Carcinogenesis of asbestos switched on by inducing cross-linkage between DNA complementary pair bases

Since the beginning of the 1980s, Dai Qianhuan predicted based upon his di-region theory that the carcinogenesis switched on by the so-called physical carcinogenic factors including radiation, asbestos and foreign matter implantation, is just initiated through the cross-linking between DNA complementary pair bases induced by them. In this note, it was evidenced with the DNA filter elution method that the oxygenase activated by asbestos induces the cross-linking between DNA inter-strands and DNA-protein with dosage correlation, in which over 80% of DNA inter-strand cross-link ratio account for the total cross-link ratio. Obviously, both of the cross-linkages are just induced by hydroxyl free radical, HO ·, because the ferrous ion increased the cross-link ratios up to several times through Fenton reaction and vitamin C inhibited the cross-link ratios with factors of 8–9 by destroying the hydroxyl radical. Non-carcinogen but with lower free radical formation energy, pyrene, by culturing with asbestos gave 3–4 times cross-link ratios than the original ratios induced by asbestos only. Estradiol, an endogenous carcinogen, as a bio-electrophilic species but with higher free radical formation energy by culturing with asbestos, gave only 1.2 time cross-link ratios than expected ones. Ferrous ion which can increase HO · concentration through Fenton reaction, increased the ratios to 2–2.5 times in the former case but only 1.2 time in the latter case. Vitamin C, a free radical scavenger, gave a powerful inhibition to the cross-linking with a factor of 8–11 in the former case but a weak inhibition with a factor of 1.2 only in the latter case. So, it is evidenced further that the cross-linkages induced by asbestos are originated from hydroxyl radical. Reasonable structures of the cross-linking products induced by asbestos or hydroxyl radical have been depicted based upon AMI MO theory. These structures have been verified further by a reasonable explanation of the mutational spectrum induced by hydroxyl radical.     

AM1 computational study on metabolites of 1,3-butadiene interstrand cross-linking DNA

The alkylating reactions of 1,2-epoxy-3,4-butene (EB) and 1,2,3,4-diepoxybutane (DEB) —the important metabolites of rodent carcinogenic 1,3-butadiene, with adenine and cytosine and interaction with fragment of DNA on major groove—have been computed. Results show that there are little differences in activation energy between EB and DEB, so it is difficult to explain the fact that the mutagenicity of DEB is greater (about 100-fold) than that of EB by the ability of alkylation. It is also known that DEB can interstrand cross-link with DNA through two times alkylating reactions, whereas EB cannot. So this may contribute to the significant different genotoxicity of the two agents. Meanwhile, DEB can interstrand cross-link with many sequences of DNA in major groove vs. two in minor groove, which increases opportunity of interstrand cross-link with DNA in major groove. This difference may be the reason of base selection of DEB mutation. The deformation of some cross-linked DNA may also contribute to this selection to some degree.     

Determination of copy number for 5S rDNA and centromeric sequence RCS2 in rice by Fiber-FISH

The copy number of 5S rDNA and centromeric sequence RCS2 was determined by extended DNA fiber based fluorescence in situ hybridization (Fiber-FISH) in rice (Oryza sativa ssp. indica cv. Guangluai No. 4) genome. In order to determine the copy number, it is necessary to know the basepair number that a given length DNA fiber contains under a microscope. Therefore, the length of two DNA fragments, in which the basepair number had been already known, was measured. The insert sequence of the tested BAC 38D17 was 136 kb and its extended DNA was 56.4 μm long, 2.41 kb/μm on average, while that of the tested BAC 44B4 was 144.5 kb in total and 55.7 μm long, 2.60 kb/μm on average under the microscope. They were very close to the theoretical value of B-DNA in the Watson-Crick DNA model, which is 2.97 kb/μm. According to the average value of basepair number per μm of the two samples mentioned above, that is, 2.51 kb/μm, it could be estimated that the copy number was about 686 for 5S rDNA and 286-1121 for the centromere sequence RCS2.     

