定向驯化对异养蛋白核小球藻硒的耐受性及富集能力的影响

硒(Se)是生物体必不可少的微量元素,硒缺乏会导致人产生克山病、大骨节病等疾病,缺硒也会给畜牧业带来巨大损失。目前的补硒产品存在硒含量和生物利用度低、安全性差等问题,而通过小球藻培养可获得生物利用度高、安全性好的有机硒,因此是非常有应用前景的补硒产品。首先,为了获得对硒的耐受性和富集能力更强的藻种,研究通过定向驯化的方式逐步提高培养基中Na₂SeO₃浓度来驯化蛋白核小球藻,并对驯化时间和驯化过程中Na₂SeO₃的浓度梯度进行了优化。结果表明,驯化后的藻种对硒的耐受性和富集能力明显提高。在5L发酵罐中,驯化后的藻株可以耐受40mg/L的Na₂SeO₃,胞内有机硒合成速率提高了175.6%。之后,在5L发酵罐中进一步优化了硒的补加方式,在异养培养过程中分批补料添加40mg/LNa₂SeO₃时,最终获得的蛋白核小球藻细胞干重达106.4g/L,有机硒含量为1227mg/kg,有机硒合成速率为1.36mg/(L·h)。研究结果与已有蛋白核小球藻异养富硒文献报道的最高细胞密度75g/L和最高有机硒含量560mg/kg相比分别提高了41.9%和119.1%。上述结果表明,通过定向驯化的方法,可大大提高蛋白核小球藻对硒的耐受性和富集能力。     

Microcalorimetric studies of the action of Na2SeO3 on the growth of Halobacterium halobium R1

The effect of Na₂SeO₃ on the growth of Halobacterium halobium R1 was investigated by means of microcalorimetry at 37°C. The biological response to toxicants is observed as the inhibition of the rate constant of growth of living cells. A low concentration of Na₂SeO₃ stimulated the growth of H. halobium R1, and a high concentration of Na₂SeO₃ inhibited the growth of H. halobium R1. Toxicity may be expressed as the half-inhibition concentration (IC₅₀). The rate constants of growth (k) and the concentrations of Na₂SeO₃ (c) shows a linear relationship: k=1.790 × 10⁻⁶ − 2.27 × 10⁻³ c. The value of IC₅₀ obtained from the accompanying figure of I-c is 679 μg/mL.     

马利亚霉菌(Mariannaeasp.) HJ合成金属硒化物及机理研究

[背景] 金属硒化物因其优异的光电和催化特性,近年来在半导体、电化学及抗癌等领域成为了研究热点。相较于传统的化学还原法,生物合成金属硒化物具有环境友好、耗能较低等优势。然而,目前有关生物合成金属硒化合物的微生物资源较少且相关合成机理尚不明晰。[目的] 利用马利亚霉菌（Mariannaea sp.） HJ合成了3种金属硒化物并对其合成机理进行了初步探索。[方法] 利用X射线衍射（X-Ray Diffraction,XRD）和傅里叶转换红外线光谱（Fourier Transform Infrared Spectroscopy,FTIR）对菌株HJ合成的金属硒化物进行了初步的表征,考察了纳米材料合成过程中总巯基含量、总抗氧化性能及自由基含量变化,并且验证了转运蛋白DMT1在金属硒化物合成中所起的关键性作用。[结果] XRD结果表明菌株HJ能够在Bi³⁺、Pb²⁺、Co²⁺与SeO₃^2-作用下分别合成Bi₄Se₃、PbSe和CoSe₂纳米颗粒,其合成的最优pH条件分别为6.0、7.0、8.0。FTIR结果表明,合成的金属硒化物表面含有氨基、羧基、羟基等官能团。3种金属硒化物的合成反应体系与空白对照组相比,总巯基含量明显下降,而总抗氧化性能却有所提高,这表明巯基等酶促体系或氨基酸金属蛋白类的非酶促体系可能参与了SeO₃^2-的还原过程。苄基异硫脲盐酸盐屏蔽实验表明,转运蛋白DMT1在SeO₃^2-转运和金属硒化物分泌过程中起到关键作用。此外,Bi³⁺、Pb²⁺和Co²⁺的加入使得菌株HJ产生氧化应激反应,在胞外分泌了大量的过氧化氢、羟基自由基和超氧自由基,而上述自由基可通过诱导热激效应的方式增强金属离子或纳米颗粒的转运过程。[结论] 利用马利亚霉菌（Mariannaeasp.） HJ合成了Bi₄Se₃、PbSe和CoSe₂纳米颗粒,为研究金属硒化物的生物合成及机理提供了一定的理论参考。     

