Prenatal diagnosis of twin zygosity by DNA ‘fingerprint’ analysis

A twin pregnancy with one hydrocephalic fetus with oligohydramnios is presented. Sonographic evaluation could not exclude monochorionicity. Before considering selective feticide, blood samples from both fetuses were examined for DNA ‘fingerprint’ analysis. The different banding patterns of the blood samples established dizygosity. This procedure is suggested in cases where sonography fails to determine chorionicity.     

Degradation of phenol in an upflow anaerobic sludge blanket (UASB) reactor at ambient temperature

IntroductionPhenolistherawmaterialforthecommercialproductionofawidevarietyofresins ,includingphenolicresinsasconstructionmaterialsforautomobilesandappliances,expoxyresinsasadhesives ,andpolyamideforvariousapplications.Inaddition ,phenolsareoftenfoundinwas…     

环境污染条件下生物体内DNA损伤的生物标记物研究进展

在正常条件下生物体内的基因组是稳定的;但在环境污染条件下,其DNA容易遭受伤害,主要损伤形式为:碱基改变、脱碱基位点、碱基错配、插入或缺失片段、嘧啶联合、DNA加合物、DNA链断裂、甲基化损伤、DNA链内和链间交联等. 这些受损的DNA对生物细胞产生遗传毒性或细胞毒性. 利用生物标记物进行DNA损伤的检测和定量分析是可行的方法. 本文重点介绍了一些典型的DNA损伤(如DNA加合物、断裂、DNA序列改变等)的生物标记物及其检测方法;认为这些生物标记物在环境污染物的早期诊断和评价方面具有广阔的应用前景. 参32     

Mechanisms of ultraviolet disinfection and chlorination of Escherichia coli: Culturability, membrane permeability, metabolism, and genetic damage

Traditional culture methods may underestimate the tolerance of microorganisms to disinfectants because of the existence of viable but nonculturable or sublethally injured cells after disinfection.The selection of a strict method is crucial for the evaluation of disinfection performance.The actions of 2 typical disinfectants–ultraviolet(UV)and chlorine–on the fecal indicator Escherichia coli were investigated by the detection of culturability,membrane permeability,metabolic activity,deoxyribonucleic acid(DNA),and messenger ribonucleic acid(m RNA).During UV disinfection,the irreversible damages in the cell membrane and cellular adenosine triphosphate(ATP)were negligible at low UV doses(80 m J/cm~2).However,membrane permeability was damaged at low doses of chlorine(5 mg/L),leading to leakage of cellular ATP.Our study showed that a slight lesion in DNA was detected even at high doses of UV(400 m J/cm~2)and chlorine(5 mg/L)treatments.The decay of m RNA was more rapid than that of DNA.The degradation level of m RNA depended on the choice of target genes.After exposure to 50 m J/cm~2UV dose or 5 mg/L chlorine for30 min,the DNA damage repair function(Rec A m RNA)was inhibited.The m RNA involved in the DNA damage repair function can be a potential indicator of bacterial viability.     

北京大气PM_(2.5)对A549细胞炎性因子及DNA损伤的毒性

为探讨大气PM_(2.5)及其不同组分对人肺上皮细胞A549的毒性作用及其剂量-反应关系,将前期采集的PM_(2.5)颗粒物用不同方法进一步制备PM_(2.5)水溶性组分、PM_(2.5)脂溶性组分和PM_(2.5)单纯颗粒物,将制备的PM_(2.5)颗粒物及其组分以不同浓度(10,50,100,200,400μg/m L)对A549细胞染毒,用MTS法分别在染毒6,10,24,48,72h后测定细胞活力,染毒24h后用ELISA及RT-QPCR法测定炎性因子IL-6和TNF-α表达量,AP位点计数法测定细胞DNA损伤情况.结果表明:除PM_(2.5)水溶性组分外,其余染毒样本高浓度染毒时始终对细胞生长表现出抑制作用,其中低浓度染毒时可在较短时间对细胞生长表现出抑制作用,染毒时间较长时抑制作用减弱或消失,PM_(2.5)水溶性组分对细胞生长抑制作用并不显著;除PM_(2.5)水溶性组分外,其余染毒样本都显著升高了IL-6m RNA的相对表达量和IL-6蛋白的分泌,除PM_(2.5)脂溶性组分外,其余染毒样本都显著升高了TNF-αm RNA的相对表达量;除PM_(2.5)水溶性组分外,其余染毒样本都显著提高了DNA碱基缺失程度.总的来说,PM_(2.5)水溶性组分在抑制细胞活力、造成炎性损伤及DNA损伤方面作用相对较小,而PM_(2.5)所产生的毒性作用并不仅限于其所吸附的复杂成分,其中作为载体的固体核心颗粒对机体可能造成的毒性作用也不容忽视.     

Screening pesticides for their ability to damage bacterial DNA

Abstract

Twenty‐six pesticides and pesticide degradation products were screened (125 μg ‐ 2000 μg) for their ability to induce unrepairable damage to bacterial DNA. Three repair test systems were utilized in this study, the Salmonella typhimurium (TA1538/TA1978), the E. coli K‐12 (Pol A₁ ⁺/Pol^‐ ₁) and the E. coli WP₂ (WP₂, WP₂ uvrA, WP67, CM611 and CM571). Aldicarb (1000 μg), benomyl (250 μg), 2‐aminobenzimidazole (2000 μg), captan (125 μg), fenazalor (500 μg), 5,6‐dichloro‐2‐trifluoromethylbenzimida‐zole (NC‐2983) (250 μg), isothymol (250 μg), maleic hydrazide(1000 μg), pentachloronitrobenzene (1000 μg) were DNA‐damaglng to one or more bacterial test systems. Isothymol and NC‐2983 affected all three test systems. Chlorinated hydrocarbon insec ticides, some being recognized as carcinogens, did not: produce a zone of inhibition in any of the tester strains possibly due to their poor solubility and diffusion in the agar overlay. It was concluded that these tests can be performed along with bacterial reversion tests to complement each other as short‐term screening tests for potential carcinogens and mutagens.     