Phenomenon and mechanism of “secondary damage” during repair of DNA damage in irradiated mammalian cells

By means of comet assay, a study of kinetics curve of DNA damage repair in irradiated SX-9 cells that came from mouse breast cancer proceeded. It was found that while the initial DNA damages had nearly been repaired, DNA damages arose for the second time, then they were repaired again. As a result, a phenomenon of “secondary damage” was found during the repair of DNA damages in irradiated SX-9 cells. Further research illuminated that 3-aminobenzamide (3AB), which is an inhibitor of poly (ADP-ribose) polymerase, could change the proceeding of “secondary damage”. For this reason it is poasible that there exists some inner relationship between the phenomenon of “secondary damage” and the function of poly(ADP-ribose) polymerase.     

青岛中港微型生物膜中污损细菌群落初探

本研究拟通过揭示微型生物膜中的细菌群落特征,为污损生物学和海洋腐蚀学的微观研究提供参考和启示。试验采用3种方法:传统稀释涂布法、流式细胞仪计数法以及16srDNA扩增子高通量测序对青岛中港冬季海水以及玻璃板中微型生物膜中的污损细菌群落结构进行分析。试验结果表明冬季海水表层和底层中细菌的浓度分别为3.0×10~8 CFU/L和6.9×10~7 CFU/L,表层和底层玻璃板微型生物膜中的细菌浓度分别为7.2×10~7 CFU/L和2.1×10~7 CFU/L,海水和玻璃板中的细菌以Proteobacteria和Bacteroidetes为主。另外,海水中细菌群落结构组成与玻璃板微型生物膜中细菌具有较高的一致性。本研究利用不同的试验方法揭示微型生物膜中的细菌群落特征,对深入研究生物污损,从根本上解决海洋生物污损问题、制定绿色防污措施具有重要的现实意义。     

The Earth’s inner core deviation to plate movement

Under the premise of the Earth’s layer structure and mass distribution, the solid inner core cannot be stable in the center of the sphere. Thus, the deviation of the inner core is towards the sphere’s center and the liquid outer core must have asymmetrical thermal convection. Based on the two suggestions, a concise and self-consistent global motion model can be built. The model consists of the following cycle: an asymmetrical thermal convection structure in the outer core led by the dislocation of the inner core → the plume is in special upwell position because of differential activation → the formation and split of lithosphere → the split plates drift and assemble in a new location, the mass of which causes the inner core to deviate towards this direction again → the new asymmetry is formed. As this circulation continues, a definite and periodical motion emerges/forms. Its nonlinear features result in the Earth’s motion with simple mechanisms but complex behavior. Ultraterrestrial events may disturb or even interrupt this movement but would not significantly affect the cycle.     

Expression of human cytomegalovirus DNA polymerase in insect cells using baculovirus expression system: Purification and biochemical characterizations

Human Cytomegalovirus (HCMV) DNA polymerase gene was overexpressed in insect cells using the baculovirus transfer system. A6. 2 kb HCMV Rsr II-EcoRI DNA fragment with intact HCMV pol gene coding sequence was engineered into NheI site of vector pBlueBac under the control of polyhedrin promoter of Autographa californica nuclear polyhedrosis virus (AcNPV). Recombinant AcNPV carried HCMV pol gene was generated by cotransfection of Spodoptera frugiperta cell (SF21) with AcNPV DNA and baculovirus transfer vector with HCMV pol gene. Infection of SF21 cell with recombinant virus lead to the expression of 140 kD peptide of HCMV specific DNA polymerase at the level approximately 2 mg per 10⁸ cells. The polypeptide was purified from the infected SF21 cells by a series of column chromatography to homogeneity. The purified enzyme had a molecular weight of 140 kD and reacted with antiserum specific for HCMV DNA polymerase. It exhibited both 3′–5′ and 5′–3′ exonuclease activities. This enzyme is also sensitive to phosphono acetate. Ye Linbai: born in Feb. 1948. Professor. Current research interest is in Vitology and Molecular Biology Supported by Public Health Service Grants CA21773, CA15036 and AI12717 from the National Institutes of Health     

A nut-type ultrasonic motor and its application in the focus system

This article reports a screw-driven polyhedron linear ultrasonic motor （USM） of nut-type. It is comprised of a stator, which is a threaded metal nut bonded with piezoelectric plates on its external surface, and a rotor with external threads, which engage with the internal threads of the stator. A traveling wave in the plane is stimulated on the stator when harmonic electric signals are applied to the piezoelectric plates. The traveling wave drives the rotor to rotate, and the threads transform the rotation into a linear motion. The lens can be fixed in the rotor and realize the integrated design of the auto focus system. This structure can omit retarder and directly drive to obtain a high accuracy of positioning, and it is shockproof. A mini AF cellular phone module （8.5 mm×8.5 mm×5.9 mm） driven by this motor was made and an image resolution of 3-5 MP was obtained in the module prototypes of the cellular phone.     

