Mechanism of comutagenesis of sodium arsenite withn-methyl-n-nitrosourea

Arsenic compounds are known carcinogens. Although many carcinogens are also mutagens, we have previously shown that sodium arsenite is not mutagenic at either the Na⁺/K⁺ ATPase orhprt locus in Chinese hamster V79 cells. It can, however, enhance UV-mutagenesis. We now confirm the nonmutagenicity of sodium arsenite in line G12, a pSV2gpt-transformed V79 (hprt ⁻) cell line, which is able to detect multilocus deletions in addition to point mutations and small deletions. The lack of arsenic mutagenicity has led to studies emphasizing its comutagenicity. Sodium arsenite at relatively nontoxic concentrations (5 μM for 24 h or 10 μM for 3 h) is comutagenic withN-methyl-N-nitrosourea (MMU) at thehprt locus in V79 cells. Using a nick translation assay, which measures DNA strand breaks by incorporating radioactive deoxyribonucleoside monophosphate at their 3′OH ends in permeabilized cells, we found that much more incorporation was seen in cells treated with MNU (4 mM, 15 min) followed by 3-h incubation with 10 μM sodium arsenite compared with cells exposed to the same MNU treatment followed by 3-h incubation without sodium arsenite. This result shows that in the presence of arsenite, strand breaks resulting from MNU or its repair accumulate over a 3-h period. We suggest that the repair of MNU-induced DNA lesions may be inhibited by arsenite either by affecting the incorporation of dNMPs into the MNU-damaged DNA template or by interfering with the ligation step.     

Ultrastructural immunocytochemical localization of fibronectin deposition during corneal endothelial wound repair. Evidence for cytoskeletal involvement

The distribution of the extracellular matrix (ECM) protein, fibronectin (FN), has been examined ultrastructurally in noninjured and injured rat corneal endothelium in vivo and in vitro by immunoperoxidase cytochemistry. In noninjured endothelia, FN was observed within the rough endoplasmic reticulum (RER) cisternae but not along the cell-Descemet's membrane (DM) interface. Twenty-four and 48 h after a circular freeze injury, immunoperoxidase reaction product was detected at the cell-DM interface as well as within cytoplasmic vesicles and intercellular spaces. By 1 and 2 wk post-injury, a line of reaction product could still be demonstrated at the cell-DM interface and evidence for newly deposited basement membrane material was observed in this region. In order to understand whether fibronectin deposition during wound repair was dependent on cytoskeletal influences, organ culture experiments were performed in which the media was supplemented with either 10(-8) M colchicine or 2.5 X 10(-3) M cytochalasin B. Without inhibitors, injured corneas cultured for 24 h had FN deposition at the cell-DM interface similar to the in vivo results. Corneas cultured in the presence of cytochalasin B also showed FN deposition at the cell-DM interface. However, when injured endothelia were cultured in the presence of colchicine, no reaction product was observed at the cell-DM interface, although it could be detected intracellularly within RER. Incubating the tissues in the presence of puromycin abolished all extracellular and intracellular staining. These results indicate that during wound repair, corneal endothelial cells produce fibronectin and deposit it upon Descemet's membrane by a mechanism that may be mediated by microtubules.     

以点杂交方法对人β型血珠蛋白基因的染色质结构与功能进行的研究

我采用点杂交的方法,对人β型血珠蛋白基因簇的染色质结构与基因转录活性之间的关系进行了研究。以对DNase Ⅰ消化的敏感性作为染色质的结构参数,我将β型血珠蛋白基因簇中11个区域之间以及其与不表达基因区(乳糖白蛋白和免疫球蛋白不变区λ轻链基因)的染色结构进行比较。实验的细胞系统为K 562红白血病细胞与人胚皮肤细胞株(HES)。所获得的结果提示,在细胞核内,表达基因的染色质结构疏松,对DNase Ⅰ消化的敏感性远较不表达基因区的为高。此外,本文还对有关点杂交的方法学问题进行了较为详尽的讨论。     