The adsorption of short single-stranded DNA oligomers to mineral surfaces

We studied the adsorption of short single-stranded deoxyribonucleic acid (ssDNA) oligomers, of approximately 30 nucleotides (nt) in length, of varying sequence, adenine + guanine + cytosine (AGC) content, and propensity to form secondary structure, to equal surface area samples of olivine, pyrite, calcite, hematite, and rutile in 0.1 M NaCl, 0.05 M pH 8.1 KHCO₃ buffer. Although the mineral surfaces have widely varying points of zero charge, under these conditions they show remarkably similar adsorption of ssDNA regardless of oligomer characteristics. Mineral surfaces appear to accommodate ssDNA comparably, or ssDNA oligomers of this length are able to find binding sites of comparable strength and density due to their flexibility, despite the disparate surface properties of the different minerals. This may partially be due charge shielding by the ionic strength of the solutions tested, which are typical of many natural environments. These results may have some bearing on the adsorption and accumulation of biologically derived nucleic acids in sediments as well as the abiotic synthesis of nucleic acids before the origin of life.     

Determination of hypoxia and dietary copper mediated sub-lethal toxicity in carp, Cyprinus carpio, at different levels of biological organisation

Hypoxic events frequently occur in the aquatic environment in association with micro pollutants, including heavy metals. Only a few studies are however available on the uptake and biological responses of heavy metals under hypoxic conditions. To elucidate the phenomenon, mirror carp Cyprinus carpio L. (16.13-16.22 g) were exposed chronically to dietary copper (Cu; 250 and 500 mg kg dry wt.⁻¹) for 30 d under normoxic (8.25 mg O₂ L⁻¹) and hypoxic (∼3 mg O₂ L⁻¹) conditions and adopting an integrated approach, sub-lethal biomarker responses were determined at different levels of biological organisation. Level of oxidative DNA damage (as determined by modified Comet assay) showed strong significant difference following exposure to dietary Cu level under normoxic (1.6-fold) as well as under hypoxic condition at both Cu levels (2.1 and 2.5-folds respectively). Significant difference was also observed for haematological parameters (i.e. increased red and white blood cells, haematocrit value and haemoglobin concentration). Quantitative histology revealed alterations in tissues (i.e. liver and gills) for hypoxic and all dietary Cu treatment groups under both normoxic and hypoxic conditions suggesting a compensatory response to these organs (p < 0.05). The order of Cu accumulation in tissues (as determined by ICP-OES) was liver > intestine > kidney > gill. Interestingly, SGR under both normoxic and hypoxic conditions reduced with elevating Cu levels (p = 0.019). Overall, the results provide evidence for enhanced toxicological responses in fish following exposure to Cu either alone or in combination with hypoxic condition and lends support to the evolving viewpoint that many water quality guidelines should be revisited in terms of new ecotoxicological criteria.     

Cadmium-induced DNA damage and mutations in Arabidopsis plantlet shoots identified by DNA fingerprinting

Random amplified polymorphic DNA (RAPD) test is a feasible method to evaluate the toxicity of environmental pollutants on vegetal organisms. Herein, Arabidopsis thaliana (Arabidopsis) plantlets following Cadmium (Cd) treatment for 26 d were screened for DNA genetic alterations by DNA fingerprinting. Four primers amplified 20-23 mutated RAPD fragments in 0.125-3.0 mg L⁻¹ Cd-treated Arabidopsis plantlets, respectively. Cloning and sequencing analysis of eight randomly selected mutated fragments revealed 99-100% homology with the genes of VARICOSE-Related, SLEEPY1 F-box, 40S ribosomal protein S3, phosphoglucomutase, and noncoding regions in Arabidopsis genome correspondingly. The results show the ability of RAPD analysis to detect significant genetic alterations in Cd-exposed seedlings. Although the exact functional importance of the other mutated bands is unknown, the presence of mutated loci in Cd-treated seedlings, prior to the onset of significant physiological effects, suggests that these altered loci are the early events in Cd-treated Arabidopsis seedlings and would greatly improve environmental risk assessment.     

Use of sensitive methods for detection of DNA damage on human lymphocytes exposed to p,p′-DDT: Comet assay and new criteria for scoring micronucleus test

Wide distribution, stability and long persistence in the environment of dichlorodiphenyltrichloroethane (DDT), probably the best-known and most useful insecticide in the world, imposes the need for further examination of the effect of this chemical on human health and especially on the human genome. In this study, peripheral blood human lymphocytes from a healthy donor were exposed to 0.025 mg/L concentration of p,p′-DDT at different time periods (1, 2, 24 and 48 h). For the assessment of genotoxic effect, the new criteria for scoring micronucleus test and alkaline comet assay were used. Both methods showed that p,p′-DDT induces DNA damage in low concentration used in this research. Results of micronucleus test showed a statistically significant (p < 0.05) genotoxic effect of p,p′-DDT on human lymphocytes compared with corresponding control and a different exposure time. A comet assay also showed increased DNA damage caused in p,p′-DDT-exposed human lymphocytes than in corresponding control cells for the tail length. Results obtained by measuring the level of DNA migration and incidence of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) indicate the sensitivity of these tests and their application in detection of primary genome damage after long-term exposure to establish the effect of p,p′-DDT on human genome.