亚硒酸钠对巨噬细胞内游离Ca2+和Mg2+的影响

用单细胞阳离子测定系统研究了SeO^2-₃对巨噬细胞内游离Ca²⁺和Mg²⁺的影响.实验结果表明:SeO^2-₃高于10^-4mol/L时,有显著的细胞毒性.SeO^2-₃对细胞的毒性作用使细胞内游离Ca²⁺和Mg²⁺的浓度升高但Ca²⁺浓度的升高速率比Mg²⁺快.还有,高于10^-4mol/L的SeO^2-₃对红细胞膜上的Ca²⁺-ATP酶活性有明显抑制作用.     

Mg2+和SeO2-3对鸡胚软骨细胞受模拟微重力不良影响的拮抗

在回转模拟微重力条件下,研究了鸡胚负重软骨细胞骨架的微管系统和碱性磷酸酶活性两项指标的变化,以及1 mg/L亚硒酸钠和5 mmol/L Mg²⁺对这些指标的影响.流式细胞仪对微管含量的测定显示回转后微管蛋白含量的减少,说明微管系统受到不良影响.碱性磷酸酶活性比对照组明显降低,表明模拟微重力能降低软骨细胞的钙化能力.如果在回转前加入SeO^2-₃和Mg²⁺,发现SeO^2-₃可以在一定程度上拮抗模拟微重力引起的微管蛋白及碱性磷酸酶活性改变,而Mg²⁺基本上可以完全拮抗模拟微重力对这两项指标的不良影响.     

渗透胁迫和离子毒害对转Bt基因棉幼苗生长及离子分布的影响

研究了渗透和盐胁迫处理对转Bt基因抗虫棉(Gossypium hirsutum) 99B种子的萌发和幼苗生长的影响,以及幼苗不同器官离子吸收和分配的差异。结果表明:渗透和盐胁迫均对转Bt基因抗虫棉幼苗的生长有抑制作用,其中PEG的抑制作用最强,而3种盐的抑制程度以CaCl₂>NaCl>Na₂SO₄,且在Na⁺含量相同时,Cl^-的毒害大于SO₄^2-。渗透胁迫下使根、茎和叶中的Na⁺和Cl^-含量提高,K⁺、Ca²⁺、SO₄^2-含量和K⁺/Na⁺、Ca²⁺/Na⁺和SO₄^2-/Cl^-比值降低,且地上部的变化幅度大于地下部的,其中以PEG的影响最为显著,其次是CaCl₂,Na₂SO₄处理最弱。这些说明,转Bt基因抗虫棉99B的耐盐性较弱。     

On the relationship between the number of negative slope regions in the voltage-current curve of the Hodgkin-Huxley model and its parameter values

 We consider the problem of the existence of a negative slope region (NSR) in the voltage-current curve of the neuronal membrane and the relationship between this phenomenon and the membrane parameters. For the Hodgkin-Huxley model it is proposed to determine the dependence of the number of NSR on the values of the maximal sodium (gˉ_Na) and potassium (gˉ_K) conductances. The method is suggested for constructing the boundaries on the (gˉ_Na, gˉ_K) plane, in passing through which the number of NSR changes to 1. Using the method we partition the (gˉ_Na, gˉ_K) plane into the regions corresponding to the curves with the different number of NSR. This number can be changed from 0 to 2 in changing the values of gˉ_Na and gˉ_K over the physiologically possible range. Received: 9 December 1993/Accepted in revised form: 10 February 1995     