The recognition of adakite-type gneisses in the North Dabie Mountain and its implication to ultrahigh pressure metamorphic geology

Adakite[1] is an igneous rock generated by the partial melting of subducting ocean slab in an island arc setting. Meanwhile, residuals of the slab-melting are transformed to eclogite or garnet-amphibolite during a slab subduction process. This note reports the adakite-type grey gneisses discovered in the North Dabie Mountain. These adakitic rocks are characterized by high Al2O3 (14.97%-17.56%), Sr (545-941 μg/g), Sr/Y (44.71-218.98), La/Yb (16.70-97.50), and low Y (3.32-12.19?μg/g), Yb (0.31-1.34 μg/g), Sc (0.92-4.2 μg/g), as well as positive Sr anomaly but absent or positive Eu anomaly. They are different from the low aluminum grey gneisses with low Al2O3 (13.73%-14.38%), Sr (262-409μg/g), Sr/Y (6.46-15.97) and La/Yb (1.90-22.60), high Y(21.79- 36.24 μg/g), Yb (2.00-5.59 μg/g) , Sc (8.30%-12.70%), and negative Sr, Eu anomaly in the North Dabie Mountain. The adakite-type grey gneisses in the North Dabie Mountain were most probably formed by the partial melting of subducting ocean slab before the Indo-Chinese epoch. We suggest that the subducting of an ocean slab took place prior to the incorporation between the Yangtze and North China plates in the Indo-Chinese epoch, resulting in formation of adakite-type igneous rocks and some ultrahigh pressure eclogites related to the subducted ocean slab. Therefore, besides the ultrahigh pressure metamorphism of the Indo-Chinese epoch, some ultrahigh pressure ecoglits were most likely formed prior to the Indo-Chinese epoch in the Dabie Mountain. In addition, the adakite-type grey gneisses with arc origin occur in the North Dabie Mountain, inferring that the Yangtze plate subducted down below the North China plate and the main suture zone between the two plates was likely along the southern side to the North Dabie Mountain.     

The DNA concentration effect on DNA radiation damage induced by <Superscript>7</Superscript>Li ions and γ rays

To evaluate the influence of the DNA concentration in the aqueous solution on DNA radiation damage, the plasmid DNA in the presence or absence of Mannitol （scavenger of free radical OH.） was irradiated by ^7Li ions and γ rays at various DNA concentrations. Gel electrophoresis analysis revealed that the DNA damage of single and double strand breaks induced by irradiation became more severe at lower DNA concentration. In the condition of γ-ray irradiation, most of double strand breaks （DSB） damage was neutralized and less associated with DNA concentration in the presence of mannitol. However, under ^7Li irradiation, DSB damage could not be cleared by mannitol but was gradually aggravated with decreasing DNA concentrations. These findings imply that under low-LET irradiation, most of the DSB damage is generated by free radical OH·diffusion, and thus may be counteracted by scavengers, while at higher-LET irradiation, quite a fraction of DSB induction is caused by direct ionizing energy deposition of heavy ions, which cannot be eliminated. This work also indicates that the proportion between free radical damage and direct ionizing damage is s constant which is independent of DNA concentration when the DNA concentration is under a certain value （50ng/μL）. Our study sheds light on the un- derlying mechanisms in the DNA radiation damage process.     

Determination of DNA single-strand breaks by low-energy heavy ion and analysis of dose-effect curves

Calf thymus DNA was exposed to low-energy heavy ions (N+) and 60Co-γ-rays, and the dose-effect on DNA single-strand breaks (SSB) has been investigated. The results indicate that the dose-effect curve by N+ irradiation is different from that of conventional ionizing radiation. While the curve from γ-irradiation follows exponential type, the effect curve produced by N+ ion is of "saddle type". The yield of DNASSB per dose unit per DNA unit remained at a certain level under different doses of γ-rays. In contrast, the DNASSB at low dosage region of N+ showed an obvious peak before it decreased rapidly to a lower level.     

