection of mutations in TSC1 to analyze the DNA extracted from a chorion villus sample in order to perform a prenatal diagnosis for TSC. The fetus was found not to have inherited the deleterious mutation and the DHPLC diagnosis was confirmed by haplotype analysis. This represents the first DHPLC-based prenatal diagnosis of a genetic disease. Copyright © 2001 John Wiley & Sons, Ltd.     

Prenatal exclusion of Hurler's disease by leucocyte α-L-iduronidase assay

Hurler's disease was excluded in a fetus at 23 weeks' gestation by demonstrating normal iduronidase activity in fetal leucocytes following failure of amniotic cell culture after amnic-centesis at 16 and 19 weeks' gestation. The diagnosis was confirmed in the neonate.     

编码水稻胞外核苷酸双磷酸酶基因的表达特性

NTPDase(Nucleoside triphosphate-diphosphohydrolase,核苷酸双磷酸酶)在哺乳动物巾分布于细胞膜,可以水解细胞外ATP和其他核苷酸,从而调控胞外核苷酸代谢及信号应答.根据水稻NTPD基因序列设计引物,在不同胁迫条件样品中,通过PCR扩增水稻NTPD基因,结果表明水稻NTPD基因表达水平在盐胁迫样品中升高.为进一步分析水稻NTPD基因的功能,构建了该基因的过量表达、RNA干涉载体,通过根癌农杆菌介导转入水稻品种日本晴并获得阳性植株.半定量RT-PCR分析达到过量表达和干涉日的.在盐胁迫条件下,TO代过表达转基因植株表现出一定程度的抗胁迫能力,RNA干涉转基因植株的生长则受到抑制.图7表3参20     

Uptake of cadmium into cells in culture∗

Cadmium has been recognized as pollutant of the environment for many years and numerous studies on its toxic effects have been carried out. Little, however, is known about its metabolic behaviour e.g. why the metal is accumulated so extremely rapidly into the organs of men and animals. Since the study of the individual metabolic steps is very difficult in vivo cell cultures may be used to obtain first indications of what happens in the whole animal.

We used CHO cells in monolayer culture to study the conditions under which the uptake of cadmium occurs. From serumfree medium the metal is accumulated rapidly in the cells. The uptake is inhibited very strongly by the presence of serum or albumin. Accumulation occurs against a concentration gradient and is dependent on the incubation temperature. Below 10°C no cadmium uptake is seen. Several substances which are known to affect cell metabolism have been used to influence cadmium accumulation. Neither inhibitors of energy production nor microtubule or microfilament disruptors showed any substantial effect. In contrast SH‐group blocking agents markedly reduced cadmium uptake.

The results show that cadmium uptake does not occur by passive diffusion but by some active mechanism.     

Biowaste effects on soil and native plants in a semiarid ecosystem

Many soils of the Mediterranean region with a semiarid climate are subjected to progressive degradation as a result of water erosion. Biosolids and municipal solid wastes (MSW) were surface-applied once at three rates (40, 80, and 120 Mg ha(-1)) to different plots in a degraded semiarid ecosystem. The study was conducted to determine the effects of such applications on soil chemical properties and native vegetation over a three-year period. Soil N, P, and K initially increased with increasing biowaste application rates, but then decreased over time. Levels of Zn and Cu were higher in MSW than biosolid-treated plots, and increased in both years after application. Concentrations of soil Cd, Pb, Ni, and Cr did not change as a result of biowaste amendment in the study period. The growth of native plants was enhanced by the addition of biowastes. Total plant canopy and plant biomass increased significantly and remained higher in all treatments than in the control plot over the three-year period. The species richness of native plants decreased with increasing biowaste rates. Differences in the development of native plant communities between treatments were observed, and were more remarkable three years after biowaste application. Tissue N, P, K, Zn, and Cu levels increased with the biowaste application rate, but concentrations of tissue Pb, Cd, Ni, and Cr did not increase significantly. Biowastes applied at the rate of 80 Mg ha(-1) gave rise to the most favorable soil and native vegetation results while avoiding environmental risks.     

USE OF STAGE DATA TO CHARACTERIZE HYDROLOGIC CONDITIONS IN AN URBANIZING ENVIRONMENT1

ABSTRACT: This paper presents the results of a study on the use of continuous stage data to describe the relation between urban development and three aspects of hydrologic condition that are thought to influence stream ecosystems—overall stage variability, stream flashiness, and the duration of extreme‐stage conditions. This relation is examined using data from more than 70 watersheds in three contrasting environmental settings—the humid Northeast (the metropolitan Boston, Massachusetts, area); the very humid Southeast (the metropolitan Birmingham, Alabama, area); and the semiarid West (the metropolitan Salt Lake City, Utah, area). Results from the Birmingham and Boston studies provide evidence linking increased urbanization with stream flashiness. Fragmentation of developed land cover patches appears to ameliorate the effects of urbanization on overall variability and flashiness. There was less success in relating urbanization and streamflow conditions in the Salt Lake City study. A related investigation of six North Carolina sites with long term discharge and stage data indicated that hydrologic condition metrics developed using continuous stage data are comparable to flow based metrics, particularly for stream flashiness measures.     

Maximum rates of nitrate removal in a denitrification wall

Denitrification walls are constructed by mixing a carbon source such as sawdust into soils through which ground water passes. These systems can reduce nitrate inputs to receiving waters by enhancing denitrification. Maximum rates of nitrate removal by denitrification need to be determined for design purposes. To determine maximum rates of nitrate removal we added excess nitrate (50 mg N L(-1)) to a trench up-gradient of a denitrification wall during a 9-d trial. Bromide (100 g L(-1)) was also added as a conservative tracer. Movement of nitrate and bromide was measured from shallow wells and soil samples were removed for measurements of denitrification, carbon availability, nitrate, and other microbial parameters. Rates of nitrate removal, determined from the ratio of NO3-N to Br and ground water flow, averaged 1.4 g N m(-3) of wall d(-1) and were markedly greater than denitrification rates determined using the acetylene block technique (average: 0.11 g N m(-3) of wall d(-1)). These nitrate removal rates were generally lower than reported in other denitrification walls. Denitrification rates increased when nitrate was added to the laboratory incubations, indicating that despite large nitrate inputs in the field, denitrification remained limited by nitrate. This limitation was partially attributed to nitrate predominantly moving through zones of greater hydraulic conductivity or in the mobile fraction of the ground water and slow diffusion to the immobile fraction where denitrifiers were active.     

LANDSAT MID-INFRARED DATA AND GIS IN REGIONAL SURFACE SOIL-MOISTURE ASSESSMENT1

ABSTRACT: Landsat satellite Thematic Mapper (TM) data were used to assess regional soil moisture conditions. The mid-infrared (MIR) data of TM band 7 were overlain onto four principal land-use categories (Agricultural/Irrigated, Urban/Clearings, Forest/ Wetlands, Water) using a geographic information system (GIS). M data were used to assess four qualitative surface soil-moisture conditions (water/very wet, wet, moist, and dry) within each land-use category of a 208,354 ha southwestern Florida study area. The MIR response was inversely related to the qualitative surface soil-moisture content. Integration of Landsat TM MIR data with land use through GIS appears to be a useful technique for high-resolution regional soil moisture assessment, and further research to reline this technique is recommended.