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Synergistic and non-specific nucleic acid production by T7 RNA polymerase and Bsu DNA polymerase catalyzed by single-stranded polynucleotides

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Synthetic and Systems Biotechnology 2018年第2期3卷 130-134页

作者： nicholas J.Emery Sagardip Majumder Allen p.Liu Department of Mechanical Engineering University of MichiganAnn ArborMIUnited States Cellular and Molecular Biology program University of MichiganAnn ArborMIUnited States Department of Biomedical Engineering University of MichiganAnn ArborMIUnited States Biophysics program University of MichiganAnn ArborMIUnited States Department of Biomedical Engineering Boston UniversityBostonMAUnited States.

point-of-care molecular diagnostic tests show great promise for providing accurate,timely results in lowinfrastructure healthcare settings and at *** design space for these tests is limited by a variety of possible background reactions,which often originate from relatively weak promiscuous activities of the enzymes used for nucleic acid *** this background signal is amplified alongside the signal of the intended biomarker,the dynamic range of the test can be severely ***,a detailed knowledge of potential side reactions arising from enzyme promiscuity can improve rational design of point-of-care molecular diagnostic *** this end,we report a previously unknown synergistic reaction between T7 RNA polymerase and Bsu DNA polymerase that produces nucleic acid in the presence of single-stranded DNA or *** reaction occurs in the absence of any previously reported substrates for either polymerases and compromises a theoretical microRNA amplification scheme utilizing these polymerases.

关键词： catalyzed utilizing rational

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New methods for identification of disinfection byproducts of toxicological relevance: progress and future directions

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Journal of Environmental Sciences 2021年第1期33卷 151-159页

作者： nicholas J.p.Wawryk Caley B.Craven Lindsay K.Jmaiff Blackstock Xing-Fang Li Division of Analytical and Environmental Toxicology Department of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of AlbertaEdmontonABT6G 2G3Canada

Disinfection byproducts(DBps)represent a ubiquitous source of chemical exposure in disinfected *** over 700 DBps have been identified,the drivers of toxicity remain poorly ***,ever evolving water treatment practices have led to a continually growing list of *** of analytical technologies have enabled the identification of new classes of DBps and the quantification of these chemically diverse sets of *** we summarize advances in new workflows for DBp analysis,including sample preparation,chromatographic separation with mass spectrometry(MS)detection,and data *** aid in the selection of techniques for future studies,we discuss necessary considerations for each step in the *** review focuses on how each step of a workflow can be optimized to capture diverse classes of DBps within a single ***,we highlight new MS-based approaches that can be powerful for identifying novel DBps of toxicological *** discuss current challenges and provide perspectives on future research directions with respect to studying new DBps of toxicological *** analytical technologies continue to advance,new strategies will be increasingly used to analyze complex DBps produced in different treatment processes with the aim to identify potential drivers of toxicity.

关键词： Water disinfection byproducts(DBps) Emerging contaminants Mass spectrometry Chromatography Nontargeted analysis Toxicity drivers

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Neurotoxicity and transcriptome changes in embryonic zebrafish induced by halobenzoquinone exposure

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Journal of Environmental Sciences 2022年第7期34卷 129-140页

作者： Xue Yang Chang Wang Lihua Yang Qi Zheng Qiongyu Liu nicholas J.p.Wawryk Xing-Fang Li Hubei Key Laboratory of Environmental and Health Effects of persistent Toxic Substances School of Environment and HealthJianghan UniversityWuhan 430056China Division of Analytical and Environmental Toxicology Department of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of AlbertaEdmontonAlbertaT6G 2G3Canada South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center Guangdong Provincial Key Laboratory of Marine Resources and Coastal EngineeringSchool of Marine SciencesSun Yat-Sen UniversityZhuhai 519082China

Halobenzoquinones(HBQs) are emerging disinfection byproducts(DBps) with a widespread presence in drinking water that exhibit much higher cytotoxicity than regulated DBps. However, the developmental neurotoxicity of HBQs has not been studied in vivo. In this work, we studied the neurotoxicity of HBQs on zebrafish embryos, after exposure to varying concentrations(0-8 μmol/L) of three HBQs, 2,5-dichloro-1,4-benzoquinone(2,5-DCBQ), 2,6-dichloro-1,4-benzoquinone(2,6-DCBQ), and 2,5-dibromo-1,4-benzoquinone(2,5-DBBQ) for 4 to 120 hr post fertilization(hpf). HBQ exposure significantly decreased the locomotor activity of larvae, accompanied by significant reduction of neurotransmitters(dopamine and γ-aminobutyric acid) and acetylcholinesterase activity. Furthermore, the expression of genes involved in neuronal morphogenesis( gfap, α1-tubulin, mbp, and syn-2 α) were downregulated by 4.4-, 5.2-, 3.0-, and 4.5-fold in the 5 μmol/L 2,5-DCBQ group and 2.0-, 1.6-, 2.1-, and 2.3-fold in the 5 μmol/L 2,5-DBBQ group, respectively. Transcriptomic analysis revealed that HBQ exposure affected the signaling pathways of neural development. This study demonstrates the significant neurotoxicity of HBQs in embryonic zebrafish and provides molecular evidence for understanding the potential mechanisms of HBQ neurotoxicity.

