Mutagenesis of benzo[a]pyrene diol epoxide in yeast: requirement for DNA polymerase zeta and involvement of DNA polymerase eta

Benzo[a]pyrene is a potent environmental carcinogen, which can be metabolized in cells to the DNA damaging agent anti-benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (anti-BPDE). We hypothesize that mutations induced by BPDE DNA adducts are mainly generated through an error-prone translesion synthesis that requires a specialized DNA polymerase (Pol). Using an in vivo mutagenesis assay in the yeast model system, we have examined the potential roles of Pol(zeta) and Pol(eta) in (+/-)-anti-BPDE-induced mutagenesis. In cells proficient in mutagenesis, (+/-)-anti-BPDE induced 85% base substitutions with predominant G --> C followed by G --> T transversions, 9% deletions of 1-3 nucleotides, and 6% insertions of 1-3 nucleotides. In rad30 mutant cells lacking Pol(eta), (+/-)-anti-BPDE-induced mutagenesis was reduced and accompanied by a moderate decrease in base substitutions and more significant decrease in deletions and insertions of 1-3 nucleotides. In rev3 mutant cells lacking Pol(zeta), (+/-)-anti-BPDE-induced mutagenesis was mostly abolished, leading to a great decrease in both base substitutions and deletions/insertions of 1-3 nucleotides. In contrast, large deletions/insertions were significantly increased in cells lacking Pol(zeta). Consistent with the in vivo results, purified yeast Pol(zeta) performed limited translesion synthesis opposite (+)- and (-)-trans-anti-BPDE-N(2)-dG DNA adducts with predominant G incorporation opposite the lesion. These results show that (+/-)-anti-BPDE-induced mutagenesis in yeast requires Pol(zeta) and partially involves Pol(eta) and suggest that Pol(zeta) directly participates in nucleotide insertions opposite the lesion, while Pol(eta) significantly contributes to deletions and insertions of 1-3 nucleotides.     

Increase of intracellular Ca(2+) during ischemia/reperfusion injury of heart is mediated by cyclic ADP-ribose

While the molecular mechanisms by which oxidants cause cytotoxicity are still poorly understood, disruption of Ca(2+) homeostasis appears to be one of the critical alterations during the oxidant-induced cytotoxic process. Here, we examined the possibility that oxidative stress may alter the metabolism of cyclic ADP-ribose (cADPR), a potent Ca(2+)-mobilizing second messenger in the heart. Isolated heart perfused by Langendorff technique was subjected to ischemia/reperfusion injury and endogenous cADPR level was determined using a specific radioimmunoassay. Following ischemia/reperfusion injury, a significant increase in intracellular cADPR level was observed. The elevation of cADPR content was closely correlated with the increase in ADP-ribosyl cyclase activity. Inclusion of oxygen free radical scavengers, 2,2,6,6-tetramethyl-1-piperidinyloxy and mannitol, in the reperfusate prevented the ischemia/reperfusion-induced increases in cADPR level and the ADP-ribosyl cyclase activity. Exposure of isolated cardiomyocytes to t-butyl hydroperoxide increased the ADP-ribosyl cyclase activity, cADPR level, and intracellular Ca(2+) concentration ([Ca(2+)](i)) and consequently resulting in cell lethal damage. The oxidant-induced elevation of [Ca(2+)](i) as well as cell lethal damage was blocked by a cADPR antagonist, 8-bromo-cADPR. These results provide evidence for involvement of cADPR and its producing enzyme in alteration of Ca(2+) homeostasis during the ischemia/reperfusion injury of the heart.     

Effects of La3+ on growth,transformation, and gene expression of Escherichia coli

Rare earth elements have been emitted into the environment largely as fertilizer components. This has caused much fear about whether they would influence our environment, especially on the metabolism and genetics of microorganisms. In this article, the trivalent ion of a rare earth element, lanthanum, was studied for the effects on growth, transformation, and gene expression of Escherichia coli. The results showed that La³⁺ at concentrations from 50 to 150 μg/mL stimulated the endogenic metabolism and ectogenic metabolism, but had few effects on gene expression. La³⁺ at lower concentrations from 0.5 to 30 μg/mL inhibit intensively E. coli-absorbing external DNA, decreasing the transformation efficiency. It is also supported by observations using transmission electron microscopy. Our results are significant in understanding the function of rare earth elements to microorganisms and assessing the risk of application of rare earth compounds.     

江西省野生维管植物名录

为了全面了解江西省野生维管植物资源的现状并为开展相应的生物多样性保护实践提供基础资料, 本文在前人的基础上, 通过进一步的文献收集、标本考证、数据分析, 并结合作者的野外调查结果, 按照维管植物最新的分类系统整理出江西省野生维管植物名录, 并根据标本信息核实了每个物种县市级的分布信息。结果显示, 江西省共有野生维管植物214科1,253属4,761种(含种下等级), 其中石松类与蕨类植物35科103属444种, 裸子植物5科21属36种, 被子植物174科1,129属4,281种。石松类与蕨类植物中, 物种数最多的前5个科为鳞毛蕨科(84种)、水龙骨科(55种)、凤尾蕨科(49种)、蹄盖蕨科(47种)和金星蕨科(47种), 最多的属是鳞毛蕨属(Dryopteris, 40种); 裸子植物中物种数最多的科和属为松科(7属12种)和松属(Pinus, 5种); 被子植物是江西省野生维管植物的最主要组成部分, 分别占科、属、种总数的81.3%、90.1%和89.9%, 其中种数最多的前5个科分别是禾本科(315种)、菊科(241种)、蔷薇科(228种)、唇形科(200种)和豆科(197种), 前6个属依次是薹草属(Carex, 71种)、悬钩子属(Rubus, 66种)、冬青属(Ilex, 56种)、蓼属(Persicaria, 40种)、珍珠菜属(Lysimachia, 37种)和杜鹃花属(Rhododendron, 37种)。江西省维管植物调查呈现出地区不均衡性, 建议增加后续调查工作的广度和深度, 尤其在武夷山脉、赣南山区、鄱阳湖湿地等生物多样性较高或较特殊的地区进行补充采集。同时, 本文呼吁加强专科专属的分类学研究, 为后续名录的修订与完善奠定基础。     

