Genome Stability vs. Deprivation or Enrichment of the Geomagnetic Field

Summary This study suggested that cell-cycle kinetics, DNA replication and DNA repair react to magnetic fields differently. During their culture growth cycle, which lasted about five days, Friend erythroleukemia cells were either kept in the absence of magnetic fields in a magnetically shielded room or irradiated in a solenoid with 70 μT at 50 Hz plus 45 μT DC of the Earth: some cells grew without inducer of in vitro differentiation; others were induced to differentiate hemoglobin through dimethylsulfoxide. It emerged that, during a single culture growth cycle, while proliferation was slightly accelerated by the magnetic-field irradiation achieved in the solenoid both in undifferentiating and dimethylsulfoxide-differentiating cells, DNA replication did not appear to significantly depend on the magnetic-field deprivation achieved in the magnetically shielded room. However, as a result of a 318-day long magnetic-field irradiation in the solenoid, DNA replication remained unchanged in undifferentiating cells and partially inhibited in dimethylsulfoxide-differentiating cells. Following the same long magnetic-field irradiation in the solenoid, the amount of labelled repair patches in the parental DNA strands was slightly reduced.     

Spatial relationships and matrilineal kinship in African savanna elephant (Loxodonta africana) clans

African savanna elephants, Loxodonta africana, live in stable family groups consisting of adult females and their dependent offspring. During the dry season, clans consisting of several family groups typically share a common home range. We compared spatial relationships and mitochondrial DNA (mtDNA) haplotypes among 14 adult female elephants within 3 clans during the dry season in northern Zimbabwe. Spatial relationships were studied by radio-tracking. Home-range similarity was quantified by correlating the estimated utilization distributions of all pairs of elephants. Clans were identified by cluster analysis of the home-range similarity values. All three clans contained at least two of the five mtDNA haplotypes that were found, indicating that clan members are not necessarily matrilineally related. Within clans, home ranges of elephants with the same haplotype were not significantly more similar to each other than those of elephants with different haplotypes. Most elephants within each clan used their shared home ranges independently of each other: the distribution of distances between their positions at any given time did not differ from the distribution expected by chance. However, 8 out of the 26 within-clan pairs exhibited long-term coordination of space use by remaining within known hearing distance of each others low-frequency calls significantly more often than expected by chance. At least four of these coordinated pairs consisted of animals in different family groups. Elephants in three of the four different-family pairs whose movements were coordinated had different haplotypes. Further research is needed to determine the relationship between these coordinated movements and conventionally defined bond-group behavior.Electronic Supplementary Material Supplementary material is available in the online version of this article at .Communicated by C. Nunn     

Prenatal identification of a Y-chromosome deletion by Y-specific single copy DNA probes

A sex chromosome deletion was identified in the course of prenatal diagnosis for maternal age. Ultrasound pictures revealed male fetal sex and a comparison with the father's Y chromosome suggested that the altered chromosome might be a de novo deletion of the Y chromosome. DNA hybridization with five human Y-specific probes shows that, among the Y-specific sequences recognized by the probes, only two of them are absent. The normal infant, at birth, was mosaic 46, XYq- /46,XY.     

Exposure of Marine Mammals to Genotoxic Environmental Contaminants: Application of the 32P-Postlabeling Assay for Measuring DNA-Xenobiotic Adducts♯

There is little information on exposure of marine mammals to genotoxic environmental contaminants. The 32P-postlabeling assay has been successfully used to assess exposure to genotoxic polycyclic aromatic compounds in fish and humans. In the present study, a preliminary investigation showed that polycyclic aromatic compound-like DNA adducts were present in hepatic tissues of harbor seals (Phoca vitulina richardsi) exposed to petroleum following the Exxon Valdez oil spill. However, for marine mammals, effects from changes in tissue condition on DNA recovery and quality is of concern, because tissue samples are often collected from animals that have been dead for unknown periods of time. To assess the effects of postmortem thermal history on DNA recovery from tissue and on DNA adduct quantitation, samples of harbor porpoise (Phocoena phocoena) hepatic tissue were incubated for up to 10 d at 4 and 30 °C. Only traces (<4 g) of hepatic DNA were recovered from 200 mg of tissue after incubation at 30 °C for 36 h. At 4 °C, DNA (50–130 g) was recovered from tissue incubated for up to 6 d; whereas DNA recovery at 10 d was minimal. Chromatograms of 32P-labeled DNA digests of liver tissue held at 4 and 30 °C and salmon sperm DNA held at 30 °C for 2 d had comparable profiles, suggesting that alteration of DNA bases had occurred during incubation of porpoise liver tissue. Moreover, the chromatograms of DNA extracted from liver tissues of harbor porpoises caught incidentally in a northwest Atlantic fishery, packed in ice and sampled several days later also exhibited similar altered DNA structures. Although, altered DNA structures that can interfere with the DNA adduct quantitation were present in autolyzed tissue, changes in the 32P-postlabeling chromatography conditions can decrease the interference. Moreover, in a study with tissues taken from California sea lions (Zalophus californianus) immediately postmortem and stored at –80 °C until processing, DNA structures associated with tissue breakdown were not observed. The DNA from sea lions, however, had putative age-dependent hepatic DNA modifications, which have a distinctive profile, and must be considered when evaluating exposure of marine mammals to polycyclic aromatic compounds. Overall, the findings showed that with attention to the postmortem thermal history of the tissue samples hepatic DNA adducts, as measured by 32P-postlabeling, have the potential to serve as a biological indicator of exposure of marine mammals to environmental genotoxic compounds.     