大鼠前脂细胞质膜Na+/H+交换同时参与细胞的增殖和分化

以原代培养的大鼠前脂细胞为模型 ,以 2′ ,7′ bis ( 2 carboxyethyl) 5 ( 6 ) carboxyfluorescein (BCECF)作为检测胞内pH(pHi)的荧光探针 ,测定不同生长因子刺激下胞内pH的变化 ,证明大鼠肾周前脂细胞质膜存在Na⁺/H⁺交换活性 ,胎牛血清(FCS)能快速激活Na⁺/H⁺交换 ,导致pHi升高 (约 0 .2pH单位 ) ,并引起DNA合成 .Ethyl isopropyl amiloride (EIPA)抑制Na⁺/H⁺交换与DNA合成 .在无血清条件下 ,胰岛素不刺激DNA合成但引起细胞分化 ,表现为胞内脂滴积累和 3 磷酸 甘油脱氢酶(G₃ PDH酶 )活性增强 ,同时激活Na⁺/H⁺交换活性导致pHi升高 ;EIPA既抑制胰岛素对Na⁺/H⁺交换的激活 ,也抑制G₃ PDH酶活性增强 .结果证明 :Na⁺/H⁺交换的激活不仅与大鼠前脂细胞增殖相关 ,同时也是细胞分化的早期事件 .     

黄河口湿地柽柳灌丛土壤盐渍化特征

为探究黄河三角洲湿地柽柳灌丛下土壤的盐渍化特征,在黄河三角洲国家级自然保护区(37°35''-12''N,118°33''-119°20''E)黄河入海口附近,根据长势基本一致的原则分别在碱蓬群落、柽柳群落和芦苇群落各选3株柽柳,采集柽柳灌丛下土壤样品,分析土壤盐分和盐碱化参数的空间分布以及距基茎不同距离处研究对象(土壤总盐(TS)、电导率(EC)、pH、交换性钠百分率(ESP))和环境因子(Na⁺、K⁺、Ca²⁺、Mg²⁺、Cl^-、HCO₃^-、SO^2-₄)之间的关系。结果表明：(1)研究区土壤为弱碱化盐土,离子含量由高到低依次为Cl^->Na⁺>SO^2-₄ >Ca²⁺>Mg²⁺>HCO^-₃>K⁺。除pH在土壤表层数值最低外,表层土壤TS、EC、ESP和盐分离子大于深层土壤,显示表聚性。(2)土壤盐分和盐碱化参数空间分布总体为：在柽柳基茎周围形成"盐谷"、"碱谷"效应, Na⁺、Mg²⁺、Cl^-表现为"盐谷",K⁺ 、SO^2-₄ 、Ca²⁺ 表现为"盐岛"。(3)在整个土壤剖面中,与TS、EC相关性最强的阴阳离子为Mg²⁺、Cl^-,从灌丛中心到灌丛间裸地Ca²⁺、SO^2-₄与TS、EC的相关性逐渐减弱,Mg²⁺、Cl^-与TS、EC的相关性逐渐增强。Ca²⁺和SO^2-₄与pH表现为较强的负相关性;与ESP相关性最强的阴离子为HCO^-₃,与之相关性最强的阳离子为Na⁺和K⁺,并且Na⁺和K⁺与ESP的相关性表现出从灌丛中心向外逐渐增强。(4)土壤盐渍化主要受控于Na⁺,从灌丛下到灌丛间裸地Cl^-对盐渍化程度的影响逐渐增加,SO^2-₄的影响逐渐降低。     

超氧歧化酶(SOD)对人红细胞膜的保护作用

前文曾报道,红细胞膜经外源O₂^-_·自由基(碱性条件下,邻苯三酚自氧化产生)处理后有明显损伤作用。在此基础上,本文研究猪红细胞SOD对人红细胞膜氧化损伤的保护作用。结果表明,外源SOD对人红细胞膜有明显保护作用,表现为膜流动性和膜重封闭能力保持正常水平,膜蛋白交联作用有显著减小。     

亚硒酸钠诱导SW480细胞凋亡过程中活性氧的作用

为探讨亚硒酸钠诱导人结肠癌SW480细胞凋亡的机理,将荧光探针2′,7′－二氯荧光黄乙二脂（2′,7′－DCFH－DA）、罗丹明123（rhodamine123）负载人结肠癌细胞,利用多光子成像系统测定胞内活性氧（ROS）、线粒体跨膜电位（△Ψm）的变化。结果发现（1）Na2SeO3作用SW480细胞,可导致细胞凋亡和胞内的ROS增加。SOD、过氧化氢酶可降低凋亡率并抑制ROS的增加。（2）线粒体电子传递链抑制剂鲁藤酮及氰化钠可抑制OS增加。（3）Na2SeO3可导致线粒体的跨膜电位的下降。表明Na2SeO3作用细胞可导致来源于线粒体的ROS增加,ROS介导亚硒酸钠诱导细胞凋亡。     