The Wilms Tumor Gene,Wt1, Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans

Azoospermia is one of the major reproductive disorders which cause male infertility in humans; however, the etiology of this disease is largely unknown. In the present study, six missense mutations of WT1 gene were detected in 529 human patients with non-obstructive azoospermia (NOA), indicating a strong association between WT1 mutation and NOA. The Wilms tumor gene, Wt1, is specifically expressed in Sertoli cells (SCs) which support spermatogenesis. To examine the functions of this gene in spermatogenesis, Wt1 was deleted in adult testis using Wt1^flox and Cre-ER^TM mice strains. We found that inactivation of Wt1 resulted in massive germ cell death and only SCs were present in most of the seminiferous tubules which was very similar to NOA in humans. In investigating the potential mechanism for this, histological studies revealed that the blood–testis barrier (BTB) was disrupted in Wt1 deficient testes. In vitro studies demonstrated that Wt1 was essential for cell polarity maintenance in SCs. Further studies found that the expression of cell polarity associated genes (Par6b and E-cadherin) and Wnt signaling genes (Wnt4, Wnt11) were downregulated in Wt1 deficient SCs, and that the expression of Par6b and E-cadherin was regulated by Wnt4. Our findings suggest that Wt1 is important in spermatogenesis by regulating the polarity of SCs via Wnt signaling pathway and that WT1 mutation is one of the genetic causes of NOA in humans.     

Phenotypic and genetic differences between opaque and translucent colonies of Vibrio alginolyticus

Many pathogens undergo phase variation between rugose and smooth colony morphology or between opaque and translucent colony morphology, which is mainly due to the variation in the surface polysaccharides. In this study, Vibrio alginolyticus ZJ-51 displayed phase variation between opaque, rugose colonies (Op) and translucent, smooth colonies (Tr). Unlike the vibrios reported previously, Tr cells of ZJ-51 enhanced biofilm formation and motility, but they did not differ from Op cells in the quantity of surface polysaccharides produced. Real time PCR was used to analyze the expression of the genes involved in polysaccharide biosynthesis, flagellar synthesis, and the AI-2 quorum-sensing system. The results revealed that the K-antigen capsule gene cluster (which consists of homologs to the cpsA-K in Vibrio parahaemolyticus) and O-antigen polysaccharide gene cluster (which contains homologs to the wza-wzb-wzc) were significantly more transcribed in Tr cells. The AI-2 quorum-sensing genes showed enhanced expression in the Tr variant which also exhibited greater expression of genes associated with polar flagellar biosynthesis. These results suggest that colony phase variation might affect the virulence and survival ability in the stressful environment inhabited by V. alginolyticus.     

长薄鳅胚胎发育的观察

1999年5月,作者首次通过人工繁殖获得长薄鳅的受精卵,并对其胚胎发育特征进行了连续的观察。卵膜无粘性,吸水后卵周间隙明显膨胀扩大,卵膜径3.67-4.00 mm,平均3.79mm,属漂流性卵。胚胎发育可分为26个时期,初孵仔鱼长5.0mm左右,通体透明无色素。     

Changes in winter snow depth affects photosynthesis and physiological characteristics of biological soil crusts in the Tengger Desert

Water availability is a major limiting factor in desert ecosystems. However, a winter snowfall role in the growth of biological soil crusts is still less investigated. Here, four snow treatments were designed to evaluate the effects of snow depth on photosynthesis and physiological characteristics of biological soil crusts. Results showed that snow strongly affected the chlorophyll fluorescence properties. The increased snow depth led to increased contents of photosynthetic pigments and soluble proteins. However, all biological soil crusts also exhibited a decline in malondialdehyde and soluble sugar contents as snow increased. Results demonstrated that different biological soil crusts exhibited different responses to snow depth treatment due to differences in their morphological characteristics and microhabitat. In addition, interspecies differentiation in response to snow depth treatment might affect the survival of some biological soil crusts. Further, this influence might lead to changes in the structural composition and functional communities of biological soil crusts.

     

Databases and Web Tools for Cancer Genomics Study

Publicly-accessible resources have promoted the advance of scientific discovery. The era of genomics and big data has brought the need for collaboration and data sharing in order to make effective use of this new knowledge. Here, we describe the web resources for cancer genomics research and rate them on the basis of the diversity of cancer types, sample size, omics data comprehensiveness, and user experience. The resources reviewed include data repository and analysis tools; and we hope such introduction will promote the awareness and facilitate the usage of these resources in the cancer research community.