Purification of total DNA extracted from activated sludge

Purification of the total DNA extracted from activated sludge samples was studied. The effects of extraction buffers and lysis treatments (lysozyme, sodium dodecyl sulfate (SDS), sonication, mechanical mill and thermal shock) on yield and purity of the total DNA extracted from activated sludge were investigated. It was found that SDS and mechanical mill were the most effective ways for cell lysis, and both gave the highest DNA yields, while by SDS and thermal shock, the purest DNA extract could be obtained. The combination of SDS with other lysis treatment, such as sonication and thermal shock, could apparently increase the DNA yields but also result in severe shearing. For the purification of the crude DNA extract, polyvinyl polypyrrolidone was used for the removal of humic contaminants. Cetyltrimethyl ammonium bromide, potassium acetate and phenol/chloroform were used to remove proteins and polysaccharides from crude DNA. Crude DNA was further purified by isopropanol precipitation. Thus, a suitable protocol was proposed for DNA extraction, yielding about 49.9 mg (total DNA)/g volatile suspended solids, and the DNA extracts were successfully used in PCR amplifications for 16S rDNA and 16S rDNA V3 region. The PCR products of 16S rDNA V3 region allowed the DGGE analysis (denatured gradient gel electrophoresis) to be possible.     

经DNA改组的植酸酶纯化和酶学性质

经DNA改组的重组植酸酶通过有机膜超滤、DEAE -SepharoseF .F离子交换层析两步纯化 ,其纯度可达90 %左右 .SDS -PAGE分析表明 ,重组植酸酶的分子量Mr 约为 85 0 0 0 .酶学实验结果表明 :该酶反应最适pH为 4 .5 ,最适温度为 4 0℃ ,Km为 0 .11mmolL-1,Vmax为 96 4mmolL-1min-1,植酸酶活性不依赖任何金属离子的存在 .向酶液中分别添加MgSO4、阿拉伯糖、果糖、半乳糖、乳糖、海藻糖、蔗糖及葡萄糖 (10 0 g/L)后 ,植酸酶在 90℃处理 10min的活性比对照增加了 1～ 3倍 ,说明这些物质为酶保护剂 ;在 37℃下以植酸钠为底物的SDS对酶活性具有强烈的抑制作用 ;金属离子如Cu2 ,Zn2 ,K 以及EDTA对酶活性具有较弱抑制作用 ;金属离子如Mn2 ,Mg2 、Fe2 、Ba2 、Ca2 、Co2 低浓度时对酶活性有促进作用 ,当Mg2 浓度增加到 10mmolL-1时 ,则对酶活性起抑制效应 .图 4表 4参 19     

Slow Growth and Lack of Recovery in Overfished Holothurians on the Great Barrier Reef: Evidence from DNA Fingerprints and Repeated Large-Scale Surveys

Abstract:  Commercially fished holothurians have important functions in nutrient recycling, which increases the benthic productivity of coral reef ecosystems. Thus, removal of these animals through fishing may reduce the overall productivity of affected coral reefs. To investigate the potential for recovery of overfished holothurian (  Holothuria nobilis ) stocks on the Great Barrier Reef (GBR), we (1) conducted field surveys on 23 reefs after fishery closure, (2) modeled total virgin biomass and compared it with the total amount fished, and (3) estimated individual growth rates with a DNA fingerprinting technique. Two years after fishery closure, no recovery of H. nobilis stocks on reefs previously open to fishing was observed. Densities on reefs protected from fishing since the onset of the fishery in the mid 1980s remained about four times higher than on fished reefs. Based on density estimates and geographic information system data on the habitat area of each reef, we calculated that the virgin biomass (in the main fished area between 12° and 19°S) was about 5500 t and is now about 2500 t. The reduction is on the same order of magnitude as the total amount fished until 1999 (approximately 2500 t). The DNA analysis of repeated samples on three locations indicated high recapture rates of fingerprinted and released individuals of H. nobilis . Fitting growth curves with Francis's growth function indicated that medium-sized individuals (1 kg) grew 35–533 g /year, whereas large animals (2.5 kg) consistently shrank. Small animals (<500 g) were rarely observed. In combination, these data indicate that production of H. nobilis stocks is very low, presumably with low mortality, low recruitment, and slow individual growth rates. Consistent with anecdotal evidence, recovery of H. nobilis stocks on the GBR may take several decades, and we suggest a highly conservative management plan to protect both the stocks and the ecosystem.     

榨菜线粒体DNA雄性不育相关片段克隆及序列分析

研究了榨菜(Brassica juncea var．tumida Tsen and Lee)细胞质雄性不育性(CMS)与线粒体基因组的关系,以榨菜细胞质雄性不育系及其保持系线粒体DNA(mtDNA)为模板,设计合成引物进行PCR扩增,从榨菜胞质雄性不育系mtDNA上扩增得到与雄性不育相关的特异片段T1170,点杂交分析表明,T1170仅与榨菜胞质雄性不育系mtDNA有杂交信号,而保持系的mtDNA则无,序列测定该片段全长1173bp,编码389个氨基酸,其中疏水氨基酸占35％。图4参9。