中国的红树林

我国红树林主要分布在热带及亚热带沿海盐滩和河川出口的冲积盐性土上,包括24个科,47种,属于东方群系。从广西钦州湾开始经北海港,沿雷州半岛的海安分为两支,一支往南经海南岛的儋县新英港,向东经琼山县的东寨港和文昌县的清澜港,沿着东海岸至崖县的三亚港,又折向北到达东方县的八所港;另一支向东北经湛江的赤坎,阳江的海陵岛,台山的上下川岛,中山的三灶岛,新会的崖门,再沿海岸往北至福建的福鼎和台湾的新竹港。根据我国红树林的主要构成种类、外貌、生境,大致可分为3大群系和10个群落。(1) 矮灌木群系——这个群系多见于纬度稍高的海滩前缘,包括海榄雌群落和桐花树群落。这些群落在演替上处于前期阶段。(2) 高大稠密灌木群系——这个群系由于气温、年降雨量的差异和土壤性质的不同,组成这个群系的种类也有不同,不过都是以红树族的种类为建群种,它包括4个群落:红树群落、红海兰群落和角果木群落,这些群落在演替上属于中期阶段。(3) 乔木群系——这个群系主要分布在海南岛东北和东部海岸,所在地一般地势较高,前缘地带每日潮涨仍有海水浸淹。后缘地带只有每月大潮时才有海水到达,土壤比较固结,它包括木榄群落、海莲和尖瓣海莲群落、海桑群落及半红树林水椰群落。这些群落在演替上是处于后期阶段。组成我国红树林的种类大多数都具有较高的经济价值,如角果木是优质单宁的原料,有些种类经过发酵处理成为高营养的饲料和肥料。此外,沿海浮游生物的产量有红树林比无红树林的地带高达7倍,因此加强对红树林的经营和保护应采取积极态度和有效措施。     

中国的红树林

我国红树林主要分布在热带及亚热带沿海盐滩和河川出口的冲积盐性土上,包括24个科,47种,属于东方群系。从广西钦州湾开始经北海港,沿雷州半岛的海安分为两支,一支往南经海南岛的儋县新英港,向东经琼山县的东寨港和文昌县的清澜港,沿着东海岸至崖县的三亚港,又折向北到达东方县的八所港;另一支向东北经湛江的赤坎,阳江的海陵岛,台山的上下川岛,中山的三灶岛,新会的崖门,再沿海岸往北至福建的福鼎和台湾的新竹港。根据我国红树林的主要构成种类、外貌、生境,大致可分为3大群系和10个群落。(1) 矮灌木群系——这个群系多见于纬度稍高的海滩前缘,包括海榄雌群落和桐花树群落。这些群落在演替上处于前期阶段。(2) 高大稠密灌木群系——这个群系由于气温、年降雨量的差异和土壤性质的不同,组成这个群系的种类也有不同,不过都是以红树族的种类为建群种,它包括4个群落:红树群落、红海兰群落和角果木群落,这些群落在演替上属于中期阶段。(3) 乔木群系——这个群系主要分布在海南岛东北和东部海岸,所在地一般地势较高,前缘地带每日潮涨仍有海水浸淹。后缘地带只有每月大潮时才有海水到达,土壤比较固结,它包括木榄群落、海莲和尖瓣海莲群落、海桑群落及半红树林水椰群落。这些群落在演替上是处于后期阶段。组成我国红树林的种类大多数都具有较高的经济价值,如角果木是优质单宁的原料,有些种类经过发酵处理成为高营养的饲料和肥料。此外,沿海浮游生物的产量有红树林比无红树林的地带高达7倍,因此加强对红树林的经营和保护应采取积极态度和有效措施。     

Opposing effects of heparin with TGF-beta or aFGF during repair of a mechanical wound of human endothelium. Influence of cAMP on cell migration.

The effects on vascular wound repair in vitro of aFGF and TGF-beta, growth factors having opposite influences on endothelial cell growth and angiogenesis, were studied using as a model a mechanical lesion of confluent endothelium. Modulation by heparin of the activities of these growth factors during the repair process was also examined. Whereas heparin alone inhibited repair by lowering both cell proliferation and cell migration, TGF-beta alone mainly inhibited cell proliferation. When added together, TGF-beta and heparin exerted a combined inhibitory effect resulting in a residual lesion 50% larger than in controls. aFGF alone accelerated lesion coverage and this effect was enhanced by 40% over control values when heparin was added with aFGF. This acceleration was slightly (less than 10%) but consistently diminished by TGF-beta. Cell density in confluent unwounded areas was increased by 40% in the presence of aFGF, but TGF-beta diminished cell density by 20%. A small (30%) increase in intracellular cAMP was measured whenever aFGF was present during the repair process. In comparison, intracellular cAMP inducing agents (forskolin, dbcAMP) accelerated cell migration by 20% during lesion recovery without affecting cell proliferation or density. The present results show that the inhibitory effects of TGF-beta during vascular wound repair are opposed by aFGF. Furthermore, heparin (or heparan sulfates in vivo) modulates growth factors having activating or inhibiting functions and thus plays a regulatory role during the repair process. cAMP-inducing substances other than growth factors are able to accelerate cell migration.     