A 4 V Na+ Intercalation Material in a New Na‐Ion Cathode Family

Large‐scale electrochemical energy storage is a critical factor in the development of renewable energy sources to enable their intermittent power to become dispatchable. In this context, Na‐ion batteries are seen as promising alternatives to Li‐ion batteries, but their advancement requires the discovery of new materials, their electrochemical properties, and a better understanding of structure–property relationships that underpin the electrochemistry. This study presents a new class of Na⁺ insertion materials for Na‐ion batteries. By virtue of its moderately inductive polyanionic framework, the air and moisture stable selenite Na₂Co₂(SeO₃)₃ displays a highly suitable redox potential of ≈ 4 V versus Na/Na⁺ based on the Co²⁺/Co³⁺ couple, rendering it compatible with conventional liquid organic electrolytes. A microwave hydrothermal synthesis route is developed for the rapid synthesis of nanostructured Na₂Co₂(SeO₃)₃ and its conductive graphene oxide composite. The electrochemistry and structural evolution of Na₂Co₂(SeO₃)₃ determined on cycling the cathode in a Na battery was investigated by operando X‐ray diffraction, X‐ray photoelectron spectroscopy, and temperature dependent magnetic susceptibility measurements. These studies reveal good structural and electrochemical reversibility.     

Reactive oxygen species from mitochondria mediate SW480 cells apoptosis induced by Na2SeO3

A number of selenium compounds have been found to inhibit tumorigenesis in a variety of animal and cell models. In order to explore the molecular mechanism involved in the anticarcinogenesis activity of selenium, we examined the effects of sodium selenite on cell viabilty, generation of reactive oxygen species (ROS), and mitochondrial transmembrane potential (Δω _m ) in human colonic carcinoma cells SW480. The result from MTT test showed that sodium selenite reduced cell viability. Morophologic and flow cytometric results indicated that Na₂SeO₃ induced the apoptosis of SW480 cells. Na₂SeO₃ increased the generation of intracellular ROS, whereas BAPTA-AM, rotenone, and NaCN completely inhibited the increase of ROS induced by Na₂SeO₃. Na₂SeO₃ also caused the disruption of Δω _m . The intracellular ROS increase and apoptosis induced by Na₂SeO₃ were significantly decreased by superoxide dismutase (SOD), catalase. These data suggest that the ROS mediate apoptosis induced by Na₂SeO₃ and mitochondria may be a major source of Na₂SeO₃-induced ROS.     

Sodium Selenite Prevents Paraquat-Induced Neurotoxicity in Zebrafish

Considering the antioxidant properties of sodium selenite (Na₂SeO₃) and the involvement of oxidative stress events in paraquat-induced neurotoxicity, this study investigated the protective effect of dietary Na₂SeO₃ on biochemical and behavioral parameters of zebrafish exposed to paraquat (PQ). Fish were pretreated with a Na₂SeO₃ diet for 21 days and then PQ (20 mg/kg) was administered intraperitoneally with six injections for 16 days. In the novel tank test, the Na₂SeO₃ diet prevented the locomotor impairments, as well as the increase in the time spent in the top area of the tank, and the exacerbation of freezing episodes. In the preference for conspecifics and in the mirror-induced aggression (MIA) tasks, Na₂SeO₃ prevented the increase in the latency to enter the area closer to conspecifics and the agonistic behavior of PQ-treated animals, respectively. Na₂SeO₃ prevented the increase of carbonylated protein (CP), reactive oxygen species (ROS), and nitrite/nitrate (NOx) levels, as well as the decrease in non-protein thiols (NPSH) levels. Regarding the antioxidant enzymatic defenses, Na₂SeO₃ prevented the increase in catalase (CAT) and glutathione peroxidase (GPx) activities caused by PQ. Altogether, dietary Na₂SeO₃ improves behavioral and biochemical function impaired by PQ treatment in zebrafish, by modulating not only redox parameters, but also anxiety- and aggressive-like phenotypes in zebrafish.     