Gene product expression of cyclin D<Subscript>2</Subscript> and p16 during the transition from cardiac myocyte hyperplasia to hypertrophy

The current study was to investigate mRNA expression of cyclin D₂ and p16 during the transition from cardiac myocyte hyperplasia to hypertrophy. Cultured cardiac myocytes (CM) and fibroblasts (FC) obtained from 1-day-old Sparague-Dawley rats were used in this study. We have determined (1) hyperplasia by cell growth curve and fluorescence activated cell sorting (FACS); and (2) ultrastructure by electron microscope observation; and (3) expresions of cyclin D₂ mRNA and p16 mRNA by using in situ hybridization and image analysis. The results were shown (1) Results of cell growth curve and FACS analysis showed CM could proliferate in the first 3 cultured days (4 days in postnatal development). But the ability decreased quickly, concomitant with the differentiation. (2) The ultrastructure of CM showed the large amount of myofilaments and mitochondrion and FC showed moderate amount of rough endoplasmic reticulum. (3) The expression of cyclin D₂ mRNA in 3−, 4−, 5−day CM group was 0.89 times (p<0.05), 0.80 times (p<0.05) and 0.56 times (p<0.01) of that in 1-day group respectively. P16 mRNA in 2−, 3−, 4−, 5−day CM group were 1.63 times (p<0.01), 1.72 times (p<0.01), 1.99 times (p<0.01) and 2.84 times (p<0.01) of that in 1−day group respectively. It can be concluded that cultured neonatal rat cardiac myocytes could proliferate during the first 3 cultured days, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D₂ and p16 have the key roles during the transition from myocyte hyperplasia to hypertrophy. Biography: Zhang Yu-xia (1974-), female, Master, research direction: cardiovascular pathology.     

Mechanism of oxidative damage to DNA by Fe-loaded MCM-41 irradiated with visible light

The mechanism of oxidative damage to deoxyribonucleic acid (DNA) by iron-containing mesoporous molecular sieves (MCM-41) irradiated with visible light was elucidated. Fe-loaded MCM-41 (Fe/MCM-41) was used as a photocatalyst and the damage to calf thymus DNA caused by hydrogen peroxide (H2O2) was studied. The damage and extent of oxidation of DNA were measured by high-performance liquid chromatography (HPLC) and intermediate products were detected by HPLC/electrospray ionization tandem mass spectrometry. Electron spin resonance was used to detect changes in reactive oxygen species and peroxidase catalytic spectrophotometry was used to determine the concentration of H2O2. The results indicated that Fe/MCM-41 efficiently activated H2O2 in solution at pH 4.0-8.0 under irradiation with visible light. The photocatalytic system degraded DNA most effectively at pH 5.0-6.0 but also operated at pH 8.0. At pH 4.2, the degree of DNA damage reached 25.65% after 5 h and the kinetic constant was 5.89×10 2 min 1. Damage to DNA was predominantly caused by hydroxyl radicals generated in the system. The mechanism of DNA damage is of potential concern to human health because it can occur in neutral solutions irradiated by visible light.     

Detection of DNA methylation changes during seed germination in rapeseed （Brassica napus）

DNA methylation is a common yet important modi- fication of DNA in eukaryotic organisms. DNA methy- lation, especially methylation of cytosine (m5C), have both epigenetic and mutagenic effects on various cellu- lar activities such as differential gene exp…     

Sensitive Determination of DNA by RLS Enhancement of Metal Ions

0　IntroductionThequantitativeanalysisofnucleicacids,especiallythemi cro determinationofnucleicacids,isbecomingmoreandmoreimportantinmanybiologicalstudies.Recently ,apromisingspectraltechnique ,whichwasbasedonthemeasurementofen hancedresonancelightscattering (RLS) [1 ,2 ] ,hasgivenrisetostronginterestbyanalystsandbiochemistsfornucleicacidsandproteinassay[3 9] .Uptonow ,manykindscompoundshavebeenfoundRLSenhancementwhilebindingtoDNA ,andallthesecompoundsarecharacterizedofpositivechargewhicha…     