关键词： Disinfection byproducts Halobenzoquinone Neurotoxicity Zebrafish embryos

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NOD2/CARD15 , ATG16L1 and IL23R gene polymorphisms and childhood-onset of Crohn’s disease

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World Journal of Gastroenterology 2010年第14期16卷 1753-1758页

作者： Maria Gazouli Ioanna pachoula Ioanna panayotou Gerassimos Mantzaris George Chrousos nicholas p Anagnou Eleftheria Roma-Giannikou Department of Biology School of Medicine University of Athens Athens 11527 Greece Laboratory of Cell and Gene Therapy Center of Basic Research Ⅱ Foundation for Biomedical Research of the Academy of Athens (IIBEAA) Athens 11527 Greece First Department of pediatrics "Aghia Sophia" Children's Hospital School of Medicine University of Athens Athens 11527 Greece Department of Gastroenterology Evagelismos Hospital Athens 11521 Greece

AIM: To assess whether the polymorphisms of NOD2/ CARD15 , autophagy-related 16-like 1 (ATG16L1 ), and interleukin-23 receptor (IL23R ) genes play a more critical role in the susceptibility of childhood-onset than in adult-onset Crohn’s disease (CD). METHODS: polymorphisms R702W, G908R, and 3020insC of NOD2/CARD15 ; rs2241880 A/G of ATG16L1 , and rs11209026 (R381Q) of IL23R gene were assessed in 110 childhood-onset CD, 364 adult-onset CD, and 539 healthy individuals. Analysis of polymorphisms R702W, G908R, and 3020insC of NOD2/CARD15 genotyping was performed by allele specific polymerase chain reaction (pCR) or by pCR-restriction fragment length polymor-phism analysis. The polymorphisms rs2241880 A/G of the ATG16L1 , and rs11209026 (R381Q) of the IL23R gene in the children’s cohort were genotyped by pCR and melting curve analysis whereas adult group genotyping was performed using the Affymetrix Genome-Wide Human SNp Array 5.0 (500K). RESULTS: The 3020insC allele in NOD2/CARD15 was significantly higher in childhood than in adult-onset CD (p = 0.0067). Association with at least 1 NOD2/CARD15 variant was specific for ileal disease (with or without co- lonic involvement). Even if the frequency of G allele of the rs2241880 ATG16L1 polymorphism was increased in both paediatric and adult CD patients compared to con- trols (p = 0.017 and p = 0.001, respectively), no difference was observed between the childhood and the adult cohort. The rare Q allele of IL23R rs11209026 polymorphism was underrepresented in both paediatric and adult CD cases (p = 0.0018 and p = 0.04, respectively) and no difference was observed between the childhood and the adult cohort. The presence of the rs2241880 ATG16L1 and rs11209026 IL23R polymorphisms did not influence disease phenotype. CONCLUSION: polymorphism 3020insC in NOD2/ CARD15 occurs statistically significantly more often in patients with childhood-onset CD than in patients with adult-onset CD. The ATG16L1 and IL23R variants are associated

关键词： Genetics Childhood-onset Inflammatory bowel disease Crohn’s disease Genetic susceptibility NOD2/CARD15 ATG16L1 IL23R polymorphisms

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Effects of Exhaust Gas Recirculation on the Homogeneous Charge Combustion process of n-Heptane at Different Load Conditions

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Transactions of Tianjin University 2005年第4期11卷 262-267页

作者：刘德新冯洪庆郑进才 MILLER David L CERNANSKY nicholas p State Key Laboratory of Engines School of Mechanical Engineering Tianjin University Tianjin 300072 China Department of Mechanical Engineering Drexel University Philadelphia PA19104-2875 USA

Effects of exhaust gas recirculation （EGR） on homogeneous charge combustion of n-heptane was studied through simulation and experiment. Experiments were carried out in a single cylinder, four-stroke, air cooled engine and a single cylinder, two-stroke, water cooled engine. In the four-stroke engine, experiments of the effects of EGR were examined using heated N2 addition as a surrogate for external EGR and modifying engine to increase internal EGR. The ignition timing was sensitive to EGR due to thermal and chemical effects. EGR or extra air is a key factor in eliminating knock during mid-load conditions. For higher load operation the only way to avoid knock is to control reaction timing through the use of spark ignition. Experimental and modeling results from the two-stroke engine show that auto-ignition can be avoided by increasing the engine speed. The two-stroke engine experiments indicate that high levels of internal EGR can enable spark ignition at lean conditions. At higher load conditions, increasing the engine speed is an effective method to control transition from homogeneous charge compression ignition （HCCI） operation to non-HCCI operation and successful spark ignition of a highly dilute mixture can avoid serious knock.

关键词： homogeneous charge combustion exhaust gas recirculation n-heptane auto-ignition

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