SELENIUM: AN ESSENTIAL ELEMENT FOR GROWTH OF THE COASTAL MARINE DIATOM THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE)1,2

An obligate requirement for selenium is demonstrated in axenic culture of the coastal marine diatom Thalassiosira pseudonana (clone 3H) (Hust.) Hasle and Heimdal grown in artificial seawater medium. Selenium deficiency was characterized by a reduction in growth rate and eventually by a cessation of cell division. The addition of 10⁻¹⁰ M Na₂SeO₃ to nutrient enriched artificial seawater resulted in excellent growth of T. pseudonana and only a slight inhibition of growth occurred at Na₂SeO₃ concentrations of 10⁻³ and 10⁻² M. By contrast, Na₂SeO₄ failed to support growth of T. pseudonana when supplied at concentrations less than 10⁻⁷ M and the growth rate at this concentration was only one quarter of the maximum growth rate. The addition of 10⁻³ and 10⁻² M Na₂SeO₄ to the culture medium was toxic and cell growth was completely inhibited. Eleven trace elements were tested for their ability to replace the selenium requirement by this alga find all were without effect. In selenium-deficient and selenium-starved cultures of T. pseudonana cell volume increased as much as 10-fold as a result of an increase in cell length (along the pervalvar axis) but cell width was constant. It is concluded that selenium is an indispensable element for the growth of T. pseudonana and it should be included as a nutrient enrichment to artificial seawater medium when culturing this alga.     

The influence of selenium and caffeine on chemical carcinogenesis in rats,mutagenesis in bacteria,and unscheduled DNA synthesis in human lymphocytes

The influence of sodium selenite (Na₂SeO₃) and caffeine on chemical carcinogenesis induced in rats by diethylnitrosamine (DEN), N-nitrosomorpholine (NM), andN-methyl-N-nitro-N-nitrosoguanidine (MNNG) was investigated. A dose-dependent inhibitory effect of Na₂SeO₃ (l–10 ppm) on hepatocarcinogenesis induced by DEN was demonstrated. Na₂SeO₃ also increased the latency period for stomach tumor formation in rats treated with MNNG. Combined treatment of rats with Na₂SeO₃ plus vitamin C added to the diet resulted in a slight inhibition of NM-induced liver carcinogenesis. Supplementation of diet with Na₂SeO₃ plus butylated hydroxytoluene, vitamin C, and vitamin E did not reveal any additive inhibitory effect compared to the inhibitory effect of Na₂SeO₃ given alone. Caffeine (600 rag/L) reduced the number of liver tumors induced in rats by DEN. Preliminary experiments have indicated that combined treatment of rats with selenium and caffeine could result in more effective inhibition of DEN-induced liver carcinogenesis. Further experiments are being conducted to study the influence of selenium and caffeine on mutagenic activity of 1-methyl-l-nitrosourea (MNU) inSalmonella typhimurium TA 1535. The pretreatment of bacteria cells with Na₂SeO₃ (3–10 p.g/mL) increased the mutagenic response of bacteria to MNU. A synergistic stimulation of mutagenic activity of MNU was observed in bacteria pretreated simultaneously with Na₂SeO₃ and caffeine. In addition the influence of Na₂SeO₃ on UDS induced by DEN in human lymphocytes was investigated. The trace element inhibited the UDS up to 82%. The possible role of potentiation by NazSeO3 of the cell killing effect of DEN in inhibition of liver carcinogenesis was discussed.     

The incorporation of Na2 75SeO3 into sheep erythrocytes

Injection of Na₂ ⁷⁵SeO₃ into untreated, phlebotomized, and phenylhydrazine treated sheet has shown that the rate of⁷⁵SeO₃-incorporation into erythrocytes is dependent on the degree of stimulation. Analysis of labeled erythrocytes by gel filtration and twodimensional electrophoresis has indicated that the transient labeling of a hemoglobin-like peptide is the only protein labeled in addition to glutathione peroxidase.     

Se-mediated domain-domain communication in Band 3 of human erythrocytes

Na₂SeO₃ could affect the anion flux of Band 3 of inside-out erythrocyte membrane vesicles (IOVs). Such effect was believed to be based on the interaction of SH groups of Band 3 with Na₂SeO₃. This effect could be eliminated when the cytoplasmic domain of Band 3 was proteolytically removed by trypsin. This suggested that SH groups in the cytoplasmic domain were involved in such interaction. Measurement of the pH dependence of intrinsic fluorescence intensity provided evidence that conformational changes of Band 3 occurred as a consequence of interaction with selenite. KI quenching of intrinsic fluorescence of Band 3 could also show that there was a conformational change in the cytoplasmic domain of Band 3 after reaction with Na₂SeO₃. Such conformational change in turn could be transmitted to the membrane domain of Band 3 monitored by quenching of intrinsic fluorescence of Band 3 using hypocrellin B (HB) (a photosensitive pigment obtained from a parasitic fungus growing in Yunnan, China). It is suggested that the cytoplasmic domain of Band 3 is not necessary for its anion flux, but is essential for the regulation (e.g., by Se) of its active site located at the membrane domain, and hence, it may provide evidence of communication between the cytoplasmic domain and the membrane domain of Band 3.     