东南亚和云南爬行动物区系的一致性及其起源和演化

云南与川、黔、贵等邻近省区都属于东南亚的一部分。依据爬行动物分布范围广的特点,把云南爬行动物区系起源和演化与东南亚甚至南亚的爬行动物作为一个整体进行研究,可能为解决东南亚及云南爬行动物某些类群的起源这一共同性问题,提供一些有用的资料。 本文将印度半岛、东南亚及其邻近岛屿现生爬行动物与世界范围的相同科级阶元的分布进行比较,并以古地质、古地理演变资料推论科级阶元同祖先起源地的大致范围。在此基础上,再将印度半岛、中南半岛和邻近岛屿,云南高原及邻近地区爬行动物的科属种进行比较,以探讨地区间差异的规律性。运用新构造运动和古气候变化观点阐明其物种或类群的迁移规律及其变化原因。     

DNA sequence changes in mutations induced by ultraviolet light in the gpt gene on the chromosome of Escherichia coli uvr+ and urvA cells

Summary Sequence changes in mutations induced by ultraviolet light are reported for the chromosomal Escherichia coli gpt gene in almost isogenic E. coli uvr ⁺ and excision-deficient uvrA cells. Differences between the mutagenic spectra are ascribed to preferential removal of photoproducts in the transcribed strand by excision repair in uvr ⁺ cells. This conclusion is confirmed by analysis of published results for genes in both uvr ⁺ and uvr ^– cells, showing a similar selective removal of mutagenic products from the transcribed strand of the E. coli lacI gene and of the lambda phage cl repressor gene. Comparison of these data with published results for ultraviolet mutagenesis of gpt on a chromosome in Chinese hamster ovary cells showed that a mutagenic hot spot in mammalian cells is not present in E. coli; the possibility is suggested that the hot spot might arise from localized lack of excision repair. Otherwise, mutagenesis in hamster cells appeared similar to that in E. coli uvr ⁺ cells, except there appears to be a smaller fraction of single-base additions and deletions (frameshifts) in mammalian than in bacterial cells. Phenotypes of 6-thioguanine-resistant E. coli showed there is a gene (or genes) other than gpt involved in the utilization of thioguanine by bacteria.     

Evidence of abortive recombination in ruv mutants of Escherichia coli K12

Summary Genetic recombination in Escherichia coli was investigated by measuring the effect of mutations in ruv and rec genes on F-prime transfer and mobilization of nonconjugative plasmids. Mutation of ruv was found to reduce the recovery of F-prime transconjugants in crosses with recB recC sbcA strains by about 30-fold and with recB recC sbcB sbcC strains by more than 300-fold. Conjugative plasmids lacking any significant homology with the chromosome were transferred normally to these ruv mutants. Mobilization of the plasmid cloning vectors pHSG415, pBR322, pACYC184 and pUC18 were reduced by 20- to 100-fold in crosses with ruv rec ⁺ sbc ⁺ strains, depending on the plasmid used. Recombinant plasmids carrying ruv ⁺ were transferred efficiently. With both F-prime transfer and F-prime cointegrate mobilization, the effect of ruv was suppressed by inactivating recA. It is proposed that the failure to recover transconjugants in ruv recA ⁺strains is due to abortive recombination and that the ruv genes define activities which function late in recombination to help convert recombination intermediates into viable products.     

The mechanism of extrachromosomal homologous DNA recombination in plant cells

Summary By cotransfecting plasmids carrying particular mutations in the -glucuronidase (GUS) gene into Nicotiana plumbaginifolia protoplasts and by monitoring the recombination rates using a recently developed transient assay, we were able to obtain insights into the mechanism of extrachromosomal recombination operating in plant cells. An exchange of flanking markers takes place in over 90% of the recombination events. In most of the remaining cases two consecutive, independent single crossover events occur. These events involve the same DNA substrate and lead to two successive exchanges of flanking markers, thus mimicking a presumed double crossover intermediate. A comparison of the outcome of our experiments with the predictions of two recombination models originally proposed for mammalian cells indicates that extrachromosomal recombination in plant cells is best described by the single strand annealing model. According to this model all recombination events result in an exchange of flanking markers. Our results rule out the double strand break repair model which predicts that flanking markers are exchanged in only half of all events.