Graphite-polypyrrole coated 316L stainless steel as bipolar plates for proton exchange membrane fuel cells

316L stainless steel (SS 316L) is quite attractive as bipolar plates in proton exchange membrane fuel cells (PEMFC). In this study, graphite-polypyrrole was coated on SS 316L by the method of cyclic voltammetry. The surface morphology and chemical composition of the graphite-polypyrrole composite coating were investigated by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). A simulated working environment of PEMFC was applied for testing the corrosion properties of graphite-polypyrrole coated SS 316L. The current densities in the simulated PEMFC anode and cathode conditions are around 3×10-9 and 9×10-5 A·cm-2, respectively. In addition, the interfacial contact resistance (ICR) was also investigated. The ICR value of graphite-polypyrrole coated SS 316L is much lower than that of bare SS 316L. Therefore, graphite-polypyrrole coated SS 316L indicates a great potential for the application in PEMFC.     

Construction of DNA damage response gene pprI functiondeficient and function-complementary mutants in Deinococcus radiodurans

PprI, a DNA damage response factor from the extraordinary radioresistant bacterium Deinococcus radiodurans, plays a central regulatory role in multiple DNA damage repair. In this study, a fusion DNA fragment carrying kanamycin resistance gene with the D. radiodurans groEL promoter was cloned by PCR amplification and reversely inserted into the pprl locus in the genome of the wild-type strain R1. The resultingpprl-deficient strain, designated YR1,was very sensitive to ionizing radiation. Meanwhile, the recombinant DNA fragment was cloned into the shuttle vector pRADZ3, and resulted in plasmid pRADK with kanamycin resistance in D. radiodurans. The fragments containing complete pprl gene and 3‘-terminal deletion pprIΔ were cloned into plasmid pRADK. The resulted plasmids designated pRADKpprI and pRADKpprIΔ were then transformed to YR1. Results show that YR1 carrying pRADKpprI was able to fully restore the extreme radioresistance to the same level as the wild-type D. raiodurans R1, whereas YR1 pRADKpprIΔ failed to do so. Construction of DNA repair switch PprI function-deficient and function-complementary mutants in D. radiodurans is not only useful to elucidating the relationship between domains and functions of PprI protein, but also opens the door to the further studies of the biological functions of PprI protein in vivo.     

The role of CopG mediated DNA bending on the regulation of the σ^54-dependent promoters in E. coli

In prokaryotic cells, although the σ54 RNA poly- merase can stably bind to the σ54-dependent promoter without the enhancer-binding proteins (EBPs), it re- mains as a closed complex which is silent for transcrip- tion[1]. When binding to the enhancer-lik…     

Urotensin Ⅱ increases endothelin production by vascular smooth muscle cells in rats

The aim of this study was to observe the effects of urotensin Ⅱ (UII) on production of endothelin (ET) in rat aortic vascular smooth muscle cells (VSMC). Cultured VSMCs incubated with various concentrations of UII were used to measure the VSMC ³H-TdR incorporation, the amount of ET mRNA and ET production in VSMCs. In this work we found that UII (10^-10—10^-8 mol/L) promoted VSMC ³H-TdR incorporation (47%—83%, P < 0.01) and increased the amount of ET mRNA by 17.1% (P < 0.05) to 112.8% (P < 0.01), respectively, in a concentration dependent manner compared with control. After 4 and 8 h incubation, 10^-10—10^-8 mol/L of UII elevated the ET synthesis and release in a concentration dependent manner. After 4 h incubation, the content of ET in medium was 4.9, 5.36 and 7.12 pg/mL (P < 0.01). After 8 h incubation, the ET content released from VSMCs was 12.6, 12.07 and 17.17 pg/mL (P < 0.01). In addition, it was found that BQ₁₂₃, a specific ET_A receptor antagonist, obviously decreased the VSMC DNA synthesis induced by UII. The results of this study showed that UII could stimulate the ET mRNA expression and ET production in VSMC. The effects of UII on VSMC DNA synthesis were partly mediated by ET autocrine pathway. It suggests that the interaction between UII and ET plays an important biological regulating role as endogenous active peptides.