SELENIUM: AN ESSENTIAL ELEMENT FOR GROWTH OF THE COASTAL MARINE DIATOM THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE)1,2

An obligate requirement for selenium is demonstrated in axenic culture of the coastal marine diatom Thalassiosira pseudonana (clone 3H) (Hust.) Hasle and Heimdal grown in artificial seawater medium. Selenium deficiency was characterized by a reduction in growth rate and eventually by a cessation of cell division. The addition of 10⁻¹⁰ M Na₂eO₃ to nutrient enriched artifical seawater resulted in excellent growth of T. pseudonana and only a slight inhibition of growth occurred at Na₂SeO₃ concentrations of 10⁻³ and 10_-2 M. By contrast, Na₂SeO₄ failed to support growth of T. pseudonana when supplied at concentrations less than 10⁻⁷ M and the growth rate at this concentration was only one quarter of the maximum growth rate. The addition of 10⁻³ and 10⁻² M Na₂SeO₄ to the culture medium was toxic and cell growth was completely inhibited. Eleven trace elements were tested for their ability to replace the selenium requirement by this alga and all were without effect. In selenium-deficient and selenium-starved cultures of T. pseudonana cell volume increased as much as 10-fold as a result of an increase in cell length (along the pervalvar axis) but cell width was constant. It is concluded that selenium is an indispensable element for the growth of T. pseudonana and it should be included as a nutrient enrichment to artificial seawater medium when culturing this alga.     

Additive coclastogenicity of sodium selenite and caffeine in CHO cells treated withN-methyl-N′-Nitro-N-Nitrosoguanidine

The clastogenic effect ofN-methyl-N′-nitro-N-nitrosoguanidine (MNNG) in Chinese hamster ovary (CHO) cells and its modulation by Na₂SeO₃ and caffeine were studied by metaphase analysis of chromosome aberrations (CA) as well as by measuring the formation and repair of single-strand (ss) DNA breaks employing hydroxylapatite chromatography. Treatment of CHO cells with MNNG (1.25 or 2.5 × 10-5M) for 3 h caused CA in 11 and 19% of metaphases scored, respectively. Pretreatment of cells with Na₂SeO₃ (1–5 μg/mL) or caffeine (0.2–2.0 mg/mL) for 2 h resulted in a 2–3.5-fold increase of CA frequency. Addition of both modulators during the mutagen exposure tended to cause a slight inhibition of clastogenic activity of MNNG (1.25 × 10⁻⁵ M) or had no effect on CA number when MNNG was used at a concentration of 2.5 × 10−5M. Posttreatment of CHO cells with Na₂SeO₃ for 20 h after MNNG was ineffective in influencing the number of metaphases with CA, whereas, at these conditions, caffeine enhanced up to 6-7-fold the clastogenic activity of MNNG. Addition of both modulators during the whole experiment, 2 h pretreatment included, resulted in a further significant increase of CA frequency up to the total pulverization of chromosomes in all metaphases scored. The coclastogenic effect of caffeine was greater in this case. The enhancement of chromosome-damaging activity of MNNG by selenite and caffeine was better expressed when this carcinogen was applied at the higher concentration used. An additive coclastogenic effect was observed in CHO cells treated simultaneously with Na₂SeO₃ and caffeine plus MNNG. In addition, the treatment of CHO cells with MNNG (5 × 10⁻⁶ M) caused a rapid increase of ssDNA breaks number reaching maximal values after 30–45 min. However, up to 50–60% of MNNG-induced ssDNA breaks were repaired during the first 60–150 min after the mutagen exposure. The 2 h pretreatment of CHO cells with Na₂SeO₃ (2 μg/mL) or the addition of this trace element after MNNG had no effect on formation and repair of MNNG-induced ssDNA breaks. The coclastogenic effect of Na₂SeO₃ in CHO cells treated with MNNG was not directly linked to the induction and disappearance of ssDNA breaks measured by hydroxylapatite chromatography.